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Sample records for high-affinity system km

  1. Characterization of high-affinity (/sup 3/H)ouabain binding in the rat central nervous system

    Energy Technology Data Exchange (ETDEWEB)

    Hauger, R.; Luu, H.M.; Meyer, D.K.; Goodwin, F.K.; Paul, S.M.

    1985-06-01

    The characteristics of (/sup 3/H)ouabain binding were examined in various areas of rat brain. In the striatum, Scatchard analysis revealed a single class of high-affinity binding sites with an apparent binding affinity (KD) of 10.4 +/- 0.9 nM and an estimated binding capacity (Bmax) of 7.6 +/- 1.9 pmol/mg protein. Similar monophasic Scatchard plots were found in the brainstem, cerebellum, hypothalamus, and frontal cerebral cortex. (/sup 3/H)Ouabain binding to rat brain was sodium- and ATP-dependent and strongly inhibited by potassium. Proscillariden A was the most potent cardiac glycoside tested in inhibiting specific (/sup 3/H)ouabain binding to brain membranes, and the rank order of inhibitory potencies for a series of cardiac glycosides was similar to that previously reported for inhibition of heart Na,K-ATPase. To assess whether the high-affinity binding sites for (/sup 3/H)ouabain were localized to neuronal or nonneuronal membranes, the effect of discrete kainic acid lesions on striatal (/sup 3/H)ouabain binding was examined. Kainic acid lesions of the striatum reduced (/sup 3/H)ouabain binding to striatal homogenates by 79.6 +/- 1.6%. This suggests that the high-affinity (/sup 3/H)ouabain binding sites measured in our experiments are localized to neuronal elements. Thus, the high-affinity binding of (/sup 3/H)ouabain to brain membranes may selectively label a neuronal form or conformation of Na,K-ATPase.

  2. PERANCANGAN SISTEM PERPIPAAN KM. NUSANTARA (PIPING SYSTEM

    Directory of Open Access Journals (Sweden)

    Aulia Windyandari

    2013-10-01

    Full Text Available Sistem perpipaan merupakan sistem komplek yang didesain seefektif dan  seefisien mungkin untuk memenuhi kebutuhan dalam kapal ,crew ,muatan dan menjaga keamanan kapal baik saat berlayar ataupun berlabuh. Secara umum sistem pipa dapat diartikan sebagai  bagian utama suatu sistem yang menghubungkan titik dimana fluida di simpan ke titik pengeluaran semua pipa baik untuk memindahkan tenaga atau pemompaan harus dipertimbangkan secara teliti karena keamanan dari sebuah kapal akan tergantung pada susunan perpipaaan seperti halnya pada perlengkapan kapal lainnya Paper ini akan menguraikan tahap-tahap yang harus dilakukan serta pertimbangan-pertimbangan matematis yang diambil  oleh seorang ship engineer  dalam merancang suatu system perpipaan pada kapal KM. Nusantara. Hasil akhir dari paper ini adalah sebuah desain system perpipaan pada pada sebuah kapal,yaitu KM Nusantara, dengan mempertimbangkan system perpipaan yang paling efektif dalam pengoperasiannya.

  3. Solutions for 80 km DWDM systems

    DEFF Research Database (Denmark)

    Dochhan, Annika; Griesser, Helmut; Eiselt, Nicklas

    2016-01-01

    Currently discussed solutions for 80 km DWDM transmission targeting inter-data center connections at 100G and 400G line rates are reviewed. PDM-64QAM, PAM4, and discrete multi-tone transmission (DMT) are investigated, while the focus lies on directly detected solutions. For DMT, the vestigial...

  4. N-Acetyl-2-Aminofluorene (AAF) Processing in Adult Rat Hepatocytes in Primary Culture Occurs by High-Affinity Low-Velocity and Low-Affinity High-Velocity AAF Metabolite-Forming Systems.

    Science.gov (United States)

    Koch, Katherine S; Moran, Tom; Shier, W Thomas; Leffert, Hyam L

    2018-05-01

    N-acetyl-2-aminofluorene (AAF) is a procarcinogen used widely in physiological investigations of chemical hepatocarcinogenesis. Its metabolic pathways have been described extensively, yet little is known about its biochemical processing, growth cycle expression, and pharmacological properties inside living hepatocytes-the principal cellular targets of this hepatocarcinogen. In this report, primary monolayer adult rat hepatocyte cultures and high specific-activity [ring G-3 H]-N-acetyl-2-aminofluorene were used to extend previous observations of metabolic activation of AAF by highly differentiated, proliferation-competent hepatocytes in long-term cultures. AAF metabolism proceeded by zero-order kinetics. Hepatocytes processed significant amounts of procarcinogen (≈12 μg AAF/106 cells/day). Five ring-hydroxylated and one deacetylated species of AAF were secreted into the culture media. Extracellular metabolite levels varied during the growth cycle (days 0-13), but their rank quantitative order was time invariant: 5-OH-AAF > 7-OH-AAF > 3-OH-AAF > N-OH-AAF > aminofluorene (AF) > 1-OH-AAF. Lineweaver-Burk analyses revealed two principal classes of metabolism: System I (high-affinity and low-velocity), Km[APPARENT] = 1.64 × 10-7  M and VMAX[APPARENT] = 0.1 nmol/106 cells/day and System II (low-affinity and high-velocity), Km[APPARENT] = 3.25 × 10-5  M and VMAX[APPARENT] = 1000 nmol/106 cells/day. A third system of metabolism of AAF to AF, with Km[APPARENT] and VMAX[APPARENT] constants of 9.6 × 10-5  M and 4.7 nmol/106 cells/day, was also observed. Evidence provided in this report and its companion paper suggests selective roles and intracellular locations for System I- and System II-mediated AAF metabolite formation during hepatocarcinogenesis, although some of the molecules and mechanisms responsible for multi-system processing remain to be fully defined.

  5. Remote (250 km Fiber Bragg Grating Multiplexing System

    Directory of Open Access Journals (Sweden)

    Manuel Lopez-Amo

    2011-09-01

    Full Text Available We propose and demonstrate two ultra-long range fiber Bragg grating (FBG sensor interrogation systems. In the first approach four FBGs are located 200 km from the monitoring station and a signal to noise ratio of 20 dB is obtained. The second improved version is able to detect the four multiplexed FBGs placed 250 km away, offering a signal to noise ratio of 6–8 dB. Consequently, this last system represents the longest range FBG sensor system reported so far that includes fiber sensor multiplexing capability. Both simple systems are based on a wavelength swept laser to scan the reflection spectra of the FBGs, and they are composed by two identical-lengths optical paths: the first one intended to launch the amplified laser signal by means of Raman amplification and the other one is employed to guide the reflection signal to the reception system.

  6. 2,2'-Dithiobis(N-ethyl-spermine-5-carboxamide) is a high affinity, membrane-impermeant antagonist of the mammalian polyamine transport system.

    Science.gov (United States)

    Huber, M; Pelletier, J G; Torossian, K; Dionne, P; Gamache, I; Charest-Gaudreault, R; Audette, M; Poulin, R

    1996-11-01

    We have synthesized 2,2'-dithiobis(N-ethyl-spermine-5-carboxamide) (DESC), its thiol monomer (MESC), and the mixed MESC-cysteamine disulfide (DEASC) as potential inhibitors of polyamine transport in mammalian cells. DESC was the most potent antagonist of spermine transport in ZR-75-1 human breast cancer cells, with Ki values of 5. 0 +/- 0.7, 80 +/- 31, and 16 +/- 3 microM for DESC, MESC, and DEASC, respectively. DESC also strongly blocked putrescine and spermidine uptake in ZR-75-1 cells (Ki = 1.6 +/- 0.5 and 2.7 +/- 1.1 microM, respectively). While DESC and MESC were purely competitive inhibitors of putrescine transport, DEASC was a mixed competitive/noncompetitive antagonist. Remarkably, DESC was virtually impermeant in ZR-75-1 cells despite its low Ki toward polyamine transport. The marked difference in affinity between DESC and MESC was essentially due to the tail-to-tail juxtaposition of two spermine-like structures, suggesting that dimeric ligands of the polyamine transporter might simultaneously interact with more than one binding site. While DESC strongly decreased the initial rate of [3H]spermidine transport, even a 40-fold molar excess of antagonist could not completely abolish intracellular spermidine accumulation. Moreover, as little as 0.3 microM spermidine fully restored growth in ZR-75-1 cells treated with an inhibitor of polyamine biosynthesis in the presence of 50 microM DESC, thus emphasizing the importance of uptake of trace amounts of exogenous polyamines. Thus, reducing the exogenous supply of polyamines with a potent competitive inhibitor may be kinetically inadequate to block replenishment of the polyamine pool in polyamine-depleted tumor cells that display high transport capacity. These results demonstrate that polyamine analogues cross-linked into a dimeric structure such as DESC interact with high affinity with the mammalian polyamine carrier without being used as substrates. These novel properties provide a framework for the design of

  7. Computerized radiation control system in the Chernobyl 30-km zone

    International Nuclear Information System (INIS)

    Vasil'chenko, D.L.; Pis'mak, A.D.; Livertovskij, G.P.; Chikhladze, V.S.

    1992-01-01

    Computerized radiation control system is intended for monitoring radiation in the zone of radiation-hazardous objects for providing nuclear power station services with current information on radiactive situation in the zone of observation and the doses of radiation taken by the personnel. Described are tasks and functions of the system, as well as its structure. It is noted that unique conditions of the Chernobyl' 30-km zone open up opportunities for experiments and testing new models of equipment. 2 figs

  8. Enterprise KM System: IT based Tool for Nuclear Malaysia

    International Nuclear Information System (INIS)

    Mohamad Safuan Sulaiman; Siti Nurbahyah Hamdan; Mohd Dzul Aiman Aslan

    2014-01-01

    Implementation of right and suitable tool for enterprise Knowledge Management (KM) system to an organization is not an easy task. Everything needs to be taken into account before its implementation come true. One of them is to ensure full cooperation is given by the whole entire organization to succeed the knowledge sharing culture utilizing the tool. From selection of potential tools until the implementation and deployment strategies, these shall be thoroughly and carefully organized. A study of choosing the suitable tools and those strategies has been done in Nuclear Malaysia as resulted from Process Oriented Knowledge Management (POKM) project. As far as enterprise KM system is concerned, Microsoft Share Point technology is one of the potential tools in this context. This paper articulates approach and methodology of choosing the technology including its planning, deployment and implementation strategies. (author)

  9. Expression and distributeion of 'high affinity' glutamate transporters GLT1, GLAST, EAAC and of GCPII in the rat peripheral nervous system

    Czech Academy of Sciences Publication Activity Database

    Carozzi, V. A.; Canta, A.; Oggioni, N.; Ceresa, C.; Marmiroli, P.; Konvalinka, Jan; Zoia, Ch.; Bossi, M.; Ferrarese, C.; Tredici, G.; Cavaletti, G.

    2008-01-01

    Roč. 213, č. 4 (2008), s. 539-546 ISSN 0021-8782 Institutional research plan: CEZ:AV0Z40550506 Keywords : glutamate * glutamate transporters * immunoblotting * immunohistochemistry * peripheral nervous system Subject RIV: CE - Biochemistry Impact factor: 2.063, year: 2008

  10. High affinity hemoglobin and Parkinson's disease.

    Science.gov (United States)

    Graham, Jeffrey; Hobson, Douglas; Ponnampalam, Arjuna

    2014-12-01

    Parkinson's disease (PD) is a neurodegenerative disorder characterized by the loss of dopaminergic neurons in the substantia nigra (SN) region of the midbrain. Oxidative damage in this region has been shown to play an important role in the pathogenesis of this disease. Human neurons have been discovered to contain hemoglobin, with an increased concentration seen in the neurons of the SN. High affinity hemoglobin is a clinical entity resulting from mutations that create a functional increase in the binding of hemoglobin to oxygen and an inability to efficiently unload it to tissues. This can result in a number of metabolic compensatory changes, including an elevation in circulating hemoglobin and an increase in the molecule 2,3-diphosphoglycerate (2,3-DPG). Population based studies have revealed that patients with PD have elevated hemoglobin as well as 2,3-DPG levels. Based on these observations, we hypothesize that the oxidative damage seen in PD is related to an underlying high affinity hemoglobin subtype. Copyright © 2014 Elsevier Ltd. All rights reserved.

  11. The relational database system of KM3NeT

    Science.gov (United States)

    Albert, Arnauld; Bozza, Cristiano

    2016-04-01

    The KM3NeT Collaboration is building a new generation of neutrino telescopes in the Mediterranean Sea. For these telescopes, a relational database is designed and implemented for several purposes, such as the centralised management of accounts, the storage of all documentation about components and the status of the detector and information about slow control and calibration data. It also contains information useful during the construction and the data acquisition phases. Highlights in the database schema, storage and management are discussed along with design choices that have impact on performances. In most cases, the database is not accessed directly by applications, but via a custom designed Web application server.

  12. Kinetics and autoradiography of high affinity uptake of serotonin by primary astrocyte cultures

    International Nuclear Information System (INIS)

    Katz, D.M.; Kimelberg, H.K.

    1985-01-01

    Primary astrocyte cultures prepared from the cerebral cortices of neonatal rats showed significant accumulation of serotonin (5-hydroxytryptamine; [ 3 H]-5-HT). At concentrations in the range of 0.01 to 0.7 microM [ 3 H]-5-HT, this uptake was 50 to 85% Na+ dependent and gave a Km of 0.40 +/- 0.11 microM [ 3 H]-5-HT and a Vmax of 6.42 +/- 0.85 (+/- SEM) pmol of [ 3 H]-5-HT/mg of protein/4 min for the Na+-dependent component. In the absence of Na+ the uptake was nonsaturable. Omission of the monoamine oxidase inhibitor pargyline markedly reduced the Na+-dependent component of [ 3 H]-5-HT uptake but had a negligible effect on the Na+-independent component. This suggest significant oxidative deamination of serotonin after it has been taken up by the high affinity system, followed by release of its metabolite. The authors estimated that this system enabled the cells to concentrate [ 3 H]-5-HT up to 44-fold at an external [ 3 H]-5-HT concentration of 10(-7) M. Inhibition of [ 3 H]-5-HT uptake by a number of clinically effective antidepressants was also consistent with a specific high affinity uptake mechanism for 5-HT, the order of effectiveness of inhibition being chlorimipramine greater than fluoxetine greater than imipramine = amitriptyline greater than desmethylimipramine greater than iprindole greater than mianserin. Uptake of [ 3 H]-5-HT was dependent on the presence of Cl- as well as Na+ in the medium, and the effect of omission of both ions was nonadditive. Varying the concentration of K+ in the media from 1 to 50 mM had a limited effect on [ 3 H]-5-HT uptake

  13. North American Mesoscale Forecast System (NAM) [12 km

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The North American Mesoscale Forecast System (NAM) is one of the major regional weather forecast models run by the National Centers for Environmental Prediction...

  14. Editorial: Practical applications of KM systems for organizational learning

    Directory of Open Access Journals (Sweden)

    Murali Raman

    2013-12-01

    Full Text Available Modern-day organizations are subject to continuous change. To remain relevant and competitive, it is imperative that organizations cultivate and foster learning capabilities. This special issue focus on examining practical applications of knowledge management systems in support of organizational learning efforts.

  15. Underwater acoustic positioning system for the SMO and KM3NeT - Italia projects

    Energy Technology Data Exchange (ETDEWEB)

    Viola, S.; Barbagallo, G.; Cacopardo, G.; Calí, C.; Cocimano, R.; Coniglione, R.; Costa, M.; Cuttone, G.; D' Amato, C.; D' Amato, V.; D' Amico, A.; De Luca, V.; Del Tevere, F.; Distefano, C.; Ferrera, F.; Gmerk, A.; Grasso, R.; Imbesi, M.; Larosa, G.; Lattuada, D. [INFN - Laboratori Nazionali del Sud, via Santa Sofia 62, 95125 Catania (Italy); and others

    2014-11-18

    In the underwater neutrino telescopes, the positions of the Cherenkov light sensors and their movements must be known with an accuracy of few tens of centimetres. In this work, the activities of the SMO and KM3NeT-Italia teams for the development of an acoustic positioning system for KM3NeT-Italia project are presented. The KM3NeT-Italia project foresees the construction, within two years, of 8 towers in the view of the several km{sup 3}-scale neutrino telescope KM3NeT.

  16. Underwater acoustic positioning system for the SMO and KM3NeT - Italia projects

    International Nuclear Information System (INIS)

    Viola, S.; Barbagallo, G.; Cacopardo, G.; Calí, C.; Cocimano, R.; Coniglione, R.; Costa, M.; Cuttone, G.; D'Amato, C.; D'Amato, V.; D'Amico, A.; De Luca, V.; Del Tevere, F.; Distefano, C.; Ferrera, F.; Gmerk, A.; Grasso, R.; Imbesi, M.; Larosa, G.; Lattuada, D.

    2014-01-01

    In the underwater neutrino telescopes, the positions of the Cherenkov light sensors and their movements must be known with an accuracy of few tens of centimetres. In this work, the activities of the SMO and KM3NeT-Italia teams for the development of an acoustic positioning system for KM3NeT-Italia project are presented. The KM3NeT-Italia project foresees the construction, within two years, of 8 towers in the view of the several km 3 -scale neutrino telescope KM3NeT

  17. Na+-Dependent High-Affinity Nitrate, Phosphate and Amino Acids Transport in Leaf Cells of the Seagrass Posidonia oceanica (L. Delile

    Directory of Open Access Journals (Sweden)

    Lourdes Rubio

    2018-05-01

    Full Text Available Posidonia oceanica (L. Delile is a seagrass, the only group of vascular plants to colonize the marine environment. Seawater is an extreme yet stable environment characterized by high salinity, alkaline pH and low availability of essential nutrients, such as nitrate and phosphate. Classical depletion experiments, membrane potential and cytosolic sodium measurements were used to characterize the high-affinity NO3−, Pi and amino acids uptake mechanisms in this species. Net uptake rates of both NO3− and Pi were reduced by more than 70% in the absence of Na+. Micromolar concentrations of NO3− depolarized mesophyll leaf cells plasma membrane. Depolarizations showed saturation kinetics (Km = 8.7 ± 1 μM NO3−, which were not observed in the absence of Na+. NO3− induced depolarizations at increasing Na+ also showed saturation kinetics (Km = 7.2 ± 2 mM Na+. Cytosolic Na+ measured in P. oceanica leaf cells (17 ± 2 mM Na+ increased by 0.4 ± 0.2 mM Na+ upon the addition of 100 μM NO3−. Na+-dependence was also observed for high-affinity l-ala and l-cys uptake and high-affinity Pi transport. All together, these results strongly suggest that NO3−, amino acids and Pi uptake in P. oceanica leaf cells are mediated by high-affinity Na+-dependent transport systems. This mechanism seems to be a key step in the process of adaptation of seagrasses to the marine environment.

  18. ISOLATION AND CHARACTERIZATION OF THE HIGH-AFFINITY K+-TRANSLOCATING ATPASE FROM RHODOBACTER-SPHAEROIDES

    NARCIS (Netherlands)

    ABEE, T; SIEBERS, A; ALTENDORF, K; KONINGS, WN

    1992-01-01

    Cells of the purple nonsulfur bacterium Rhodobacter sphaeroides express a high-affinity K+ uptake system when grown in media with low K+ concentrations. A vanadate-sensitive, K+-stimulated and Mg2+-stimulated ATPase was purified from membranes of these cells by solubilization with

  19. Marshall Space Flight Center Propulsion Systems Department (PSD) Knowledge Management (KM) Initiative

    Science.gov (United States)

    Caraccioli, Paul; Varnedoe, Tom; Smith, Randy; McCarter, Mike; Wilson, Barry; Porter, Richard

    2006-01-01

    NASA Marshall Space Flight Center's Propulsion Systems Department (PSD) is four months into a fifteen month Knowledge Management (KM) initiative to support enhanced engineering decision making and analyses, faster resolution of anomalies (near-term) and effective, efficient knowledge infused engineering processes, reduced knowledge attrition, and reduced anomaly occurrences (long-term). The near-term objective of this initiative is developing a KM Pilot project, within the context of a 3-5 year KM strategy, to introduce and evaluate the use of KM within PSD. An internal NASA/MSFC PSD KM team was established early in project formulation to maintain a practitioner, user-centric focus throughout the conceptual development, planning and deployment of KM technologies and capabilities within the PSD. The PSD internal team is supported by the University of Alabama's Aging Infrastructure Systems Center of Excellence (AISCE), lntergraph Corporation, and The Knowledge Institute. The principle product of the initial four month effort has been strategic planning of PSD KNI implementation by first determining the "as is" state of KM capabilities and developing, planning and documenting the roadmap to achieve the desired "to be" state. Activities undertaken to suppoth e planning phase have included data gathering; cultural surveys, group work-sessions, interviews, documentation review, and independent research. Assessments and analyses have beon pedormed including industry benchmarking, related local and Agency initiatives, specific tools and techniques used and strategies for leveraging existing resources, people and technology to achieve common KM goals. Key findings captured in the PSD KM Strategic Plan include the system vision, purpose, stakeholders, prioritized strategic objectives mapped to the top ten practitioner needs and analysis of current resource usage. Opportunities identified from research, analyses, cultural1KM surveys and practitioner interviews include

  20. Share point 2013 Implementation Strategy for Supporting KM System Requirements in Nuclear Malaysia

    International Nuclear Information System (INIS)

    Mohamad Safuan Sulaiman; Siti Nurbahyah Hamdan; Abdul Muin Abdul Rahman

    2015-01-01

    Knowledge Management system (KMS or KM System) is an important tool for knowledge intensive organization such as Nuclear Malaysia. In June 2010, MS Share Point 2007 was deployed as a tool for KM System in Nuclear Malaysia and was functioning correctly until the end of 2013, whereby the system failed due to software malfunction and inability of the infrastructure to support its continuous operation and usage expansion. This led to difficulties for users to access their operational data and information, hence hampering access to one of the most important tool for KM System in Nuclear Malaysia. However, recently a newer and updated version of the system for example Share point 2013 was deployed to meet the same objectives. Learning from previous failures, the tool has been analyzed at various stages of technical and management reviews. The implementation of this newer version has been designed to overcome most of the deficiencies faced by the older version, both from the software and infrastructure point of views. The tool has performed very well ever since its commissioning from December 2014 till today. As it is still under warranty till March 2016, minimum maintenance issues have been experienced and any problems have been rectified promptly. This paper describes the implementation strategy in preparing the design information of software and hardware architecture of the new tool to overcome the problems of older version, in order to provide a better platform for KM System in Nuclear Malaysia. (author)

  1. The fourth dimension in immunological space: how the struggle for nutrients selects high-affinity lymphocytes.

    Science.gov (United States)

    Wensveen, Felix M; van Gisbergen, Klaas P J M; Eldering, Eric

    2012-09-01

    Lymphocyte activation via the antigen receptor is associated with radical shifts in metabolism and changes in requirements for nutrients and cytokines. Concomitantly, drastic changes occur in the expression of pro-and anti-apoptotic proteins that alter the sensitivity of lymphocytes to limiting concentrations of key survival factors. Antigen affinity is a primary determinant for the capacity of activated lymphocytes to access these vital resources. The shift in metabolic needs and the variable access to key survival factors is used by the immune system to eliminate activated low-affinity cells and to generate an optimal high-affinity response. In this review, we focus on the control of apoptosis regulators in activated lymphocytes by nutrients, cytokines, and costimulation. We propose that the struggle among individual clones that leads to the formation of high-affinity effector cell populations is in effect an 'invisible' fourth signal required for effective immune responses. © 2012 John Wiley & Sons A/S.

  2. Molecular basis of a high affinity murine interleukin-5 receptor.

    OpenAIRE

    Devos, R; Plaetinck, G; Van der Heyden, J; Cornelis, S; Vandekerckhove, J; Fiers, W; Tavernier, J

    1991-01-01

    The mouse interleukin-5 receptor (mIL-5R) consists of two components one of which, the mIL-5R alpha-chain, binds mIL-5 with low affinity. Recently we demonstrated that monoclonal antibodies (Mabs) recognizing the second mIL-5R beta-chain, immunoprecipitate a p130-140 protein doublet which corresponds to the mIL-3R and the mIL-3R-like protein, the latter chain for which so far no ligand has been identified. In this study we show that a high affinity mIL-5R can be reconstituted on COS1 cells by...

  3. The Trigger and Data Acquisition System for the KM3NeT-Italia towers

    Directory of Open Access Journals (Sweden)

    Favaro M.

    2016-01-01

    Full Text Available KM3NeT-Italia is an INFN project supported with Italian PON fundings for building the core of the Italian node of the KM3NeT neutrino telescope. The detector, made of 700 10′′ Optical Modules (OMs lodged along 8 vertical structures called towers, will be deployed starting from fall 2015 at the KM3NeT-Italy site, about 80 km off Capo Passero, Italy, 3500 m deep. The all data to shore approach is used to reduce the complexity of the submarine detector, demanding for an on-line trigger integrated in the data acquisition system running in the shore station, called TriDAS. Due to the large optical background in the sea from 40K decays and bioluminescence, the throughput from the underwater detector can range up to 30 Gbps. This puts strong constraints on the design and performances of the TriDAS and of the related network infrastructure. In this contribution the technology behind the implementation of the TriDAS infrastructure is reviewed, focusing on the relationship between the various components and their performances. The modular design of the TriDAS, which allows for its scalability up to a larger detector than the 8-tower configuration is also discussed.

  4. Rapid transfer of short-lived radioisotopes via a 2. 4 km rabbit system

    Energy Technology Data Exchange (ETDEWEB)

    Burgerjon, J J; Gelbart, Z; Lau, V; Lehnart, D; Lenz, J; Pate, B D; Ruth, T J; Sprenger, H P; van Oers, N S.C.

    1984-09-01

    A 2.4 km long pipeline between a cyclotron and a hospital is used for the rapid transfer of short-lived radiopharmaceuticals. The vials containing the pharmaceuticals are placed inside capsules (rabbits) that are blown through a tube by means of compressed air. Travel times as short as 2 min are achieved, which makes the system suitable for the transfer of /sup 15/O, which has a 2 min half-life. The construction and test results of the system are described along with a computer model, developed to explain some properties of the system. 7 references, 15 figures, 2 tables.

  5. High affinity, ligand specific uptake of complexed copper-67 by brain tissue incubated in vitro

    International Nuclear Information System (INIS)

    Barnea, A.; Hartter, D.E.

    1987-01-01

    Copper is an essential metal that is highly concentrated in the brain. The blood, the sole source of tissue Cu, contains 16-20 μM Cu, of which >95% is complexed to proteins and 2 was 10 times greater than that of CuAlbumin or Cu(II). Within the range of 0.2-150μM Cu, multiple uptake sites for CuHis were apparent. Increasing the molar ratio of His:Cu had a differential effect on Cu uptake: enhancing uptake at [Cu] 1 μM. Thus, using a His:Cu ratio of 1000, they observed a high affinity process exhibiting saturating and half saturating values of 5 μM and 1.5 μM Cu, respectively; using a His:Cu ratio of 2, they observed a low affinity process exhibiting saturating and half-saturating values of 100 μM and 40 μM Cu, respectively. Both processes required thermic but not metabolic energy, suggestive of facilitated diffusion. Considering the blood brain barrier for proteins, CuHis appears to be the major substrate for Cu uptake by neuronal tissue. They demonstrate the existence of a ligand specific, high affinity (apparent Km about 1.5 μM Cu) uptake process for CuHis in the brain, operative at the physiological concentration range of CuHis and histidine

  6. Power and Submarine Cable Systems for the KM3NeT kilometre cube Neutrino Telescope

    CERN Document Server

    Sedita, M; Hallewell, G

    2009-01-01

    The KM3NeT EU-funded consortium, pursuing a cubic kilometre scale neutrino telescope in the Mediterranean Sea, is developing technical solutions for the construction of this challenging project, to be realized several kilometres below the sea level. In this framework a proposed DC/DC power system has been designed, maximizing reliability and minimizing difficulties and expensive underwater activities. The power conversion, delivery, transmission and distribution network will be described with particular attention to: the main electro-optical cable, on shore and deep sea power conversion, the subsea distribution network and connection systems, together with installation and maintenance issues.

  7. Opioid receptor subtypes mediating the noise-induced decreases in high-affinity choline uptake in the rat brain.

    Science.gov (United States)

    Lai, H; Carino, M A

    1992-07-01

    Acute (20 min) exposure to 100-dB white noise elicits a naltrexone-sensitive decrease in sodium-dependent high-affinity choline uptake in the frontal cortex and hippocampus of the rat. In the present study, the subtypes of opioid receptors involved were investigated by pretreating rats with microinjection of specific opioid-receptor antagonists into the lateral cerebroventricle before noise exposure. We found that the noise-induced decrease in high-affinity choline uptake in the hippocampus was blocked by pretreatment with either mu-, delta-, or kappa-opioid-receptor antagonists, whereas the effect of noise on frontal cortical high-affinity choline uptake was blocked by a mu- and delta- but not by a kappa-antagonist. These data further confirm the role of endogenous opioids in mediating the effects of noise on central cholinergic activity and indicate that different neural mechanisms are involved in the effects of noise on the frontal cortical and hippocampal cholinergic systems.

  8. The Trigger and Data Acquisition System for the KM3NeT-Italy neutrino telescope

    Science.gov (United States)

    Chiarusi, T.; Favaro, M.; Giacomini, F.; Manzali, M.; Margiotta, A.; Pellegrino, C.

    2017-10-01

    KM3NeT-Italy is an INFN project that will develop the central part of a submarine cubic-kilometer neutrino telescope in the Ionian Sea, at about 80 km from the Sicilian coast (Italy). It will use hundreds of distributed optical modules to measure the Cherenkov light emitted by high-energy muons, whose signal-to-noise ratio is quite disfavoured. In this contribution the Trigger and Data Acquisition System (TriDAS) developed for the KM3NeT-Italy detector is presented. The “all data to shore” approach is adopted to reduce the complexity of the submarine detector: at the shore station the TriDAS collects, processes and filters all the data coming from the detector, storing triggered events to a permanent storage for subsequent analysis. Due to the large optical background in the sea from 40K decays and bioluminescence, the throughput from the sea can range up to 30 Gbps. This puts strong constraints on the performances of the TriDAS processes and the related network infrastructure.

  9. R&D for an innovative acoustic positioning system for the KM3NeT neutrino telescope

    Science.gov (United States)

    Ameli, F.; Ardid, M.; Bertin, V.; Bonori, M.; Bou-Cabo, M.; Calì, C.; D'Amico, A.; Giovanetti, G.; Imbesi, M.; Keller, P.; Larosa, G.; Llorens, C. D.; Masullo, R.; Randazzo, N.; Riccobene, G.; Speziale, F.; Viola, S.; KM3NeT Consortium

    2011-01-01

    An innovative Acoustic Positioning System for the km3-scale neutrino telescope has been designed and is under realization within the KM3NeT Consortium. Compared to the Acoustic Positioning Systems used for the km3 demonstrators, ANTARES and NEMO Phase 1, this new system is based on the “all data to shore” concept and it will permit the enhancement of detector positioning performances, reduction of costs and its use as real-time monitor of environmental acoustic noise.

  10. Stability of hydrocarbon systems at thermobaric conditions corresponding to depth down to 50 km

    Science.gov (United States)

    Kutcherov, V.; Kolesnikov, A.; Mukhina, E.; Serovaiskii, A.

    2017-12-01

    Most of the theoretical models show that crude oil stability is limited by the depth of 6-8 km (`oil window'). Commercial discovery of crude oil deposits on the depth more than 10 km in the different petroleum basins worldwide casts doubt on the validity of the above-mentioned theoretical calculations. Therefore, the question at which depth complex hydrocarbon systems could be stable is important not only from fundamental research point of view but has a great practical application. To answer this question a hydrocarbon mixture was investigated under thermobaric conditions corresponding to the conditions of the Earth's lower crust. Experiments were conducted by means of Raman Mössbauer spectroscopy. The results obtained show that the complex hydrocarbon systems could be stable and remain their qualitative and quantitative composition at temperature 320-450 °C and pressure 0.7-1.4 GPa. The oxidizing resistance of hydrocarbon system was tested in the modelled the Earth's crust surrounding. The hydrocarbon system stability at the presence of Fe2O3 strongly confirms that the Earth's crust oxygen fugacity does not influence on petroleum composition. The data obtained broaden our knowledge about the possible range of depths for crude oil and natural gas deposits in the Earth's crust and give us the possibility to revise the depth of petroleum deposits occurrence.

  11. ZrFsy1, a high-affinity fructose/H+ symporter from fructophilic yeast Zygosaccharomyces rouxii.

    Directory of Open Access Journals (Sweden)

    Maria José Leandro

    Full Text Available Zygosaccharomyces rouxii is a fructophilic yeast than can grow at very high sugar concentrations. We have identified an ORF encoding a putative fructose/H(+ symporter in the Z. rouxii CBS 732 genome database. Heterologous expression of this ORF in a S. cerevisiae strain lacking its own hexose transporters (hxt-null and subsequent kinetic characterization of its sugar transport activity showed it is a high-affinity low-capacity fructose/H(+ symporter, with Km 0.45 ± 0.07 mM and Vmax 0.57 ± 0.02 mmol h(-1 (gdw(-1. We named it ZrFsy1. This protein also weakly transports xylitol and sorbose, but not glucose or other hexoses. The expression of ZrFSY1 in Z. rouxii is higher when the cells are cultivated at extremely low fructose concentrations (<0.2% and on non-fermentable carbon sources such as mannitol and xylitol, where the cells have a prolonged lag phase, longer duplication times and change their microscopic morphology. A clear phenotype was determined for the first time for the deletion of a fructose/H(+ symporter in the genome where it occurs naturally. The effect of the deletion of ZrFSY1 in Z. rouxii cells is only evident when the cells are cultivated at very low fructose concentrations, when the ZrFsy1 fructose symporter is the main active fructose transporter system.

  12. Detection of Waterborne Viruses Using High Affinity Molecularly Imprinted Polymers.

    Science.gov (United States)

    Altintas, Zeynep; Gittens, Micah; Guerreiro, Antonio; Thompson, Katy-Anne; Walker, Jimmy; Piletsky, Sergey; Tothill, Ibtisam E

    2015-07-07

    Molecularly imprinted polymers (MIPs) are artificial receptor ligands which can recognize and specifically bind to a target molecule. They are more resistant to chemical and biological damage and inactivation than antibodies. Therefore, target specific-MIP nanoparticles are aimed to develop and implemented to biosensors for the detection of biological toxic agents such as viruses, bacteria, and fungi toxins that cause many diseases and death due to the environmental contamination. For the first time, a molecularly imprinted polymer (MIP) targeting the bacteriophage MS2 as the template was investigated using a novel solid-phase synthesis method to obtain the artificial affinity ligand for the detection and removal of waterborne viruses through optical-based sensors. A high affinity between the artificial ligand and the target was found, and a regenerative MIP-based virus detection assay was successfully developed using a new surface plasmon resonance (SPR)-biosensor which provides an alternative technology for the specific detection and removal of waterborne viruses that lead to high disease and death rates all over the world.

  13. Ballistic Performance of Porous Ceramic Thermal Protection Systems at 9 km/s

    Science.gov (United States)

    Miller, Joshua E.; Bohl, W. E.; Foreman, C. D.; Christiansen, Eric L.; Davis, B. A.

    2009-01-01

    Porous-ceramic, thermal-protection-systems are used heavily in current reentry vehicles like the Orbiter, and they are currently being proposed for the next generation of manned spacecraft, Orion. These materials insulate the structural components and sensitive electronic components of a spacecraft against the intense thermal environments of atmospheric reentry. Furthermore, these materials are also highly exposed to space environmental hazards like meteoroid and orbital debris impacts. This paper discusses recent impact testing up to 9 km/s on ceramic tiles similar to those used on the Orbiter. These tiles have a porous-batting of nominally 8 lb/cubic ft alumina-fiber-enhanced-thermal-barrier (AETB8) insulating material coated with a damage-resistant, toughened-unipiece-fibrous-insulation (TUFI) layer.

  14. Ballistic Performance of Porous-Ceramic, Thermal Protection Systems to 9 km/s

    Science.gov (United States)

    Miller, Joshua E.; Bohl, William E.; Foreman, Cory D.; Christiansen, Eric C.; Davis, Bruce A.

    2010-01-01

    Porous-ceramic, thermal protection systems are used heavily in current reentry vehicles like the Orbiter, and they are currently being proposed for the next generation of US manned spacecraft, Orion. These materials insulate the structural components and sensitive components of a spacecraft against the intense thermal environments of atmospheric reentry. These materials are also highly exposed to solid particle space environment hazards. This paper discusses recent impact testing up to 9.65 km/s on ceramic tiles similar to those used on the Orbiter. These tiles are a porous-ceramic insulator of nominally 8 lb/ft(exp 3) alumina-fiber-enhanced-thermal-barrier (AETB8) coated with a damage-resistant, toughened-unipiece-fibrous-insulation/reaction-cured-glass layer (TUFI/RCG).

  15. Development of an acoustic transceiver for the KM3NeT positioning system

    Energy Technology Data Exchange (ETDEWEB)

    Larosa, G., E-mail: giula@doctor.upv.es [Universitat Politèctnica de València, Institut d’Investigació per a la Gestiò Integrada de Zones Costaneres (IGIC), C/Paranimf 1, 46730 Gandia, València (Spain); Ardid, M.; Llorens, C.D.; Bou-Cabo, M.; Martínez-Mora, J.A.; Adrián-Martínez, S. [Universitat Politèctnica de València, Institut d’Investigació per a la Gestiò Integrada de Zones Costaneres (IGIC), C/Paranimf 1, 46730 Gandia, València (Spain)

    2013-10-11

    In this paper we describe an acoustic transceiver developed for the KM3NeT positioning system. The acoustic transceiver is composed of a commercial free flooded transducer, which works mainly in the 20–40 kHz frequency range and withstands high pressures (up to 500 bars). A sound emission board was developed that is adapted to the characteristics of the transducer and meets all requirements: low power consumption, high intensity of emission, low intrinsic noise, arbitrary signals for emission and the capacity of acquiring the receiving signals with very good timing precision. The results of the different tests made with the transceiver in the laboratory and shallow sea water are described, as well as, the activities for its integration in the Instrumentation Line of the ANTARES neutrino telescope and in a NEMO tower for the in situ tests.

  16. High-Affinity Accumulation of Chloroquine by Mouse Erythrocytes Infected with Plasmodium berghei

    Science.gov (United States)

    Fitch, Coy D.; Yunis, Norman G.; Chevli, Rekha; Gonzalez, Yolanda

    1974-01-01

    Washed erythrocytes infected with chloroquine-susceptible (CS) or with chloroquine-resistant (CR) P. berghei were used in model systems in vitro to study the accumulation of chloroquine with high affinity. The CS model could achieve distribution ratios (chloroquine in cells: chloroquine in medium) of 100 in the absence of substrate. 200—300 in the presence of 10 mM pyruvate or lactate, and over 600 in the presence of 1 mM glucose or glycerol. In comparable studies of the CR model, the distribution ratios were 100 in the absence of substrate and 300 or less in the presence of glucose or glycerol. The presence of lactate stimulated chloroquine accumulation in the CR model, whereas the presence of pyruvate did not. Lactate production from glucose and glycerol was undiminished in the CR model, and ATP concentrations were higher than in the CS model. Cold, iodoacetate, 2,4-dinitrophenol, or decreasing pH inhibited chloroquine accumulation in both models. These findings demonstrate substrate involvement in the accumulation of chloroquine with high affinity. In studies of the CS model, certain compounds competitively inhibited chloroquine accumulation, while others did not. This finding is attributable to a specific receptor that imposes structural constraints on the process of accumulation. For chloroquine analogues, the position and length of the side chain, the terminal nitrogen atom of the side chain, and the nitrogen atom in the quinoline ring are important determinants of binding to this receptor. PMID:4600044

  17. 85 km Long Reach PON System Using a Reflective SOA-EA Modulator and Distributed Raman Fiber Amplification

    DEFF Research Database (Denmark)

    Tafur Monroy, Idelfonso; Öhman, Filip; Yvind, Kresten

    2006-01-01

    We report on a bidirectional 85 km long reach PON system supported by distributed fiber Raman amplification with a record 7.5 Gb/s remote carrier modulated upstream signal by employing a reflective SOA-EA monolithically integrated circuit......We report on a bidirectional 85 km long reach PON system supported by distributed fiber Raman amplification with a record 7.5 Gb/s remote carrier modulated upstream signal by employing a reflective SOA-EA monolithically integrated circuit...

  18. Exploiting the high-affinity phosphonate-hydroxyapatite nanoparticle interaction for delivery of radiation and drugs

    International Nuclear Information System (INIS)

    Ong, Hooi Tin; Loo, Joachim S. C.; Boey, Freddy Y. C.; Russell, Stephen J.; Ma Jan; Peng, Kah-Whye

    2008-01-01

    Hydroxyapatite is biocompatible and used in various biomedical applications. Here, we generated hydroxyapatite nanoparticles (HNPs) of various sizes (40-200 nm) and demonstrated that they can be stably loaded with drugs or radioisotopes by exploiting the high-affinity HA-(poly)phosphonate interaction. Clinically available phosphonates, clodronate, and Tc-99m-methylene-diphosphonate (Tc-99m-MDP), were efficiently loaded onto HNPs within 15 min. Biodistribution of radiolabeled HNP-MDP-Tc99m in mice was monitored non-invasively using microSPECT-CT. Imaging and dosimetry studies indicated that the HNPs, regardless of size, were quickly taken up by Kupffer cells in the liver after systemic administration into mice. Clodronate loaded onto HNPs remained biologically active and were able to result in selective depletion of Kupffer cells. This method of drug or isotope loading on HA is fast and easy as it eliminates the need for additional surface modifications of the nanoparticles

  19. A high affinity Ca2(+)-ATPase on the surface membrane of Leishmania donovani promastigote

    International Nuclear Information System (INIS)

    Ghosh, J.; Ray, M.; Sarkar, S.; Bhaduri, A.

    1990-01-01

    A Ca2(+)-dependent ATP-hydrolytic activity was detected in the crude membrane ghost of the promastigote or vector form of the protozoal parasite Leishmania donovani, the pathogen responsible for kala azar. The Ca2(+)-ATPase was purified to apparent homogeneity after solubilization with deoxycholate. The enzyme consists of two subunits of Mr = 51,000 and 57,000 and has an apparent molecular weight of 215,000 +/- 12,000. The enzyme activity is exclusively dependent on Ca2+, and the pure enzyme can hydrolyze 1.6 mumol of ATP/min/mg of protein. The apparent Km for Ca2+ is 35 nM, which is further reduced to 12 nM in the presence of heterologous calmodulin. The enzyme is sensitive to vanadate, but is insensitive to oligomycin and ouabain. The enzyme is strongly associated with the plasma membrane and has its catalytic site oriented toward the cytoplasmic face. The enzyme spans across the plasma membrane as surface labeling with radioiodine shows considerable radioactivity in the completely purified enzyme. The localization and orientation of this high affinity, calmodulin-sensitive Ca2(+)-ATPase suggest some role of this enzyme in Ca2+ movement in the life cycle of this protozoal parasite

  20. A high affinity Ca2(+)-ATPase on the surface membrane of Leishmania donovani promastigote

    Energy Technology Data Exchange (ETDEWEB)

    Ghosh, J.; Ray, M.; Sarkar, S.; Bhaduri, A. (Indian Institute of Chemical Biology, Calcutta (India))

    1990-07-05

    A Ca2(+)-dependent ATP-hydrolytic activity was detected in the crude membrane ghost of the promastigote or vector form of the protozoal parasite Leishmania donovani, the pathogen responsible for kala azar. The Ca2(+)-ATPase was purified to apparent homogeneity after solubilization with deoxycholate. The enzyme consists of two subunits of Mr = 51,000 and 57,000 and has an apparent molecular weight of 215,000 +/- 12,000. The enzyme activity is exclusively dependent on Ca2+, and the pure enzyme can hydrolyze 1.6 mumol of ATP/min/mg of protein. The apparent Km for Ca2+ is 35 nM, which is further reduced to 12 nM in the presence of heterologous calmodulin. The enzyme is sensitive to vanadate, but is insensitive to oligomycin and ouabain. The enzyme is strongly associated with the plasma membrane and has its catalytic site oriented toward the cytoplasmic face. The enzyme spans across the plasma membrane as surface labeling with radioiodine shows considerable radioactivity in the completely purified enzyme. The localization and orientation of this high affinity, calmodulin-sensitive Ca2(+)-ATPase suggest some role of this enzyme in Ca2+ movement in the life cycle of this protozoal parasite.

  1. Insights from the Fungus Fusarium oxysporum Point to High Affinity Glucose Transporters as Targets for Enhancing Ethanol Production from Lignocellulose

    Science.gov (United States)

    Ali, Shahin S.; Nugent, Brian; Mullins, Ewen; Doohan, Fiona M.

    2013-01-01

    Ethanol is the most-widely used biofuel in the world today. Lignocellulosic plant biomass derived from agricultural residue can be converted to ethanol via microbial bioprocessing. Fungi such as Fusarium oxysporum can simultaneously saccharify straw to sugars and ferment sugars to ethanol. But there are many bottlenecks that need to be overcome to increase the efficacy of microbial production of ethanol from straw, not least enhancement of the rate of fermentation of both hexose and pentose sugars. This research tested the hypothesis that the rate of sugar uptake by F. oxysporum would enhance the ethanol yields from lignocellulosic straw and that high affinity glucose transporters can enhance ethanol yields from this substrate. We characterized a novel hexose transporter (Hxt) from this fungus. The F. oxysporum Hxt represents a novel transporter with homology to yeast glucose signaling/transporter proteins Rgt2 and Snf3, but it lacks their C-terminal domain which is necessary for glucose signalling. Its expression level decreased with increasing glucose concentration in the medium and in a glucose uptake study the Km(glucose) was 0.9 mM, which indicated that the protein is a high affinity glucose transporter. Post-translational gene silencing or over expression of the Hxt in F. oxysporum directly affected the glucose and xylose transport capacity and ethanol yielded by F. oxysporum from straw, glucose and xylose. Thus we conclude that this Hxt has the capacity to transport both C5 and C6 sugars and to enhance ethanol yields from lignocellulosic material. This study has confirmed that high affinity glucose transporters are ideal candidates for improving ethanol yields from lignocellulose because their activity and level of expression is high in low glucose concentrations, which is very common during the process of consolidated processing. PMID:23382943

  2. Insights from the fungus Fusarium oxysporum point to high affinity glucose transporters as targets for enhancing ethanol production from lignocellulose.

    Directory of Open Access Journals (Sweden)

    Shahin S Ali

    Full Text Available Ethanol is the most-widely used biofuel in the world today. Lignocellulosic plant biomass derived from agricultural residue can be converted to ethanol via microbial bioprocessing. Fungi such as Fusarium oxysporum can simultaneously saccharify straw to sugars and ferment sugars to ethanol. But there are many bottlenecks that need to be overcome to increase the efficacy of microbial production of ethanol from straw, not least enhancement of the rate of fermentation of both hexose and pentose sugars. This research tested the hypothesis that the rate of sugar uptake by F. oxysporum would enhance the ethanol yields from lignocellulosic straw and that high affinity glucose transporters can enhance ethanol yields from this substrate. We characterized a novel hexose transporter (Hxt from this fungus. The F. oxysporum Hxt represents a novel transporter with homology to yeast glucose signaling/transporter proteins Rgt2 and Snf3, but it lacks their C-terminal domain which is necessary for glucose signalling. Its expression level decreased with increasing glucose concentration in the medium and in a glucose uptake study the Km((glucose was 0.9 mM, which indicated that the protein is a high affinity glucose transporter. Post-translational gene silencing or over expression of the Hxt in F. oxysporum directly affected the glucose and xylose transport capacity and ethanol yielded by F. oxysporum from straw, glucose and xylose. Thus we conclude that this Hxt has the capacity to transport both C5 and C6 sugars and to enhance ethanol yields from lignocellulosic material. This study has confirmed that high affinity glucose transporters are ideal candidates for improving ethanol yields from lignocellulose because their activity and level of expression is high in low glucose concentrations, which is very common during the process of consolidated processing.

  3. A High Affinity Adenosine Kinase from Anopheles gambiae

    Science.gov (United States)

    Cassera, María B.; Ho, Meng-Chiao; Merino, Emilio F.; Burgos, Emmanuel S.; Rinaldo-Matthis, Agnes; Almo, Steven C.; Schramm, Vern L.

    2011-01-01

    Genome analysis revealed a mosquito orthologue of adenosine kinase in Anopheles gambiae (AgAK; the most important vector for the transmission of Plasmodium falciparum in Africa). P. falciparum are purine auxotrophs and do not express an adenosine kinase but rely on their hosts for purines. AgAK was kinetically characterized and found to have the highest affinity for adenosine (Km 8.1 nM) of any known adenosine kinase. AgAK is specific for adenosine at the nucleoside site but several nucleotide triphosphate phosphoryl donors are tolerated. The AgAK crystal structure with a bound bisubstrate analogue Ap4A (2.0 Å resolution) reveals interactions for adenosine, ATP and the geometry for phosphoryl transfer. The polyphosphate charge is partly neutralized by a bound Mg2+ ion and an ion pair to a catalytic site Arg. The AgAK structure consists of a large catalytic core in a three-layered α/β/α sandwich, and a small cap domain in contact with adenosine. The specificity and tight-binding for adenosine arises from hydrogen bond interactions of Asn14, Leu16, Leu40, Leu133, Leu168, Phe168 and Thr171 and the backbone of Ile39 and Phe168 with the adenine ring as well as through hydrogen bond interactions between Asp18, Gly64 and Asn68 and the ribosyl 2′- and 3′-hydroxyl groups. The structure is more similar to human adenosine kinase (48% identity) than to AK from Toxoplasma gondii (31% identity). With this extraordinary affinity for AgAK, adenosine is efficiently captured and converted to AMP at near the diffusion limit, suggesting an important role of this enzyme to maintain the adenine nucleotide pool. mRNA analysis verifies that AgAK transcripts are produced in the adult insects. PMID:21247194

  4. Design and performance simulation of 532 nm Rayleigh-Mie Doppler lidar system for 5-50 km wind measurement

    Science.gov (United States)

    Shen, Fahua; Wang, Bangxin; Shi, Wenjuan; Zhuang, Peng; Zhu, Chengyun; Xie, Chenbo

    2018-04-01

    A novel design of the 532 nm Rayleigh-Mie Doppler lidar receiving system is carried out. The use of polarization isolation technology to effectively improve the receiving system optical reception efficiency, suppress the background noise, not only improves the system wind field detection accuracy, while achieving a high-accuracy temperature measurement. The wind speed and temperature measurement principle of the system are discussed in detail, and the triple Fabry-Perot etalon parameters are optimized. Utilizing the overall design parameters of the system, the system detection performance is simulated. The simulation results show that from 5 to 50 km altitude with vertical resolution of 0.1 km@5 ∼20 km, 0.5 km@20 ∼40 km, 1 km@40 ∼50 km, by using the laser with single pulse energy of 600 mJ, repetition frequency of 50 Hz and the receiving telescope with aperture of 0.8 m, with 2min integration time and in ±50 m/s radial wind speed range, the radial wind speed measurement accuracies of our designed lidar in the day and night are better than 2.6 m/s and 0.9 m/s respectively, and its performance is obviously superior to that of traditional system 5.6 m/s and 1.4 m/s wind speed accuracies; with 10min integration time and in 210 ∼280 K temperature range, the temperature measurement accuracies of the system in the day and night are better than 3.4 K and 1.2 K respectively; since the wind speed sensitivities of the Mie and Rayleigh scattering signals are not exactly the same, in ±50 m/s radial wind speed range, the wind speed bias induced by Mie signal is less than 1 m/s in the temperature range of 210-290 K and in the backscatter ratio range of 1-1.5 for pair measurement.

  5. 85 km long reach PON system using a reflective SOA-EA modulator and distributed Raman fiber amplification

    NARCIS (Netherlands)

    Tafur Monroy, I.; Öhman, F.; Yvind, K.; Kjaer, R.; Peucheret, C.; Koonen, A.M.J.; Jeppesen, P.

    2006-01-01

    We report on a bidirectional 85 km long reach PON system supported by distributed fiber Raman amplification with a record 7.5 Gb/s remote carrier modulated upstream signal by employing a reflective SOA-EA monolithically integrated circuit.

  6. Antioxidant mechanism of milk mineral-high-affinity iron binding.

    Science.gov (United States)

    Allen, K; Cornforth, D

    2007-01-01

    Milk mineral (MM), a by-product of whey processing, is an effective antioxidant in meat systems, but the antioxidant mechanism has not been established. MM has been postulated to chelate iron and prevent iron-catalysis of lipid oxidation. The objective of this research was to examine this putative mechanism. MM was compared to sodium tripolyphosphate (STPP), calcium phosphate monobasic (CPM), and calcium pyrophosphate (CPP) to determine iron-binding capacity, sample solubility, and eluate soluble phosphorus after treating samples with a ferrous chloride standard. Scanning electron microscopy with energy-dispersive X-ray analysis was used to localize minerals on iron-treated MM particle surfaces. Histochemical staining for calcium was performed on raw and cooked ground beef samples with added MM. MM bound more iron per gram (P compounds, and was much less soluble (P iron across the MM particle surface, directly demonstrating iron binding to MM particles. Unlike other common chelating agents, such as STPP and citrate, histochemical staining demonstrated that MM remained insoluble in ground beef, even after cooking. The ability of MM to bind iron and remain insoluble may enhance its antioxidant effect by removing iron ions from solution. However, MM particles must be small and well distributed in order to adequately bind iron throughout the food system.

  7. Transmission of 2.5 Gbit/s Spectrum-sliced WDM System for 50 km Single-mode Fiber

    Science.gov (United States)

    Ahmed, Nasim; Aljunid, Sayed Alwee; Ahmad, R. Badlisha; Fadil, Hilal Adnan; Rashid, Mohd Abdur

    2011-06-01

    The transmission of a spectrum-sliced WDM channel at 2.5 Gbit/s for 50 km of single mode fiber using an system channel spacing only 0.4 nm is reported. We have investigated the system performance using NRZ modulation format. The proposed system is compared with conventional system. The system performance is characterized as the bit-error-rate (BER) received against the system bit rates. Simulation results show that the NRZ modulation format performs well for 2.5 Gbit/s system bit rates. Using this narrow channel spectrum-sliced technique, the total number of multiplexed channels can be increased greatly in WDM system. Therefore, 0.4 nm channel spacing spectrum-sliced WDM system is highly recommended for the long distance optical access networks, like the Metro Area Network (MAN), Fiber-to-the-Building (FTTB) and Fiber-to-the-Home (FTTH).

  8. Carbonate-sensitive phytotransferrin controls high-affinity iron uptake in diatoms

    Science.gov (United States)

    McQuaid, Jeffrey B.; Kustka, Adam B.; Oborník, Miroslav; Horák, Aleš; McCrow, John P.; Karas, Bogumil J.; Zheng, Hong; Kindeberg, Theodor; Andersson, Andreas J.; Barbeau, Katherine A.; Allen, Andrew E.

    2018-03-01

    In vast areas of the ocean, the scarcity of iron controls the growth and productivity of phytoplankton. Although most dissolved iron in the marine environment is complexed with organic molecules, picomolar amounts of labile inorganic iron species (labile iron) are maintained within the euphotic zone and serve as an important source of iron for eukaryotic phytoplankton and particularly for diatoms. Genome-enabled studies of labile iron utilization by diatoms have previously revealed novel iron-responsive transcripts, including the ferric iron-concentrating protein ISIP2A, but the mechanism behind the acquisition of picomolar labile iron remains unknown. Here we show that ISIP2A is a phytotransferrin that independently and convergently evolved carbonate ion-coordinated ferric iron binding. Deletion of ISIP2A disrupts high-affinity iron uptake in the diatom Phaeodactylum tricornutum, and uptake is restored by complementation with human transferrin. ISIP2A is internalized by endocytosis, and manipulation of the seawater carbonic acid system reveals a second-order dependence on the concentrations of labile iron and carbonate ions. In P. tricornutum, the synergistic interaction of labile iron and carbonate ions occurs at environmentally relevant concentrations, revealing that carbonate availability co-limits iron uptake. Phytotransferrin sequences have a broad taxonomic distribution and are abundant in marine environmental genomic datasets, suggesting that acidification-driven declines in the concentration of seawater carbonate ions will have a negative effect on this globally important eukaryotic iron acquisition mechanism.

  9. Enhanced membrane pore formation through high-affinity targeted antimicrobial peptides.

    Directory of Open Access Journals (Sweden)

    Christopher J Arnusch

    Full Text Available Many cationic antimicrobial peptides (AMPs target the unique lipid composition of the prokaryotic cell membrane. However, the micromolar activities common for these peptides are considered weak in comparison to nisin, which follows a targeted, pore-forming mode of action. Here we show that AMPs can be modified with a high-affinity targeting module, which enables membrane permeabilization at low concentration. Magainin 2 and a truncated peptide analog were conjugated to vancomycin using click chemistry, and could be directed towards specific membrane embedded receptors both in model membrane systems and whole cells. Compared with untargeted vesicles, a gain in permeabilization efficacy of two orders of magnitude was reached with large unilamellar vesicles that included lipid II, the target of vancomycin. The truncated vancomycin-peptide conjugate showed an increased activity against vancomycin resistant Enterococci, whereas the full-length conjugate was more active against a targeted eukaryotic cell model: lipid II containing erythrocytes. This study highlights that AMPs can be made more selective and more potent against biological membranes that contain structures that can be targeted.

  10. Early signs of pathological cognitive aging in mice lacking high-affinity nicotinic receptors.

    Directory of Open Access Journals (Sweden)

    Eleni eKonsolaki

    2016-04-01

    Full Text Available In order to address pathological cognitive decline effectively, it is critical to adopt early preventive measures in individuals considered at risk. It is therefore essential to develop approaches that identify such individuals before the onset of irreversible dementia. Α deficient cholinergic system has been consistently implicated as one of the main factors associated with a heightened vulnerability to the aging process. In the present study we used mice lacking high affinity nicotinic receptors (β2-/-, which have been proposed as an animal model of accelerated/premature cognitive aging. Our aim was to identify behavioural signs that could serve as indicators or predictors of impending cognitive decline. We used test batteries in order to assess cognitive functions and additional tasks to investigate spontaneous behaviours, such as species-specific activities and exploration/locomotion in a novel environment. Our data confirm and extend the hypothesis that β2-/- animals exhibit age-related cognitive impairments, manifested in both spatial learning and recognition memory tasks. In addition, we reveal deficits in spontaneous behaviour and habituation processes earlier in life. To our knowledge, this is the first study to perform an extensive behavioural examination of an animal model of premature cognitive aging, and our results suggest that β2-nAChR dependent cognitive deterioration progressively evolves from initial subtle behavioural changes to global dementia due to the combined effect of the neuropathology and aging.

  11. The electronics readout and data acquisition system of the KM3NeT neutrino telescope node

    Energy Technology Data Exchange (ETDEWEB)

    Real, Diego [IFIC, Instituto de Física Corpuscular, CSIC-Universidad de Valencia, C/Catedrático José Beltrán, 2, 46980 Paterna (Spain); Collaboration: KM3NeT Collaboration

    2014-11-18

    The KM3NeT neutrino telescope will be composed by tens of thousands of glass spheres, called Digital Optical Module (DOM), each of them containing 31 PMTs of small photocathode area (3'). The readout and data acquisition system of KM3NeT have to collect, treat and send to shore, in an economic way, the enormous amount of data produced by the photomultipliers and at the same time to provide time synchronization between each DOM at the level of 1 ns. It is described in the present article the Central Logic Board, that integrates the Time to Digital Converters and the White Rabbit protocol used for the DOM synchronization in a transparent way, the Power Board used in the DOM, the PMT base to readout the photomultipliers and the respective collecting boards, the so called Octopus Board.

  12. The electronics readout and data acquisition system of the KM3NeT neutrino telescope node

    International Nuclear Information System (INIS)

    Real, Diego

    2014-01-01

    The KM3NeT neutrino telescope will be composed by tens of thousands of glass spheres, called Digital Optical Module (DOM), each of them containing 31 PMTs of small photocathode area (3'). The readout and data acquisition system of KM3NeT have to collect, treat and send to shore, in an economic way, the enormous amount of data produced by the photomultipliers and at the same time to provide time synchronization between each DOM at the level of 1 ns. It is described in the present article the Central Logic Board, that integrates the Time to Digital Converters and the White Rabbit protocol used for the DOM synchronization in a transparent way, the Power Board used in the DOM, the PMT base to readout the photomultipliers and the respective collecting boards, the so called Octopus Board

  13. Discovery of PF-06928215 as a high affinity inhibitor of cGAS enabled by a novel fluorescence polarization assay.

    Science.gov (United States)

    Hall, Justin; Brault, Amy; Vincent, Fabien; Weng, Shawn; Wang, Hong; Dumlao, Darren; Aulabaugh, Ann; Aivazian, Dikran; Castro, Dana; Chen, Ming; Culp, Jeffrey; Dower, Ken; Gardner, Joseph; Hawrylik, Steven; Golenbock, Douglas; Hepworth, David; Horn, Mark; Jones, Lyn; Jones, Peter; Latz, Eicke; Li, Jing; Lin, Lih-Ling; Lin, Wen; Lin, David; Lovering, Frank; Niljanskul, Nootaree; Nistler, Ryan; Pierce, Betsy; Plotnikova, Olga; Schmitt, Daniel; Shanker, Suman; Smith, James; Snyder, William; Subashi, Timothy; Trujillo, John; Tyminski, Edyta; Wang, Guoxing; Wong, Jimson; Lefker, Bruce; Dakin, Leslie; Leach, Karen

    2017-01-01

    Cyclic GMP-AMP synthase (cGAS) initiates the innate immune system in response to cytosolic dsDNA. After binding and activation from dsDNA, cGAS uses ATP and GTP to synthesize 2', 3' -cGAMP (cGAMP), a cyclic dinucleotide second messenger with mixed 2'-5' and 3'-5' phosphodiester bonds. Inappropriate stimulation of cGAS has been implicated in autoimmune disease such as systemic lupus erythematosus, thus inhibition of cGAS may be of therapeutic benefit in some diseases; however, the size and polarity of the cGAS active site makes it a challenging target for the development of conventional substrate-competitive inhibitors. We report here the development of a high affinity (KD = 200 nM) inhibitor from a low affinity fragment hit with supporting biochemical and structural data showing these molecules bind to the cGAS active site. We also report a new high throughput cGAS fluorescence polarization (FP)-based assay to enable the rapid identification and optimization of cGAS inhibitors. This FP assay uses Cy5-labelled cGAMP in combination with a novel high affinity monoclonal antibody that specifically recognizes cGAMP with no cross reactivity to cAMP, cGMP, ATP, or GTP. Given its role in the innate immune response, cGAS is a promising therapeutic target for autoinflammatory disease. Our results demonstrate its druggability, provide a high affinity tool compound, and establish a high throughput assay for the identification of next generation cGAS inhibitors.

  14. Discovery of PF-06928215 as a high affinity inhibitor of cGAS enabled by a novel fluorescence polarization assay

    Energy Technology Data Exchange (ETDEWEB)

    Hall, Justin; Brault, Amy; Vincent, Fabien; Weng, Shawn; Wang, Hong; Dumlao, Darren; Aulabaugh, Ann; Aivazian, Dikran; Castro, Dana; Chen, Ming; Culp, Jeffrey; Dower, Ken; Gardner, Joseph; Hawrylik, Steven; Golenbock, Douglas; Hepworth, David; Horn, Mark; Jones, Lyn; Jones, Peter; Latz, Eicke; Li, Jing; Lin, Lih-Ling; Lin, Wen; Lin, David; Lovering, Frank; Niljanskul, Nootaree; Nistler, Ryan; Pierce, Betsy; Plotnikova, Olga; Schmitt, Daniel; Shanker, Suman; Smith, James; Snyder, William; Subashi, Timothy; Trujillo, John; Tyminski, Edyta; Wang, Guoxing; Wong, Jimson; Lefker, Bruce; Dakin, Leslie; Leach, Karen (UMASS, MED); (Pfizer)

    2017-09-21

    Cyclic GMP-AMP synthase (cGAS) initiates the innate immune system in response to cytosolic dsDNA. After binding and activation from dsDNA, cGAS uses ATP and GTP to synthesize 2', 3' -cGAMP (cGAMP), a cyclic dinucleotide second messenger with mixed 2'-5' and 3'-5' phosphodiester bonds. Inappropriate stimulation of cGAS has been implicated in autoimmune disease such as systemic lupus erythematosus, thus inhibition of cGAS may be of therapeutic benefit in some diseases; however, the size and polarity of the cGAS active site makes it a challenging target for the development of conventional substrate-competitive inhibitors. We report here the development of a high affinity (KD = 200 nM) inhibitor from a low affinity fragment hit with supporting biochemical and structural data showing these molecules bind to the cGAS active site. We also report a new high throughput cGAS fluorescence polarization (FP)-based assay to enable the rapid identification and optimization of cGAS inhibitors. This FP assay uses Cy5-labelled cGAMP in combination with a novel high affinity monoclonal antibody that specifically recognizes cGAMP with no cross reactivity to cAMP, cGMP, ATP, or GTP. Given its role in the innate immune response, cGAS is a promising therapeutic target for autoinflammatory disease. Our results demonstrate its druggability, provide a high affinity tool compound, and establish a high throughput assay for the identification of next generation cGAS inhibitors.

  15. N-Oxide analogs of WAY-100635 : new high affinity 5-HT (1A) receptor antagonists

    NARCIS (Netherlands)

    Oberwinkler - Marchais, Sandrine; Nowicki, B; Pike, VW; Halldin, C; Sandell, J; Chou, YH; Gulyas, B; Brennum, LT; Farde, L; Wikstrom, H V

    2005-01-01

    WAY-100635 [N-(2-(1-(4-(2-methoxyphenyl)piperazinyl)ethyl))-N-(2-pyridinyl)cyclohexanecarboxamide] 1 and its O-des-methyl derivative DWAY 2 are well-known high affinity 5-HT1A receptor antagonists. which when labeled with carbon-II (beta(+): t(1/2) 20.4min) in the carbonyl group are effective

  16. High affinity calmodulin target sequence in the signalling molecule PI 3-kinase

    DEFF Research Database (Denmark)

    Fischer, R; Julsgart, J; Berchtold, M W

    1998-01-01

    M-binding peptide derived from the p110gamma isoform interacts with CaM in a calcium-dependent way. Using gel shift analysis and fluorescence spectrophotometry we discovered that the peptide forms a high affinity complex with CaM. Titration experiments using dansylated CaM gave an affinity constant of 5 n...

  17. The role of CH/π interactions in the high affinity binding of streptavidin and biotin.

    Science.gov (United States)

    Ozawa, Motoyasu; Ozawa, Tomonaga; Nishio, Motohiro; Ueda, Kazuyoshi

    2017-08-01

    The streptavidin-biotin complex has an extraordinarily high affinity (Ka: 10 15 mol -1 ) and contains one of the strongest non-covalent interactions known. This strong interaction is widely used in biological tools, including for affinity tags, detection, and immobilization of proteins. Although hydrogen bond networks and hydrophobic interactions have been proposed to explain this high affinity, the reasons for it remain poorly understood. Inspired by the deceased affinity of biotin observed for point mutations of streptavidin at tryptophan residues, we hypothesized that a CH/π interaction may also contribute to the strong interaction between streptavidin and biotin. CH/π interactions were explored and analyzed at the biotin-binding site and at the interface of the subunits by the fragment molecular orbital method (FMO) and extended applications: PIEDA and FMO4. The results show that CH/π interactions are involved in the high affinity for biotin at the binding site of streptavidin. We further suggest that the involvement of CH/π interactions at the subunit interfaces and an extended CH/π network play more critical roles in determining the high affinity, rather than involvement at the binding site. Copyright © 2017 Elsevier Inc. All rights reserved.

  18. Blockage of High-Affinity Choline Transporter Increases Visceral Hypersensitivity in Rats with Chronic Stress

    Directory of Open Access Journals (Sweden)

    Chen Zhao

    2018-01-01

    Full Text Available Background. Visceral hypersensitivity is a common feature of irritable bowel syndrome. Cholinergic system involves in the development of visceral hypersensitivity, and high-affinity choline transporter (CHT1 is of crucial importance in choline uptake system. However, involvement of CHT1 in visceral hypersensitivity remains unknown. The research aimed to study the CHT1 expression in dorsal root ganglions (DRGs and the role of CHT1 in visceral hypersensitivity. Methods. Repetitive water avoidance stress (WAS was used to induce visceral hypersensitivity in rats. Colorectal distension (CRD was determined, and the abdominal withdrawal reflex (AWR and threshold intensity data were recorded to measure the visceral sensitivity. After intraperitoneal injection of hemicholinium-3 (HC-3, the specific inhibitor of CHT1, CRD data were also recorded. The CHT1 expression of DRGs was investigated by Western blotting, immunohistochemistry, and quantitative RT-PCR. Acetylcholine levels in the DRGs were detected by the assay kit. Results. Repetitive WAS increased the AWR score of CRD at high distension pressure and decreased the mean threshold of rats. The CHT1 expression and acetylcholine concentration of DRG were significantly increased in WAS rats. After the administration of HC-3, the AWR score in WAS group was significantly increased at higher distension pressure while the threshold intensity was significantly reduced compared to the normal saline group. Acetylcholine concentration was significantly lower than the normal saline rats. Conclusion. Our research firstly reports that CHT1 is overexpressed in noninflammatory visceral hypersensitivity, and blockage of CHT1 can enhance the visceral hypersensitivity. CHT1 may play an inhibitory role in visceral hypersensitivity.

  19. Blockage of High-Affinity Choline Transporter Increases Visceral Hypersensitivity in Rats with Chronic Stress

    Science.gov (United States)

    2018-01-01

    Background Visceral hypersensitivity is a common feature of irritable bowel syndrome. Cholinergic system involves in the development of visceral hypersensitivity, and high-affinity choline transporter (CHT1) is of crucial importance in choline uptake system. However, involvement of CHT1 in visceral hypersensitivity remains unknown. The research aimed to study the CHT1 expression in dorsal root ganglions (DRGs) and the role of CHT1 in visceral hypersensitivity. Methods Repetitive water avoidance stress (WAS) was used to induce visceral hypersensitivity in rats. Colorectal distension (CRD) was determined, and the abdominal withdrawal reflex (AWR) and threshold intensity data were recorded to measure the visceral sensitivity. After intraperitoneal injection of hemicholinium-3 (HC-3), the specific inhibitor of CHT1, CRD data were also recorded. The CHT1 expression of DRGs was investigated by Western blotting, immunohistochemistry, and quantitative RT-PCR. Acetylcholine levels in the DRGs were detected by the assay kit. Results Repetitive WAS increased the AWR score of CRD at high distension pressure and decreased the mean threshold of rats. The CHT1 expression and acetylcholine concentration of DRG were significantly increased in WAS rats. After the administration of HC-3, the AWR score in WAS group was significantly increased at higher distension pressure while the threshold intensity was significantly reduced compared to the normal saline group. Acetylcholine concentration was significantly lower than the normal saline rats. Conclusion Our research firstly reports that CHT1 is overexpressed in noninflammatory visceral hypersensitivity, and blockage of CHT1 can enhance the visceral hypersensitivity. CHT1 may play an inhibitory role in visceral hypersensitivity. PMID:29849603

  20. The mediator-modulator brain system in minks exposed to radiation factors of the Chernobyl NPP 10-km zone

    International Nuclear Information System (INIS)

    Mishunina, T.M.; Kalinskaya, L.N.; Pil'kevich, L.I.; Kononenko, V.Ya.; Ryasenko, V.I.; Bogdanova, T.I.

    1996-01-01

    The paper is devoted to the study of the activity of the important metabolizing mediators/modulators of the central nervous system: adenosine, angiotensin and GABA as well as specific bonding of 14 C-GABA by synaptic membranes of brain structures in minks who were in the 10-km zone of the Chernobyl NPP from 0.5 months to 3 years. Adenosine deaminase, angiotensin converting enzyme and glutamate decarboxylase activities endured significant changes in the hypothalamus, striatum, hippocampus, cerebellum, cerebral, cortex, medulla oblongata and midbrain of minks; the direction and the extent of manifestation of those changes depend on the term of animals' stay in the zone. The GABA reception in the cerebral cortex and medulla oblongata increased significantly and progressively with the prolongation of exposure to radiation. It is concluded that the variety of changes, their regional-specificity and dependence on exposure of animals to the radiation factors of the Chernobyl NPP 10-km zone may induce development of serious complications not only in nervous but in some other systems

  1. Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms.

    Science.gov (United States)

    Yang, Danlin; Singh, Ajit; Wu, Helen; Kroe-Barrett, Rachel

    2017-04-17

    Label-free optical biosensors are powerful tools in drug discovery for the characterization of biomolecular interactions. In this study, we describe the use of four routinely used biosensor platforms in our laboratory to evaluate the binding affinity and kinetics of ten high-affinity monoclonal antibodies (mAbs) against human proprotein convertase subtilisin kexin type 9 (PCSK9). While both Biacore T100 and ProteOn XPR36 are derived from the well-established Surface Plasmon Resonance (SPR) technology, the former has four flow cells connected by serial flow configuration, whereas the latter presents 36 reaction spots in parallel through an improvised 6 x 6 crisscross microfluidic channel configuration. The IBIS MX96 also operates based on the SPR sensor technology, with an additional imaging feature that provides detection in spatial orientation. This detection technique coupled with the Continuous Flow Microspotter (CFM) expands the throughput significantly by enabling multiplex array printing and detection of 96 reaction sports simultaneously. In contrast, the Octet RED384 is based on the BioLayer Interferometry (BLI) optical principle, with fiber-optic probes acting as the biosensor to detect interference pattern changes upon binding interactions at the tip surface. Unlike the SPR-based platforms, the BLI system does not rely on continuous flow fluidics; instead, the sensor tips collect readings while they are immersed in analyte solutions of a 384-well microplate during orbital agitation. Each of these biosensor platforms has its own advantages and disadvantages. To provide a direct comparison of these instruments' ability to provide quality kinetic data, the described protocols illustrate experiments that use the same assay format and the same high-quality reagents to characterize antibody-antigen kinetics that fit the simple 1:1 molecular interaction model.

  2. Carbon-11-labelling of a novel, trishomocubane-derived, high affinity and selectivity DAT ligand

    International Nuclear Information System (INIS)

    Dolle, F.; Le Helleix, St.; Peyronneau, M.A.; Saba, W.; Tournier, N.; Valette, H.; Banister, S.; Kassiou, M.

    2011-01-01

    Complete text of publication follows: Objectives: Parkinson's disease, schizophrenia, attention deficit disorder and drug abuse are related to abnormalities within the brain's dopaminergic system. The neuronal dopamine transporter (DAT) plays a key role in regulating the synaptic concentration of dopamine and thus dopamine neurotransmission in the brain. Since the DAT can be considered as a marker of the integrity and number of the presynaptic striatal dopamine-producing neurons, considerable efforts have been spent in recent years on the design and development of DAT-selective radioligands for use in Positron Emission Tomography (PET) studies. Notably, the tropane PE2I and its fluorinated analogue LBT-999 were identified as having high affinity and selectivity for the DAT over the norepinephrine transporter (NET) and the serotonin transporter (SERT). Besides tropanes, only a few bicyclic frameworks, e.g. bicyclo[2.2.2]octanes, have delivered compounds with high affinity for the DAT. Recently, novel poly-carbocyclic DAT ligands with selectivity over the NET and the SERT were reported. The lead compound of this series (1, N-methyl-N-(3-fluoro) benzyl-pentacyclo[5.4.0.0 2, 6 .0 3, 10 .0 5, 9 ] undec-8-ylamine, Ki = 1.2 nM, ≥ 8300-fold selectivity over NET and SERT) was selected as a potential candidate for imaging the DAT with PET and isotopically labelled with carbon-11 using [ 11 C]methyl triflate. Methods: The trishomocubane derivatives 1 (reference) and 2 (precursor for labelling with carbon-11) were prepared from commercially available Cookson's diketone in 6 and 7 steps, respectively. Carbon-11 labelling of 1 was performed using a TRACERLab FX-C Pro synthesizer (GEMS) and comprises (1) trapping at -10 C of [ 11 C]MeOTf in acetone (0.4 mL) containing the nor-derivative 2 (0.6-0.9 mg, free base) and aq. 3N NaOH (8 μL); (2) heating at 110 C for 2 min; (3) concentration to dryness and taking up the residue in 1.0 mL of the HPLC mobile phase; (4) purification

  3. Design and first tests of an acoustic positioning and detection system for KM3NeT

    Science.gov (United States)

    Simeone, F.; Ameli, F.; Ardid, M.; Bertin, V.; Bonori, M.; Bou-Cabo, M.; Calì, C.; D'Amico, A.; Giovanetti, G.; Imbesi, M.; Keller, P.; Larosa, G.; Llorens, C. D.; Masullo, R.; Randazzo, N.; Riccobene, G.; Speziale, F.; Viola, S.; KM3NeT Consortium

    2012-01-01

    In a deep-sea neutrino telescope it is mandatory to locate the position of the optical sensors with a precision of about 10 cm. To achieve this requirement, an innovative Acoustic Positioning System (APS) has been designed in the frame work of the KM3NeT neutrino telescope. The system will also be able to provide an acoustic guide during the deployment of the telescope’s components and seafloor infrastructures (junction boxes, cables, etc.). A prototype of the system based on the successful acoustic systems of ANTARES and NEMO is being developed. It will consist of an array of hydrophones and a network of acoustic transceivers forming the Long Baseline. All sensors are connected to the telescope data acquisition system and are in phase and synchronised with the telescope master clock. Data from the acoustic sensors, continuously sampled at 192 kHz, will be sent to shore where signal recognition and analysis will be carried out. The design and first tests of the system elements will be presented. This new APS is expected to have better precision compared to the systems used in ANTARES and NEMO, and can also be used as a real-time monitor of acoustic sources and environmental noise in deep sea.

  4. Selective high-affinity polydentate ligands and methods of making such

    Energy Technology Data Exchange (ETDEWEB)

    Denardo, Sally J.; Denardo, Gerald L.; Balhorn, Rodney L.

    2018-02-06

    This invention provides novel polydentate selective high affinity ligands (SHALs) that can be used in a variety of applications in a manner analogous to the use of antibodies. SHALs typically comprise a multiplicity of ligands that each bind different region son the target molecule. The ligands are joined directly or through a linker thereby forming a polydentate moiety that typically binds the target molecule with high selectivity and avidity.

  5. Quantifying high-affinity binding of hydrophobic ligands by isothermal titration calorimetry.

    Science.gov (United States)

    Krainer, Georg; Broecker, Jana; Vargas, Carolyn; Fanghänel, Jörg; Keller, Sandro

    2012-12-18

    A fast and reliable quantification of the binding thermodynamics of hydrophobic high-affinity ligands employing a new calorimetric competition experiment is described. Although isothermal titration calorimetry is the method of choice for a quantitative characterization of intermolecular interactions in solution, a reliable determination of a dissociation constant (K(D)) is typically limited to the range 100 μM > K(D) > 1 nM. Interactions displaying higher or lower K(D) values can be assessed indirectly, provided that a suitable competing ligand is available whose K(D) falls within the directly accessible affinity window. This established displacement assay, however, requires the high-affinity ligand to be soluble at high concentrations in aqueous buffer and, consequently, poses serious problems in the study of protein binding involving small-molecule ligands dissolved in organic solvents--a familiar case in many drug-discovery projects relying on compound libraries. The calorimetric competition assay introduced here overcomes this limitation, thus allowing for a detailed thermodynamic description of high-affinity receptor-ligand interactions involving poorly water-soluble compounds. Based on a single titration of receptor into a dilute mixture of the two competing ligands, this competition assay provides accurate and precise values for the dissociation constants and binding enthalpies of both high- and moderate-affinity ligands. We discuss the theoretical background underlying the approach, demonstrate its practical application to metal ion chelation and high-affinity protein-inhibitor interactions, and explore its potential and limitations with the aid of simulations and statistical analyses.

  6. Amyloid-beta binds catalase with high affinity and inhibits hydrogen peroxide breakdown.

    OpenAIRE

    Milton, N G

    1999-01-01

    Amyloid-beta (Abeta) specifically bound purified catalase with high affinity and inhibited catalase breakdown of H(2)O(2). The Abeta-induced catalase inhibition involved formation of the inactive catalase Compound II and was reversible. CatalaseAbeta interactions provide rapid functional assays for the cytotoxic domain of Abeta and suggest a mechanism for some of the observed actions of Abeta plus catalase in vitro.

  7. Reconstitution of high-affinity opioid agonist binding in brain membranes

    Energy Technology Data Exchange (ETDEWEB)

    Remmers, A.E.; Medzihradsky, F. (Univ. of Michigan Medical School, Ann Arbor (United States))

    1991-03-15

    In synaptosomal membranes from rat brain cortex, the {mu} selective agonist ({sup 3}H)dihydromorphine in the absence of sodium, and the nonselective antagonist ({sup 3}H)naltrexone in the presence of sodium, bound to two populations of opioid receptor sites with K{sub d} values of 0.69 and 8.7 nM for dihydromorphine, and 0.34 and 5.5 nM for naltrexone. The addition of 5 {mu}M guanosine 5{prime}-({gamma}-thio)triphosphate (GTP({gamma}S)) strongly reduced high-affinity agonist but not antagonist binding. Exposure of the membranes to high pH reduced the number of GTP({gamma}-{sup 35}S) binding sites by 90% and low K{sub m}, opioid-sensitive GTPase activity by 95%. In these membranes, high-affinity agonist binding was abolished and modulation of residual binding by GTP({gamma}S) was diminished. Alkali treatment of the glioma cell membranes prior to fusion inhibited most of the low K{sub m} GTPase activity and prevented the reconstitution of agonist binding. The results show that high-affinity opioid agonist binding reflects the ligand-occupied receptor - guanine nucleotide binding protein complex.

  8. Single-experiment displacement assay for quantifying high-affinity binding by isothermal titration calorimetry.

    Science.gov (United States)

    Krainer, Georg; Keller, Sandro

    2015-04-01

    Isothermal titration calorimetry (ITC) is the gold standard for dissecting the thermodynamics of a biomolecular binding process within a single experiment. However, reliable determination of the dissociation constant (KD) from a single titration is typically limited to the range 100 μM>KD>1 nM. Interactions characterized by a lower KD can be assessed indirectly by so-called competition or displacement assays, provided that a suitable competitive ligand is available whose KD falls within the directly accessible window. However, this protocol is limited by the fact that it necessitates at least two titrations to characterize one high-affinity inhibitor, resulting in considerable consumption of both sample material and time. Here, we introduce a fast and efficient ITC displacement assay that allows for the simultaneous characterization of both a high-affinity ligand and a moderate-affinity ligand competing for the same binding site on a receptor within a single experiment. The protocol is based on a titration of the high-affinity ligand into a solution containing the moderate-affinity ligand bound to the receptor present in excess. The resulting biphasic binding isotherm enables accurate and precise determination of KD values and binding enthalpies (ΔH) of both ligands. We discuss the theoretical background underlying the approach, demonstrate its practical application to metal ion chelation, explore its potential and limitations with the aid of simulations and statistical analyses, and elaborate on potential applications to protein-inhibitor interactions. Copyright © 2014 Elsevier Inc. All rights reserved.

  9. Isolation of Anti-Ricin Protective Antibodies Exhibiting High Affinity from Immunized Non-Human Primates

    Directory of Open Access Journals (Sweden)

    Tal Noy-Porat

    2016-03-01

    Full Text Available Ricin, derived from the castor bean plant Ricinus communis, is one of the most potent and lethal toxins known, against which there is no available antidote. To date, the use of neutralizing antibodies is the most promising post-exposure treatment for ricin intoxication. The aim of this study was to isolate high affinity anti-ricin antibodies that possess potent toxin-neutralization capabilities. Two non-human primates were immunized with either a ricin-holotoxin- or subunit-based vaccine, to ensure the elicitation of diverse high affinity antibodies. By using a comprehensive set of primers, immune scFv phage-displayed libraries were constructed and panned. A panel of 10 antibodies (five directed against the A subunit of ricin and five against the B subunit was isolated and reformatted into a full-length chimeric IgG. All of these antibodies were found to neutralize ricin in vitro, and several conferred full protection to ricin-intoxicated mice when given six hours after exposure. Six antibodies were found to possess exceptionally high affinity toward the toxin, with KD values below pM (koff < 1 × 10−7 s−1 that were well correlated with their ability to neutralize ricin. These antibodies, alone or in combination, could be used for the development of a highly-effective therapeutic preparation for post-exposure treatment of ricin intoxication.

  10. Proadifen-sensitive high affinity binding of 3H-alaproclate to liver membranes

    International Nuclear Information System (INIS)

    Ross, S.B.

    1987-01-01

    3 H-alaproclate, a selective 5 h ydroxytryptamine uptake inhibitor, was found to bind to microsomal membranes from the rat liver with high affinity (K D -=3 nM) and large capacity (B max about 2 nmol/g liver). This binding was stereoselective since S-( - )-alaproclate was 30 times more potent than the R-( + )-enantiomer to displace the 3 H-labelled racemate. Proadifen (SKF 525A), an inhibitor of cytochrome P-450, displaced the 3 H-alaproclate binding with the same, high affinity (K i =3 nM) as alaproclate itself. Repeated treatment with phenobarbital sodium (5x75 mg/kg intraperitoneally) increased the number of alaproclate binding sites 7-8 times without changing the affinity. However, most of the phenobarbital induced 3 H-alaproclate binding was not displaceable by proadifen, showing the presence of at least two different high affinity binding sites. The possible involvement of cytochrome P-450 in the alaproclate binding is discussed. (author)

  11. Proadifen-sensitive high affinity binding of /sup 3/H-alaproclate to liver membranes

    Energy Technology Data Exchange (ETDEWEB)

    Ross, S.B.

    1987-01-01

    /sup 3/H-alaproclate, a selective 5/sub h/ydroxytryptamine uptake inhibitor, was found to bind to microsomal membranes from the rat liver with high affinity (K/sub D/-=3 nM) and large capacity (B/sub max/ about 2 nmol/g liver). This binding was stereoselective since S-( - )-alaproclate was 30 times more potent than the R-( + )-enantiomer to displace the /sup 3/H-labelled racemate. Proadifen (SKF 525A), an inhibitor of cytochrome P-450, displaced the /sup 3/H-alaproclate binding with the same, high affinity (K/sub i/=3 nM) as alaproclate itself. Repeated treatment with phenobarbital sodium (5x75 mg/kg intraperitoneally) increased the number of alaproclate binding sites 7-8 times without changing the affinity. However, most of the phenobarbital induced /sup 3/H-alaproclate binding was not displaceable by proadifen, showing the presence of at least two different high affinity binding sites. The possible involvement of cytochrome P-450 in the alaproclate binding is discussed.

  12. 100-Gb/s Transmission Over a 2520-km Integrated MCF System Using Cladding-Pumped Amplifiers

    DEFF Research Database (Denmark)

    Castro, Carlos; Jain, Saurabh; De Man, Erik

    2017-01-01

    A 10.5-Tb/s optical transmission (15 x 100 Gb/s QPSK channels per core) over 2520 km of multicore fiber is achieved using an integrated multicore transmission link consisting of directly spliced multicore components, such as fan-in/fan-out fiber couplers, a 60-km trench-assisted seven-core hexago...

  13. The Trigger and Data Acquisition System for the 8 tower subsystem of the KM3NeT detector

    Energy Technology Data Exchange (ETDEWEB)

    Manzali, M., E-mail: matteo.manzali@cnaf.infn.it [INFN CNAF, Bologna (Italy); Università degli Studi di Ferrara, Ferrara (Italy); Chiarusi, T. [INFN BO, Bologna (Italy); Favaro, M. [INFN BO, Bologna (Italy); INFN CNAF, Bologna (Italy); Università degli Studi di Ferrara, Ferrara (Italy); Giacomini, F. [INFN CNAF, Bologna (Italy); Margiotta, A.; Pellegrino, C. [INFN BO, Bologna (Italy); Dipartimento di Fisica e Astronomia, Università degli Studi di Bologna, Bologna (Italy)

    2016-07-11

    KM3NeT is a deep-sea research infrastructure being constructed in the Mediterranean Sea. It will host a large Cherenkov neutrino telescope that will collect photons emitted along the path of the charged particles produced in neutrino interactions in the vicinity of the detector. The philosophy of the DAQ system of the detector foresees that all data are sent to shore after a proper sampling of the photomultiplier signals. No off-shore hardware trigger is implemented and a software selection of the data is performed with an on-line Trigger and Data Acquisition System (TriDAS) to reduce the large throughput due to the environmental light background. A first version of the TriDAS has been developed to operate a prototype detection unit deployed in March 2013 in the abyssal site of Capo Passero (Sicily, Italy), about 3500 m deep. A revised and improved version has been developed to meet the requirements of the final detector, using new tools and modern design solutions. First installation and scalability tests have been performed at the Bologna Common Infrastructure and results comparable to what expected have been observed.

  14. Glucose uptake and growth of glucose-limited chemostat cultures of Aspergillus niger and a disruptant lacking MstA, a high-affinity glucose transporter

    DEFF Research Database (Denmark)

    Jørgensen, Thomas R; vanKuyk, Patricia A; Poulsen, Bjarne R

    2007-01-01

    This is a study of high-affinity glucose uptake in Aspergillus niger and the effect of disruption of a high-affinity monosaccharide-transporter gene, mstA. The substrate saturation constant (K(s)) of a reference strain was about 15 microM in glucose-limited chemostat culture. Disruption of mst......-affinity uptake system of A. niger. The mstA disruptant and a reference strain were cultivated in glucose-limited chemostat cultures at low, intermediate and high dilution rate (D=0.07 h(-1), 0.14 h(-1) and 0.20 h(-1)). Mycelium harvested from steady-state cultures was subjected to glucose uptake assays...

  15. a Borehole Seismic System for Active and Passive Seimsic Studies to 3 KM at Ptrc's Aquistore Project

    Science.gov (United States)

    Schmitt, D. R.; Nixon, C.; Kofman, R.; White, D. J.; Worth, K.

    2015-12-01

    We have constructed a downhole seismic recording system for application to depths of nearly 3 km and temperatures up to 135 °C at Aquistore, an independent research and monitoring project in which liquid CO2 is being stored in a brine and sandstone water formation. The key component to this system is a set of commercially available slim-hole 3-C sondes carrying 15 Hz geophones deployable in open and cased boreholes with diameters as small as 57 mm. The system is currently hosted on a 4-conductor wireline with digital information streamed to the surface recording unit. We have further incorporated these sondes into a mobile passive monitoring unit that includes a number of redundancies such as a multiple Tbyte network accessible RAID hard-drive system (NAS) and a self-designed uninterruptible power supply. The system can be remotely controlled via the internet. The system is currently deployed covering a range of depths from 2850 m to 2910 m. Ambient temperatures at this depth are approximately 110 °C with onboard tool temperatures running at 115 °C. Data is continuously streamed to the NAS for archiving, approximately 11 GBytes of data is recorded per day at the sampling period of 0.5 ms. The lack of noise at this depth allows short data snippets to be flagged with a simple amplitude threshold criteria. The greatly reduced data volume of the snippets allows for ready access via the internet to the system for ongoing quality control. Spurious events, mostly small amplitude tube waves originating at or near the surface, are readily discounted. Active seismic measurements are carried out simultaneously but these require that an appropriately accurate independent GPS based time synchronization be used. Various experiences with event detection, orientation of sondes using both explosives and seismic vibrator, potential overheating of the surface electronics, and issues related to loss of shore power provide for a detailed case study. Aquistore, managed by the

  16. Simplified demultiplexing scheme for two PDM-IM/DD systems utilizing a single Stokes analyzer over 25-km SMF.

    Science.gov (United States)

    Pan, Yan; Yan, Lianshan; Yi, Anlin; Jiang, Lin; Pan, Wei; Luo, Bin; Zou, Xihua

    2017-10-15

    We propose a four-linear state of polarization multiplexed intensity modulation and direct detection (IM/DD) scheme based on two orthogonal polarization division multiplexing (PDM) on-off keying systems. We also experimentally demonstrate a simple demultiplexing algorithm for this scheme by utilizing only a single Stokes analyzer. At the rate of 4×10  Gbit/s, the experimental results show that the power penalty of the proposed scheme is about 1.5 dB, compared to the single PDM-IM/DD for back-to-back (B2B) transmission. Compared to B2B, just about 1.7 dB power penalty is required after 25 km Corning LEAF optical fiber transmission. Meanwhile, the performance of the polarization tracking is evaluated, and the results show that the BER fluctuation is less than 0.5 dB with a polarization scrambling rate up to 708.75 deg/s.

  17. Tropical Cyclones in the 7km NASA Global Nature Run for use in Observing System Simulation Experiments

    Science.gov (United States)

    Reale, Oreste; Achuthavarier, Deepthi; Fuentes, Marangelly; Putman, William M.; Partyka, Gary

    2018-01-01

    The National Aeronautics and Space Administration (NASA) Nature Run (NR), released for use in Observing System Simulation Experiments (OSSEs), is a 2-year long global non-hydrostatic free-running simulation at a horizontal resolution of 7 km, forced by observed sea-surface temperatures (SSTs) and sea ice, and inclusive of interactive aerosols and trace gases. This article evaluates the NR with respect to tropical cyclone (TC) activity. It is emphasized that to serve as a NR, a long-term simulation must be able to produce realistic TCs, which arise out of realistic large-scale forcings. The presence in the NR of the realistic, relevant dynamical features over the African Monsoon region and the tropical Atlantic is confirmed, along with realistic African Easterly Wave activity. The NR Atlantic TC seasons, produced with 2005 and 2006 SSTs, show interannual variability consistent with observations, with much stronger activity in 2005. An investigation of TC activity over all the other basins (eastern and western North Pacific, North and South Indian Ocean, and Australian region), together with relevant elements of the atmospheric circulation, such as, for example, the Somali Jet and westerly bursts, reveals that the model captures the fundamental aspects of TC seasons in every basin, producing realistic number of TCs with realistic tracks, life spans and structures. This confirms that the NASA NR is a very suitable tool for OSSEs targeting TCs and represents an improvement with respect to previous long simulations that have served the global atmospheric OSSE community. PMID:29674806

  18. Effect of quinolinic acid in the nucleus basalis magnocellularis on cortical high-affinity choline uptake

    Energy Technology Data Exchange (ETDEWEB)

    Metcalf, R.H.; Boegman, R.J.; Quirion, R.; Riopelle, R.J.; Ludwin, S.K.

    1987-08-01

    A transient 45% increase in cortical high-affinity choline uptake (HACU) was observed after an injection of quinolinic acid (QUIN) into the nucleus basalis magnocellularis (nbM) of the rat. This was followed by a steady decline in choline uptake, which resulted in a 46% decrease by day 7. Specific (/sup 3/H)hemicholinium-3 binding to coronal brain sections showed a similar pattern following injections of QUIN into the nbM. The increase in cortical HACU elicited by QUIN appeared to be dose dependent.

  19. Characteristics of high affinity and low affinity adenosine binding sites in human cerebral cortex

    International Nuclear Information System (INIS)

    John, D.; Fox, I.V.

    1986-01-01

    The binding characteristics of human brain cortical membrane fractions were evaluated to test the hypothesis that there are A 1 and A 2 adenosine binding sites. The ligands used were 2-chloro(8- 3 H) adenosine and N 6 -(adenine-2, 8- 3 H) cyclohexayladenosine. Binding of chloroadenosine to human brain cortical membranes was time dependent, reversible and concentration dependent. The kinetic constant determinations from binding studies of the adenosine receptor are presented. Utilizing tritium-cyclohexyladenosine as ligand the authors observed evidence for a high affinity binding site in human brain cortical membranes with a kd of 5 nM

  20. Novel and high affinity fluorescent ligands for the serotonin transporter based on (s)-citalopram

    DEFF Research Database (Denmark)

    Kumar, Vivek; Rahbek-Clemmensen, Troels; Billesbølle, Christian B

    2014-01-01

    Novel rhodamine-labeled ligands, based on (S)-citalopram, were synthesized and evaluated for uptake inhibition at the human serotonin, dopamine, and norepinephrine transporters (hSERT, hDAT, and hNET, respectively) and for binding at SERT, in transiently transfected COS7 cells. Compound 14 demons...... demonstrated high affinity binding and selectivity for SERT (K i = 3 nM). Visualization of SERT, using confocal laser scanning microscopy, validated compound 14 as a novel tool for studying SERT expression and distribution in living cells....

  1. High affinity binding of [3H]cocaine to rat liver microsomes

    International Nuclear Information System (INIS)

    El-Maghrabi, E.A.; Calligaro, D.O.; Eldefrawi, M.E.

    1988-01-01

    ] 3 H]cocaine bound reversible, with high affinity and stereospecificity to rat liver microsomes. Little binding was detected in the lysosomal, mitochondrial and nuclear fractions. The binding kinetics were slow and the kinetically calculated K/sub D/ was 2 nM. Induction of mixed function oxidases by phenobarbital did not produce significant change in [ 3 H]cocaine binding. On the other hand, chronic administration of cocaine reduced [ 3 H]cocaine binding drastically. Neither treatment affected the affinity of the liver binding protein for cocaine. Microsomes from mouse and human livers had less cocaine-binding protein and lower affinity for cocaine than those from rat liver. Binding of [ 3 H]cocaine to rat liver microsomes was insensitive to monovalent cations and > 10 fold less sensitive to biogenic amines than the cocaine receptor in rat striatum. However, the liver protein had higher affinity for cocaine and metabolites except for norcocaine. Amine uptake inhibitors displaced [ 3 H]cocaine binding to liver with a different rank order of potency than their displacement of [ 3 H]cocaine binding to striatum. This high affinity [ 3 H]cocaine binding protein in liver is not likely to be monooxygenase, but may have a role in cocaine-induced hepatotoxicity

  2. A high affinity monoclonal antibody recognizing the light chain of human coagulating factor VII.

    Science.gov (United States)

    Sarial, Sheila; Asadi, Farzad; Jeddi-Tehrani, Mahmood; Hadavi, Reza; Bayat, Ali Ahmad; Mahmoudian, Jafar; Taghizadeh-Jahed, Masoud; Shokri, Fazel; Rabbani, Hodjattallah

    2012-12-01

    Factor VII (FVII) is a serine protease-coagulating element responsible for the initiation of an extrinsic pathway of clot formation. Here we generated and characterized a high affinity monoclonal antibody that specifically recognizes human FVII. Recombinant human FVII (rh-FVII) was used for the production of a monoclonal antibody using BALB/c mice. The specificity of the antibody was determined by Western blot using plasma samples from human, mouse, sheep, goat, bovine, rabbit, and rat. Furthermore, the antibody was used to detect transiently expressed rh-FVII in BHK21 cell line using Western blot and sandwich ELISA. A mouse IgG1 (kappa chain) monoclonal antibody clone 1F1-B11 was produced against rh-FVII. The affinity constant (K(aff)) of the antibody was calculated to be 6.4×10(10) M(-1). The antibody could specifically recognize an epitope on the light chain of hFVII, with no reactivity with factor VII from several other animals. In addition, transiently expressed rh-FVII in BHK21 cells was recognized by 1F1-B11. The high affinity as well as the specificity of 1F1-B11 for hFVII will facilitate the affinity purification of hFVII and also production of FVII deficient plasma and minimizes the risk of bovine FVII contamination when fetal bovine serum-supplemented media are used for production and subsequent purification of rh-FVII.

  3. Acylated heptapeptide binds albumin with high affinity and application as tag furnishes long-acting peptides.

    Science.gov (United States)

    Zorzi, Alessandro; Middendorp, Simon J; Wilbs, Jonas; Deyle, Kaycie; Heinis, Christian

    2017-07-17

    The rapid renal clearance of peptides in vivo limits this attractive platform for the treatment of a broad range of diseases that require prolonged drug half-lives. An intriguing approach for extending peptide circulation times works through a 'piggy-back' strategy in which peptides bind via a ligand to the long-lived serum protein albumin. In accordance with this strategy, we developed an easily synthesized albumin-binding ligand based on a peptide-fatty acid chimera that has a high affinity for human albumin (K d =39 nM). This ligand prolongs the elimination half-life of cyclic peptides in rats 25-fold to over seven hours. Conjugation to a peptide factor XII inhibitor developed for anti-thrombotic therapy extends the half-life from 13 minutes to over five hours, inhibiting coagulation for eight hours in rabbits. This high-affinity albumin ligand could potentially extend the half-life of peptides in human to several days, substantially broadening the application range of peptides as therapeutics.

  4. High-affinity receptors for bombesin-like peptides in normal guinea pig lung membranes

    International Nuclear Information System (INIS)

    Lach, E.; Trifilieff, A.; Landry, Y.; Gies, J.P.

    1991-01-01

    The binding of the radiolabeled bombesin analogue [ 125 I-Tyr 4 ]bombesin to guinea-pig lung membranes was investigated. Binding of [ 125 I-Tyr 4 ]bombesin was specific, saturable, reversible and linearly related to the protein concentration. Scatchard analysis of equilibrium binding data at 25C indicated the presence of a single class of non-interacting binding sites for bombesin (B max = 7.7 fmol/mg protein). The value of the equilibrium dissociation constant (K D = 90 pM) agrees with a high-affinity binding site. Bombesin and structurally related peptides such as [ 125 I-Tyr 4 ]bombesin, neuromedin B and neuromedin C inhibited the binding of [ 125 I-Tyr 4 ]bombesin in an order of potencies as follows: [ 125 I-Tyr 4 ]bombesin > bombesin ≥ neuromedin C much-gt neuromedin B. These results indicate that guinea-pig lung membranes possess a single class of bombesin receptors with a high affinity for bombesin and a lower one for neuromedin B

  5. Reconstitution of high affinity α2 adrenergic agonist binding by fusion with a pertussis toxin substrate

    International Nuclear Information System (INIS)

    Kim, M.H.; Neubig, R.R.

    1986-01-01

    High affinity α 2 adrenergic agonist binding is thought to occur via a coupling of the α 2 receptor with N/sub i/, the inhibitory guanyl nucleotide binding protein. Human platelet membranes pretreated at pH 11.5 exhibit a selective inactivation of agonist binding and N/sub i/. To further study the mechanism of agonist binding, alkali treated membranes (ATM) were mixed with membranes pretreated with 10 μM phenoxybenzamine to block α 2 receptors (POB-M). The combined membrane pellet was incubated in 50% polyethylene glycol (PEG) to promote membrane-membrane fusion and assayed for binding to the α 2 agonist [ 3 H]UK 14,304 (UK) and the antagonist [ 3 H] yohimbine. PEG treatment resulted in a 2-4 fold enhancement of UK binding whereas yohimbine binding was unchanged. No enhancement of UK binding was observed in the absence of PEG treatment. The reconstitution was dependent on the addition of POB-M. They found that a 1:1 ratio of POB-M:ATM was optimal. Reconstituted binding was inhibited by GppNHp. Fusion of rat C6 glioma cell membranes, which do not contain α 2 receptors, also enhanced agonist binding to ATM. Fusion of C6 membranes from cells treated with pertussis toxin did not enhance [ 3 H] UK binding. These data show that a pertussis toxin sensitive membrane component, possibly N/sub i/, can reconstitute high affinity α 2 agonist binding

  6. Acylated heptapeptide binds albumin with high affinity and application as tag furnishes long-acting peptides

    Science.gov (United States)

    Zorzi, Alessandro; Middendorp, Simon J.; Wilbs, Jonas; Deyle, Kaycie; Heinis, Christian

    2017-07-01

    The rapid renal clearance of peptides in vivo limits this attractive platform for the treatment of a broad range of diseases that require prolonged drug half-lives. An intriguing approach for extending peptide circulation times works through a `piggy-back' strategy in which peptides bind via a ligand to the long-lived serum protein albumin. In accordance with this strategy, we developed an easily synthesized albumin-binding ligand based on a peptide-fatty acid chimera that has a high affinity for human albumin (Kd=39 nM). This ligand prolongs the elimination half-life of cyclic peptides in rats 25-fold to over seven hours. Conjugation to a peptide factor XII inhibitor developed for anti-thrombotic therapy extends the half-life from 13 minutes to over five hours, inhibiting coagulation for eight hours in rabbits. This high-affinity albumin ligand could potentially extend the half-life of peptides in human to several days, substantially broadening the application range of peptides as therapeutics.

  7. Isolation and characterization of high affinity aptamers against DNA polymerase iota.

    Science.gov (United States)

    Lakhin, Andrei V; Kazakov, Andrei A; Makarova, Alena V; Pavlov, Yuri I; Efremova, Anna S; Shram, Stanislav I; Tarantul, Viacheslav Z; Gening, Leonid V

    2012-02-01

    Human DNA-polymerase iota (Pol ι) is an extremely error-prone enzyme and the fidelity depends on the sequence context of the template. Using the in vitro systematic evolution of ligands by exponential enrichment (SELEX) procedure, we obtained an oligoribonucleotide with a high affinity to human Pol ι, named aptamer IKL5. We determined its dissociation constant with homogenous preparation of Pol ι and predicted its putative secondary structure. The aptamer IKL5 specifically inhibits DNA-polymerase activity of the purified enzyme Pol ι, but did not inhibit the DNA-polymerase activities of human DNA polymerases beta and kappa. IKL5 suppressed the error-prone DNA-polymerase activity of Pol ι also in cellular extracts of the tumor cell line SKOV-3. The aptamer IKL5 is useful for studies of the biological role of Pol ι and as a potential drug to suppress the increase of the activity of this enzyme in malignant cells.

  8. Solution structure of the Grb2 SH2 domain complexed with a high-affinity inhibitor

    International Nuclear Information System (INIS)

    Ogura, Kenji; Shiga, Takanori; Yokochi, Masashi; Yuzawa, Satoru; Burke, Terrence R.; Inagaki, Fuyuhiko

    2008-01-01

    The solution structure of the growth factor receptor-bound protein 2 (Grb2) SH2 domain complexed with a high-affinity inhibitor containing a non-phosphorus phosphate mimetic within a macrocyclic platform was determined by nuclear magnetic resonance (NMR) spectroscopy. Unambiguous assignments of the bound inhibitor and intermolecular NOEs between the Grb2 SH2 domain and the inhibitor was accomplished using perdeuterated Grb2 SH2 protein. The well-defined solution structure of the complex was obtained and compared to those by X-ray crystallography. Since the crystal structure of the Grb2 SH2 domain formed a domain-swapped dimer and several inhibitors were bound to a hinge region, there were appreciable differences between the solution and crystal structures. Based on the binding interactions between the inhibitor and the Grb2 SH2 domain in solution, we proposed a design of second-generation inhibitors that could be expected to have higher affinity

  9. Haemoglobin Pierre-Benite--a high affinity variant associated with relative polycythaemia.

    Science.gov (United States)

    Beard, M E; Potter, H C; Spearing, R L; Brennan, S O

    2001-12-01

    This is the second reported example of Hb Pierre--Benite (beta90 Glu-->Asp). This mutation is associated with increased oxygen affinity and polycythaemia. No instability was found and there was no charge shift detected by cellulose acetate electrophoresis at pH 8.3. The mutation was however, clearly indicated by electrospray ionization mass spectrometry (ESI MS), which showed an abnormal beta chain with a 14 Da decrease in mass. Blood volume studies documented a relative rather than a true polycythaemia and this finding has been reported in at least two other high affinity haemoglobin variants--Hb Heathrow and Hb Rahere. This finding led to delay in diagnosis because high oxygen affinity variants are conventionally considered to cause a true polycythaemia.

  10. New immunogenic form for vasopressin: production of high-affinity antiserum and RIA for plasmatic AVP

    International Nuclear Information System (INIS)

    Rougon-Rappuzi, G.; Delaage, M.A.; Conte-Devolx, B.; Millet, Y.

    1977-01-01

    A highly sensitive and specific radioimmunoassay (RIA) for arginine-vasopressin (AVP) was developped and applied to the measurement of AVP in human plasma. High-affinity antivasopressin antibodies with limited association constant heterogeneity have been induced by immunizing rabbits with Lysine-vasopressine (LVP) coupled to a human immunoglobulin (IgA). Replacing air drying of acetone-petroleum ether extracts by lyophilisation increased significantly the yields of AVP. Equilibrium dialysis was used for separating bound and free antigen, thus reducing the total time required for the assay to 48 hours. Only 1 ml of plasma was required for routine determinations due to a sensitivity threshold better than 0.5 pg/ml. Plasma AVP levels of normal subjects and of patients with inappropriate ADH secretion (SIADH) were determined during different hydratation states and following nicotin of ethanol infusions. (orig.) [de

  11. Effects of anticonvulsants in vivo on high affinity choline uptake in vitro in mouse hippocampal synaptosomes.

    Science.gov (United States)

    Miller, J. A.; Richter, J. A.

    1985-01-01

    The effects of several anticonvulsant drugs on sodium-dependent high affinity choline uptake (HACU) in mouse hippocampal synaptosomes was investigated. HACU was measured in vitro after in vivo administration of the drug to mice. HACU was inhibited by drugs which have in common the ability to facilitate gamma-aminobutyric acid (GABA) transmission, pentobarbitone, phenobarbitone, barbitone, diazepam, chloridiazepoxide, and valproic acid. Dose-response relationships were determined for these drugs and the drugs' potencies at inhibiting HACU correlated well with their anticonvulsant potencies. Clonazepam, ethosuximide, carbamazepine, and barbituric acid had no effect on HACU in the doses used while phenytoin and trimethadione stimulated HACU. These results suggest that certain anticonvulsants may elicit a part of their anticonvulsant activity by modulating cholinergic neurones. This effect may be mediated through a GABA mechanism. PMID:3978310

  12. Production and Identification of High Affinity Monoclonal Antibodies Against Pesticide Carbofuran

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    To produce high-affinity monoclonal antibodies against pesticide carbofuran, and the develop immunochemical assays for people's health and environmental protection, the hapten 4-[[(2,3-dihydro-2,2-dimethyl-7-benzofuranyloxy) carbonyl]-amino]-butanoic acid (BFNB) of carbofuran was synthesized and Balb/c mice were immunized by the hapten-carrier (BFNB-bovine serum albumin, BFNB-BSA) conjugates. The splenocytes of immunized mice were fused with Sp2/0 cells and the cultural supernatants of hybridoma cells were screened by the indirect enzyme-linked immunoabsorbent assay (ELISA), based on BFNB-ovoalbumin conjugates (BFNB-OVA). Purified monoclonal antibody (McAb) was obtained from fluids of ascites, deposited by octanoic acid and ammonium sulfate. The affinity and the specificity of McAb were characterized by ELISA or indirect competitive ELISA. A hybridoma cell line (5D3) secreting anti-carbofuran McAb had been established. The titer of culture medium and ascites was up to 1:2.048 × 103 and 1:1.024 × 106, respectively, and the subtype of the McAb was IgG1. The affinity constant of the McAb was about 2.54 × 109 L mol-1, with an IC50 value of 1.18 ng mL-1 and a detection limit of 0.01 ng mL-1. Cross-reactivity studies showed that the McAb was quiet specific for carbofuran, as among the four analogous compounds, they were all hardly recognized (4.59 × 10-4% for 2,3-dihydro-2,2-dimethyl-7-benzofuranol and less than 3.0 × 10-4% for others). The prepared McAb had a very high affinity and specificity,and it could be used to develop ELISA for rapid determination of carbofuran.

  13. High Affinity IgE-Fc Receptor alpha and gamma Subunit Interactions

    International Nuclear Information System (INIS)

    Rashid, A.; Housden, J. E. M.; Sabban, S.; Helm, B.

    2014-01-01

    Objective: To explore the relationships between the subunits (alpha, beta and gamma) of the high affinity IgE receptor (Fc and RI) and its ability to mediate transmembrane signaling. Study Design: Experimental study. Place and Duration of Study: Department of Molecular Biology and Biotechnology, University of Sheffield, UK, from 2008 to 2009. Methodology: The approach employed was to create a chimera (human alpha-gamma-gamma) using the extracellular (EC) domain of the human high affinity IgE receptor. The alpha subunit (huFc and RIalpha) of IgE receptor was spliced onto the rodent gamma TM and cytoplasmic domain (CD). This was transfected into the Rat Basophilic Leukemia cell line in order to assess the possibility of selectively activating cells transfected with this single pass construct for antigen induced mediator release. Results: The RBLs cell lines transfected with the huFc and RIalpha/gamma/gamma cDNA constructs were assessed for the cell surface expression of the huFc and RIalpha subunit and the response to the antigenic stimulus by looking for degranulation and intracellular Ca2+ mobilisation. The results obtained showed the absence of huFc and RIalpha subunit expression on the surface of transfected cells as seen by flowcytometric studies, beta-hexosaminidase assays and intracellular calcium mobilisation studies. Conclusion: In the present study the grounds for non-expression of huFc and RIalpha/gamma/gamma cDNA remains elusive but may be due to the fact that the human-rodent chimeric receptors are assembled differently than the endogenous rodent receptors as seen in study in which COS 7 cells were transfected with human/rat chimeric complexes. (author)

  14. Selection of DNA aptamers against epidermal growth factor receptor with high affinity and specificity

    International Nuclear Information System (INIS)

    Wang, Deng-Liang; Song, Yan-Ling; Zhu, Zhi; Li, Xi-Lan; Zou, Yuan; Yang, Hai-Tao; Wang, Jiang-Jie; Yao, Pei-Sen; Pan, Ru-Jun; Yang, Chaoyong James; Kang, De-Zhi

    2014-01-01

    Highlights: • This is the first report of DNA aptamer against EGFR in vitro. • Aptamer can bind targets with high affinity and selectivity. • DNA aptamers are more stable, cheap and efficient than RNA aptamers. • Our selected DNA aptamer against EGFR has high affinity with K d 56 ± 7.3 nM. • Our selected DNA aptamer against EGFR has high selectivity. - Abstract: Epidermal growth factor receptor (EGFR/HER1/c-ErbB1), is overexpressed in many solid cancers, such as epidermoid carcinomas, malignant gliomas, etc. EGFR plays roles in proliferation, invasion, angiogenesis and metastasis of malignant cancer cells and is the ideal antigen for clinical applications in cancer detection, imaging and therapy. Aptamers, the output of the systematic evolution of ligands by exponential enrichment (SELEX), are DNA/RNA oligonucleotides which can bind protein and other substances with specificity. RNA aptamers are undesirable due to their instability and high cost of production. Conversely, DNA aptamers have aroused researcher’s attention because they are easily synthesized, stable, selective, have high binding affinity and are cost-effective to produce. In this study, we have successfully identified DNA aptamers with high binding affinity and selectivity to EGFR. The aptamer named TuTu22 with K d 56 ± 7.3 nM was chosen from the identified DNA aptamers for further study. Flow cytometry analysis results indicated that the TuTu22 aptamer was able to specifically recognize a variety of cancer cells expressing EGFR but did not bind to the EGFR-negative cells. With all of the aforementioned advantages, the DNA aptamers reported here against cancer biomarker EGFR will facilitate the development of novel targeted cancer detection, imaging and therapy

  15. Antibody Binding Selectivity: Alternative Sets of Antigen Residues Entail High-Affinity Recognition.

    Directory of Open Access Journals (Sweden)

    Yves Nominé

    Full Text Available Understanding the relationship between protein sequence and molecular recognition selectivity remains a major challenge. The antibody fragment scFv1F4 recognizes with sub nM affinity a decapeptide (sequence 6TAMFQDPQER15 derived from the N-terminal end of human papilloma virus E6 oncoprotein. Using this decapeptide as antigen, we had previously shown that only the wild type amino-acid or conservative replacements were allowed at positions 9 to 12 and 15 of the peptide, indicating a strong binding selectivity. Nevertheless phenylalanine (F was equally well tolerated as the wild type glutamine (Q at position 13, while all other amino acids led to weaker scFv binding. The interfaces of complexes involving either Q or F are expected to diverge, due to the different physico-chemistry of these residues. This would imply that high-affinity binding can be achieved through distinct interfacial geometries. In order to investigate this point, we disrupted the scFv-peptide interface by modifying one or several peptide positions. We then analyzed the effect on binding of amino acid changes at the remaining positions, an altered susceptibility being indicative of an altered role in complex formation. The 23 starting variants analyzed contained replacements whose effects on scFv1F4 binding ranged from minor to drastic. A permutation analysis (effect of replacing each peptide position by all other amino acids except cysteine was carried out on the 23 variants using the PEPperCHIP® Platform technology. A comparison of their permutation patterns with that of the wild type peptide indicated that starting replacements at position 11, 12 or 13 modified the tolerance to amino-acid changes at the other two positions. The interdependence between the three positions was confirmed by SPR (Biacore® technology. Our data demonstrate that binding selectivity does not preclude the existence of alternative high-affinity recognition modes.

  16. Selection of DNA aptamers against epidermal growth factor receptor with high affinity and specificity

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Deng-Liang [The First Clinical Medical College of Fujian Medical University, Fuzhou (China); Department of Neurosurgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou (China); Song, Yan-Ling; Zhu, Zhi; Li, Xi-Lan; Zou, Yuan [State Key Laboratory for Physical Chemistry of Solid Surfaces, Key Laboratory for Chemical Biology of Fujian Province, Key Laboratory of Analytical Chemistry, and Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005 (China); Yang, Hai-Tao; Wang, Jiang-Jie [The First Clinical Medical College of Fujian Medical University, Fuzhou (China); Yao, Pei-Sen [Department of Neurosurgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou (China); Pan, Ru-Jun [The First Clinical Medical College of Fujian Medical University, Fuzhou (China); Yang, Chaoyong James, E-mail: cyyang@xmu.edu.cn [State Key Laboratory for Physical Chemistry of Solid Surfaces, Key Laboratory for Chemical Biology of Fujian Province, Key Laboratory of Analytical Chemistry, and Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005 (China); Kang, De-Zhi, E-mail: kdzy99988@163.com [The First Clinical Medical College of Fujian Medical University, Fuzhou (China); Department of Neurosurgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou (China)

    2014-10-31

    Highlights: • This is the first report of DNA aptamer against EGFR in vitro. • Aptamer can bind targets with high affinity and selectivity. • DNA aptamers are more stable, cheap and efficient than RNA aptamers. • Our selected DNA aptamer against EGFR has high affinity with K{sub d} 56 ± 7.3 nM. • Our selected DNA aptamer against EGFR has high selectivity. - Abstract: Epidermal growth factor receptor (EGFR/HER1/c-ErbB1), is overexpressed in many solid cancers, such as epidermoid carcinomas, malignant gliomas, etc. EGFR plays roles in proliferation, invasion, angiogenesis and metastasis of malignant cancer cells and is the ideal antigen for clinical applications in cancer detection, imaging and therapy. Aptamers, the output of the systematic evolution of ligands by exponential enrichment (SELEX), are DNA/RNA oligonucleotides which can bind protein and other substances with specificity. RNA aptamers are undesirable due to their instability and high cost of production. Conversely, DNA aptamers have aroused researcher’s attention because they are easily synthesized, stable, selective, have high binding affinity and are cost-effective to produce. In this study, we have successfully identified DNA aptamers with high binding affinity and selectivity to EGFR. The aptamer named TuTu22 with K{sub d} 56 ± 7.3 nM was chosen from the identified DNA aptamers for further study. Flow cytometry analysis results indicated that the TuTu22 aptamer was able to specifically recognize a variety of cancer cells expressing EGFR but did not bind to the EGFR-negative cells. With all of the aforementioned advantages, the DNA aptamers reported here against cancer biomarker EGFR will facilitate the development of novel targeted cancer detection, imaging and therapy.

  17. A rhodium(III) complex for high-affinity DNA base-pair mismatch recognition

    Science.gov (United States)

    Junicke, Henrik; Hart, Jonathan R.; Kisko, Jennifer; Glebov, Oleg; Kirsch, Ilan R.; Barton, Jacqueline K.

    2003-01-01

    A rhodium(III) complex, rac-[Rh(bpy)2phzi]3+ (bpy, 2,2′-bipyridine; phzi, benzo[a]phenazine-5,6-quinone diimine) has been designed as a sterically demanding intercalator targeted to destabilized mismatched sites in double-helical DNA. The complex is readily synthesized by condensation of the phenazine quinone with the corresponding diammine complex. Upon photoactivation, the complex promotes direct strand scission at single-base mismatch sites within the DNA duplex. As with the parent mismatch-specific reagent, [Rh(bpy)2(chrysi)]3+ [chrysene-5,6-quinone diimine (chrysi)], mismatch selectivity depends on the helix destabilization associated with mispairing. Unlike the parent chrysi complex, the phzi analogue binds and cleaves with high affinity and efficiency. The specific binding constants for CA, CC, and CT mismatches within a 31-mer oligonucleotide duplex are 0.3, 1, and 6 × 107 M−1, respectively; site-specific photocleavage is evident at nanomolar concentrations. Moreover, the specificity, defined as the ratio in binding affinities for mispaired vs. well paired sites, is maintained. The increase in affinity is attributed to greater stability in the mismatched site associated with stacking by the heterocyclic aromatic ligand. The high-affinity complex is also applied in the differential cleavage of DNA obtained from cell lines deficient in mismatch repair vs. those proficient in mismatch repair. Agreement is found between photocleavage by the mismatch-specific probes and deficiency in mismatch repair. This mismatch-specific targeting, therefore, offers a potential strategy for new chemotherapeutic design. PMID:12610209

  18. Peptides in headlock – a novel high-affinity and versatile peptide-binding nanobody for proteomics and microscopy

    Science.gov (United States)

    Braun, Michael B.; Traenkle, Bjoern; Koch, Philipp A.; Emele, Felix; Weiss, Frederik; Poetz, Oliver; Stehle, Thilo; Rothbauer, Ulrich

    2016-01-01

    Nanobodies are highly valuable tools for numerous bioanalytical and biotechnical applications. Here, we report the characterization of a nanobody that binds a short peptide epitope with extraordinary affinity. Structural analysis reveals an unusual binding mode where the extended peptide becomes part of a β-sheet structure in the nanobody. This interaction relies on sequence-independent backbone interactions augmented by a small number of specificity-determining side chain contacts. Once bound, the peptide is fastened by two nanobody side chains that clamp it in a headlock fashion. Exploiting this unusual binding mode, we generated a novel nanobody-derived capture and detection system. Matrix-coupled nanobody enables the fast and efficient isolation of epitope-tagged proteins from prokaryotic and eukaryotic expression systems. Additionally, the fluorescently labeled nanobody visualizes subcellular structures in different cellular compartments. The high-affinity-binding and modifiable peptide tag of this system renders it a versatile and robust tool to combine biochemical analysis with microscopic studies. PMID:26791954

  19. Peptides in headlock--a novel high-affinity and versatile peptide-binding nanobody for proteomics and microscopy.

    Science.gov (United States)

    Braun, Michael B; Traenkle, Bjoern; Koch, Philipp A; Emele, Felix; Weiss, Frederik; Poetz, Oliver; Stehle, Thilo; Rothbauer, Ulrich

    2016-01-21

    Nanobodies are highly valuable tools for numerous bioanalytical and biotechnical applications. Here, we report the characterization of a nanobody that binds a short peptide epitope with extraordinary affinity. Structural analysis reveals an unusual binding mode where the extended peptide becomes part of a β-sheet structure in the nanobody. This interaction relies on sequence-independent backbone interactions augmented by a small number of specificity-determining side chain contacts. Once bound, the peptide is fastened by two nanobody side chains that clamp it in a headlock fashion. Exploiting this unusual binding mode, we generated a novel nanobody-derived capture and detection system. Matrix-coupled nanobody enables the fast and efficient isolation of epitope-tagged proteins from prokaryotic and eukaryotic expression systems. Additionally, the fluorescently labeled nanobody visualizes subcellular structures in different cellular compartments. The high-affinity-binding and modifiable peptide tag of this system renders it a versatile and robust tool to combine biochemical analysis with microscopic studies.

  20. System approach for evaluation of air pollution toxic compounds in the 30-km area of nuclear power plants

    International Nuclear Information System (INIS)

    Shevtsova, O.V.; Zhigunova, L.N.; Makovskaya, N.A.; Pavlovich, E.L.

    2012-01-01

    The article shows the importance of a systematic approach to address environmental problems that arise during the construction of nuclear power plants, and identified the need to consider the transformation and biotransformation of primary pollutants and monitoring secondary pollutants. The basic pathways of pollutants in the air a 30-km zone of nuclear power plants established. The content of primary and secondary air pollutants identified. The evaluation of general toxic risk from primary pollutant and the calculation of the carcinogenic risk of secondary pollutants entering the body by inhalation are carried out. (authors)

  1. System approach for evaluation of soil contamination by nitrogen containing pollutants in the 30-km zone power nuclear plant

    International Nuclear Information System (INIS)

    Shevtsova, O.V.; Zhigunova, L.N.; Makovskaya, N.A.; Pavlovich, E.L.

    2013-01-01

    This article presents the results of studies of the nitrogen-containing chemical pollutants of the soil 30 km zone nuclear power plant in order to evaluate the chemical carcinogenic risk to public health. As a result of this work the least amount of NDMA fixed at 0,83 × 10 -4 mg/kg, the highest - 128,29 × 10 -4 mg/kg, which is 0,83 and 128,3 %, respectively, of Madman established in the Russian Federation and Republic of Kazakhstan, the content of NDMA precursors - nitrite ions varies from 0,07 to 4,43 mg/kg. Found that most of the compounds reported in agricultural land. Calculate the total carcinogenic risk for the population living in the region - the set value corresponds to the maximum risk tolerance, i. e. the upper limit of acceptable risk. These levels are subject to continuous monitoring. (authors)

  2. Design, Synthesis, and Biological Evaluation of Small, High-Affinity Siglec-7 Ligands: Toward Novel Inhibitors of Cancer Immune Evasion.

    Science.gov (United States)

    Prescher, Horst; Frank, Martin; Gütgemann, Stephan; Kuhfeldt, Elena; Schweizer, Astrid; Nitschke, Lars; Watzl, Carsten; Brossmer, Reinhard

    2017-02-09

    Natural killer cells are able to directly lyse tumor cells, thereby participating in the immune surveillance against cancer. Unfortunately, many cancer cells use immune evasion strategies to avoid their eradication by the immune system. A prominent escape strategy of malignant cells is to camouflage themselves with Siglec-7 ligands, thereby recruiting the inhibitory receptor Siglec-7 expressed on the NK cell surface which subsequently inhibits NK-cell-mediated lysis. Here we describe the synthesis and evaluation of the first, high-affinity low molecular weight Siglec-7 ligands to interfere with cancer cell immune evasion. The compounds are Sialic acid derivatives and bind with low micromolar K d values to Siglec-7. They display up to a 5000-fold enhanced affinity over the unmodified sialic acid scaffold αMe Neu5Ac, the smallest known natural Siglec-7 ligand. Our results provide a novel immuno-oncology strategy employing natural immunity in the fight against cancers, in particular blocking Siglec-7 with low molecular weight compounds.

  3. VNARs: An Ancient and Unique Repertoire of Molecules That Deliver Small, Soluble, Stable and High Affinity Binders of Proteins

    Directory of Open Access Journals (Sweden)

    Caroline Barelle

    2015-09-01

    Full Text Available At 420 million years, the variable domain of New Antigen Receptors or VNARs are undoubtedly the oldest (and smallest antigen binding single domains identified in the vertebrate kingdom. Their role as an integral part of the adaptive immune system of sharks has been well established and has served to provide a greater understanding of the evolution of humoral immunity; their cellular components and processes as well as the underlying genetic organization and molecular control mechanisms. Intriguingly, unlike the variable domain of the camelid heavy chain antibodies or VHH, VNARs do not conform to all of the characteristic properties of classical antibodies with an ancestral origin that clearly distinguishes them from true immunoglobulin antibodies. However, this uniqueness of their origin only adds to their potential as next generation therapeutic biologics with their structural and functional attributes and commercial freedom all enhancing their profile and current success. In fact their small size, remarkable stability, molecular flexibility and solubility, together with their high affinity and selectivity for target, all reinforce the potential of these domains as drug candidates. The purpose of this review is to provide an overview of the existing basic biology of these unique domains, to highlight the drug-like properties of VNARs and describe current progress in their journey towards the clinic.

  4. Visual and Plasmon Resonance Absorption Sensor for Adenosine Triphosphate Based on the High Affinity between Phosphate and Zr(IV)

    OpenAIRE

    Qi, Wenjing; Liu, Zhongyuan; Zhang, Wei; Halawa, Mohamed Ibrahim; Xu, Guobao

    2016-01-01

    Zr(IV) can form phosphate and Zr(IV) (?PO3 2??Zr4+?) complex owing to the high affinity between Zr(IV) with phosphate. Zr(IV) can induce the aggregation of gold nanoparticles (AuNPs), while adenosine triphosphate(ATP) can prevent Zr(IV)-induced aggregation of AuNPs. Herein, a visual and plasmon resonance absorption (PRA)sensor for ATP have been developed using AuNPs based on the high affinity between Zr(IV)with ATP. AuNPs get aggregated in the presence of certain concentrations of Zr(IV). Aft...

  5. New Synthesis and Tritium Labeling of a Selective Ligand for Studying High-affinity γ-Hydroxybutyrate (GHB) Binding Sites

    Science.gov (United States)

    Vogensen, Stine B.; Marek, Aleš; Bay, Tina; Wellendorph, Petrine; Kehler, Jan; Bundgaard, Christoffer; Frølund, Bente; Pedersen, Martin H.F.; Clausen, Rasmus P.

    2013-01-01

    3-Hydroxycyclopent-1-enecarboxylic acid (HOCPCA, 1) is a potent ligand for the high-affinity GHB binding sites in the CNS. An improved synthesis of 1 together with a very efficient synthesis of [3H]-1 is described. The radiosynthesis employs in situ generated lithium trimethoxyborotritide. Screening of 1 against different CNS targets establishes a high selectivity and we demonstrate in vivo brain penetration. In vitro characterization of [3H]-1 binding shows high specificity to the high-affinity GHB binding sites. PMID:24053696

  6. Fc-Binding Ligands of Immunoglobulin G: An Overview of High Affinity Proteins and Peptides

    Directory of Open Access Journals (Sweden)

    Weonu Choe

    2016-12-01

    Full Text Available The rapidly increasing application of antibodies has inspired the development of several novel methods to isolate and target antibodies using smart biomaterials that mimic the binding of Fc-receptors to antibodies. The Fc-binding domain of antibodies is the primary binding site for e.g., effector proteins and secondary antibodies, whereas antigens bind to the Fab region. Protein A, G, and L, surface proteins expressed by pathogenic bacteria, are well known to bind immunoglobulin and have been widely exploited in antibody purification strategies. Several difficulties are encountered when bacterial proteins are used in antibody research and application. One of the major obstacles hampering the use of bacterial proteins is sample contamination with trace amounts of these proteins, which can invoke an immune response in the host. Many research groups actively develop synthetic ligands that are able to selectively and strongly bind to antibodies. Among the reported ligands, peptides that bind to the Fc-domain of antibodies are attractive tools in antibody research. Besides their use as high affinity ligands in antibody purification chromatography, Fc-binding peptides are applied e.g., to localize antibodies on nanomaterials and to increase the half-life of proteins in serum. In this review, recent developments of Fc-binding peptides are presented and their binding characteristics and diverse applications are discussed.

  7. Structural insights into a high affinity nanobody:antigen complex by homology modelling.

    Science.gov (United States)

    Skottrup, Peter Durand

    2017-09-01

    Porphyromonas gingivalis is a major periodontitis-causing pathogens. P. gingivalis secrete a cysteine protease termed RgpB, which is specific for Arg-Xaa bonds in substrates. Recently, a nanobody-based assay was used to demonstrate that RgpB could represent a novel diagnostic target, thereby simplifying. P. gingivalis detection. The nanobody, VHH7, had a high binding affinity and was specific for RgpB, when tested towards the highly identical RgpA. In this study a homology model of VHH7 was build. The complementarity determining regions (CDR) comprising the paratope residues responsible for RgpB binding were identified and used as input to the docking. Furthermore, residues likely involved in the RgpB epitope was identified based upon RgpB:RgpA alignment and analysis of residue surface accessibility. CDR residues and putitative RgpB epitope residues were used as input to an information-driven flexible docking approach using the HADDOCK server. Analysis of the VHH7:RgpB model demonstrated that the epitope was found in the immunoglobulin-like domain and residue pairs located at the molecular paratope:epitope interface important for complex stability was identified. Collectively, the VHH7 homology model and VHH7:RgpB docking supplies knowledge of the residues involved in the high affinity interaction. This information could prove valuable in the design of an antibody-drug conjugate for specific RgpB targeting. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Characterization of a high affinity cocaine binding site in rat brain

    International Nuclear Information System (INIS)

    Calligaro, D.; Eldefrawi, M.

    1986-01-01

    Binding of [ 3 H]cocaine to synaptic membranes from whole rat brain was reversible and saturable. Nonlinear regression analysis of binding isotherms indicated two binding affinities: one with k/sub d/ = 16 nM, B/sub max/ = 0.65 pmoles/mg protein and the other with K/sub d/ = 660 nM, B/sub max/ = 5.1 pmoles/mg protein. The high-affinity binding of [ 3 H]cocaine was sensitive to the actions of trypsin and chymotrypsin but not carboxypeptidase, and was eliminated by exposure of the membranes to 95 0 C for 5 min. Specific binding at 2 nM was higher at pH 8.8 than at pH 7.0. Binding of [ 3 H]cocaine (15 nM) was inhibited by increasing concentrations of Na + ions. Several cocaine analogues, neurotransmitter uptake inhibitors and local anesthetics displaced specific [ 3 H]cocaine binding at 2 nM with various potencies. The cocaine analogue (-)-norcocaine was the most potent (IC 50 = 10 nM), while the local anesthetic tetracaine was the least potent in inhibiting [ 3 H]cocaine binding. Several biogenic amine uptake inhibitors, including tricyclic antidepressants and phencyclidine, had IC 50 values below μM concentrations

  9. High-affinity binding of two molecules of cysteine proteinases to low-molecular-weight kininogen.

    Science.gov (United States)

    Turk, B.; Stoka, V.; Björk, I.; Boudier, C.; Johansson, G.; Dolenc, I.; Colic, A.; Bieth, J. G.; Turk, V.

    1995-01-01

    Human low-molecular-weight kininogen (LK) was shown by fluorescence titration to bind two molecules of cathepsins L and S and papain with high affinity. By contrast, binding of a second molecule of cathepsin H was much weaker. The 2:1 binding stoichiometry was confirmed by titration monitored by loss of enzyme activity and by sedimentation velocity experiments. The kinetics of binding of cathepsins L and S and papain showed the two proteinase binding sites to have association rate constants kass,1 = 10.7-24.5 x 10(6) M-1 s-1 and kass,2 = 0.83-1.4 x 10(6) M-1 s-1. Comparison of these kinetic constants with previous data for intact LK and its separated domains indicate that the faster-binding site is also the tighter-binding site and is present on domain 3, whereas the slower-binding, lower-affinity site is on domain 2. These results also indicate that there is no appreciable steric hindrance for the binding of proteinases between the two binding sites or from the kininogen light chain. PMID:8528085

  10. Cyclic GMP-AMP Containing Mixed Phosphodiester Linkages Is An Endogenous High Affinity Ligand for STING

    Science.gov (United States)

    Zhang, Xu; Shi, Heping; Wu, Jiaxi; Zhang, Xuewu; Sun, Lijun; Chen, Chuo; Chen, Zhijian J.

    2013-01-01

    The presence of microbial or self DNA in the cytoplasm of mammalian cells is a danger signal detected by the DNA sensor cyclic-GMP-AMP (cGAMP) synthase (cGAS), which catalyzes the production of cGAMP that in turn serves as a second messenger to activate innate immune responses. Here we show that endogenous cGAMP in mammalian cells contains two distinct phosphodiester linkages, one between 2′-OH of GMP and 5′-phosphate of AMP, and the other between 3′-OH of AMP and 5′-phosphate of GMP. This molecule, termed 2′3′-cGAMP, is unique in that it binds to the adaptor protein STING with a much greater affinity than cGAMP molecules containing other combinations of phosphodiester linkages. The crystal structure of STING bound to 2′3′-cGAMP revealed the structural basis of this high-affinity binding and a ligand-induced conformational change in STING that may underlie its activation. PMID:23747010

  11. Identification of a High Affinity Nucleocapsid Protein Binding Element from The Bovine Leukemia Virus Genome

    Science.gov (United States)

    Yildiz, F. Zehra; Babalola, Kathleen; Summers, Michael F.

    2012-01-01

    Retroviral genome recognition is mediated by interactions between the nucleocapsid (NC) domain of the virally encoded Gag polyprotein and cognate RNA packaging elements that, for most retroviruses, appear to reside primarily within the 5′-untranslated region (5′-UTR) of the genome. Recent studies suggest that a major packaging determinant of Bovine Leukemia Virus (BLV), a member of the human T-cell leukemia virus (HTLV)/BLV family and a non-primate animal model for HTLV-induced leukemogenesis, resides within the gag open reading frame. We have prepared and purified the recombinant BLV NC protein and conducted electrophoretic mobility shift and isothermal titration calorimetry studies with RNA fragments corresponding to these proposed packaging elements. The gag-derived RNAs did not exhibit significant affinity for NC, suggesting an alternate role in packaging. However, an 83-nucleotide fragment of the 5′-UTR that resides just upstream of the gag start codon binds NC stoichiometrically and with high affinity (Kd = 136 ± 21 nM). These nucleotides were predicted to form tandem hairpin structures, and studies with smaller fragments indicate that the NC binding site resides exclusively within the distal hairpin (residues G369- U399, Kd = 67 ± 8 nM at physiological ionic strength). Unlike all other structurally characterized retroviral NC binding RNAs, this fragment is not expected to contain exposed guanosines, suggesting that RNA binding may be mediated by a previously uncharacterized mechanism. PMID:22846919

  12. Identification of high-affinity calmodulin-binding proteins in rat liver

    International Nuclear Information System (INIS)

    Hanley, R.M.; Dedman, J.R.; Shenolikar, S.

    1987-01-01

    The Ca 2+ -dependent binding of [ 125 I] calmodulin (CaM) to hepatic proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was utilized to identify CaM binding or acceptor proteins or CAPs. Two proteins of apparent molecular weight of 60,000 (CAP-60) and 45,000 (CAP-45) comprised > 80% of the Ca 2+ -dependent CaM binding in rat liver cytosol. CAP-60 and CAP-45 were partially purified by a variety of chromatographic steps, including affinity chromatography on CaM Sepharose. CAP-60 possessed a native molecular size of 400,000, indicating it to be the CaM-binding subunit of a larger oligomeric complex. In contrast, CAP-45 was monomeric as judged by gel filtration. Neither CAP-60 nor CAP-45 possessed chromatographic properties consistent with known CaM-dependent enzymes reported in the literature. Two-dimensional peptide mapping provided convincing evidence that CAP-60 and CAP-45 were unrelated to other well-characterized CAPs, namely Ca 2+ (CaM)-dependent protein kinase II, calcineurin, or the CaM-dependent cyclic nucleotide phosphodiesterase. The relative abundance and high affinity for CaM could suggest that these novel target proteins, CAP-60 and CAP-45, represent a dominant pathway for CaM action in the mammalian liver

  13. Humic Acid Confers HIGH-AFFINITY K+ TRANSPORTER 1-Mediated Salinity Stress Tolerance in Arabidopsis.

    Science.gov (United States)

    Khaleda, Laila; Park, Hee Jin; Yun, Dae-Jin; Jeon, Jong-Rok; Kim, Min Gab; Cha, Joon-Yung; Kim, Woe-Yeon

    2017-12-31

    Excessive salt disrupts intracellular ion homeostasis and inhibits plant growth, which poses a serious threat to global food security. Plants have adapted various strategies to survive in unfavorable saline soil conditions. Here, we show that humic acid (HA) is a good soil amendment that can be used to help overcome salinity stress because it markedly reduces the adverse effects of salinity on Arabidopsis thaliana seedlings. To identify the molecular mechanisms of HA-induced salt stress tolerance in Arabidopsis, we examined possible roles of a sodium influx transporter HIGH-AFFINITY K+ TRANSPORTER 1 (HKT1). Salt-induced root growth inhibition in HKT1 overexpressor transgenic plants (HKT1-OX) was rescued by application of HA, but not in wild-type and other plants. Moreover, salt-induced degradation of HKT1 protein was blocked by HA treatment. In addition, the application of HA to HKT1-OX seedlings led to increased distribution of Na+ in roots up to the elongation zone and caused the reabsorption of Na+ by xylem and parenchyma cells. Both the influx of the secondary messenger calcium and its cytosolic release appear to function in the destabilization of HKT1 protein under salt stress. Taken together, these results suggest that HA could be applied to the field to enhance plant growth and salt stress tolerance via post-transcriptional control of the HKT1 transporter gene under saline conditions.

  14. High affinity soluble ILT2 receptor: a potent inhibitor of CD8(+) T cell activation.

    Science.gov (United States)

    Moysey, Ruth K; Li, Yi; Paston, Samantha J; Baston, Emma E; Sami, Malkit S; Cameron, Brian J; Gavarret, Jessie; Todorov, Penio; Vuidepot, Annelise; Dunn, Steven M; Pumphrey, Nicholas J; Adams, Katherine J; Yuan, Fang; Dennis, Rebecca E; Sutton, Deborah H; Johnson, Andy D; Brewer, Joanna E; Ashfield, Rebecca; Lissin, Nikolai M; Jakobsen, Bent K

    2010-12-01

    Using directed mutagenesis and phage display on a soluble fragment of the human immunoglobulin super-family receptor ILT2 (synonyms: LIR1, MIR7, CD85j), we have selected a range of mutants with binding affinities enhanced by up to 168,000-fold towards the conserved region of major histocompatibility complex (MHC) class I molecules. Produced in a dimeric form, either by chemical cross-linking with bivalent polyethylene glycol (PEG) derivatives or as a genetic fusion with human IgG Fc-fragment, the mutants exhibited a further increase in ligand-binding strength due to the avidity effect, with resident half-times (t(1/2)) on the surface of MHC I-positive cells of many hours. The novel compounds antagonized the interaction of CD8 co-receptor with MHC I in vitro without affecting the peptide-specific binding of T-cell receptors (TCRs). In both cytokine-release assays and cell-killing experiments the engineered receptors inhibited the activation of CD8(+) cytotoxic T lymphocytes (CTLs) in the presence of their target cells, with subnanomolar potency and in a dose-dependent manner. As a selective inhibitor of CD8(+) CTL responses, the engineered high affinity ILT2 receptor presents a new tool for studying the activation mechanism of different subsets of CTLs and could have potential for the development of novel autoimmunity therapies.

  15. Physiological epidermal growth factor concentrations activate high affinity receptors to elicit calcium oscillations.

    Directory of Open Access Journals (Sweden)

    Béatrice Marquèze-Pouey

    Full Text Available Signaling mediated by the epidermal growth factor (EGF is crucial in tissue development, homeostasis and tumorigenesis. EGF is mitogenic at picomolar concentrations and is known to bind its receptor on high affinity binding sites depending of the oligomerization state of the receptor (monomer or dimer. In spite of these observations, the cellular response induced by EGF has been mainly characterized for nanomolar concentrations of the growth factor, and a clear definition of the cellular response to circulating (picomolar concentrations is still lacking. We investigated Ca2+ signaling, an early event in EGF responses, in response to picomolar doses in COS-7 cells where the monomer/dimer equilibrium is unaltered by the synthesis of exogenous EGFR. Using the fluo5F Ca2+ indicator, we found that picomolar concentrations of EGF induced in 50% of the cells a robust oscillatory Ca2+ signal quantitatively similar to the Ca2+ signal induced by nanomolar concentrations. However, responses to nanomolar and picomolar concentrations differed in their underlying mechanisms as the picomolar EGF response involved essentially plasma membrane Ca2+ channels that are not activated by internal Ca2+ store depletion, while the nanomolar EGF response involved internal Ca2+ release. Moreover, while the picomolar EGF response was modulated by charybdotoxin-sensitive K+ channels, the nanomolar response was insensitive to the blockade of these ion channels.

  16. Physiological epidermal growth factor concentrations activate high affinity receptors to elicit calcium oscillations.

    Science.gov (United States)

    Marquèze-Pouey, Béatrice; Mailfert, Sébastien; Rouger, Vincent; Goaillard, Jean-Marc; Marguet, Didier

    2014-01-01

    Signaling mediated by the epidermal growth factor (EGF) is crucial in tissue development, homeostasis and tumorigenesis. EGF is mitogenic at picomolar concentrations and is known to bind its receptor on high affinity binding sites depending of the oligomerization state of the receptor (monomer or dimer). In spite of these observations, the cellular response induced by EGF has been mainly characterized for nanomolar concentrations of the growth factor, and a clear definition of the cellular response to circulating (picomolar) concentrations is still lacking. We investigated Ca2+ signaling, an early event in EGF responses, in response to picomolar doses in COS-7 cells where the monomer/dimer equilibrium is unaltered by the synthesis of exogenous EGFR. Using the fluo5F Ca2+ indicator, we found that picomolar concentrations of EGF induced in 50% of the cells a robust oscillatory Ca2+ signal quantitatively similar to the Ca2+ signal induced by nanomolar concentrations. However, responses to nanomolar and picomolar concentrations differed in their underlying mechanisms as the picomolar EGF response involved essentially plasma membrane Ca2+ channels that are not activated by internal Ca2+ store depletion, while the nanomolar EGF response involved internal Ca2+ release. Moreover, while the picomolar EGF response was modulated by charybdotoxin-sensitive K+ channels, the nanomolar response was insensitive to the blockade of these ion channels.

  17. Specific, high affinity receptors for insulin-like growth factor II in the rat kidney glomerulus

    International Nuclear Information System (INIS)

    Haskell, J.F.; Pillion, D.J.; Meezan, E.

    1988-01-01

    Rat renal glomeruli were isolated by a technique involving kidney perfusion with a solution containing magnetic iron oxide particles, followed by homogenization, sieving, and concentration over a strong magnet. Isolated glomeruli were treated with 1% Triton X-100 to solubilize plasma membrane components, while insoluble basement membrane components were removed by centrifugation. [ 125 I]Insulin-like growth factor II (IGF-II) binding to this preparation was competitively inhibited by increasing amounts of unlabeled IGF-II, with 50% inhibition at an IGF-II concentration of 1 ng/ml. [ 125 I]IGF-II was covalently cross-linked with disuccinimidyl suberate to its receptor in rat renal glomeruli and a specific high mol wt (255,000) band could be identified on autoradiograms of dodecyl sulfate-polyacrylamide gels. [ 125 I]IGF-II binding and cross-linking to this band was inhibited by a polyclonal antibody against the type II IGF receptor. These results demonstrate for the first time that the isolated rat renal glomerulus contains a high affinity receptor for IGF-II

  18. Affinity Crystallography: A New Approach to Extracting High-Affinity Enzyme Inhibitors from Natural Extracts.

    Science.gov (United States)

    Aguda, Adeleke H; Lavallee, Vincent; Cheng, Ping; Bott, Tina M; Meimetis, Labros G; Law, Simon; Nguyen, Nham T; Williams, David E; Kaleta, Jadwiga; Villanueva, Ivan; Davies, Julian; Andersen, Raymond J; Brayer, Gary D; Brömme, Dieter

    2016-08-26

    Natural products are an important source of novel drug scaffolds. The highly variable and unpredictable timelines associated with isolating novel compounds and elucidating their structures have led to the demise of exploring natural product extract libraries in drug discovery programs. Here we introduce affinity crystallography as a new methodology that significantly shortens the time of the hit to active structure cycle in bioactive natural product discovery research. This affinity crystallography approach is illustrated by using semipure fractions of an actinomycetes culture extract to isolate and identify a cathepsin K inhibitor and to compare the outcome with the traditional assay-guided purification/structural analysis approach. The traditional approach resulted in the identification of the known inhibitor antipain (1) and its new but lower potency dehydration product 2, while the affinity crystallography approach led to the identification of a new high-affinity inhibitor named lichostatinal (3). The structure and potency of lichostatinal (3) was verified by total synthesis and kinetic characterization. To the best of our knowledge, this is the first example of isolating and characterizing a potent enzyme inhibitor from a partially purified crude natural product extract using a protein crystallographic approach.

  19. High-affinity cannabinoid binding site in brain: A possible marijuana receptor

    International Nuclear Information System (INIS)

    Nye, J.S.

    1988-01-01

    The mechanism by which delta 9 tetrahydrocannabinol (delta 9 THC), the major psychoactive component of marijuana or hashish, produces its potent psychological and physiological effects is unknown. To find receptor binding sites for THC, we designed a water-soluble analog for use as a radioligand. 5'-Trimethylammonium-delta 8 THC (TMA) is a positively charged analog of delta- 8 THC modified on the 5' carbon, a portion of the molecule not important for its psychoactivity. We have studied the binding of [ 3 H]-5'-trimethylammonium-delta- 8 THC ([ 3 H]TMA) to rat neuronal membranes. [ 3 H]TMA binds saturably and reversibly to brain membranes with high affinity to apparently one class of sites. Highest binding site density occurs in brain, but several peripheral organs also display specific binding. Detergent solubilizes the sites without affecting their pharmacologial properties. Molecular sieve chromatography reveals a bimodal peak of [ 3 H]TMA binding activity of approximately 60,000 daltons apparent molecular weight

  20. Inhibition of high affinity choline uptake by N-allyl-3-quinuclidinol

    International Nuclear Information System (INIS)

    Asermely, K.E.; O'Neill, J.J.

    1986-01-01

    The peripheral actions of N-allyl-3-quinuclidinol (N-Al-3-OHQ) on high affinity choline uptake (HAChU) on rat phrenic nerve diaphragm are described. Endplate regions (EPA) identified by the Koelle histochemical techniques for acetylcholinesterase, were dissected from adult rat hemidiaphragms and placed in cold Krebs solution (pH-7.35). All measurements of HAChU were at 37 0 C in buffers containing tritium choline (5 μM 0.124 μC/mmole) at intervals of 1, 2, 4, 8, 15 and 30 min. Tissues were washed 3x, digested in 1N NaOH and counted for tritium in Chaikoff's solution. All data are expressed as pmole Ch/g wet weight. Comparison between EPA and non-EPA tissue demonstrate HAChU and slow choline diffusion, respectively. Steady state is observed in 15 min. N-Al-3-OHQ produces 15% inhibition at 5 x 10 -5 M compared with 50% inhibition on brain synaptosomes. At 5 x 10 -4 M N-Al-3-OHQ, 30% inhibition is observed. Attempts to deplete ACh by pre-stimulation with high K + -ion (25 mM) were unsuccessful; tissue 3 H-choline uptake appeared to oscillate over a 30 min period

  1. Inhibition of high affinity choline uptake by N-allyl-3-quinuclidinol

    Energy Technology Data Exchange (ETDEWEB)

    Asermely, K.E.; O' Neill, J.J.

    1986-03-01

    The peripheral actions of N-allyl-3-quinuclidinol (N-Al-3-OHQ) on high affinity choline uptake (HAChU) on rat phrenic nerve diaphragm are described. Endplate regions (EPA) identified by the Koelle histochemical techniques for acetylcholinesterase, were dissected from adult rat hemidiaphragms and placed in cold Krebs solution (pH-7.35). All measurements of HAChU were at 37/sup 0/C in buffers containing tritium choline (5 ..mu..M 0.124 ..mu..C/mmole) at intervals of 1, 2, 4, 8, 15 and 30 min. Tissues were washed 3x, digested in 1N NaOH and counted for tritium in Chaikoff's solution. All data are expressed as pmole Ch/g wet weight. Comparison between EPA and non-EPA tissue demonstrate HAChU and slow choline diffusion, respectively. Steady state is observed in 15 min. N-Al-3-OHQ produces 15% inhibition at 5 x 10/sup -5/ M compared with 50% inhibition on brain synaptosomes. At 5 x 10/sup -4/ M N-Al-3-OHQ, 30% inhibition is observed. Attempts to deplete ACh by pre-stimulation with high K/sup +/-ion (25 mM) were unsuccessful; tissue /sup 3/H-choline uptake appeared to oscillate over a 30 min period.

  2. Synthesis of site-heterologous haptens for high-affinity anti-pyraclostrobin antibody generation.

    Science.gov (United States)

    Mercader, Josep V; Agulló, Consuelo; Abad-Somovilla, Antonio; Abad-Fuentes, Antonio

    2011-03-07

    The design and synthesis of functional chemical derivatives of small organic molecules is usually a key step for the intricate production of a variety of bioconjugates. In this respect, the derivatization site at which the spacer arm is introduced in immunizing conjugates constitutes a highly critical parameter for the generation of high-affinity and selective antibodies. However, due to the usual complexity of the required synthetic procedures, the appropriate comparison of alternative tethering positions has often been neglected. In the present study, meticulous strategies were followed to prepare synthetic derivatives of pyraclostrobin with the same linkers located at diverse rationally-chosen sites. Activity appraisal of antibodies and bioconjugates was carried out by bidimensional competitive direct and indirect immunoassays, and a superior performance of two of the three synthesized haptens was found. Finally, a detailed analysis of the conformations of the target molecule and the synthesized haptens in aqueous solution was done using computer assisted molecular modeling techniques. This study suggested that the lower titers and affinities of one set of antibodies are most probably due to conformational effects of the spacer arm in the immunizing bioconjugate.

  3. Constitutive expression of a putative high-affinity nitrate transporter in Nicotiana plumbaginifolia: evidence for post-transcriptional regulation by a reduced nitrogen source.

    Science.gov (United States)

    Fraisier, V; Gojon, A; Tillard, P; Daniel-Vedele, F

    2000-08-01

    The NpNRT2.1 gene encodes a putative inducible component of the high-affinity nitrate (NO3-) uptake system in Nicotiana plumbaginifolia. Here we report functional and physiological analyses of transgenic plants expressing the NpNRT2.1 coding sequence fused to the CaMV 35S or rolD promoters. Irrespective of the level of NO3- supplied, NO3- contents were found to be remarkably similar in wild-type and transgenic plants. Under specific conditions (growth on 10 mM NO3-), the steady-state NpNRT2. 1 mRNA level resulting from the deregulated transgene expression was accompanied by an increase in 15NO3- influx measured in the low concentration range. This demonstrates for the first time that the NRT2.1 sequence codes a limiting element of the inducible high-affinity transport system. Both 15NO3- influx and mRNA levels decreased in the wild type after exposure to ammonium, in agreement with previous results from many species. Surprisingly, however, influx was also markedly decreased in transgenic plants, despite stable levels of transgene expression in independent transformants after ammonium addition. We conclude that the conditions associated with the supply of a reduced nitrogen source such as ammonium, or with the generation of a further downstream metabolite, probably exert a repressive effect on NO3- influx at both transcriptional and post-transcriptional levels.

  4. Novel high-affinity and selective biaromatic 4-substituted ¿-hydroxybutyric acid (GHB) analogues as GHB ligands

    DEFF Research Database (Denmark)

    Høg, Signe; Wellendorph, Petrine; Nielsen, Birgitte

    2008-01-01

    Gamma-hydroxybutyrate (GHB) is a metabolite of gamma-aminobutyric acid (GABA) and has been proposed to function as a neurotransmitter or neuromodulator. GHB is used in the treatment of narcolepsy and is a drug of abuse. GHB binds to both GABA(B) receptors and specific high-affinity GHB sites...

  5. Are basophil histamine release and high affinity IgE receptor expression involved in asymptomatic skin sensitization?

    DEFF Research Database (Denmark)

    Jensen, Bettina Margrethe; Assing, K; Jensen, Lone Hummelshøj

    2006-01-01

    Immunoglobulin (Ig)E-sensitized persons with positive skin prick test, but no allergy symptoms, are classified as being asymptomatic skin sensitized (AS). The allergic type 1 disease is dependant on IgE binding to the high affinity IgE-receptor (FcepsilonRI) expressed on basophils and mast cells....

  6. New Synthesis and Tritium Labeling of a Selective Ligand for Studying High-Affinity γ-Hydroxybutyrate (GHB) Binding Sites

    DEFF Research Database (Denmark)

    Vogensen, Stine B.; Marek, Ales; Bay, Tina

    2013-01-01

    3-Hydroxycyclopent-1-enecarboxylic acid (HOCPCA, 1) is a potent ligand for the high-affinity GHB binding sites in the CNS. An improved synthesis of 1 together with a very efficient synthesis of [3H]-1 is described. The radiosynthesis employs in situ generated lithium trimethoxyborotritide. Screen...

  7. Absence of high-affinity calreticulin autoantibodies in patients with systemic rheumatic diseases and coeliac disease

    DEFF Research Database (Denmark)

    Jørgensen, C S; Hansen, K B; Jacobsen, Søren

    2005-01-01

    agent. Using these conditions we found a relatively high level of non-specific binding in many sera. Antibodies to proteins that are used as blocking reagents in ELISA (bovine serum albumin (BSA), ovalbumin, skimmed milk powder) are frequently present in sera, and these may cause false-positive results...

  8. Diomres (k,m): An efficient method based on Krylov subspaces to solve big, dispersed, unsymmetrical linear systems

    Energy Technology Data Exchange (ETDEWEB)

    de la Torre Vega, E. [Instituto de Investigaciones Electricas, Cuernavaca (Mexico); Cesar Suarez Arriaga, M. [Universidad Michoacana SNH, Michoacan (Mexico)

    1995-03-01

    In geothermal simulation processes, MULKOM uses Integrated Finite Differences to solve the corresponding partial differential equations. This method requires to resolve efficiently big linear dispersed systems of non-symmetrical nature on each temporal iteration. The order of the system is usually greater than one thousand its solution could represent around 80% of CPU total calculation time. If the elapsed time solving this class of linear systems is reduced, the duration of numerical simulation decreases notably. When the matrix is big (N{ge}500) and with holes, it is inefficient to handle all the system`s elements, because it is perfectly figured out by its elements distinct of zero, quantity greatly minor than N{sup 2}. In this area, iteration methods introduce advantages with respect to gaussian elimination methods, because these last replenish matrices not having any special distribution of their non-zero elements and because they do not make use of the available solution estimations. The iterating methods of the Conjugated Gradient family, based on the subspaces of Krylov, possess the advantage of improving the convergence speed by means of preconditioning techniques. The creation of DIOMRES(k,m) method guarantees the continuous descent of the residual norm, without incurring in division by zero. This technique converges at most in N iterations if the system`s matrix is symmetrical, it does not employ too much memory to converge and updates immediately the approximation by using incomplete orthogonalization and adequate restarting. A preconditioned version of DIOMRES was applied to problems related to unsymmetrical systems with 1000 unknowns and less than five terms per equation. We found that this technique could reduce notably the time needful to find the solution without requiring memory increment. The coupling of this method to geothermal versions of MULKOM is in process.

  9. Endothelial targeting of high-affinity multivalent polymer nanocarriers directed to intercellular adhesion molecule 1.

    Science.gov (United States)

    Muro, Silvia; Dziubla, Thomas; Qiu, Weining; Leferovich, John; Cui, Xiumin; Berk, Erik; Muzykantov, Vladimir R

    2006-06-01

    Targeting of diagnostic and therapeutic agents to endothelial cells (ECs) provides an avenue to improve treatment of many maladies. For example, intercellular adhesion molecule 1 (ICAM-1), a constitutive endothelial cell adhesion molecule up-regulated in many diseases, is a good determinant for endothelial targeting of therapeutic enzymes and polymer nanocarriers (PNCs) conjugated with anti-ICAM (anti-ICAM/PNCs). However, intrinsic and extrinsic factors that control targeting of anti-ICAM/PNCs to ECs (e.g., anti-ICAM affinity and PNC valency and flow) have not been defined. In this study we tested in vitro and in vivo parameters of targeting to ECs of anti-ICAM/PNCs consisting of either prototype polystyrene or biodegradable poly(lactic-coglycolic) acid polymers (approximately 200 nm diameter spheres carrying approximately 200 anti-ICAM molecules). Anti-ICAM/PNCs, but not control IgG/PNCs 1) rapidly (t1/2 approximately 5 min) and specifically bound to tumor necrosis factor-activated ECs in a dose-dependent manner (Bmax approximately 350 PNC/cell) at both static and physiological shear stress conditions and 2) bound to ECs and accumulated in the pulmonary vasculature after i.v. injection in mice. Anti-ICAM/PNCs displayed markedly higher EC affinity versus naked anti-ICAM (Kd approximately 80 pM versus approximately 8 nM) in cell culture and, probably because of this factor, higher value (185.3 +/- 24.2 versus 50.5 +/- 1.5% injected dose/g) and selectivity (lung/blood ratio 81.0 +/- 10.9 versus 2.1 +/- 0.02, in part due to faster blood clearance) of pulmonary targeting. These results 1) show that reformatting monomolecular anti-ICAM into high-affinity multivalent PNCs boosts their vascular immuno-targeting, which withstands physiological hydrodynamics and 2) support potential anti-ICAM/PNCs utility for medical applications.

  10. 99mTc(CO)3-DTMA bombesin conjugates having high affinity for the GRP receptor

    International Nuclear Information System (INIS)

    Lane, Stephanie R.; Veerendra, Bhadrasetty; Rold, Tammy L.; Sieckman, Gary L.; Hoffman, Timothy J.; Jurisson, Silvia S.; Smith, Charles J.

    2008-01-01

    Introduction: Targeted diagnosis of specific human cancer types continues to be of significant interest in nuclear medicine. 99m Tc is ideally suited as a diagnostic radiometal for in vivo tumor targeting due to its ideal physical characteristics and diverse labeling chemistries in numerous oxidation states. Methods: In this study, we report a synthetic approach toward design of a new tridentate amine ligand for the organometallic aqua-ion [ 99m Tc(H 2 O) 3 (CO) 3 ] + . The new chelating ligand framework, 2-(N,N'-Bis(tert-butoxycarbonyl)diethylenetriamine) acetic acid (DTMA), was synthesized from a diethylenetriamine precursor and fully characterized by mass spectrometry and nuclear magnetic resonance spectroscopy ( 1 H and 13 C). DTMA was conjugated to H 2 N-(X)-BBN(7-14)NH 2 , where X=an amino acid or aliphatic pharmacokinetic modifier and BBN=bombesin peptide, by means of solid phase peptide synthesis. DTMA-(X)-BBN(7-14)NH 2 conjugates were purified by reversed-phase high-performance chromatography and characterized by electrospray-ionization mass spectrometry. Results: The new conjugates were radiolabeled with [ 99m Tc(H 2 O) 3 (CO) 3 ] + produced via Isolink radiolabeling kits to produce [ 99m Tc(CO) 3 -DTMA-(X)-BBN(7-14)NH 2 ]. Radiolabeled conjugates were purified by reversed-phase high-performance chromatography. Effective receptor binding behavior was evaluated in vitro and in vivo. Conclusions: [ 99m Tc(CO) 3 -DTMA-(X)-BBN(7-14)NH 2 ] conjugates displayed very high affinity for the gastrin releasing peptide receptor in vitro and in vivo. Therefore, these conjugates hold some propensity to be investigated as molecular imaging agents that specifically target human cancers uniquely expressing the gastrin releasing peptide receptor subtypes

  11. Characterization of high affinity [3H]triazolam binding in rat brain

    International Nuclear Information System (INIS)

    Earle, M.; Concas, A.; Yamamura, H.I.

    1986-01-01

    The hypnotic Triazolam (TZ), a triazolo (1,4)-benzodiazepine, displays a short physiological half life and has been used for the treatment of insomnia related to anxiety states. Specific binding properties of this recently tritiated TZ were characterized. The authors major objectives were the direct measurement of the temperature dependence and the GABA effect on [ 3 H]TZ binding. Saturation studies showed a shift to lower affinity at 37 0 C (K/sub d/ = 0.25 +/- 0.01 nM at O 0 C; K/sub d/ = 1.46 +/- 0.03 nM at 37 0 C) while the B/sub max/ values remained unchanged (1003 +/- 37 fmoles/mg prot. at 0 0 C and 1001 +/- 43 fmoles/mg prot. at 37 0 C). Inhibition studies showed that [ 3 H]TZ binding displayed no GABA shift at 0 0 C(K/sub i/ 0.37 +/- 0.03 nM/- GABA and K/sub i/ = 0.55 +/- 0.13 nM/+GABA) but a nearly two-fold shift was apparent at 37 0 C (K/sub i/ = 2.92 +/- 0.2 nM/-GABA; K/sub i/ = 1.37 +/- 0.11 mM/+GABA). These results were also confirmed by saturation studies in the presence or absence of GABA showing a shift to higher affinity in the presence of GABA only at 37 0 C. In Ro 15-1788/[ 3 H]TZ competition experiments the presence of GABA did not affect the inhibitory potency of Ro 15-1788 on [ 3 H]TZ binding at both temperatures. In conclusion [ 3 H]TZ binding showed an extremely high affinity for benzodiazepine receptors. In contrast to reported literature, the findings suggest that TZ interacts with benzodiazepine receptors similar to other benzodiazepine agonists

  12. Cartilage Acidic Protein 2 a hyperthermostable, high affinity calcium-binding protein.

    Science.gov (United States)

    Anjos, Liliana; Gomes, Ana S; Melo, Eduardo P; Canário, Adelino V; Power, Deborah M

    2013-03-01

    Cartilage Acidic Protein 2 (CRTAC2) is a novel protein present from prokaryotes to vertebrates with abundant expression in the teleost fish pituitary gland and an isoform of CRTAC1, a chondrocyte marker in humans. The two proteins are non-integrins containing N-terminal integrin-like Ca(2+)-binding motifs and their structure and function remain to be assigned. Structural studies of recombinant sea bream (sb)CRTAC2 revealed it is composed of 8.8% α-helix, 33.4% β-sheet and 57.8% unordered protein. sbCRTAC2 bound Ca(2+) with high affinity (K(d)=1.46nM) and favourable Gibbs free energy (∆G=-12.4kcal/mol). The stoichiometry for Ca(2+) bound to sbCRTAC2 at saturation indicated six Ca(2+) ligand-binding sites exist per protein molecule. No conformational change in sbCRTAC2 occurred in the presence of Ca(2+). Fluorescence emission revealed that the tertiary structure of the protein is hyperthermostable between 25°C and 95°C and the fully unfolded state is only induced by chemical denaturing (4M GndCl). sbCRTAC has a widespread tissue distribution and is present as high molecular weight aggregates, although strong reducing conditions promote formation of the monomer. sbCRTAC2 promotes epithelial cell outgrowth in vitro suggesting it may share functional homology with mammalian CRTAC1, recently implicated in cell-cell and cell-matrix interactions. Copyright © 2012 Elsevier B.V. All rights reserved.

  13. Engineering of bispecific affinity proteins with high affinity for ERBB2 and adaptable binding to albumin.

    Directory of Open Access Journals (Sweden)

    Johan Nilvebrant

    Full Text Available The epidermal growth factor receptor 2, ERBB2, is a well-validated target for cancer diagnostics and therapy. Recent studies suggest that the over-expression of this receptor in various cancers might also be exploited for antibody-based payload delivery, e.g. antibody drug conjugates. In such strategies, the full-length antibody format is probably not required for therapeutic effect and smaller tumor-specific affinity proteins might be an alternative. However, small proteins and peptides generally suffer from fast excretion through the kidneys, and thereby require frequent administration in order to maintain a therapeutic concentration. In an attempt aimed at combining ERBB2-targeting with antibody-like pharmacokinetic properties in a small protein format, we have engineered bispecific ERBB2-binding proteins that are based on a small albumin-binding domain. Phage display selection against ERBB2 was used for identification of a lead candidate, followed by affinity maturation using second-generation libraries. Cell surface display and flow-cytometric sorting allowed stringent selection of top candidates from pools pre-enriched by phage display. Several affinity-matured molecules were shown to bind human ERBB2 with sub-nanomolar affinity while retaining the interaction with human serum albumin. Moreover, parallel selections against ERBB2 in the presence of human serum albumin identified several amino acid substitutions that dramatically modulate the albumin affinity, which could provide a convenient means to control the pharmacokinetics. The new affinity proteins competed for ERBB2-binding with the monoclonal antibody trastuzumab and recognized the native receptor on a human cancer cell line. Hence, high affinity tumor targeting and tunable albumin binding were combined in one small adaptable protein.

  14. A mix-and-read drop-based in vitro two-hybrid method for screening high-affinity peptide binders

    Science.gov (United States)

    Cui, Naiwen; Zhang, Huidan; Schneider, Nils; Tao, Ye; Asahara, Haruichi; Sun, Zhiyi; Cai, Yamei; Koehler, Stephan A.; de Greef, Tom F. A.; Abbaspourrad, Alireza; Weitz, David A.; Chong, Shaorong

    2016-01-01

    Drop-based microfluidics have recently become a novel tool by providing a stable linkage between phenotype and genotype for high throughput screening. However, use of drop-based microfluidics for screening high-affinity peptide binders has not been demonstrated due to the lack of a sensitive functional assay that can detect single DNA molecules in drops. To address this sensitivity issue, we introduced in vitro two-hybrid system (IVT2H) into microfluidic drops and developed a streamlined mix-and-read drop-IVT2H method to screen a random DNA library. Drop-IVT2H was based on the correlation between the binding affinity of two interacting protein domains and transcriptional activation of a fluorescent reporter. A DNA library encoding potential peptide binders was encapsulated with IVT2H such that single DNA molecules were distributed in individual drops. We validated drop-IVT2H by screening a three-random-residue library derived from a high-affinity MDM2 inhibitor PMI. The current drop-IVT2H platform is ideally suited for affinity screening of small-to-medium-sized libraries (103–106). It can obtain hits within a single day while consuming minimal amounts of reagents. Drop-IVT2H simplifies and accelerates the drop-based microfluidics workflow for screening random DNA libraries, and represents a novel alternative method for protein engineering and in vitro directed protein evolution. PMID:26940078

  15. BOOTES: Technological Developments and Scientific Results by a Stereoscopic System with two Stations Spaced by 240 km

    International Nuclear Information System (INIS)

    Mateo Sanguino, T. de J.; Castro-Tirado, A.J.; Ugarte Postigo, A. de; Gorosabel, J.; Fernandez Palomo, M.T.; Morena Carretero, B.A. de la; Castro Ceron, J.M.; Berna Galiano, J.A.; Pata, P.; Bernas, M.; Vitek, S.; Soldan, J.; Hudec, R.; Jelinek, M.; Kubanek, P.; McBreen, S.; Garcia Dabo, C.E.; Soria, T.; Torres Riera, J.

    2004-01-01

    An overview of the technological developments at the Burst Observer and Optical Transient Exploring System (BOOTES) is given. The most important scientific results obtained so far are the detection of an OT in the GRB 000313 error box and the non detection of optical emission simultaneous for GRB 010220. GRB 030226 (long/soft events), GRB 020531 and GRB 021201 (short/hard events). With the recent instrumental and technical developments, BOOTES multiplies its science capabilities

  16. Twins in spirit part II: DOTATATE and high-affinity DOTATATE - the clinical experience

    Energy Technology Data Exchange (ETDEWEB)

    Brogsitter, Claudia; Zoephel, Klaus; Hartmann, Holger; Kotzerke, Joerg [Technische Universitaet Dresden, Department of Nuclear Medicine, Dresden (Germany); Schottelius, Margret; Wester, Hans-Juergen [Technische Universitaet Muenchen, Pharmaceutical Radiochemistry and Department of Nuclear Medicine, Muenchen (Germany)

    2014-06-15

    Over recent decades interest in diagnosis and treatment of neuroendocrine tumours (NET) has steadily grown. The basis for diagnosis and therapy of NET with radiolabelled somatostatin (hsst) analogues is the variable overexpression of hsst receptors (hsst1-5 receptors). We hypothesized that radiometal derivatives of DOTA-iodo-Tyr{sup 3}-octreotide analogues might be excellent candidates for somatostatin receptor imaging. We therefore explored the diagnostic potential of {sup 68}Ga-DOTA-iodo-Tyr{sup 3}-octreotate [{sup 68}Ga-DOTA,3-iodo-Tyr{sup 3},Thr{sup 8}]octreotide ({sup 68}Ga-HA-DOTATATE; HA, high-affinity) compared to the established {sup 68}Ga-DOTA-Tyr{sup 3}-octreotate ({sup 68}Ga-DOTATATE) in vivo. The study included 23 patients with known somatostatin receptor-positive metastases from NETs, thyroid cancer or glomus tumours who were investigated with both {sup 68}Ga-HA-DOTATATE and {sup 68}Ga-DOTATATE. A patient-based and a lesion-based comparative analysis was carried out of normal tissue distribution and lesion detectability in a qualitative and a semiquantitative manner. {sup 68}Ga-HA-DOTATATE and {sup 68}Ga-DOTATATE showed comparable uptake in the liver (SUV{sub mean} 8.9 ± 2.2 vs. 9.3 ± 2.5, n.s.), renal cortex (SUV{sub mean} 13.3 ± 3.9 vs. 14.5 ± 3.7, n.s.) and spleen (SUV{sub mean} 24.0 ± 6.7 vs. 22.9 ± 7.3, n.s.). A somewhat higher pituitary uptake was found with {sup 68}Ga-HA-DOTATATE (SUV{sub mean} 6.3 ± 1.8 vs. 5.4 ± 2.1, p < 0.05). On a lesion-by-lesion basis a total of 344 lesions were detected. {sup 68}Ga-HA-DOTATATE demonstrated 328 lesions (95.3 % of total lesions seen), and {sup 68}Ga-DOTATATE demonstrated 332 lesions (96.4 %). The mean SUV{sub max} of all lesions was not significantly different between {sup 68}Ga-HA-DOTATATE and {sup 68}Ga-DOTATATE (17.8 ± 11.4 vs. 16.7 ± 10.7, n.s.). Our analysis demonstrated very good concordance between {sup 68}Ga-HA-DOTATATE and {sup 68}Ga-DOTATATE PET data. As the availability and use of {sup

  17. System approach for evaluation of soil contamination by nitrogen containing pollutants in the 30-km zone power nuclear plant; Sistemnyj podkhod dlya otsenki zagryazneniya okruzhayushchej sredy azotsoderzhashchimi pollyutantami v 30-km zone aehs

    Energy Technology Data Exchange (ETDEWEB)

    Shevtsova, O. V.; Zhigunova, L. N.; Makovskaya, N. A.; Pavlovich, E. L. [Ob' ' edinennyj institut ehnergeticheskikh i yadernykh issledovanij – Sosny NAN Belarusi, Minsk(Belarus)

    2013-01-15

    This article presents the results of studies of the nitrogen-containing chemical pollutants of the soil 30 km zone nuclear power plant in order to evaluate the chemical carcinogenic risk to public health. As a result of this work the least amount of NDMA fixed at 0,83 × 10{sup -4} mg/kg, the highest - 128,29 × 10{sup -4} mg/kg, which is 0,83 and 128,3 %, respectively, of Madman established in the Russian Federation and Republic of Kazakhstan, the content of NDMA precursors - nitrite ions varies from 0,07 to 4,43 mg/kg. Found that most of the compounds reported in agricultural land. Calculate the total carcinogenic risk for the population living in the region - the set value corresponds to the maximum risk tolerance, i. e. the upper limit of acceptable risk. These levels are subject to continuous monitoring. (authors)

  18. Case study: Atlantis Systems International - Using KM principles to drive productivity and performance, prevent critical knowledge loss and encourage innovation

    International Nuclear Information System (INIS)

    Melnick, B.

    2007-01-01

    Full text: 'Knowledge and information are different. You can manage information; you can't manage knowledge. However you can manage the process that converts information into knowledge. At Atlantis, we treat knowledge management as a construct of information management (product, content) and knowledge building (process, people)' In 2003 Atlantis Systems International a 28 year old Aerospace engineering company was on the brink of insolvency. While the company was a leader in providing simulation-based training products, their market share had decreased steadily over the past 3 years. This decline was due in part to uncontrollable external forces - The formation of the European Economic Union and the tragic events of September 11. Both events lead to restricted market access in the U.S. and Europe for Atlantis's products and services. In addition to these external forces, the company experienced prolonged instability at the management level that resulted in a loss of key personnel to other companies. As no formal knowledge management processes were in place for capturing Intellectual capital, these employees took their knowledge and with them when they left the firm leaving critical gaps in knowledge base of the company. For a company to become knowledge centric organization where knowledge is continuously captured and leveraged to drive innovation requires an understanding of the processes that convert information into usable knowledge. This is as much a social challenge as it is a systems or use of technology challenge. The central question the company or organization has to address is, 'how do we get people (employees and management) to willing share what they know in order to grow the company?' The type of organizational change is synonymous with building a 'learning organization', but while this is certainly implicit, the term is sufficiently ambiguous as to include almost any activity where there is some transfer of information or knowledge - It does not

  19. Dopamine inhibition of anterior pituitary adenylate cyclase is mediated through the high-affinity state of the D2 receptor

    International Nuclear Information System (INIS)

    Borgundvaag, B.; George, S.R.

    1985-01-01

    The diterpinoid forskolin stimulated adenylate cyclase activity (measured by conversion of [ 3 H]-ATP to [ 3 H]-cAMP) in anterior pituitary from male and female rats. Inhibition of stimulated adenylate cyclase activity by potent dopaminergic agonists was demonstrable only in female anterior pituitary. The inhibition of adenylate cyclase activity displayed a typically dopaminergic rank order of agonist potencies and could be completely reversed by a specific dopamine receptor antagonist. The IC 50 values of dopamine agonist inhibition of adenylate cyclase activity correlated with equal molarity with the dissociation constant of the high-affinity dopamine agonist-detected receptor binding site and with the IC 50 values for inhibition of prolactin secretion. These findings support the hypothesis that it is the high-affinity form of the D 2 dopamine receptor in anterior pituitary which is responsible for mediating the dopaminergic function of attenuating adenylate cyclase activity. 12 references, 4 figures, 1 table

  20. Formation of continental crust in a temporally linked arc magma system from 5 to 30 km depth: ~ 90 Ma plutonism in the Cascades Crystalline Core composite arc section

    Science.gov (United States)

    Ratschbacher, B. C.; Miller, J. S.; Kent, A. J.; Miller, R. B.; Anderson, J. L.; Paterson, S. R.

    2015-12-01

    Continental crust has an andesitic bulk composition with a mafic lower crust and a granodioritic upper crust. The formation of stratified continental crust in general and the vertical extent of processes active in arc crustal columns leading to the differentiation of primitive, mantle-derived melts entering the lower crust are highly debated. To investigate where in the crustal column magma mixing, fractionation, assimilation and crystal growth occur and to what extent, we study the ~ 90 Ma magmatic flare-up event of the Cascades arc, a magma plumbing system from ~ 5 to 30 km depth. We focus on three intrusive complexes, emplaced at different depths during major regional shortening in an exceptionally thick crust (≥ 55 km1) but which are temporally related: the upper crustal Black Peak intrusion (1-3 kbar at 3.7 to 11 km; ~ 86.8 to 91.7 Ma2), the mid-crustal Mt. Stuart intrusion (3.5-4.0 kbar at 13 to 15 km; 90.8 and 96.3 Ma3) and the deep crustal Tenpeak intrusion (7 to 10 kbar at 25 to 37 km; 89.7 to 92.3 Ma4). These intrusive complexes are well characterized by geochronology showing that they have been constructed incrementally by multiple magma batches over their lifespans and thus allow the monitoring and comparison of geochemical parameters over time at different depths. We use a combination of whole rock major and trace element data and isotopes combined with detailed investigation of amphibole, which has been recognized to be important in the generation of calc-alkaline rocks in arcs to test the following hypotheses: (a) compositional bimodality is produced in the lower crust, whereas upper crustal levels are dominated by mixing to form intermediate compositions, or (b) differentiation occurs throughout the crustal column with different crystallizing phases and their compositions controlling the bulk chemistry. 1. Miller et al. 2009: GSA Special Paper 456, p. 125-149 2. Shea 2014: PhD thesis, Massachusetts Institute of Technology 3. Anderson et al. 2012

  1. NEMO-SMO acoustic array: A deep-sea test of a novel acoustic positioning system for a km3-scale underwater neutrino telescope

    Science.gov (United States)

    Viola, S.; Ardid, M.; Bertin, V.; Enzenhöfer, A.; Keller, P.; Lahmann, R.; Larosa, G.; Llorens, C. D.; NEMO Collaboration; SMO Collaboration

    2013-10-01

    Within the activities of the NEMO project, the installation of a 8-floors tower (NEMO-Phase II) at a depth of 3500 m is foreseen in 2012. The tower will be installed about 80 km off-shore Capo Passero, in Sicily. On board the NEMO tower, an array of 18 acoustic sensors will be installed, permitting acoustic detection of biological sources, studies for acoustic neutrino detection and primarily acoustic positioning of the underwater structures. For the latter purpose, the sensors register acoustic signals emitted by five acoustic beacons anchored on the sea-floor. The data acquisition system of the acoustic sensors is fully integrated with the detector data transport system and is based on an “all data to shore” philosophy. Signals coming from hydrophones are continuously sampled underwater at 192 kHz/24 bit and transmitted to shore through an electro-optical cable for real-time analysis. A novel technology for underwater GPS time-stamping of data has been implemented and tested. The operation of the acoustic array will permit long-term test of sensors and electronics technologies that are proposed for the acoustic positioning system of KM3NeT.

  2. Functional characterization of the high affinity IgG Receptor : making heads and tails of FcγRI

    NARCIS (Netherlands)

    van der Poel, C.E.

    2011-01-01

    This thesis focuses on human FcγRI, a high affinity receptor for antibodies of the IgG isotype. IgG is the most abundant antibody type in blood and all currently FDA approved therapeutic antibodies are of the IgG isotype. FcγRI, a member of the activating Fcγ receptors, exists as a complex of a

  3. Benzodiazepines have high-affinity binding sites and induce melanogenesis in B16/C3 melanoma cells.

    OpenAIRE

    Matthew, E; Laskin, J D; Zimmerman, E A; Weinstein, I B; Hsu, K C; Engelhardt, D L

    1981-01-01

    We found that two markers of differentiation, tyrosinase (monophenol, dihydroxyphenylalanine:oxygen oxidoreductase, EC 1.14.18.1) activity and melanin synthesis, are induced by diazepam in B16/C3 mouse melanoma cells. We also demonstrated high-affinity binding sites for [3H]diazepam in these cells by radioreceptor assay, and we visualized binding to the cell surface by fluorescence microscopy with a benzodiazepine analog conjugated to a fluorescein-labeled protein. Our studies also showed tha...

  4. Photoaffinity labeling of mammalian α1-adrenergic receptors: identification of the ligand binding subunit with a high affinity radioiodinated probe

    International Nuclear Information System (INIS)

    Leeb-Lundberg, L.M.F.; Dickinson, K.E.J.; Heald, S.L.

    1984-01-01

    A description is given of the synthesised and characterization of a novel high affinity radioiodinated α 1 -adrenergic receptor photoaffinity probe, 4-amino-6,7-dimethoxy-2-[4-[5-(4-azido-3-[ 125 I]iodophenyl)pentanoyl]-1-piperazinyl] quinazoline. In the absence of light, this ligand binds with high affinity (K/sub d/ = 130 pm) in a reverisble and saturable manner to sites in rat hepatic plasma membranes. The binding is stereoselective and competitively inhibited by adrenergic agonists and antagonists with an α 1 -adrenergic specificity. Upon photolysis, this ligand incorporates irreversibly into plasma membranes prepared from several mammalian tissues including rat liver, rat, guinea pig, and rabbit spleen, rabbit lung, and rabbit aorta vascular smooth muscle cells, also with typical α 1 -adrenergic specificity. Autoradiograms of such membrane samples subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis reveal a major specifically labeled polypeptide at M/sub 4/ = 78,000-85,000, depending on the tissue used, in addition to some lower molecular weight peptides. Protease inhibitors, in particular EDTA, a metalloprotease inhibitor, dramatically increases the predominance of the M/sub r/ = 78,000-85,000 polypeptide while attenuating the labeling of the lower molecular weight bands. This new high affinity radioiodinated photoaffinity probe should be of great value for the molecular characterization of the α 1 -adrenergic receptor

  5. High-affinity RNA aptamers to C-reactive protein (CRP): newly developed pre-elution methods for aptamer selection

    International Nuclear Information System (INIS)

    Orito, N; Umekage, S; Sakai, E; Tanaka, T; Kikuchi, Y; Sato, K; Kawauchi, S; Tanaka, H

    2012-01-01

    We have developed a modified SELEX (systematic evolution of ligands by exponential enrichment) method to obtain RNA aptamers with high affinity to C-reactive protein (CRP). CRP is a clinical biomarker present in plasma, the level of which increases in response to infections and noninfectious inflammation. The CRP level is also an important prognostic indicator in patients with several syndromes. At present, CRP content in blood is measured immunochemically using antibodies. To develop a more sensitive method using RNA aptamers, we have attempted to obtain high-affinity RNA aptamers to CRP. We succeeded in obtaining an RNA aptamer with high affinity to CRP using a CRP-immobilized Sepharose column and pre-elution procedure. Pre-elution is a method that removes the weak binding portion from a selected RNA population by washing for a short time with buffer containing CRP. By surface plasmon-resonance (SPR) analysis, the affinity constant of this aptamer for CRP was calculated to be K D = 2.25x10 -9 (M). The secondary structure, contact sites with CRP protein, and application of this aptamer will be described.

  6. Differences between high-affinity forskolin binding sites in dopamine-riche and other regions of rat brain

    International Nuclear Information System (INIS)

    Poat, J.A.; Cripps, H.E.; Iversen, L.L.

    1988-01-01

    Forskolin labelled with [ 3 H] bound to high- and low-affinity sites in the rat brain. The high-affinity site was discretely located, with highest densities in the striatum, nucleus accumbens, olfactory tubercule, substantia nigra, hippocampus, and the molecular layers of the cerebellum. This site did not correlate well with the distribution of adenylate cyclase. The high-affinity striatal binding site may be associated with a stimulatory guanine nucleotide-binding protein. Thus, the number of sites was increased by the addition of Mg 2+ and guanylyl imidodiphosphate. Cholera toxin stereotaxically injected into rat striatum increased the number of binding sites, and no further increase was noted following the subsequent addition of guanyl nucleotide. High-affinity forskolin binding sites in non-dopamine-rich brain areas (hippocampus and cerebullum) were modulated in a qualitatively different manner by guanyl nucleotides. In these areas the number of binding sites was significantly reduced by the addition of guanyl nucleotide. These results suggest that forskolin may have a potential role in identifying different functional/structural guanine nucleotide-binding proteins

  7. Energy-dependent dissociation of ATP from high affinity catalytic sites of beef heart mitochondrial adenosine triphosphatase

    International Nuclear Information System (INIS)

    Penefsky, H.S.

    1985-01-01

    Incubation of [gamma- 32 P]ATP with a molar excess of the membrane-bound form of mitochondrial ATPase (F1) results in binding of the bulk of the radioactive nucleotide in high affinity catalytic sites (Ka = 10(12) M-1). Subsequent initiation of respiration by addition of succinate or NADH is accompanied by a profound decrease in the affinity for ATP. About one-third of the bound radioactive ATP appears to dissociate, that is, the [gamma- 32 P]ATP becomes accessible to hexokinase. The NADH-stimulated dissociation of [gamma- 32 P]ATP is energy-dependent since the stimulation is inhibited by uncouplers of oxidative phosphorylation and is prevented by respiratory chain inhibitors. The rate of the energy-dependent dissociation of ATP that occurs in the presence of NADH, ADP, and Pi is commensurate with the measured initial rate of ATP synthesis in NADH-supported oxidative phosphorylation catalyzed by the same submitochondrial particles. Thus, the rate of dissociation of ATP from the high affinity catalytic site of submitochondrial particles meets the criterion of kinetic competency under the conditions of oxidative phosphorylation. These experiments provide evidence in support of the argument that energy conserved during the oxidation of substrates by the respiratory chain can be utilized to reduce the very tight binding of product ATP in high affinity catalytic sites and to promote dissociation of the nucleotide

  8. Computational modeling and molecular imprinting for the development of acrylic polymers with high affinity for bile salts.

    Science.gov (United States)

    Yañez, Fernando; Chianella, Iva; Piletsky, Sergey A; Concheiro, Angel; Alvarez-Lorenzo, Carmen

    2010-02-05

    This work has focused on the rational development of polymers capable of acting as traps of bile salts. Computational modeling was combined with molecular imprinting technology to obtain networks with high affinity for cholate salts in aqueous medium. The screening of a virtual library of 18 monomers, which are commonly used for imprinted networks, identified N-(3-aminopropyl)-methacrylate hydrochloride (APMA.HCl), N,N-diethylamino ethyl methacrylate (DEAEM) and ethyleneglycol methacrylate phosphate (EGMP) as suitable functional monomers with medium-to-high affinity for cholic acid. The polymers were prepared with a fix cholic acid:functional monomer mole ratio of 1:4, but with various cross-linking densities. Compared to polymers prepared without functional monomer, both imprinted and non-imprinted microparticles showed a high capability to remove sodium cholate from aqueous medium. High affinity APMA-based particles even resembled the performance of commercially available cholesterol-lowering granules. The imprinting effect was evident in most of the networks prepared, showing that computational modeling and molecular imprinting can act synergistically to improve the performance of certain polymers. Nevertheless, both the imprinted and non-imprinted networks prepared with the best monomer (APMA.HCl) identified by the modeling demonstrated such high affinity for the template that the imprinting effect was less important. The fitting of adsorption isotherms to the Freundlich model indicated that, in general, imprinting increases the population of high affinity binding sites, except when the affinity of the functional monomer for the target molecule is already very high. The cross-linking density was confirmed as a key parameter that determines the accessibility of the binding points to sodium cholate. Materials prepared with 9% mol APMA and 91% mol cross-linker showed enough affinity to achieve binding levels of up to 0.4 mmol g(-1) (i.e., 170 mg g(-1)) under flow

  9. Comparison of high affinity binding of {sup 3}H-proadifen and {sup 3}H-(-)-cocaine t rat liver membranes

    Energy Technology Data Exchange (ETDEWEB)

    Ross, S.B. [Astra Arcus AB, Dept. of Neuropharmacology, Soedertaelje (Sweden)

    1995-06-01

    The characteristics of the binding of {sup 3}H-proadifen to rat liver membranes were studied and compared to those of {sup 3}H-cocaine. It was found that {sup 3}H-proadifen was bound reversibly with high affinity (K{sub D}=1.8{+-}0.5 nM) and large capacity (B{sub max}=2010{+-}340 pmol/g wet tissue) to liver membranes. The corresponding values for the {sup 3}H-cocaine binding were 3.5 nM and 1000 pmol/g wet tissue. The binding of {sup 3}H-proadifen was mainly localised to the microsomal fraction. The number of binding sites was not increased by treatment of rats with phenobarbitone. With 1 {mu}M CdCl{sub 2} in the incubation buffer it was possible to differentiate between two {sup 3}H-cocaine binding sites with K{sub d} values of 1.6 and 7.7 nM and B{sub max} values of 280 and 940 pmol/g wet liver tissue. S-(-)-Alaproclate inhibited the binding of {sup 3}H-proadifen and {sup 3}H-cocaine inhibited the binding of {sup 3}H-proadifen (IC{sub 50}=10 nM) and proadifen that of {sup 3}H-cocaine (IC{sub 50}=1 nM). There was a high correlation coefficient (r{sub r}=0.972; P<0.01; n=12) in the Spearman rank test between the inhibitory potencies of compounds examined in both systems. Beside some potent alaproclate analogues a couple of compounds had moderately high affinity (IC{sub 50}=100-500 nM): chloroquine, phenoxybenzamine, amitriptyline, ajmaline, remoxipride, imipramine and (-)-alaprenolol. CdCl{sub 2}, ZnCl{sub 2} and CuCl{sub 2} inhibited the binding of both ligands with low Hill coefficients, indicating heterogeneous binding sites. The inhibition curve of Cd{sup 2+} on the cocaine binding was biphasic with a high affinity part around 50 nM and a low affinity part at 15{mu}M. The similarity of the characteristics of the binding of these ligands with that of {sup 3}H-alaproclate is discussed. It is suggested that all three compounds bind to the same sites, although additional binding sites seem to exist for proadifen. (au) (9 refs.).

  10. Comparison of systemic cytokine responses after a long distance triathlon and a 100-km run: relationship to metabolic and inflammatory processes.

    Science.gov (United States)

    Gomez-Merino, Danielle; Drogou, Catherine; Guezennec, Charles Yannick; Burnat, Pascal; Bourrilhon, Cyprien; Tomaszewski, Armand; Milhau, Stéphane; Chennaoui, Mounir

    2006-06-01

    Suggested mechanisms for the systemic, circulating cytokinemia observed during heavy physical exertion include inflammation and energy demand. We compared cytokine levels and examined the underlying physiological mechanisms between a long-distance triathlon and a 100-km run, two endurance races of similar duration but characterized by differences in muscle strain. Blood samples were collected from 12 triathletes (34.8 +/- 1.4 yr) and 11 runners (42.4 +/- 2.2 yr) the day before and at the end of races (T1, R1), and 24 h and 7 days post-race (R2, R3). At R1, significant race-related differences were observed, with greater increases in plasma levels of interleukins (IL)-6, IL-1ra, and IL-10 in the triathletes than in the runners, while levels of the chemokine IL-8 increased solely in the runners (P long distance triathlon and a 100-km run suggested that IL-6 and IL-8 could be employed as respective markers of the intensity of the muscular activity required for substrate availability and vascular inflammation.

  11. ANALISA KEANDALAN TERHADAP LIFETIME SYSTEM PENDINGIN KAPAL IKAN KM. RUKUN ARTA SENTOSA 06 MENGGUNAKAN REFRIGERAN CO2 DAN KOMPRESI BANTU DARI ENERGI PANAS

    Directory of Open Access Journals (Sweden)

    Eko Sasmito Hadi

    2012-03-01

    Full Text Available The cooling system using CO2 refrigerant is one of the vital systems to increase the quality of hauled fishing and it is more effective and more efficient and also this system is environmentally harmless than the conventional cooling system. Reliability is one of so many criteria that had to be concerned in the process of taking the decision of invention such as planning/drafting, production, or reparation process. The aim of this research is to evaluate the design of fish hatch cooling system using CO2 refrigerant and auxiliary compression by heat power in KM. RAS (Rukun Arta Sentosa 06 using the approximation based on reliability. Some steps that had been used in this research are collecting the data, design process, and evaluation by qualitative and quantitative approach. The qualitative approach is using the Failure Mode and Effect Analysis (FMEA method and also using Fault Tree Analysis (FTA method. Otherwise, the quantitative approach is using the method of Markov Process. Analytical approach is done by make a design model of the available invention system through the block diagram reliability and decrease the reliability function of the system, and for the next step those system is to be used as the base rule of quantitative evaluation system. The report of the research tells that the severity level and the failure rate are showed in some components, such as compressor, condenser, heat exchanger and expansion valve. The analytical approach will obtain the value of MTTF (Mean Time To Failure system for 250 hours and the availability value is 0,6308 or 63,08% and the unavailability value is 0,3692 or 36,92%.

  12. Large scale use of brazing and high temperature brazing for the fabrication of the 6.4 km long vacuum system of the HERA electron storage ring

    International Nuclear Information System (INIS)

    Ballion, R.; Boster, J.; Giesske, W.; Hartwig, H.; Jagnow, D.; Kouptsidis, J.; Pape, R.; Prohl, W.; Schumann, G.; Schwartz, M.; Iversen, K.; Mucklenbeck, J.

    1989-01-01

    The 6.4 km long vacuum system for electrons in the large storage ring HERA at Hamburg consists of about 1,400 components having lengths between .14 and 12 m. The vacuum components are mainly made from variously shaped tubes of the copper alloy CuSn2. This alloy combines sufficient mechanical strength with the high thermal conductivity needed to remove the 6 MW dissipated power of the synchrotron-light. The vacuum components consist additionally of parts made from stainless steel such as flanges, chambers for pumps, beam monitors, etc. All of these parts are connected in a vacuum tight manner and on a large scale by using brazing and high temperature brazing both in a vacuum or in a reducing gas atmosphere. (orig.)

  13. Blockade of the high-affinity noradrenaline transporter (NET) by the selective 5-HT reuptake inhibitor escitalopram: an in vivo microdialysis study in mice

    Science.gov (United States)

    Nguyen, Hai T; Guiard, Bruno P; Bacq, Alexandre; David, Denis J; David, Indira; Quesseveur, Gaël; Gautron, Sophie; Sanchez, Connie; Gardier, Alain M

    2013-01-01

    BACKGROUND AND PURPOSE Escitalopram, the S(+)-enantiomer of citalopram is the most selective 5-HT reuptake inhibitor approved. Although all 5-HT selective reuptake inhibitors (SSRIs) increase extracellular levels of 5-HT ([5-HT]ext). some also enhance, to a lesser extent, extracellular levels of noradrenaline ([NA]ext). However, the mechanisms by which SSRIs activate noradrenergic transmission in the brain remain to be determined. EXPERIMENTAL APPROACH This study examined the effects of escitalopram, on both [5-HT]ext and [NA]ext in the frontal cortex (FCx) of freely moving wild-type (WT) and mutant mice lacking the 5-HT transporter (SERT−/−) by using intracerebral microdialysis. We explored the possibilities that escitalopram enhances [NA]ext, either by a direct mechanism involving the inhibition of the low- or high-affinity noradrenaline transporters, or by an indirect mechanism promoted by [5-HT]ext elevation. The forced swim test (FST) was used to investigate whether enhancing cortical [5-HT]ext and/or [NA]ext affected the antidepressant-like activity of escitalopram. KEY RESULTS In WT mice, a single systemic administration of escitalopram produced a significant increase in cortical [5-HT]ext and [NA]ext. As expected, escitalopram failed to increase cortical [5-HT]ext in SERT−/− mice, whereas its neurochemical effects on [NA]ext persisted in these mutants. In WT mice subjected to the FST, escitalopram increased swimming parameters without affecting climbing behaviour. Finally, escitalopram, at relevant concentrations, failed to inhibit cortical noradrenaline and 5-HT uptake mediated by low-affinity monoamine transporters. CONCLUSIONS AND IMPLICATIONS These experiments suggest that escitalopram enhances, although moderately, cortical [NA]extin vivo by a direct mechanism involving the inhibition of the high-affinity noradrenaline transporter (NET). PMID:22233336

  14. Topography of the high-affinity lysine binding site of plasminogen as defined with a specific antibody probe

    International Nuclear Information System (INIS)

    Miles, L.A.; Plow, E.F.

    1986-01-01

    An antibody population that reacted with the high-affinity lysine binding site of human plasminogen was elicited by immunizing rabbits with an elastase degradation product containing kringles 1-3 (EDP I). This antibody was immunopurified by affinity chromatography on plasminogen-Sepharose and elution with 0.2 M 6-aminohexanoic acid. The eluted antibodies bound [ 125 I]EDP I, [ 125 I]Glu-plasminogen, and [ 125 I]Lys-plasminogen in radioimmunoassays, and binding of each ligand was at least 99% inhibited by 0.2 M 6-aminohexanoic acid. The concentrations for 50% inhibition of [ 125 I]EDP I binding by tranexamic acid, 6-aminohexanoic acid, and lysine were 2.6, 46, and l730 μM, respectively. Similar values were obtained with plasminogen and suggested that an unoccupied high-affinity lysine binding site was required for antibody recognition. The antiserum reacted exclusively with plasminogen derivatives containing the EDP I region and did not react with those lacking an EDP I region, or with tissue plasminogen activator or prothrombin, which also contains kringles. By immunoblotting analyses, a chymotryptic degradation product of M/sub r/ 20,000 was derived from EDP I that retained reactivity with the antibody. α 2 -Antiplasmin inhibited the binding of radiolabeled EDP I, Glu-plasminogen, or Lys-plasminogen by the antiserum, suggesting that the recognized site is involved in the noncovalent interaction of the inhibitor with plasminogen. The binding of [ 125 I]EDP I to fibrin was also inhibited by the antiserum. The observations provide independent evidence for the role of the high-affinity lysine binding site in the functional interactions of plasminogen with its primary substrate and inhibitor

  15. Structural implications of hERG K+ channel block by a high-affinity minimally structured blocker

    Science.gov (United States)

    Helliwell, Matthew V.; Zhang, Yihong; El Harchi, Aziza; Du, Chunyun; Hancox, Jules C.; Dempsey, Christopher E.

    2018-01-01

    Cardiac potassium channels encoded by human ether-à-go-go–related gene (hERG) are major targets for structurally diverse drugs associated with acquired long QT syndrome. This study characterized hERG channel inhibition by a minimally structured high-affinity hERG inhibitor, Cavalli-2, composed of three phenyl groups linked by polymethylene spacers around a central amino group, chosen to probe the spatial arrangement of side chain groups in the high-affinity drug-binding site of the hERG pore. hERG current (IhERG) recorded at physiological temperature from HEK293 cells was inhibited with an IC50 of 35.6 nm with time and voltage dependence characteristic of blockade contingent upon channel gating. Potency of Cavalli-2 action was markedly reduced for attenuated inactivation mutants located near (S620T; 54-fold) and remote from (N588K; 15-fold) the channel pore. The S6 Y652A and F656A mutations decreased inhibitory potency 17- and 75-fold, respectively, whereas T623A and S624A at the base of the selectivity filter also decreased potency (16- and 7-fold, respectively). The S5 helix F557L mutation decreased potency 10-fold, and both F557L and Y652A mutations eliminated voltage dependence of inhibition. Computational docking using the recent cryo-EM structure of an open channel hERG construct could only partially recapitulate experimental data, and the high dependence of Cavalli-2 block on Phe-656 is not readily explainable in that structure. A small clockwise rotation of the inner (S6) helix of the hERG pore from its configuration in the cryo-EM structure may be required to optimize Phe-656 side chain orientations compatible with high-affinity block. PMID:29545312

  16. A rhodamine-labeled citalopram analogue as a high-affinity fluorescent probe for the serotonin transporter

    DEFF Research Database (Denmark)

    Zhang, Peng; Jørgensen, Trine Nygaard; Løland, Claus Juul

    2013-01-01

    A novel fluorescent ligand was synthesized as a high-affinity, high specificity probe for visualizing the serotonin transporter (SERT). The rhodamine fluorophore was extended from an aniline substitution on the 5-position of the dihydroisobenzofuran ring of citalopram (2, 1-(3-(dimethylamino......)propyl)-1-(4-fluorophenyl)-1,3-dihydroisobenzofuran-5-carbonitrile), using an ethylamino linker. The resulting rhodamine-labeled ligand 8 inhibited [3H]5-HT uptake in COS-7 cells (Ki = 225 nM) with similar potency to the tropane-based JHC 1-064 (1), but with higher specificity towards the SERT relative...

  17. Autoradiographic imaging and quantification of the high-affinity GHB binding sites in rodent brain using (3)H-HOCPCA

    DEFF Research Database (Denmark)

    Klein, A B; Bay, T; Villumsen, I S

    2016-01-01

    analogue, 3-hydroxycyclopent-1-enecarboxylic acid (HOCPCA) as a tritiated version ((3)H-HOCPCA) to radioactively label the specific GHB high-affinity binding site and gain further insight into the density, distribution and developmental profile of this protein. We show that, in low nanomolar concentrations...... brain development. Due to the high sensitivity of this radioligand, we were able to detect low levels of specific binding already at E15 in mouse brain, which increased progressively until adulthood. Collectively, we show that (3)H-HOCPCA is a highly sensitive radioligand, offering advantages over...

  18. Inhibition of Enterococcus faecium adherence to collagen by antibodies against high-affinity binding subdomains of Acm.

    Science.gov (United States)

    Nallapareddy, Sreedhar R; Sillanpää, Jouko; Ganesh, Vannakambadi K; Höök, Magnus; Murray, Barbara E

    2007-06-01

    Strains of Enterococcus faecium express a cell wall-anchored protein, Acm, which mediates adherence to collagen. Here, we (i) identify the minimal and high-affinity binding subsegments of Acm and (ii) show that anti-Acm immunoglobulin Gs (IgGs) purified against these subsegments reduced E. faecium TX2535 strain collagen adherence up to 73 and 50%, respectively, significantly more than the total IgGs against the full-length Acm A domain (28%) (P Acm adherence with functional subsegment-specific antibodies raises the possibility of their use as therapeutic or prophylactic agents.

  19. Cost analysis of teg-powered and solar-powered cathodic protection system for a-50 km long buried natural gas pipeline located in Sindh, Pakistan

    International Nuclear Information System (INIS)

    Shahid, M.; Inam, F.; Farooq, M.; Khan, F.N.

    2005-01-01

    Corrosion leaks are of significant concern to oil and gas industry and is considered to be the largest controllable factor in pipeline safety. Cathodic Protection (CP) is a well-established method for preventing corrosion of metallic materials. Electrical power is required and it is usually difficult and expensive to install conventional power lines in remote areas for readily available power supply. Oil/gas organizations make use of thermo-electric generators (TEG), which is relatively expensive in terms of running expenditures. Utilization of renewable energies is now being widely explored due to potential danger of running out of natural resources and dates back mid of 20th century [I]. However, use of solar powered CP system for oil/gas pipelines hasn't been encouraged much in Pakistan, probably due to lack of understanding. A project was undertaken for designing a solar powered CP system for a 52.4 km buried gas pipeline located at Sui/Sara gas fields (Latitude 27.5) of Tullow Pakistan (Dev.) Ltd. in Dharki, Sindh, Pakistan. After detailed analysis of soil condition, electrochemical testing, local climatic variation and cost analysis, it has been revealed that use of solar power is quite feasible for the above-mentioned pipeline section. Cost analysis and comparison have also favored this system since the maintenance cost of the solar-powered system is much less compared to TEG system. Installation cost of the solar system is about 1.57 times the cost of TEG; however, the maintenance cost is only -20% of that for TEG system. The higher installation cost has been estimated to be recoverable in less than one year of service. (author)

  20. High-affinity DNA-binding Domains of Replication Protein A (RPA) Direct SMARCAL1-dependent Replication Fork Remodeling*

    Science.gov (United States)

    Bhat, Kamakoti P.; Bétous, Rémy; Cortez, David

    2015-01-01

    SMARCAL1 catalyzes replication fork remodeling to maintain genome stability. It is recruited to replication forks via an interaction with replication protein A (RPA), the major ssDNA-binding protein in eukaryotic cells. In addition to directing its localization, RPA also activates SMARCAL1 on some fork substrates but inhibits it on others, thereby conferring substrate specificity to SMARCAL1 fork-remodeling reactions. We investigated the mechanism by which RPA regulates SMARCAL1. Our results indicate that although an interaction between SMARCAL1 and RPA is essential for SMARCAL1 activation, the location of the interacting surface on RPA is not. Counterintuitively, high-affinity DNA binding of RPA DNA-binding domain (DBD) A and DBD-B near the fork junction makes it easier for SMARCAL1 to remodel the fork, which requires removing RPA. We also found that RPA DBD-C and DBD-D are not required for SMARCAL1 regulation. Thus, the orientation of the high-affinity RPA DBDs at forks dictates SMARCAL1 substrate specificity. PMID:25552480

  1. High-affinity DNA-binding domains of replication protein A (RPA) direct SMARCAL1-dependent replication fork remodeling.

    Science.gov (United States)

    Bhat, Kamakoti P; Bétous, Rémy; Cortez, David

    2015-02-13

    SMARCAL1 catalyzes replication fork remodeling to maintain genome stability. It is recruited to replication forks via an interaction with replication protein A (RPA), the major ssDNA-binding protein in eukaryotic cells. In addition to directing its localization, RPA also activates SMARCAL1 on some fork substrates but inhibits it on others, thereby conferring substrate specificity to SMARCAL1 fork-remodeling reactions. We investigated the mechanism by which RPA regulates SMARCAL1. Our results indicate that although an interaction between SMARCAL1 and RPA is essential for SMARCAL1 activation, the location of the interacting surface on RPA is not. Counterintuitively, high-affinity DNA binding of RPA DNA-binding domain (DBD) A and DBD-B near the fork junction makes it easier for SMARCAL1 to remodel the fork, which requires removing RPA. We also found that RPA DBD-C and DBD-D are not required for SMARCAL1 regulation. Thus, the orientation of the high-affinity RPA DBDs at forks dictates SMARCAL1 substrate specificity. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  2. Generation and characterization of a human-mouse chimeric high-affinity antibody that detects the DYKDDDDK FLAG peptide.

    Science.gov (United States)

    Ikeda, Koki; Koga, Tomoaki; Sasaki, Fumiyuki; Ueno, Ayumi; Saeki, Kazuko; Okuno, Toshiaki; Yokomizo, Takehiko

    2017-05-13

    DYKDDDDK peptide (FLAG) is a useful tool for investigating the function and localization of proteins whose antibodies (Abs) are not available. We recently established a high-affinity monoclonal antibody (mAb) for FLAG (clone 2H8). The 2H8 Ab is highly sensitive for detecting FLAG-tagged proteins by flowcytometry and immunoprecipitation, but it can yield nonspecific signals in immunohistochemistry of mouse tissues because it is of mouse origin. In this study, we reduced nonspecific signals by generating a chimeric 2H8 Ab with Fc fragments derived from human immunoglobulin. We fused a 5' terminal cDNA fragments for the Fab region of 2H8 mAb with 3' terminal cDNA fragments for Fc region of human IgG1. We transfected both chimeric plasmids and purified the resulting human-mouse chimeric 2H8. The chimeric 2H8 Ab successfully detected FLAG-tagged proteins in flowcytometry with anti-human IgG secondary Ab with comparable sensitivity to 2H8 mAb. Importantly, chimeric 2H8 detected specific FLAG peptide signals without nonspecific signals in immunohistochemical analysis with mouse tissues. This human-mouse chimeric high-affinity anti-FLAG Ab will prove useful for future immunohistochemical analysis of mouse tissues. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. Structure-guided development of a high-affinity human Programmed Cell Death-1: Implications for tumor immunotherapy

    Energy Technology Data Exchange (ETDEWEB)

    Lázár-Molnár, Eszter; Scandiuzzi, Lisa; Basu, Indranil; Quinn, Thomas; Sylvestre, Eliezer; Palmieri, Edith; Ramagopal, Udupi A.; Nathenson, Stanley G.; Guha, Chandan; Almo, Steven C.

    2017-03-01

    Programmed Cell Death-1 (PD-1) is an inhibitory immune receptor, which plays critical roles in T cell co-inhibition and exhaustion upon binding to its ligands PD-L1 and PD-L2. We report the crystal structure of the human PD-1 ectodomain and the mapping of the PD-1 binding interface. Mutagenesis studies confirmed the crystallographic interface, and resulted in mutant PD-1 receptors with altered affinity and ligand-specificity. In particular, a high-affinity mutant PD-1 (HA PD-1) exhibited 45 and 30-fold increase in binding to PD-L1 and PD-L2, respectively, due to slower dissociation rates. This mutant (A132L) was used to engineer a soluble chimeric Ig fusion protein for cell-based and in vivo studies. HA PD-1 Ig showed enhanced binding to human dendritic cells, and increased T cell proliferation and cytokine production in a mixed lymphocyte reaction (MLR) assay. Moreover, in an experimental model of murine Lewis lung carcinoma, HA PD-1 Ig treatment synergized with radiation therapy to decrease local and metastatic tumor burden, as well as in the establishment of immunological memory responses. Our studies highlight the value of structural considerations in guiding the design of a high-affinity chimeric PD-1 Ig fusion protein with robust immune modulatory properties, and underscore the power of combination therapies to selectively manipulate the PD-1 pathway for tumor immunotherapy.

  4. Structure-guided development of a high-affinity human Programmed Cell Death-1: Implications for tumor immunotherapy

    Directory of Open Access Journals (Sweden)

    Eszter Lázár-Molnár

    2017-03-01

    Full Text Available Programmed Cell Death-1 (PD-1 is an inhibitory immune receptor, which plays critical roles in T cell co-inhibition and exhaustion upon binding to its ligands PD-L1 and PD-L2. We report the crystal structure of the human PD-1 ectodomain and the mapping of the PD-1 binding interface. Mutagenesis studies confirmed the crystallographic interface, and resulted in mutant PD-1 receptors with altered affinity and ligand-specificity. In particular, a high-affinity mutant PD-1 (HA PD-1 exhibited 45 and 30-fold increase in binding to PD-L1 and PD-L2, respectively, due to slower dissociation rates. This mutant (A132L was used to engineer a soluble chimeric Ig fusion protein for cell-based and in vivo studies. HA PD-1 Ig showed enhanced binding to human dendritic cells, and increased T cell proliferation and cytokine production in a mixed lymphocyte reaction (MLR assay. Moreover, in an experimental model of murine Lewis lung carcinoma, HA PD-1 Ig treatment synergized with radiation therapy to decrease local and metastatic tumor burden, as well as in the establishment of immunological memory responses. Our studies highlight the value of structural considerations in guiding the design of a high-affinity chimeric PD-1 Ig fusion protein with robust immune modulatory properties, and underscore the power of combination therapies to selectively manipulate the PD-1 pathway for tumor immunotherapy.

  5. Biphasic regulation of development of the high-affinity saxitoxin receptor by innervation in rat skeletal muscle

    International Nuclear Information System (INIS)

    Sherman, S.J.; Catterall, W.A.

    1982-01-01

    Specific binding of 3 H-saxitoxin (STX) was used to quantitate the density of voltage-sensitive sodium channels in developing rat skeletal muscle. In adult triceps surae, a single class of sites with a KD . 2.9 nM and a density of 21 fmol/mg wet wt was detected. The density of these high-affinity sites increased from 2.0 fmol/mg wet wt to the adult value in linear fashion during days 2-25 after birth. Denervation of the triceps surae at day 11 or 17 reduced final saxitoxin receptor site density to 10.4 or 9.2 fmol/mg wet wt, respectively, without changing KD. Denervation of the triceps surae at day 5 did not alter the subsequent development of saxitoxin receptor sites during days 5-9 and accelerated the increase of saxitoxin receptor sites during days 9-13. After day 13, saxitoxin receptor development abruptly ceased and the density of saxitoxin receptor sites declined to 11 fmol/wg wet wt. These results show that the regulation of high-affinity saxitoxin receptor site density by innervation is biphasic. During the first phase, which is independent of continuing innervation, the saxitoxin receptor density increases to 47-57% of the adult level. After day 11, the second phase of development, which is dependent on continuing innervation, gives rise to the adult density of saxitoxin receptors

  6. Specific recognition of the C-terminal end of A beta 42 by a high affinity monoclonal antibody

    DEFF Research Database (Denmark)

    Axelsen, Trine Veje; Holm, Arne; Birkelund, Svend

    2009-01-01

    The neurotoxic peptide A beta(42) is derived from the amyloid precursor protein by proteolytic cleavage and is deposited in the brain of patients suffering from Alzheimer's disease (AD). In this study we generate a high affinity monoclonal antibody that targets the C-terminal end of A beta(42......) with high specificity. By this is meant that the paratope of the antibody must enclose the C-terminal end of A beta(42) including the carboxy-group of amino acid 42, and not just recognize a linear epitope in the C-terminal part of A beta. This has been accomplished by using a unique antigen construct made...... by the Ligand Presenting Assembly technology (LPA technology). This strategy results in dimeric presentation of the free C-terminal end of A beta(42). The generated Mab A beta1.1 is indeed specific for the C-terminal end of A beta(42) to which it binds with high affinity. Mab A beta1.1 recognizes the epitope...

  7. Susceptibility to endotoxin induced uveitis is not reduced in mice deficient in BLT1, the high affinity leukotriene B4 receptor

    OpenAIRE

    Smith, J R; Subbarao, K; Franc, D T; Haribabu, B; Rosenbaum, J T

    2004-01-01

    Aim: To investigate the role of arachidonic acid derived chemotactic factor, LTB4, in the development of endotoxin induced uveitis (EIU), using mice deficient in the BLT1 gene which encodes the high affinity LTB4 receptor.

  8. G1SST, 1km blended SST

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — A daily, global Sea Surface Temperature (SST) data set is produced at 1-km (also known as ultra-high resolution) by the JPL ROMS (Regional Ocean Modeling System)...

  9. Involvement of the VDE homing endonuclease and rapamycin in regulation of the Saccharomyces cerevisiae GSH11 gene encoding the high affinity glutathione transporter.

    Science.gov (United States)

    Miyake, Tsuyoshi; Hiraishi, Hiroyuki; Sammoto, Hiroyuki; Ono, Bun-Ichiro

    2003-10-10

    The Saccharomyces cerevisiae gene HGT1/GSH11 encodes the high affinity glutathione transporter and is repressed by cysteine added to the culture medium. It has been found previously that a 5'-upstream cis-element, CCGCCACAC, is responsible for regulating GSH11 expression and that several proteins bind to this element (Miyake, T., Kanayama, M., Sammoto, H., and Ono, B. (2002) Mol. Genet. Genomics 266, 1004-1011). In this report we present evidence that the most prominent of these proteins is VDE, known previously as the homing endonuclease encoded by VMA1. We show also that GSH11 is not expressed in a VDE-deleted strain and that inability to express the GSH11 of this strain is overcome by introduction of the coding region of VDE or the entire VMA1 gene. It is also found that VDE does not cut DNA in the vicinity of the GSH11 cis-element. Rapamycin, an inhibitor of the target of rapamycin (TOR) signal-transduction system, is found to enhance expression of GSH11 in a VDE-dependent manner under conditions of sulfur starvation. These results indicate that GSH11 is regulated by a system sensitive to sulfur starvation (presumably via cysteine depletion) and a more general system involving the nutritional starvation signal mediated by the TOR system. Both systems need to be operational (inhibition of TOR and sulfur starvation) for full expression of GSH11.

  10. A pharmacological profile of the high-affinity GluK5 kainate receptor

    DEFF Research Database (Denmark)

    Møllerud, Stine; Kastrup, Jette Sandholm Jensen; Pickering, Darryl S

    2016-01-01

    -hydroxyisoxazol-4-yl)propionate (ATPA), dihydrokainate and (2 S,4 R)−4-methyl-glutamate (SYM2081) have higher affinity at GluK3 compared to GluK5. Since some studies have indicated that GluK5 is associated with various diseases in the central nervous system (e.g. schizophrenia, temporal lobe epilepsy, bipolar...

  11. Synthetic Receptors for the High-Affinity Recognition of O-GlcNAc Derivatives

    NARCIS (Netherlands)

    Rios, Pablo; Carter, Tom S; Mooibroek, Tiddo J; Crump, Matthew P; Lisbjerg, Micke; Pittelkow, Michael; Supekar, Nitin T; Boons, Geert-Jan|info:eu-repo/dai/nl/088245489; Davis, Anthony P

    2016-01-01

    The combination of a pyrenyl tetraamine with an isophthaloyl spacer has led to two new water-soluble carbohydrate receptors ("synthetic lectins"). Both systems show outstanding affinities for derivatives of N-acetylglucosamine (GlcNAc) in aqueous solution. One receptor binds the methyl glycoside

  12. Tsetse salivary gland proteins 1 and 2 are high affinity nucleic acid binding proteins with residual nuclease activity.

    Directory of Open Access Journals (Sweden)

    Guy Caljon

    Full Text Available Analysis of the tsetse fly salivary gland EST database revealed the presence of a highly enriched cluster of putative endonuclease genes, including tsal1 and tsal2. Tsal proteins are the major components of tsetse fly (G. morsitans morsitans saliva where they are present as monomers as well as high molecular weight complexes with other saliva proteins. We demonstrate that the recombinant tsetse salivary gland proteins 1&2 (Tsal1&2 display DNA/RNA non-specific, high affinity nucleic acid binding with K(D values in the low nanomolar range and a non-exclusive preference for duplex. These Tsal proteins exert only a residual nuclease activity with a preference for dsDNA in a broad pH range. Knockdown of Tsal expression by in vivo RNA interference in the tsetse fly revealed a partially impaired blood digestion phenotype as evidenced by higher gut nucleic acid, hematin and protein contents.

  13. Characterization of glucagon-like peptide-1 receptor beta-arrestin 2 interaction: a high-affinity receptor phenotype

    DEFF Research Database (Denmark)

    Jorgensen, Rasmus; Martini, Lene; Schwartz, Thue W

    2005-01-01

    To dissect the interaction between beta-arrestin ((beta)arr) and family B G protein-coupled receptors, we constructed fusion proteins between the glucagon-like peptide 1 receptor and (beta)arr2. The fusion constructs had an increase in apparent affinity selectively for glucagon, suggesting...... that (beta)arr2 interaction locks the receptor in a high-affinity conformation, which can be explored by some, but not all, ligands. The fusion constructs adopted a signaling phenotype governed by the tethered (beta)arr2 with an attenuated G protein-mediated cAMP signal and a higher maximal internalization...... of that which has previously been characterized for family A G protein-coupled receptors, suggesting similarities in the effect of (beta)arr interaction between family A and B receptors also at the molecular level....

  14. High-affinity α4β2 nicotinic receptors mediate the impairing effects of acute nicotine on contextual fear extinction.

    Science.gov (United States)

    Kutlu, Munir Gunes; Holliday, Erica; Gould, Thomas J

    2016-02-01

    Previously, studies from our lab have shown that while acute nicotine administered prior to training and testing enhances contextual fear conditioning, acute nicotine injections prior to extinction sessions impair extinction of contextual fear. Although there is also strong evidence showing that the acute nicotine's enhancing effects on contextual fear conditioning require high-affinity α4β2 nicotinic acetylcholine receptors (nAChRs), it is unknown which nAChR subtypes are involved in the acute nicotine-induced impairment of contextual fear extinction. In this study, we investigated the effects of acute nicotine administration on contextual fear extinction in knock-out (KO) mice lacking α4, β2 or α7 subtypes of nAChRs and their wild-type (WT) littermates. Both KO and WT mice were first trained and tested for contextual fear conditioning and received a daily contextual extinction session for 4 days. Subjects received intraperitoneal injections of nicotine (0.18 mg/kg) or saline 2-4 min prior to each extinction session. Our results showed that the mice that lack α4 and β2 subtypes of nAChRs showed normal contextual fear extinction but not the acute nicotine-induced impairment while the mice that lack the α7 subtype showed both normal contextual extinction and nicotine-induced impairment of contextual extinction. In addition, control experiments showed that acute nicotine-induced impairment of contextual fear extinction persisted when nicotine administration was ceased and repeated acute nicotine administrations alone did not induce freezing behavior in the absence of context-shock learning. These results clearly demonstrate that high-affinity α4β2 nAChRs are necessary for the effects of acute nicotine on contextual fear extinction. Copyright © 2015 Elsevier Inc. All rights reserved.

  15. Visual and Plasmon Resonance Absorption Sensor for Adenosine Triphosphate Based on the High Affinity between Phosphate and Zr(IV).

    Science.gov (United States)

    Qi, Wenjing; Liu, Zhongyuan; Zhang, Wei; Halawa, Mohamed Ibrahim; Xu, Guobao

    2016-10-12

    Zr(IV) can form phosphate and Zr(IV) (-PO₃ 2- -Zr 4+ -) complex owing to the high affinity between Zr(IV) with phosphate. Zr(IV) can induce the aggregation of gold nanoparticles (AuNPs), while adenosine triphosphate(ATP) can prevent Zr(IV)-induced aggregation of AuNPs. Herein, a visual and plasmon resonance absorption (PRA)sensor for ATP have been developed using AuNPs based on the high affinity between Zr(IV)with ATP. AuNPs get aggregated in the presence of certain concentrations of Zr(IV). After the addition of ATP, ATP reacts with Zr(IV) and prevents AuNPs from aggregation, enabling the detection of ATP. Because of the fast interaction of ATP with Zr(IV), ATP can be detected with a detection limit of 0.5 μM within 2 min by the naked eye. Moreover, ATP can be detected by the PRA technique with higher sensitivity. The A 520nm / A 650nm values in PRA spectra increase linearly with the concentrations of ATP from 0.1 μM to 15 μM (r = 0.9945) with a detection limit of 28 nM. The proposed visual and PRA sensor exhibit good selectivity against adenosine, adenosine monophosphate, guanosine triphosphate, cytidine triphosphate and uridine triphosphate. The recoveries for the analysis of ATP in synthetic samples range from 95.3% to 102.0%. Therefore, the proposed novel sensor for ATP is promising for real-time or on-site detection of ATP.

  16. Visual and Plasmon Resonance Absorption Sensor for Adenosine Triphosphate Based on the High Affinity between Phosphate and Zr(IV

    Directory of Open Access Journals (Sweden)

    Wenjing Qi

    2016-10-01

    Full Text Available Zr(IV can form phosphate and Zr(IV (–PO32−–Zr4+– complex owing to the high affinity between Zr(IV with phosphate. Zr(IV can induce the aggregation of gold nanoparticles (AuNPs, while adenosine triphosphate(ATP can prevent Zr(IV-induced aggregation of AuNPs. Herein, a visual and plasmon resonance absorption (PRAsensor for ATP have been developed using AuNPs based on the high affinity between Zr(IVwith ATP. AuNPs get aggregated in the presence of certain concentrations of Zr(IV. After the addition of ATP, ATP reacts with Zr(IV and prevents AuNPs from aggregation, enabling the detection of ATP. Because of the fast interaction of ATP with Zr(IV, ATP can be detected with a detection limit of 0.5 μM within 2 min by the naked eye. Moreover, ATP can be detected by the PRA technique with higher sensitivity. The A520nm/A650nm values in PRA spectra increase linearly with the concentrations of ATP from 0.1 μM to 15 μM (r = 0.9945 with a detection limit of 28 nM. The proposed visual and PRA sensor exhibit good selectivity against adenosine, adenosine monophosphate, guanosine triphosphate, cytidine triphosphate and uridine triphosphate. The recoveries for the analysis of ATP in synthetic samples range from 95.3% to 102.0%. Therefore, the proposed novel sensor for ATP is promising for real-time or on-site detection of ATP.

  17. Two high-affinity ligand binding states of uterine estrogen receptor distinguished by modulation of hydrophobic environment

    International Nuclear Information System (INIS)

    Hutchens, T.W.; Li, C.M.; Zamah, N.M.; Besch, P.K.

    1987-01-01

    The steroid binding function of soluble (cytosolic) estrogen receptors from calf uteri was evaluated under conditions known to modify the extent of hydrophobic interaction with receptor-associated proteins. Receptor preparations were equilibrated into 6 M urea buffers and control buffers by chromatography through small columns of Sephadex G-25 or by dialysis at 0.6 0 C. Equilibrium dissociation constants (K/sub d/) and binding capacities (n) of experimental and control receptor preparations were determined by 13-point Scatchard analyses using concentrations of 17β-[ 3 H]estradiol from 0.05 to 10 nM. Nonspecific binding was determined at each concentration by parallel incubations with a 200-fold molar excess of the receptor-specific competitor diethylstilbestrol. The control receptor population was consistently found to be a single class of binding sites with a high affinity for estradiol which was unaffected by G-25 chromatography, by dialysis, by dilution, or by the presence of 0.4 M KCl. However, equilibration into 6 M urea induced a discrete (10-fold) reduction in receptor affinity to reveal a second, thermodynamically stable, high-affinity binding state. The presence of 0.4 M KCl did not significantly influence the discrete change in receptor affinity induced by urea. The effects of urea on both receptor affinity and binding capacity were reversible, suggesting a lack of covalent modification. These results demonstrate nonenzymatic means by which not only the binding capacity but also the affinity of receptor for estradiol can be reversibly controlled, suggesting that high concentrations of urea might be more effectively utilized during the physicochemical characterization and purification of steroid receptor proteins

  18. Cyr61/CCN1 displays high-affinity binding to the somatomedin B(1-44 domain of vitronectin.

    Directory of Open Access Journals (Sweden)

    Ivo M B Francischetti

    2010-02-01

    Full Text Available Cyr61 is a member of the CCN (Cyr61, connective tissue growth, NOV family of extracellular-associated (matricellular proteins that present four distinct functional modules, namely insulin-like growth factor binding protein (IGFBP, von Willebrand factor type C (vWF, thrombospondin type 1 (TSP, and C-terminal growth factor cysteine knot (CT domain. While heparin sulphate proteoglycans reportedly mediate the interaction of Cyr61 with the matrix and cell surface, the role of other extracellular associated proteins has not been revealed.In this report, surface plasmon resonance (SPR experiments and solid-phase binding assays demonstrate that recombinant Cyr61 interacts with immobilized monomeric or multimeric vitronectin (VTNC with K(D in the nanomolar range. Notably, the binding site for Cyr61 was identified as the somatomedin B domain (SMTB(1-44 of VTNC, which mediates its interaction with PAI-1, uPAR, and integrin alphav beta3. Accordingly, PAI-1 outcompetes Cyr61 for binding to immobilized SMTB(1-44, and Cyr61 attenuates uPAR-mediated U937 adhesion to VTNC. In contrast, isothermal titration calorimetry shows that Cyr61 does not display high-affinity binding for SMTB(1-44 in solution. Nevertheless, competitive ELISA revealed that multimeric VTNC, heat-modified monomeric VTNC, or SMTB(1-44 at high concentrations attenuate Cyr61 binding to immobilized VTNC, while monomeric VTNC was ineffective. Therefore, immobilization of VTNC exposes cryptic epitopes that recognize Cyr61 with high affinity, as reported for a number of antibodies, beta-endorphin, and other molecules.The finding that Cyr61 interacts with the SMTB(1-44 domain suggests that VTNC represent a point of anchorage for CCN family members to the matrix. Results are discussed in the context of the role of CCN and VTNC in matrix biology and angiogenesis.

  19. High affinity antigen recognition of the dual specific variants of herceptin is entropy-driven in spite of structural plasticity.

    Directory of Open Access Journals (Sweden)

    Jenny Bostrom

    Full Text Available The antigen-binding site of Herceptin, an anti-human Epidermal Growth Factor Receptor 2 (HER2 antibody, was engineered to add a second specificity toward Vascular Endothelial Growth Factor (VEGF to create a high affinity two-in-one antibody bH1. Crystal structures of bH1 in complex with either antigen showed that, in comparison to Herceptin, this antibody exhibited greater conformational variability, also called "structural plasticity". Here, we analyzed the biophysical and thermodynamic properties of the dual specific variants of Herceptin to understand how a single antibody binds two unrelated protein antigens. We showed that while bH1 and the affinity-improved bH1-44, in particular, maintained many properties of Herceptin including binding affinity, kinetics and the use of residues for antigen recognition, they differed in the binding thermodynamics. The interactions of bH1 and its variants with both antigens were characterized by large favorable entropy changes whereas the Herceptin/HER2 interaction involved a large favorable enthalpy change. By dissecting the total entropy change and the energy barrier for dual interaction, we determined that the significant structural plasticity of the bH1 antibodies demanded by the dual specificity did not translate into the expected increase of entropic penalty relative to Herceptin. Clearly, dual antigen recognition of the Herceptin variants involves divergent antibody conformations of nearly equivalent energetic states. Hence, increasing the structural plasticity of an antigen-binding site without increasing the entropic cost may play a role for antibodies to evolve multi-specificity. Our report represents the first comprehensive biophysical analysis of a high affinity dual specific antibody binding two unrelated protein antigens, furthering our understanding of the thermodynamics that drive the vast antigen recognition capacity of the antibody repertoire.

  20. Structure-based engineering to restore high affinity binding of an isoform-selective anti-TGFβ1 antibody

    Science.gov (United States)

    Honey, Denise M.; Best, Annie; Qiu, Huawei

    2018-01-01

    ABSTRACT Metelimumab (CAT192) is a human IgG4 monoclonal antibody developed as a TGFβ1-specific antagonist. It was tested in clinical trials for the treatment of scleroderma but later terminated due to lack of efficacy. Subsequent characterization of CAT192 indicated that its TGFβ1 binding affinity was reduced by ∼50-fold upon conversion from the parental single-chain variable fragment (scFv) to IgG4. We hypothesized this result was due to decreased conformational flexibility of the IgG that could be altered via engineering. Therefore, we designed insertion mutants in the elbow region and screened for binding and potency. Our results indicated that increasing the elbow region linker length in each chain successfully restored the isoform-specific and high affinity binding of CAT192 to TGFβ1. The crystal structure of the high binding affinity mutant displays large conformational rearrangements of the variable domains compared to the wild-type antigen-binding fragment (Fab) and the low binding affinity mutants. Insertion of two glycines in both the heavy and light chain elbow regions provided sufficient flexibility for the variable domains to extend further apart than the wild-type Fab, and allow the CDR3s to make additional interactions not seen in the wild-type Fab structure. These interactions coupled with the dramatic conformational changes provide a possible explanation of how the scFv and elbow-engineered Fabs bind TGFβ1 with high affinity. This study demonstrates the benefits of re-examining both structure and function when converting scFv to IgG molecules, and highlights the potential of structure-based engineering to produce fully functional antibodies. PMID:29333938

  1. Geomagnetic Kn, Ks, Km Indices

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — A full description of the indices Kn, Ks, Km is given in a monography,"Indices Kn, Ks et Km, 1964-1967", edited in 1968 by the Centre National de la Recherche...

  2. Regulation of rat liver cytochrome P450j, a high affinity N-nitrosodimethylamine demethylase (NDMAD)

    International Nuclear Information System (INIS)

    Thomas, P.E.; Bandiera, S.; Maines, S.L.; Ryan, D.E.; Levin, W.

    1987-01-01

    Purified IgG from sera of rabbits immunized with homogeneous P450j was absorbed to produce monospecific anti-P450j. Results using anti-P450j in ELISA show that rat liver microsomal P450j content decreases between 3 and 6 wks of age in both sexes. Several xenobiotics (Aroclor 1254, mirex and 3-methylcholanthrene) repressed P450j levels when administered to male rats. In contrast, hepatic levels of P450j were induced by isoniazid, dimethylsulfoxide, pyrazole, 4-methylpyrazole, ethanol and chemically-induced diabetes. P450j levels were measurable in kidney, whereas this isozyme was barely detectable in lung, ovaries and testes; however, extra-hepatic P450j was inducible by isoniazid. Between 80-90% of microsomal NDMAD was inhibited by anti-P450j whether the microsomes were isolated from untreated rats or animals administered inducers or repressors of P450j. Results obtained with the reconstituted system suggest that the remaining microsomal NDMAD resistant to antibody inhibition is the result of the inaccessibility of a certain proportion of P450j due to interference by NADPH-P450 reductase. P450j content and NDMAD activity correlated well in microsomes from rats of all treatment groups. The evidence indicates that P450j is the primary, and possibly only, microsomal catalyst of NDMAD at substrate concentrations relevant to hepatocarcinogenesis induced by NDMA

  3. Rapid Diagnostic Assay for Intact Influenza Virus Using a High Affinity Hemagglutinin Binding Protein.

    Science.gov (United States)

    Anderson, Caitlin E; Holstein, Carly A; Strauch, Eva-Maria; Bennett, Steven; Chevalier, Aaron; Nelson, Jorgen; Fu, Elain; Baker, David; Yager, Paul

    2017-06-20

    Influenza is a ubiquitous and recurring infection that results in approximately 500 000 deaths globally each year. Commercially available rapid diagnostic tests are based upon detection of the influenza nucleoprotein, which are limited in that they are unable to differentiate by species and require an additional viral lysis step. Sample preprocessing can be minimized or eliminated by targeting the intact influenza virus, thereby reducing assay complexity and leveraging the large number of hemagglutinin proteins on the surface of each virus. Here, we report the development of a paper-based influenza assay that targets the hemagglutinin protein; the assay employs a combination of antibodies and novel computationally designed, recombinant affinity proteins as the capture and detection agents. This system leverages the customizability of recombinant protein design to target the conserved receptor-binding pocket of the hemagglutinin protein and to match the trimeric nature of hemagglutinin for improved avidity. Using this assay, we demonstrate the first instance of intact influenza virus detection using a combination of antibody and affinity proteins within a porous network. The recombinant head region binder based assays yield superior analytical sensitivity as compared to the antibody based assay, with lower limits of detection of 3.54 × 10 7 and 1.34 × 10 7 CEID 50 /mL for the mixed and all binder stacks, respectively. Not only does this work describe the development of a novel influenza assay, it also demonstrates the power of recombinant affinity proteins for use in rapid diagnostic assays.

  4. Glucocorticoid up-regulation of high-affinity interleukin 6 receptors on human epithelial cells

    International Nuclear Information System (INIS)

    Snyers, L.; De Wit, L.; Content, J.

    1990-01-01

    Interleukin 6 (IL-6) is a potent pleiotropic cytokine, known, among others, to stimulate immunoglobulin production by B cells and to trigger acute-phase protein synthesis by hepatocytes. Similar to IL-1, it is produced by monocytes and macrophages following an inflammatory challenge. Analysis of IL-6 receptor (IL-6R) expression on different human cell lines indicates that dexamethasone could up-regulate the number of IL-6R on one epithelial cell line (UAC) and on two hepatoma cell lines (HepG2 and Hep3B). This effect was confirmed by Scatchard analysis of binding experiments, using [ 35 S]methionine and [ 35 S]cysteine metabolically labeled IL-6. It was confirmed at the level of mRNA expression by Northern blot analysis. These results provide evidence for a link between IL-6 and glucocorticoids. They could represent an example of a system in which one role of glucocorticoids is to define more accurately the target of cytokines, and they could explain, at least partly, the frequently observed synergy between IL-6 and glucocorticoids, notably in the case of hepatocytes

  5. Premature Aging Phenotype in Mice Lacking High-Affinity Nicotinic Receptors: Region-Specific Changes in Layer V Pyramidal Cell Morphology.

    Science.gov (United States)

    Konsolaki, Eleni; Skaliora, Irini

    2015-08-01

    The mechanisms by which aging leads to alterations in brain structure and cognitive deficits are unclear. Α deficient cholinergic system has been implicated as one of the main factors that could confer a heightened vulnerability to the aging process, and mice lacking high-affinity nicotinic receptors (β2(-/-)) have been proposed as an animal model of accelerated cognitive aging. To date, however, age-related changes in neuronal microanatomy have not been studied in these mice. In the present study, we examine the neuronal structure of yellow fluorescent protein (YFP(+)) layer V neurons in 2 cytoarchitectonically distinct cortical regions in wild-type (WT) and β2(-/-) animals. We find that (1) substantial morphological differences exist between YFP(+) cells of the anterior cingulate cortex (ACC) and primary visual cortex (V1), in both genotypes; (2) in WT animals, ACC cells are more susceptible to aging compared with cells in V1; and (3) β2 deletion is associated with a regionally and temporally specific increase in vulnerability to aging. ACC cells exhibit a prematurely aged phenotype already at 4-6 months, whereas V1 cells are spared in adulthood but strongly affected in old animals. Collectively, our data reveal region-specific synergistic effects of aging and genotype and suggest distinct vulnerabilities in V1 and ACC neurons. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  6. High affinity (3H) β-Alanine uptake by scar margins of ferric chloride-induced epileptogenic foci in rat isocortex

    International Nuclear Information System (INIS)

    Robitaille, Y.; Sherwin, A.

    1984-01-01

    Cortical astrocytes of normal mammalian brain are endowed with a high affinity uptake system for β-Alanine which is competitively inhibited by gamma aminobutyric acid (GABA), a neurotransmitter strongly implicated in epileptogenesis. The authors evaluated ( 3 H) β-Alanine uptake by reactive astrocytes proliferating within scar of epileptogenic foci induced in rat motor cortex by microinjections of 100 mM ferric chloride. Following in vitro incubation of scar tissue with ( 3 H) β-Alanine, ultrastructural morphometry of grain patterns at 5, 30 and 120 days post injection revealed early and significant grain count increases over astroglial processes, predominantly those related to perivascular glial end-feet. Astrocytic cell body and endothelial cell counts showed a more gradual and stepwise increase. Similar data were obtained by comparing visual and edited mean astrocytic grain counts. These results suggest that the enhanced uptake of reactive astrocytes may reflect a marked decrease of inhibitory GABAergic neurons within ferric chloride-induced scars. 7 figures, 1 table

  7. KM3NeT

    CERN Multimedia

    KM3NeT is a large scale next-generation neutrino telescope located in the deep waters of the Mediterranean Sea, optimized for the discovery of galactic neutrino sources emitting in the TeV energy region.

  8. Comparison of high affinity binding of 3H-proadifen and 3H-(-)-cocaine t rat liver membranes

    International Nuclear Information System (INIS)

    Ross, S.B.

    1995-01-01

    The characteristics of the binding of 3 H-proadifen to rat liver membranes were studied and compared to those of 3 H-cocaine. It was found that 3 H-proadifen was bound reversibly with high affinity (K D =1.8±0.5 nM) and large capacity (B max =2010±340 pmol/g wet tissue) to liver membranes. The corresponding values for the 3 H-cocaine binding were 3.5 nM and 1000 pmol/g wet tissue. The binding of 3 H-proadifen was mainly localised to the microsomal fraction. The number of binding sites was not increased by treatment of rats with phenobarbitone. With 1 μM CdCl 2 in the incubation buffer it was possible to differentiate between two 3 H-cocaine binding sites with K d values of 1.6 and 7.7 nM and B max values of 280 and 940 pmol/g wet liver tissue. S-(-)-Alaproclate inhibited the binding of 3 H-proadifen and 3 H-cocaine inhibited the binding of 3 H-proadifen (IC 50 =10 nM) and proadifen that of 3 H-cocaine (IC 50 =1 nM). There was a high correlation coefficient (r r =0.972; P 50 =100-500 nM): chloroquine, phenoxybenzamine, amitriptyline, ajmaline, remoxipride, imipramine and (-)-alaprenolol. CdCl 2 , ZnCl 2 and CuCl 2 inhibited the binding of both ligands with low Hill coefficients, indicating heterogeneous binding sites. The inhibition curve of Cd 2+ on the cocaine binding was biphasic with a high affinity part around 50 nM and a low affinity part at 15μM. The similarity of the characteristics of the binding of these ligands with that of 3 H-alaproclate is discussed. It is suggested that all three compounds bind to the same sites, although additional binding sites seem to exist for proadifen. (au) (9 refs.)

  9. High affinity [3H]glibenclamide binding sites in rat neuronal and cardiac tissue: Localization and developmental characteristics

    International Nuclear Information System (INIS)

    Miller, J.A.; Velayo, N.L.; Dage, R.C.; Rampe, D.

    1991-01-01

    We examined the binding of the antidiabetic sulfonylurea [3H] glibenclamide to rat brain and heart membranes. High affinity binding was observed in adult rat forebrain (Kd = 137.3 pM, maximal binding site density = 91.8 fmol/mg of protein) and ventricle (Kd = 77.1 pM, maximal binding site density = 65.1 fmol/mg of protein). Binding site density increased approximately 250% in forebrain membranes during postnatal development but was constant in ventricular membranes. Quantitative autoradiography was used to examine the regional distribution of [3H] glibenclamide binding sites in sections from rat brain, spinal cord and heart. The greatest density of binding in adult brain was found in the substantia nigra and globus pallidus, whereas the other areas displayed heterogenous binding. In agreement with the membrane binding studies, 1-day-old rat brain had significantly fewer [3H]glibenclamide binding sites than adult brain. Additionally, the pattern of distribution of these sites was qualitatively different from that of the adult. In adult rat spinal cord, moderate binding densities were observed in spinal cord gray and displayed a rostral to caudal gradient. In adult rat heart, moderate binding densities were observed and the sites were distributed homogeneously. In conclusion, significant development of [3H]glibenclamide binding sites was seen in the brain but not the heart during postnatal maturation. Furthermore, a heterogeneous distribution of binding sites was observed in both the brain and spinal cord of adult rats

  10. ZipA binds to FtsZ with high affinity and enhances the stability of FtsZ protofilaments.

    Directory of Open Access Journals (Sweden)

    Anuradha Kuchibhatla

    Full Text Available A bacterial membrane protein ZipA that tethers FtsZ to the membrane is known to promote FtsZ assembly. In this study, the binding of ZipA to FtsZ was monitored using fluorescence spectroscopy. ZipA was found to bind to FtsZ with high affinities at three different (6.0, 6.8 and 8.0 pHs, albeit the binding affinity decreased with increasing pH. Further, thick bundles of FtsZ protofilaments were observed in the presence of ZipA under the pH conditions used in this study indicating that ZipA can promote FtsZ assembly and stabilize FtsZ polymers under unfavorable conditions. Bis-ANS, a hydrophobic probe, decreased the interaction of FtsZ and ZipA indicating that the interaction between FtsZ and ZipA is hydrophobic in nature. ZipA prevented the dilution induced disassembly of FtsZ polymers suggesting that it stabilizes FtsZ protofilaments. Fluorescein isothiocyanate-labeled ZipA was found to be uniformly distributed along the length of the FtsZ protofilaments indicating that ZipA stabilizes FtsZ protofilaments by cross-linking them.

  11. A novel lentiviral scFv display library for rapid optimization and selection of high affinity antibodies.

    Science.gov (United States)

    Qudsia, Sehar; Merugu, Siva B; Mangukiya, Hitesh B; Hema, Negi; Wu, Zhenghua; Li, Dawei

    2018-04-30

    Antibody display libraries have become a popular technique to screen monoclonal antibodies for therapeutic purposes. An important aspect of display technology is to generate an optimization library by changing antibody affinity to antigen through mutagenesis and screening the high affinity antibody. In this study, we report a novel lentivirus display based optimization library antibody in which Agtuzumab scFv is displayed on cell membrane of HEK-293T cells. To generate an optimization library, hotspot mutagenesis was performed to achieve diverse antibody library. Based on sequence analysis of randomly selected clones, library size was estimated approximately to be 1.6 × 10 6 . Lentivirus display vector was used to display scFv antibody on cell surface and flow cytometery was performed to check the antibody affinity to antigen. Membrane bound scFv antibodies were then converted to secreted antibody through cre/loxP recombination. One of the mutant clones, M8 showed higher affinity to antigen in flow cytometery analysis. Further characterization of cellular and secreted scFv through western blot showed that antibody affinity was increased by three fold after mutagenesis. This study shows successful construction of a novel antibody library and suggests that hotspot mutagenesis could prove a useful and rapid optimization tool to generate similar libraries with various degree of antigen affinity. Copyright © 2018 Elsevier Inc. All rights reserved.

  12. Cyclic GMP-AMP containing mixed phosphodiester linkages is an endogenous high-affinity ligand for STING.

    Science.gov (United States)

    Zhang, Xu; Shi, Heping; Wu, Jiaxi; Zhang, Xuewu; Sun, Lijun; Chen, Chuo; Chen, Zhijian J

    2013-07-25

    The presence of microbial or self DNA in the cytoplasm of mammalian cells is a danger signal detected by the DNA sensor cyclic-GMP-AMP (cGAMP) synthase (cGAS), which catalyzes the production of cGAMP that in turn serves as a second messenger to activate innate immune responses. Here we show that endogenous cGAMP in mammalian cells contains two distinct phosphodiester linkages, one between 2'-OH of GMP and 5'-phosphate of AMP, and the other between 3'-OH of AMP and 5'-phosphate of GMP. This molecule, termed 2'3'-cGAMP, is unique in that it binds to the adaptor protein STING with a much greater affinity than cGAMP molecules containing other combinations of phosphodiester linkages. The crystal structure of STING bound to 2'3'-cGAMP revealed the structural basis of this high-affinity binding and a ligand-induced conformational change in STING that may underlie its activation. Copyright © 2013 Elsevier Inc. All rights reserved.

  13. A soluble form of the high affinity IgE receptor, Fc-epsilon-RI, circulates in human serum.

    Directory of Open Access Journals (Sweden)

    Eleonora Dehlink

    Full Text Available Soluble IgE receptors are potential in vivo modulators of IgE-mediated immune responses and are thus important for our basic understanding of allergic responses. We here characterize a novel soluble version of the IgE-binding alpha-chain of Fc-epsilon-RI (sFcεRI, the high affinity receptor for IgE. sFcεRI immunoprecipitates as a protein of ∼40 kDa and contains an intact IgE-binding site. In human serum, sFcεRI is found as a soluble free IgE receptor as well as a complex with IgE. Using a newly established ELISA, we show that serum sFcεRI levels correlate with serum IgE in patients with elevated IgE. We also show that serum of individuals with normal IgE levels can be found to contain high levels of sFcεRI. After IgE-antigen-mediated crosslinking of surface FcεRI, we detect sFcεRI in the exosome-depleted, soluble fraction of cell culture supernatants. We further show that sFcεRI can block binding of IgE to FcεRI expressed at the cell surface. In summary, we here describe the alpha-chain of FcεRI as a circulating soluble IgE receptor isoform in human serum.

  14. High affinity γPNA sandwich hybridization assay for rapid detection of short nucleic acid targets with single mismatch discrimination.

    Science.gov (United States)

    Goldman, Johnathan M; Zhang, Li Ang; Manna, Arunava; Armitage, Bruce A; Ly, Danith H; Schneider, James W

    2013-07-08

    Hybridization analysis of short DNA and RNA targets presents many challenges for detection. The commonly employed sandwich hybridization approach cannot be implemented for these short targets due to insufficient probe-target binding strengths for unmodified DNA probes. Here, we present a method capable of rapid and stable sandwich hybridization detection for 22 nucleotide DNA and RNA targets. Stable hybridization is achieved using an n-alkylated, polyethylene glycol γ-carbon modified peptide nucleic acid (γPNA) amphiphile. The γPNA's exceptionally high affinity enables stable hybridization of a second DNA-based probe to the remaining bases of the short target. Upon hybridization of both probes, an electrophoretic mobility shift is measured via interaction of the n-alkane modification on the γPNA with capillary electrophoresis running buffer containing nonionic surfactant micelles. We find that sandwich hybridization of both probes is stable under multiple binding configurations and demonstrate single base mismatch discrimination. The binding strength of both probes is also stabilized via coaxial stacking on adjacent hybridization to targets. We conclude with a discussion on the implementation of the proposed sandwich hybridization assay as a high-throughput microRNA detection method.

  15. High-Affinity Interaction of the K-Ras4B Hypervariable Region with the Ras Active Site

    Science.gov (United States)

    Chavan, Tanmay S.; Jang, Hyunbum; Khavrutskii, Lyuba; Abraham, Sherwin J.; Banerjee, Avik; Freed, Benjamin C.; Johannessen, Liv; Tarasov, Sergey G.; Gaponenko, Vadim; Nussinov, Ruth; Tarasova, Nadya I.

    2015-01-01

    Ras proteins are small GTPases that act as signal transducers between cell surface receptors and several intracellular signaling cascades. They contain highly homologous catalytic domains and flexible C-terminal hypervariable regions (HVRs) that differ across Ras isoforms. KRAS is among the most frequently mutated oncogenes in human tumors. Surprisingly, we found that the C-terminal HVR of K-Ras4B, thought to minimally impact the catalytic domain, directly interacts with the active site of the protein. The interaction is almost 100-fold tighter with the GDP-bound than the GTP-bound protein. HVR binding interferes with Ras-Raf interaction, modulates binding to phospholipids, and slightly slows down nucleotide exchange. The data indicate that contrary to previously suggested models of K-Ras4B signaling, HVR plays essential roles in regulation of signaling. High affinity binding of short peptide analogs of HVR to K-Ras active site suggests that targeting this surface with inhibitory synthetic molecules for the therapy of KRAS-dependent tumors is feasible. PMID:26682817

  16. The RCK1 high-affinity Ca2+ sensor confers carbon monoxide sensitivity to Slo1 BK channels.

    Science.gov (United States)

    Hou, Shangwei; Xu, Rong; Heinemann, Stefan H; Hoshi, Toshinori

    2008-03-11

    Carbon monoxide (CO) is a lethal gas, but it is also increasingly recognized as a physiological signaling molecule capable of regulating a variety of proteins. Among them, large-conductance Ca(2+)- and voltage-gated K(+) (Slo1 BK) channels, important in vasodilation and neuronal firing, have been suggested to be directly stimulated by CO. However, the molecular mechanism of the stimulatory action of CO on the Slo1 BK channel has not been clearly elucidated. We report here that CO reliably and repeatedly activates Slo1 BK channels in excised membrane patches in the absence of Ca(2+) in a voltage-sensor-independent manner. The stimulatory action of CO on the Slo1 BK channel requires an aspartic acid and two histidine residues located in the cytoplasmic RCK1 domain, and the effect persists under the conditions known to inhibit the conventional interaction between CO and heme in other proteins. We propose that CO acts as a partial agonist for the high-affinity divalent cation sensor in the RCK1 domain of the Slo1 BK channel.

  17. Identification and properties of very high affinity brain membrane-binding sites for a neurotoxic phospholipase from the taipan venom

    Energy Technology Data Exchange (ETDEWEB)

    Lambeau, G.; Barhanin, J.; Schweitz, H.; Qar, J.; Lazdunski, M. (Centre de Biochimie, Nice (France))

    1989-07-05

    Four new monochain phospholipases were purified from the Oxyuranus scutellatus (taipan) venom. Three of them were highly toxic when injected into mice brain. One of these neurotoxic phospholipases, OS2, was iodinated and used in binding experiments to demonstrate the presence of two families of specific binding sites in rat brain synaptic membranes. The affinities were exceptionally high, Kd1 = 1.5 +/- 0.5 pM and Kd2 = 45 +/- 10 pM, and the maximal binding capacities were Bmax 1 = 1 +/- 0.4 and Bmax 2 = 3 +/- 0.5 pmol/mg of protein. Both binding sites were sensitive to proteolysis and demonstrated to be located on proteins of Mr 85,000-88,000 and 36,000-51,000 by cross-linking and photoaffinity labeling techniques. The binding of {sup 125}I-OS2 to synaptic membranes was dependent on Ca2+ ions and enhanced by Zn2+ ions which inhibit phospholipase activity. Competition experiments have shown that, except for beta-bungarotoxin, a number of known toxic snake or bee phospholipases have very high affinities for the newly identified binding sites. A good correlation (r = 0.80) was observed between toxicity and affinity but not between phospholipase activity and affinity.

  18. Identification and properties of very high affinity brain membrane-binding sites for a neurotoxic phospholipase from the taipan venom

    International Nuclear Information System (INIS)

    Lambeau, G.; Barhanin, J.; Schweitz, H.; Qar, J.; Lazdunski, M.

    1989-01-01

    Four new monochain phospholipases were purified from the Oxyuranus scutellatus (taipan) venom. Three of them were highly toxic when injected into mice brain. One of these neurotoxic phospholipases, OS2, was iodinated and used in binding experiments to demonstrate the presence of two families of specific binding sites in rat brain synaptic membranes. The affinities were exceptionally high, Kd1 = 1.5 +/- 0.5 pM and Kd2 = 45 +/- 10 pM, and the maximal binding capacities were Bmax 1 = 1 +/- 0.4 and Bmax 2 = 3 +/- 0.5 pmol/mg of protein. Both binding sites were sensitive to proteolysis and demonstrated to be located on proteins of Mr 85,000-88,000 and 36,000-51,000 by cross-linking and photoaffinity labeling techniques. The binding of 125 I-OS2 to synaptic membranes was dependent on Ca2+ ions and enhanced by Zn2+ ions which inhibit phospholipase activity. Competition experiments have shown that, except for beta-bungarotoxin, a number of known toxic snake or bee phospholipases have very high affinities for the newly identified binding sites. A good correlation (r = 0.80) was observed between toxicity and affinity but not between phospholipase activity and affinity

  19. A Dualistic Conformational Response to Substrate Binding in the Human Serotonin Transporter Reveals a High Affinity State for Serotonin*

    Science.gov (United States)

    Bjerregaard, Henriette; Severinsen, Kasper; Said, Saida; Wiborg, Ove; Sinning, Steffen

    2015-01-01

    Serotonergic neurotransmission is modulated by the membrane-embedded serotonin transporter (SERT). SERT mediates the reuptake of serotonin into the presynaptic neurons. Conformational changes in SERT occur upon binding of ions and substrate and are crucial for translocation of serotonin across the membrane. Our understanding of these conformational changes is mainly based on crystal structures of a bacterial homolog in various conformations, derived homology models of eukaryotic neurotransmitter transporters, and substituted cysteine accessibility method of SERT. However, the dynamic changes that occur in the human SERT upon binding of ions, the translocation of substrate, and the role of cholesterol in this interplay are not fully elucidated. Here we show that serotonin induces a dualistic conformational response in SERT. We exploited the substituted cysteine scanning method under conditions that were sensitized to detect a more outward-facing conformation of SERT. We found a novel high affinity outward-facing conformational state of the human SERT induced by serotonin. The ionic requirements for this new conformational response to serotonin mirror the ionic requirements for translocation. Furthermore, we found that membrane cholesterol plays a role in the dualistic conformational response in SERT induced by serotonin. Our results indicate the existence of a subpopulation of SERT responding differently to serotonin binding than hitherto believed and that membrane cholesterol plays a role in this subpopulation of SERT. PMID:25614630

  20. Disorders in endocrine link of adaptation system and their possible participation in development of endocrine and psichosomatical pathologies for persons working in the Chernobyl' NPP 30-km zone for a long time. Endocrine adaptopathology in the Chernobyl' NPP 30-km zone

    International Nuclear Information System (INIS)

    Cheban, A.K.

    1992-01-01

    The studies realized in the 30-km zone give an opportunity to reveal changes in the system including hypothalamus, hypothesis and adrenal cortex for persons working at the Pripyat' research and production joint enterprise for a long time. Stable stress in cortisol secretion at absence of expected synchronous increase in secretion of ACTG especially for persons with irradiation doses more than 0.25 Gy is noted. Connection between irradiation dose and adrenal gland secretion decreasing degree is revealed. Hypothetical model of disadaptosis radiation-induced variant is suggested. the conclusion on existance of common pathogenetic basis of many endocrine and psychosomatic diseases is made. 2 refs.; 10 tabs

  1. Spot 42 Small RNA Regulates Arabinose-Inducible araBAD Promoter Activity by Repressing Synthesis of the High-Affinity Low-Capacity Arabinose Transporter

    Science.gov (United States)

    Chen, Jiandong

    2016-01-01

    ABSTRACT The l-arabinose-inducible araBAD promoter (PBAD) enables tightly controlled and tunable expression of genes of interest in a broad range of bacterial species. It has been used successfully to study bacterial sRNA regulation, where PBAD drives expression of target mRNA translational fusions. Here we report that in Escherichia coli, Spot 42 sRNA regulates PBAD promoter activity by affecting arabinose uptake. We demonstrate that Spot 42 sRNA represses araF, a gene encoding the AraF subunit of the high-affinity low-capacity arabinose transporter AraFGH, through direct base-pairing interactions. We further show that endogenous Spot 42 sRNA is sufficient to repress araF expression under various growth conditions. Finally, we demonstrate this posttranscriptional repression has a biological consequence, decreasing the induction of PBAD at low levels of arabinose. This problem can be circumvented using strategies reported previously for avoiding all-or-none induction behavior, such as through constitutive expression of the low-affinity high-capacity arabinose transporter AraE or induction with a higher concentration of inducers. This work adds araF to the set of Spot 42-regulated genes, in agreement with previous studies suggesting that Spot 42, itself negatively regulated by the cyclic AMP (cAMP) receptor protein-cAMP complex, reinforces the catabolite repression network. IMPORTANCE The bacterial arabinose-inducible system is widely used for titratable control of gene expression. We demonstrate here that a posttranscriptional mechanism mediated by Spot 42 sRNA contributes to the functionality of the PBAD system at subsaturating inducer concentrations by affecting inducer uptake. Our finding extends the inputs into the known transcriptional control for the PBAD system and has implications for improving its usage for tunable gene expression. PMID:27849174

  2. High-affinity human leucocyte antigen class I binding variola-derived peptides induce CD4(+) T cell responses more than 30 years post-vaccinia virus vaccination

    DEFF Research Database (Denmark)

    Wang, M.; Tang, Sheila Tuyet; Lund, Ole

    2009-01-01

    Interferon-gamma secreting T lymphocytes against pox virus-derived synthetic 9-mer peptides were tested by enzyme-linked immunospot in peripheral blood of individuals vaccinated with vaccinia virus more than 30 years ago. The peptides were characterized biochemically as high-affinity human leucoc...

  3. High-Affinity Sites Form an Interaction Network to Facilitate Spreading of the MSL Complex across the X Chromosome in Drosophila

    NARCIS (Netherlands)

    Ramírez, Fidel; Lingg, Thomas; Toscano, Sarah; Lam, Kin Chung; Georgiev, Plamen; Chung, Ho-Ryun; Lajoie, Bryan R; de Wit, Elzo; Zhan, Ye; de Laat, Wouter; Dekker, Job; Manke, Thomas; Akhtar, Asifa

    2015-01-01

    Dosage compensation mechanisms provide a paradigm to study the contribution of chromosomal conformation toward targeting and spreading of epigenetic regulators over a specific chromosome. By using Hi-C and 4C analyses, we show that high-affinity sites (HAS), landing platforms of the male-specific

  4. Isolation and partial characterization of gypsy moth BTR-270, an anionic brush border membrane glycoconjugate that binds Bacillus thuringiensis Cry1A toxins with high affinity

    Science.gov (United States)

    Algimantas P. Valaitis; Jeremy L. Jenkins; Mi Kyong Lee; Donald H. Dean; Karen J. Garner

    2001-01-01

    BTR-270, a gypsy moth (Lymantria dispar) brush border membrane molecule that binds Bacillus thuringiensis (Bt) Cry1A toxins with high affinity, was purified by preparative gel electrophoresis. Rabbit antibodies specific for the Bt toxin-binding molecule were raised. Attempts to label BTR-270 by protein-directed techniques were...

  5. Contrast-enhanced CT with a High-Affinity Cationic Contrast Agent for Imaging ex Vivo Bovine, Intact ex Vivo Rabbit, and in Vivo Rabbit Cartilage

    OpenAIRE

    Stewart, Rachel C.; Bansal, Prashant N.; Entezari, Vahid; Lusic, Hrvoje; Nazarian, Rosalynn M.; Snyder, Brian D.; Grinstaff, Mark W.

    2013-01-01

    The high affinity of a cationic iodinated contrast agent for cartilage provides better tissue visualization, easier segmentation, higher contrast-to-noise ratios, and longer usable imaging windows and requires a lower dose of injected contrast agent compared with an anionic contrast agent.

  6. Soil carbon content and relative abundance of high affinity H2-oxidizing bacteria predict atmospheric H2 soil uptake activity better than soil microbial community composition

    NARCIS (Netherlands)

    Khdhiri, Mondher; Hesse, Laura; Popa, Maria Elena; Quiza, Liliana; Lalonde, Isabelle; Meredith, Laura K.; Röckmann, Thomas; Constant, Philippe

    2015-01-01

    Soil-atmosphere exchange of H2 is controlled by gas diffusion and the microbial production and oxidation activities in soil. Among these parameters, the H2 oxidation activity catalyzed by soil microorganisms harboring high affinity hydrogenase is the most difficult variable to parameterize because

  7. Amino propynyl benzoic acid building block in rigid spacers of divalent ligands binding to the Syk SH2 domains with equally high affinity as the natural ligand

    NARCIS (Netherlands)

    Dekker, Frank J; de Mol, Nico J; Fischer, Marcel J E; Liskamp, Rob M J; Dekker, Frank

    2003-01-01

    The construction of rigid spacers composed of amino propynyl benzoic acid building blocks is described. These spacers were used to link two phosphopeptide ligand sites towards obtaining divalent ligands with a high affinity for Syk tandem SH2 domains, which are important in signal transduction. The

  8. Synthesis of hapten and preparation of specific polyclonal antibody with high affinity for lenalidomide, the potent drug for treatment of multiple myeloma

    Directory of Open Access Journals (Sweden)

    Darwish Ibrahim A

    2012-10-01

    Full Text Available Abstract Background For therapeutic monitoring and pharmacokinetic studies of lenalidomide (LND, the potent drug for treatment of multiple myeloma (MM, a specific antibody was required for the development of a sensitive immunoassay system for the accurate determination of LND in plasma. Results In this study, a hapten of LND (N-glutaryl-LND was synthesized by introducing the glutaryl moiety, as a spacer, into the primary aromatic amine site of the LND molecular structure. The structure of the hapten (G-LND was confirmed by mass, 1H-NMR, and 13C spectrometric techniques. G-LND was coupled to each of bovine serum albumin (BSA and keyhole limpet hemocyanin (KLH proteins by ethyl-3-(3-dimethylaminopropyl carbodiimide as a coupling reagent. LND-KLH conjugate was used as an immunogen. Four female 2-3 months old New Zealand white rabbits were immunized with an emulsion of LND-KLH with Freund`s adjuvant. The immune response of the rabbits was monitored by direct enzyme-linked immunosorbent assay (ELISA using LND-BSA immobilized onto microwell plates as a solid phase. The rabbit that showed the highest antibody titer and affinity to LND was scarified and its sera were collected. The IgG fraction was isolated and purified by affinity chromatography on protein A column. The specificity of the purified antibody for LND was evaluated by indirect competitive ELISA using dexamethasone as a competitor as it is used with LND in a combination therapy. Conclusions The high affinity of the antibody (IC50 = 10 ng/mL will be useful in the development of an immunoassay system for the determination of plasma LND concentrations. Current research is going to optimize the assay conditions and validate the procedures for the routine application in clinical laboratories.

  9. Synthesis of hapten and preparation of specific polyclonal antibody with high affinity for lenalidomide, the potent drug for treatment of multiple myeloma.

    Science.gov (United States)

    Darwish, Ibrahim A; Alzoman, Nourh Z; Abuhejail, Reem M; El-Samani, Tilal E

    2012-10-26

    For therapeutic monitoring and pharmacokinetic studies of lenalidomide (LND), the potent drug for treatment of multiple myeloma (MM), a specific antibody was required for the development of a sensitive immunoassay system for the accurate determination of LND in plasma. In this study, a hapten of LND (N-glutaryl-LND) was synthesized by introducing the glutaryl moiety, as a spacer, into the primary aromatic amine site of the LND molecular structure. The structure of the hapten (G-LND) was confirmed by mass, 1H-NMR, and 13C spectrometric techniques. G-LND was coupled to each of bovine serum albumin (BSA) and keyhole limpet hemocyanin (KLH) proteins by ethyl-3-(3-dimethylaminopropyl) carbodiimide as a coupling reagent. LND-KLH conjugate was used as an immunogen. Four female 2-3 months old New Zealand white rabbits were immunized with an emulsion of LND-KLH with Freund`s adjuvant. The immune response of the rabbits was monitored by direct enzyme-linked immunosorbent assay (ELISA) using LND-BSA immobilized onto microwell plates as a solid phase. The rabbit that showed the highest antibody titer and affinity to LND was scarified and its sera were collected. The IgG fraction was isolated and purified by affinity chromatography on protein A column. The specificity of the purified antibody for LND was evaluated by indirect competitive ELISA using dexamethasone as a competitor as it is used with LND in a combination therapy. The high affinity of the antibody (IC50 = 10 ng/mL) will be useful in the development of an immunoassay system for the determination of plasma LND concentrations. Current research is going to optimize the assay conditions and validate the procedures for the routine application in clinical laboratories.

  10. KM-Appunti : 5. KM e "nuova" logica

    OpenAIRE

    Bogliolo, Domenico

    2002-01-01

    Paolo Bisogno leaved many tracks that must be catalogued. One of his research paths, Knowledge Management, is a sort of “Science of knowledge organization” if we decide to join the idea of knowledge with practical, managerial and directional components. We will try to outline here an operative and epistemologic layout of knowledge modelling and categorizing, helping us in understanding new knowledge boundaries of KM.

  11. Mutational analysis of the high-affinity zinc binding site validates a refined human dopamine transporter homology model.

    Directory of Open Access Journals (Sweden)

    Thomas Stockner

    Full Text Available The high-resolution crystal structure of the leucine transporter (LeuT is frequently used as a template for homology models of the dopamine transporter (DAT. Although similar in structure, DAT differs considerably from LeuT in a number of ways: (i when compared to LeuT, DAT has very long intracellular amino and carboxyl termini; (ii LeuT and DAT share a rather low overall sequence identity (22% and (iii the extracellular loop 2 (EL2 of DAT is substantially longer than that of LeuT. Extracellular zinc binds to DAT and restricts the transporter's movement through the conformational cycle, thereby resulting in a decrease in substrate uptake. Residue H293 in EL2 praticipates in zinc binding and must be modelled correctly to allow for a full understanding of its effects. We exploited the high-affinity zinc binding site endogenously present in DAT to create a model of the complete transmemberane domain of DAT. The zinc binding site provided a DAT-specific molecular ruler for calibration of the model. Our DAT model places EL2 at the transporter lipid interface in the vicinity of the zinc binding site. Based on the model, D206 was predicted to represent a fourth co-ordinating residue, in addition to the three previously described zinc binding residues H193, H375 and E396. This prediction was confirmed by mutagenesis: substitution of D206 by lysine and cysteine affected the inhibitory potency of zinc and the maximum inhibition exerted by zinc, respectively. Conversely, the structural changes observed in the model allowed for rationalizing the zinc-dependent regulation of DAT: upon binding, zinc stabilizes the outward-facing state, because its first coordination shell can only be completed in this conformation. Thus, the model provides a validated solution to the long extracellular loop and may be useful to address other aspects of the transport cycle.

  12. High affinity anti-TIM-3 and anti-KIR monoclonal antibodies cloned from healthy human individuals.

    Directory of Open Access Journals (Sweden)

    Stefan Ryser

    Full Text Available We report here the cloning of native high affinity anti-TIM-3 and anti-KIR IgG monoclonal antibodies (mAbs from peripheral blood mononuclear cells (PBMC of healthy human donors. The cells that express these mAbs are rare, present at a frequency of less than one per 105 memory B-cells. Using our proprietary multiplexed screening and cloning technology CellSpot™ we assessed the presence of memory B-cells reactive to foreign and endogenous disease-associated antigens within the same individual. When comparing the frequencies of antigen-specific memory B-cells analyzed in over 20 screening campaigns, we found a strong correlation of the presence of anti-TIM-3 memory B-cells with memory B-cells expressing mAbs against three disease-associated antigens: (i bacterial DNABII proteins that are a marker for Gram negative and Gram positive bacterial infections, (ii hemagglutinin (HA of influenza virus and (iii the extracellular domain of anaplastic lymphoma kinase (ALK. One of the native anti-KIR mAbs has similar characteristics as lirilumab, an anti-KIR mAb derived from immunization of humanized transgenic mice that is in ongoing clinical trials. It is interesting to speculate that these native anti-TIM-3 and anti-KIR antibodies may function as natural regulatory antibodies, analogous to the pharmacological use in cancer treatment of engineered antibodies against the same targets. Further characterization studies are needed to define the mechanisms through which these native antibodies may function in healthy and disease conditions.

  13. Functional mapping and implications of substrate specificity of the yeast high-affinity leucine permease Bap2.

    Science.gov (United States)

    Usami, Yuki; Uemura, Satsohi; Mochizuki, Takahiro; Morita, Asami; Shishido, Fumi; Inokuchi, Jin-ichi; Abe, Fumiyoshi

    2014-07-01

    Leucine is a major amino acid in nutrients and proteins and is also an important precursor of higher alcohols during brewing. In Saccharomyces cerevisiae, leucine uptake is mediated by multiple amino acid permeases, including the high-affinity leucine permease Bap2. Although BAP2 transcription has been extensively analyzed, the mechanisms by which a substrate is recognized and moves through the permease remain unknown. Recently, we determined 15 amino acid residues required for Tat2-mediated tryptophan import. Here we introduced homologous mutations into Bap2 amino acid residues and showed that 7 residues played a role in leucine import. Residues I109/G110/T111 and E305 were located within the putative α-helix break in TMD1 and TMD6, respectively, according to the structurally homologous Escherichia coli arginine/agmatine antiporter AdiC. Upon leucine binding, these α-helix breaks were assumed to mediate a conformational transition in Bap2 from an outward-open to a substrate-binding occluded state. Residues Y336 (TMD7) and Y181 (TMD3) were located near I109 and E305, respectively. Bap2-mediated leucine import was inhibited by some amino acids according to the following order of severity: phenylalanine, leucine>isoleucine>methionine, tyrosine>valine>tryptophan; histidine and asparagine had no effect. Moreover, this order of severity clearly coincided with the logP values (octanol-water partition coefficients) of all amino acids except tryptophan. This result suggests that the substrate partition efficiency to the buried Bap2 binding pocket is the primary determinant of substrate specificity rather than structural amino acid side chain recognition. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Changes in medium radioactivity and composition accompany high-affinity uptake of glutamate and aspartate by mouse brain slices

    International Nuclear Information System (INIS)

    Latzkovits, L.; Neidle, A.; Lajtha, A.

    1984-01-01

    In measurements of high affinity transport in tissue slices, the incubation medium is often treated as an ''infinitely large pool''. External substrate concentrations, even at the micromolar level, are assumed to be constant and metabolic interactions between tissue and medium are neglected. In the present report we describe experiments in which glutamic and aspartic acid uptake by mouse brain slices were studied using techniques that could test these assumptions. Cerebral hemispheres were cut into 0.1 mm sections and about 90 mg of tissue incubated in 10 ml of oxygenated medium. After 45 minutes of equilibration, radioactive substrates were added and the concentrations and specific activities of the amino acids and their metabolites in the medium were determined. During the first 10 min following substrate addition, rapid decreases in glutamic and aspartic acid concentrations in the medium were accompanied by large decreases in specific activity caused by the continuous release of these amino acids from the tissue. In addition, extensive conversion of both substrates to glutamine and the preferential accumulation of this metabolite, in the medium, was found. These results demonstrate that metabolism and release occur simultaneously with uptake during transport experiments in vitro and that these processes can take place in specific tissue compartments. It is therefore necessary to measure the tissue and medium concentration levels of amino acids along with their radioactivity in such experiments, since all three processes (transport, metabolism, and compartmentation) are interrelated in the clearance of amino acids from the incubation medium and probably from the extracellular spaces in vivo as well

  15. NK1 receptor fused to beta-arrestin displays a single-component, high-affinity molecular phenotype.

    Science.gov (United States)

    Martini, Lene; Hastrup, Hanne; Holst, Birgitte; Fraile-Ramos, Alberto; Marsh, Mark; Schwartz, Thue W

    2002-07-01

    Arrestins are cytosolic proteins that, upon stimulation of seven transmembrane (7TM) receptors, terminate signaling by binding to the receptor, displacing the G protein and targeting the receptor to clathrin-coated pits. Fusion of beta-arrestin1 to the C-terminal end of the neurokinin NK1 receptor resulted in a chimeric protein that was expressed to some extent on the cell surface but also accumulated in transferrin-labeled recycling endosomes independently of agonist stimulation. As expected, the fusion protein was almost totally silenced with respect to agonist-induced signaling through the normal Gq/G11 and Gs pathways. The NK1-beta-arrestin1 fusion construct bound nonpeptide antagonists with increased affinity but surprisingly also bound two types of agonists, substance P and neurokinin A, with high, normal affinity. In the wild-type NK1 receptor, neurokinin A (NKA) competes for binding against substance P and especially against antagonists with up to 1000-fold lower apparent affinity than determined in functional assays and in homologous binding assays. When the NK1 receptor was closely fused to G proteins, this phenomenon was eliminated among agonists, but the agonists still competed with low affinity against antagonists. In contrast, in the NK1-beta-arrestin1 fusion protein, all ligands bound with similar affinity independent of the choice of radioligand and with Hill coefficients near unity. We conclude that the NK1 receptor in complex with arrestin is in a high-affinity, stable, agonist-binding form probably best suited to structural analysis and that the receptor can display binding properties that are nearly theoretically ideal when it is forced to complex with only a single intracellular protein partner.

  16. Specific capture and detection of Staphylococcus aureus with high-affinity modified aptamers to cell surface components.

    Science.gov (United States)

    Baumstummler, A; Lehmann, D; Janjic, N; Ochsner, U A

    2014-10-01

    Slow off-rate modified aptamer (SOMAmer) reagents were generated to several Staphylococcus aureus cell surface-associated proteins via SELEX with multiple modified DNA libraries using purified recombinant or native proteins. High-affinity binding agents with sub-nanomolar Kd 's were obtained for staphylococcal protein A (SpA), clumping factors (ClfA, ClfB), fibronectin-binding proteins (FnbA, FnbB) and iron-regulated surface determinants (Isd). Further screening revealed several SOMAmers that specifically bound to Staph. aureus cells from all strains that were tested, but not to other staphylococci or other bacteria. SpA and ClfA SOMAmers proved useful for the selective capture and enrichment of Staph. aureus cells, as shown by culture and PCR, leading to improved limits of detection and efficient removal of PCR inhibitors. Detection of Staph. aureus cells was enhanced by several orders of magnitude when the bacterial cell surface was coated with SOMAmers followed by qPCR of the SOMAmers. Furthermore, fluorescence-labelled SpA SOMAmers demonstrated their utility as direct detection agents in flow cytometry. Significance and impact of the study: Monitoring for microbial contamination of food, water, nonsterile products or the environment is typically based on culture, PCR or antibodies. Aptamers that bind with high specificity and affinity to well-conserved cell surface epitopes represent a promising novel type of reagents to detect bacterial cells without the need for culture or cell lysis, including for the capture and enrichment of bacteria present at low cell densities and for the direct detection via qPCR or fluorescent staining. © 2014 Soma Logic, Inc. published by John Wiley & Sons Ltd On behalf of the society for Applied Microbiology.

  17. Pharmacokinetics and biodistribution of a radioiodine labeled peptidomimetic ligand for high-affinity nerve growth factor receptors

    Energy Technology Data Exchange (ETDEWEB)

    Jung, K. H.; Kim, D. H.; Paik, J. Y.; Koh, B. H.; Bae, J. S.; Choe, Y. S.; Lee, K. H.; Kim, B. T. [Samsung Medical Center, Seoul (Korea, Republic of)

    2005-07-01

    Some of the obstacles for the clinical application of whole nerve growth factor (NGF) may be overcome by utilizing small molecule mimetics. We thus investigated the in vivo pharmacokinetics and biodistribution of a small cyclic peptide derived from NGF-[C(92-96)] with high receptor binding affinity. I-125 C(92-96) was labeled with the Bolton-Hunter method, and binding to TrkA/IgG chimeric protein was confirmed on a polyacrylamide gel after cross-linking. Pharmacokinetic analysis was performed in normal ICR mice intravenously injected with 0.5 MBq I-125 C(92-96) containing varying doses of C(92-96). Biodistribution studies were done at 6 h after injection. Cross-linkage analysis confirmed binding of I-125 C(92-96) to the high affinity NGF receptor, TrkA. Intravenously injected I-125 C(92-96) was cleared from the blood in a biexponential manner with an early T1/2{alpha} of 5.2 min and late T1/2{beta} of 121.3 min. Log blood-concentration decreased over time with a k-slope of 0.0025, clearance of 11.8{+-}0.5 ml/min, T1/2 of 4.1{+-}0.4 hr, and volume of distribution of 69.7{+-}4.6 ml. The pattern of elimination from the blood remained essentially unchanged regardless of the dose of added C(92-96), with dose-proportionate increases in AUCs and peak concentrations consistent with linear pharmacokinetics. Biodistribution studies demonstrated high kidney activity suggesting renal excretion of I-125 C(92-96). There were moderate levels of accumulation in the spleen, lungs and liver, followed by the myocardium and skeletal muscle, whereas brain uptake was low (< 0.2 %ID/gm). Intravenously administered C(92-96) follows linear pharmacokinetics, and is cleared from the circulation at a rate comparable to whole NGF despite its substantially smaller size. Although intravenous C(92-96) does not adequately reach brain tissue, clinically relevant doses can achieve major organ accumulation levels that may be sufficient to elicit biologic responses through NGF receptors.

  18. Pharmacokinetics and biodistribution of a radioiodine labeled peptidomimetic ligand for high-affinity nerve growth factor receptors

    International Nuclear Information System (INIS)

    Jung, K. H.; Kim, D. H.; Paik, J. Y.; Koh, B. H.; Bae, J. S.; Choe, Y. S.; Lee, K. H.; Kim, B. T.

    2005-01-01

    Some of the obstacles for the clinical application of whole nerve growth factor (NGF) may be overcome by utilizing small molecule mimetics. We thus investigated the in vivo pharmacokinetics and biodistribution of a small cyclic peptide derived from NGF-[C(92-96)] with high receptor binding affinity. I-125 C(92-96) was labeled with the Bolton-Hunter method, and binding to TrkA/IgG chimeric protein was confirmed on a polyacrylamide gel after cross-linking. Pharmacokinetic analysis was performed in normal ICR mice intravenously injected with 0.5 MBq I-125 C(92-96) containing varying doses of C(92-96). Biodistribution studies were done at 6 h after injection. Cross-linkage analysis confirmed binding of I-125 C(92-96) to the high affinity NGF receptor, TrkA. Intravenously injected I-125 C(92-96) was cleared from the blood in a biexponential manner with an early T1/2α of 5.2 min and late T1/2β of 121.3 min. Log blood-concentration decreased over time with a k-slope of 0.0025, clearance of 11.8±0.5 ml/min, T1/2 of 4.1±0.4 hr, and volume of distribution of 69.7±4.6 ml. The pattern of elimination from the blood remained essentially unchanged regardless of the dose of added C(92-96), with dose-proportionate increases in AUCs and peak concentrations consistent with linear pharmacokinetics. Biodistribution studies demonstrated high kidney activity suggesting renal excretion of I-125 C(92-96). There were moderate levels of accumulation in the spleen, lungs and liver, followed by the myocardium and skeletal muscle, whereas brain uptake was low (< 0.2 %ID/gm). Intravenously administered C(92-96) follows linear pharmacokinetics, and is cleared from the circulation at a rate comparable to whole NGF despite its substantially smaller size. Although intravenous C(92-96) does not adequately reach brain tissue, clinically relevant doses can achieve major organ accumulation levels that may be sufficient to elicit biologic responses through NGF receptors

  19. Protoporphyrinogen oxidase: high affinity tetrahydrophthalimide radioligand for the inhibitor/herbicide-binding site in mouse liver mitochondria.

    Science.gov (United States)

    Birchfield, N B; Casida, J E

    1996-01-01

    Protoporphyrinogen oxidase (protox), the last common enzyme in heme and chlorophyll biosynthesis, is the target of several classes of herbicides acting as inhibitors in both plants and mammals. N-(4-Chloro-2-fluoro-5-(propargyloxy)phenyl)-3,4,5,6-tetrahydro phthalimide (a potent protox inhibitor referred to as THP) was synthesized as a candidate radioligand ([3H]-THP) by selective catalytic reduction of 3,6-dihydrophthalic anhydride (DHPA) with tritium gas followed by condensation in 45% yield with 4-chloro-2-fluoro-5-(propargyloxy)aniline. Insertion of tritium at the 3 and 6 carbons of DHPA as well as the expected 4 and 5 carbons resulted in high specific activity [3H]THP (92 Ci/mmol). This radioligand undergoes rapid, specific, saturable, and reversible binding to the inhibitor/herbicide binding site of the protox component of cholate-solubilized mouse liver mitochondria with an apparent Kd of 0.41 nM and Bmax of 0.40 pmol/mg of protein. In the standard assay, mouse preparation (150 micrograms of protein) and [3H]THP (0.5 nM) are incubated in 500 microL of phosphate buffer at pH 7.2 for 15 min at 25 degrees C followed by addition of ammonium sulfate and filtration with glass fiber filters. The potencies of five nitrodiphenyl ethers and two other herbicides as inhibitors of [3H]THP binding correlate well with those for inhibition of protox activity (r2 = 0.97, n = 7), thus validating the binding assay as relevant to enzyme inhibition. It is also suitable to determine in vivo block as illustrated by an approximately 50% decrease in [3H]THP binding in liver mitochondria from mice treated ip with oxyfluorfen at 4 mg/kg. This is the first report of a binding assay for protox in mammals. The high affinity and specific activity of [3H]THP facilitate quantitation of protox and therefore research on a sensitive inhibition site for porphyrin biosynthesis.

  20. Development of a new high-affinity human antibody with antitumor activity against solid and blood malignancies.

    Science.gov (United States)

    Sioud, Mouldy; Westby, Phuong; Vasovic, Vlada; Fløisand, Yngvar; Peng, Qian

    2018-04-16

    mAbs have emerged as a promising strategy for the treatment of cancer. However, in several malignancies, no effective antitumor mAbs are yet available. Identifying therapeutic mAbs that recognize common tumor antigens could render the treatment widely applicable. Here, a human single-chain variable fragment (scFv) antibody library was sequentially affinity selected against a panel of human cancer cell lines and an antibody fragment (named MS5) that bound to solid and blood cancer cells was identified. The MS5 scFv was fused to the human IgG1 Fc domain to generate an antibody (MS5-Fc fusion) that induced antibody-dependent cellular cytotoxicity and phagocytosis of cancer cells by macrophages. In addition, the MS5-Fc antibody bound to primary leukemia cells and induced antibody-dependent cellular cytotoxicity. In the majority of analyzed cancer cells, the MS5-Fc antibody induced cell surface redistribution of the receptor complexes, but not internalization, thus maximizing the accessibility of the IgG1 Fc domain to immune effector cells. In vitro stability studies showed that the MS5-Fc antibody was stable after 6 d of incubation in human serum, retaining ∼60% of its initial intact form. After intravenous injections, the antibody localized into tumor tissues and inhibited the growth of 3 different human tumor xenografts (breast, lymphoma, and leukemia). These antitumor effects were associated with tumor infiltration by macrophages and NK cells. In the Ramos B-cell lymphoma xenograft model, the MS5-Fc antibody exhibited a comparable antitumor effect as rituximab, a chimeric anti-CD20 IgG1 mAb. These results indicate that human antibodies with pan-cancer abilities can be generated from phage display libraries, and that the engineered MS5-Fc antibody could be an attractive agent for further clinical investigation.-Sioud, M., Westby, P., Vasovic, V., Fløisand, Y., Peng, Q. Development of a new high-affinity human antibody with antitumor activity against solid and

  1. In vitro and in vivo evaluation of new radiolabeled neurotensin(8-13) analogues with high affinity for NT1 receptors

    International Nuclear Information System (INIS)

    Garayoa, Elisa Garcia-; Allemann-Tannahill, Lesley; Blaeuenstein, Peter; Willmann, Martine; Carrel-Remy, Nathalie; Tourwe, Dirk; Iterbeke, Koen; Conrath, Peter; Schubiger, P. August

    2001-01-01

    The potential utility of neurotensin (NT) in cancer diagnosis and therapy is limited by its rapid degradation. New stabilized analogues were synthesized, labeled with [ 99m Tc] and screened in vitro and in vivo. High affinity and rapid internalization were obtained in binding assays. Despite their longer human plasma half-lives, a rapid degradation was observed with low concentrations as used in biodistribution tests. The tumor uptake rates were rather low but tumor/blood ratios increased according to the stability raise

  2. On Improving 4-km Mesoscale Model Simulations

    Science.gov (United States)

    Deng, Aijun; Stauffer, David R.

    2006-03-01

    A previous study showed that use of analysis-nudging four-dimensional data assimilation (FDDA) and improved physics in the fifth-generation Pennsylvania State University National Center for Atmospheric Research Mesoscale Model (MM5) produced the best overall performance on a 12-km-domain simulation, based on the 18 19 September 1983 Cross-Appalachian Tracer Experiment (CAPTEX) case. However, reducing the simulated grid length to 4 km had detrimental effects. The primary cause was likely the explicit representation of convection accompanying a cold-frontal system. Because no convective parameterization scheme (CPS) was used, the convective updrafts were forced on coarser-than-realistic scales, and the rainfall and the atmospheric response to the convection were too strong. The evaporative cooling and downdrafts were too vigorous, causing widespread disruption of the low-level winds and spurious advection of the simulated tracer. In this study, a series of experiments was designed to address this general problem involving 4-km model precipitation and gridpoint storms and associated model sensitivities to the use of FDDA, planetary boundary layer (PBL) turbulence physics, grid-explicit microphysics, a CPS, and enhanced horizontal diffusion. Some of the conclusions include the following: 1) Enhanced parameterized vertical mixing in the turbulent kinetic energy (TKE) turbulence scheme has shown marked improvements in the simulated fields. 2) Use of a CPS on the 4-km grid improved the precipitation and low-level wind results. 3) Use of the Hong and Pan Medium-Range Forecast PBL scheme showed larger model errors within the PBL and a clear tendency to predict much deeper PBL heights than the TKE scheme. 4) Combining observation-nudging FDDA with a CPS produced the best overall simulations. 5) Finer horizontal resolution does not always produce better simulations, especially in convectively unstable environments, and a new CPS suitable for 4-km resolution is needed. 6

  3. Slc5a8, a Na+-coupled high-affinity transporter for short-chain fatty acids, is a conditional tumour suppressor in colon that protects against colitis and colon cancer under low-fibre dietary conditions.

    Science.gov (United States)

    Gurav, Ashish; Sivaprakasam, Sathish; Bhutia, Yangzom D; Boettger, Thomas; Singh, Nagendra; Ganapathy, Vadivel

    2015-07-15

    Mammalian colon harbours trillions of bacteria under physiological conditions; this symbiosis is made possible because of a tolerized response from the mucosal immune system. The mechanisms underlying this tolerogenic phenomenon remain poorly understood. In the present study we show that Slc5a8 (solute carrier gene family 5a, member 8), a Na(+)-coupled high-affinity transporter in colon for the bacterial fermentation product butyrate, plays a critical role in this process. Among various immune cells in colon, dendritic cells (DCs) are unique not only in their accessibility to luminal contents but also in their ability to induce tolerogenic phenotype in T-cells. We found that DCs exposed to butyrate express the immunosuppressive enzymes indoleamine 2,3-dioxygenase 1 (IDO1) and aldehyde dehydrogenase 1A2 (Aldh1A2), promote conversion of naive T-cells into immunosuppressive forkhead box P3(+) (FoxP3(+)) Tregs (regulatory T-cells) and suppress conversion of naive T-cells into pro-inflammatory interferon (IFN)-γ-producing cells. Slc5a8-null DCs do not induce IDO1 and Aldh1A2 and do not generate Tregs or suppress IFN-γ-producing T-cells in response to butyrate. We also provide in vivo evidence for an obligatory role for Slc5a8 in suppression of IFN-γ-producing T-cells. Furthermore, Slc5a8 protects against colitis and colon cancer under conditions of low-fibre intake but not when dietary fibre intake is optimal. This agrees with the high-affinity nature of the transporter to mediate butyrate entry into cells. We conclude that Slc5a8 is an obligatory link between dietary fibre and mucosal immune system via the bacterial metabolite butyrate and that this transporter is a conditional tumour suppressor in colon linked to dietary fibre content. © 2015 Authors; published by Portland Press Limited.

  4. Characterization of an AtCCX5 gene from Arabidopsis thaliana that involves in high-affinity K+ uptake and Na+ transport in yeast

    International Nuclear Information System (INIS)

    Zhang, Xinxin; Zhang, Min; Takano, Tetsuo; Liu, Shenkui

    2011-01-01

    Highlights: → The AtCCX5 protein coding a putative cation calcium exchanger was characterized. → AtCCX5 expressed in yeast was localized in the plasma membrane and nuclear periphery. → AtCCX5 protein did not show the same transport properties as the CAXs. → AtCCX5 protein involves in mediating high-affinity K + uptake in yeast. → AtCCX5 protein also involves in Na + transport in yeast. -- Abstract: The gene for a putative cation calcium exchanger (CCX) from Arabidopsis thaliana, AtCCX5, was cloned and its function was analyzed in yeast. Green fluorescent protein-tagged AtCCX5 expressed in yeast was localized in the plasma membrane and nuclear periphery. The yeast transformants expressing AtCCX5 were created and their growth in the presence of various cations (K + , Na + , Ca 2+ , Mg 2+ , Fe 2+ , Cu 2+ , Co 2+ , Cd 2+ , Mn 2+ , Ba 2+ , Ni 2+ , Zn 2+ , and Li + ) were analyzed. AtCCX5 expression was found to affect the response to K + and Na + in yeast. The AtCCX5 transformant also showed a little better growth to Zn 2+ . The yeast mutant 9.3 expressing AtCCX5 restored growth of the mutant on medium with low K + (0.5 mM), and also suppressed its Na + sensitivity. Ion uptake experiments showed that AtCCX5 mediated relatively high-affinity K + uptake and was also involved in Na + transport in yeast. Taken together, these findings suggest that the AtCCX5 is a novel transport protein involves in mediating high-affinity K + uptake and Na + transport in yeast.

  5. In vitro and in vivo evaluation of new radiolabeled neurotensin(8-13) analogues with high affinity for NT1 receptors

    Energy Technology Data Exchange (ETDEWEB)

    Garayoa, Elisa Garcia-; Allemann-Tannahill, Lesley; Blaeuenstein, Peter; Willmann, Martine; Carrel-Remy, Nathalie; Tourwe, Dirk; Iterbeke, Koen; Conrath, Peter; Schubiger, P. August E-mail: schubiger@psi.ch

    2001-01-01

    The potential utility of neurotensin (NT) in cancer diagnosis and therapy is limited by its rapid degradation. New stabilized analogues were synthesized, labeled with [{sup 99m}Tc] and screened in vitro and in vivo. High affinity and rapid internalization were obtained in binding assays. Despite their longer human plasma half-lives, a rapid degradation was observed with low concentrations as used in biodistribution tests. The tumor uptake rates were rather low but tumor/blood ratios increased according to the stability raise.

  6. A potential therapy for chordoma via antibody-dependent cell-mediated cytotoxicity employing NK or high-affinity NK cells in combination with cetuximab.

    Science.gov (United States)

    Fujii, Rika; Schlom, Jeffrey; Hodge, James W

    2018-05-01

    OBJECTIVE Chordoma is a rare bone tumor derived from the notochord and is resistant to conventional therapies such as chemotherapy, radiotherapy, and targeting therapeutics. Expression of epidermal growth factor receptor (EGFR) in a large proportion of chordoma specimens indicates a potential target for therapeutic intervention. In this study the authors investigated the potential role of the anti-EGFR antibody cetuximab in immunotherapy for chordoma. METHODS Since cetuximab is a monoclonal antibody of the IgG1 isotype, it has the potential to mediate antibody-dependent cell-mediated cytotoxicity (ADCC) employing natural killer (NK) cells as effectors. Polymorphisms in the CD16 allele expressed on NK cells have been shown to influence the degree of ADCC of tumor cells, with the high-affinity valine (V)/V allele being responsible for more lysis than the V/phenylalanine (F) or FF allele. Unfortunately, however, only approximately 10% of the population expresses the VV allele on NK cells. An NK cell line, NK-92, has now been engineered to endogenously express IL-2 and the high-affinity CD16 allele. These irradiated high-affinity (ha)NK cells were analyzed for lysis of chordoma cells with and without cetuximab, and the levels of lysis observed in ADCC were compared with those of NK cells from donors expressing the VV, VF, and FF alleles. RESULTS Here the authors demonstrate for the first time 1) that cetuximab in combination with NK cells can mediate ADCC of chordoma cells; 2) the influence of the NK CD16 polymorphism in cetuximab-mediated ADCC for chordoma cell lysis; 3) that engineered haNK cells-that is, cells transduced to express the CD16 V158 FcγRIIIa receptor-bind cetuximab with similar affinity to normal NK cells expressing the high-affinity VV allele; and 4) that irradiated haNK cells induce ADCC with cetuximab in chordoma cells. CONCLUSIONS These studies provide rationale for the use of cetuximab in combination with irradiated haNK cells for therapy for

  7. High-Affinity Methanotrophy Informed by Genome-Wide Analysis of Upland Soil Cluster Alpha (USCα) from Axel Heiberg Island, Canadian High Arctic

    Science.gov (United States)

    Rusley, C.; Onstott, T. C.; Lau, M.

    2017-12-01

    Methane (CH4) is a potent greenhouse gas whose proper budgeting is vital to climate predictions. Recent studies have identified upland Arctic mineral cryosols as consistent CH4 sinks, drawing CH4 from both the atmosphere and underlying anaerobic soil layers. Global atmospheric CH4 uptake is proposed to be mediated by high-affinity methanotrophs based on the detection of the marker gene pmoA (particulate methane monooxygenase beta subunit). However, a lack of pure cultures and scarcity of genomic information have hindered our understanding of their metabolic capabilities and versatility. Together with the missing genetic linkage between its pmoA and 16S ribosomal RNA (rRNA) gene, the factors that control the distribution and magnitude of high-affinity methanotrophy in the Arctic permafrost-affected region have remained elusive. Using 21 metagenomic datasets of surface soils obtained from long-term core incubation experiments,1 this bioinformatics study aimed to reconstruct the draft genome of the Upland Soil Cluster α-proteobacteria (USCα), the high-affinity methanotroph previously detected in the samples,2 and to determine its phylogeny and metabolic requirements. We obtained a genome bin containing the high-affinity form of the USCα-like pmoA gene. The 3.03 Mbp assembly is 91.6% complete with a unique set of single-copy marker genes. The 16S rRNA gene fragment of USCα belongs to the α-proteobacterial family Beijerinckiaceae. Genome annotation indicates possible formaldehyde oxidation via tetrahydromethanopterin-linked C1 transfer pathways, acetate utilization, carbon fixation via the Calvin-Benson-Bassham cycle, and glycogen production. Notably, the key enzymes for formaldehyde assimilation via the serine and ribulose monophosphate pathways are missing. The presence of genes encoding nitrate reductase and hemoglobin suggests adaptation to low O2 under water-logged conditions. Since USCα has versatile carbon metabolisms, it may not be an obligate methanotroph

  8. A high-affinity, dimeric inhibitor of PSD-95 bivalently interacts with PDZ1-2 and protects against ischemic brain damage

    DEFF Research Database (Denmark)

    Bach, Anders*; Clausen, Bettina H; Møller, Magda

    2012-01-01

    Inhibition of the ternary protein complex of the synaptic scaffolding protein postsynaptic density protein-95 (PSD-95), neuronal nitric oxide synthase (nNOS), and the N-methyl-d-aspartate (NMDA) receptor is a potential strategy for treating ischemic brain damage, but high-affinity inhibitors are ...... of Tat-N-dimer (3 nmol/g) to mice subjected to focal cerebral ischemia reduces infarct volume with 40% and restores motor functions. Thus, Tat-N-dimer is a highly efficacious neuroprotective agent with therapeutic potential in stroke....

  9. The ketamine analogue methoxetamine and 3- and 4-methoxy analogues of phencyclidine are high affinity and selective ligands for the glutamate NMDA receptor.

    Directory of Open Access Journals (Sweden)

    Bryan L Roth

    Full Text Available In this paper we determined the pharmacological profiles of novel ketamine and phencyclidine analogues currently used as 'designer drugs' and compared them to the parent substances via the resources of the National Institute of Mental Health Psychoactive Drug Screening Program. The ketamine analogues methoxetamine ((RS-2-(ethylamino-2-(3-methoxyphenylcyclohexanone and 3-MeO-PCE (N-ethyl-1-(3-methoxyphenylcyclohexanamine and the 3- and 4-methoxy analogues of phencyclidine, (1-[1-(3-methoxyphenylcyclohexyl]piperidine and 1-[1-(4-methoxyphenylcyclohexyl]piperidine, were all high affinity ligands for the PCP-site on the glutamate NMDA receptor. In addition methoxetamine and PCP and its analogues displayed appreciable affinities for the serotonin transporter, whilst the PCP analogues exhibited high affinities for sigma receptors. Antagonism of the NMDA receptor is thought to be the key pharmacological feature underlying the actions of dissociative anaesthetics. The novel ketamine and PCP analogues had significant affinities for the NMDA receptor in radioligand binding assays, which may explain their psychotomimetic effects in human users. Additional actions on other targets could be important for delineating side-effects.

  10. Specificity of Bacillus thuringiensis endotoxins is correlated with the presence of high-affinity binding sites in the brush border membrane of target insect midguts

    International Nuclear Information System (INIS)

    Hofmann, C.; Vanderbruggen, H.; Hoefte, H.; Van Rie, J.; Jansens, S.; Van Mellaert, H.

    1988-01-01

    Binding studies were performed with two 125 I-labeled Bacillus thuringiensis δ-endotoxins on brush border membrane vesicles prepared from the larval midgut of the tobacco hornworm Manduca sexta or the cabbage butterfly Pieris brassicae. One δ-endotoxin, Bt2-protoxin, is a 130-kDa recombinant crystalline protein from B. thuringiensis subsp. berliner. It kills larvae of both insect species. The active Bt2-toxin is a 60-kDa proteolytic fragment of the Bt2-protoxin. It binds saturably and with high affinity to brush border membrane vesicles from the midgut of both species. The other δ-endotoxin, Bt4412-protoxin, is a 136-kDa crystalline protein from B. thuringiensis subsp. thuringiensis, which is highly toxic for P. brassicae, but not for M. sexta larvae. Bt4412-toxin, obtained after proteolytic activation of Bt4412-protoxin, shows high-affinity saturable binding to P. brassicae vesicles but not to M. sexta vesicles. The correlation between toxicity and specific binding is further strengthened by competition studies. Other B. thuringiensis δ-endotoxins active against M. sexta compete for binding of 125 I-labeled Bt2-toxin to M. sexta vesicles, whereas toxins active against dipteran or coleopteran larvae do not compete. Bt2-toxin and Bt4412-toxin bind to different sites on P. brassicae vesicles

  11. Septide and neurokinin A are high-affinity ligands on the NK-1 receptor: evidence from homologous versus heterologous binding analysis.

    Science.gov (United States)

    Hastrup, H; Schwartz, T W

    1996-12-16

    The three main tachykinins, substance P, neurokinin A (NKA), and neurokinin B, are believed to be selective ligands for respectively the NK-1, NK-2 and NK-3 receptors. However, NKA also has actions which cannot be mediated through its normal NK-2 receptor and the synthetic peptide [pGlu6,Pro9]-Substance P9-11--called septide--is known to have tachykinin-like actions despite its apparent lack of binding to any known tachykinin receptor. In the cloned NK-1 receptor expressed in COS-7 cells NKA and septide as expected were poor competitors for radiolabeled substance P. However, by using radiolabeled NKA and septide directly, it was found that both peptides in homologous binding assays as well as in competition against each other in fact bound to the NK-1 receptor with high affinity: Kd values of 0.51 +/- 0.15 nM (NKA) and 0.55 +/- 0.03 nM (septide). It is concluded that NKA and septide are high-affinity ligands for the NK-1 receptor but that they are poor competitors for substance P, which in contrast competes very well for binding with both NKA and septide.

  12. High-affinity, noninhibitory pathogenic C1 domain antibodies are present in patients with hemophilia A and inhibitors

    Science.gov (United States)

    Batsuli, Glaivy; Deng, Wei; Healey, John F.; Parker, Ernest T.; Baldwin, W. Hunter; Cox, Courtney; Nguyen, Brenda; Kahle, Joerg; Königs, Christoph; Li, Renhao; Lollar, Pete

    2016-01-01

    Inhibitor formation in hemophilia A is the most feared treatment-related complication of factor VIII (fVIII) therapy. Most inhibitor patients with hemophilia A develop antibodies against the fVIII A2 and C2 domains. Recent evidence demonstrates that the C1 domain contributes to the inhibitor response. Inhibitory anti-C1 monoclonal antibodies (mAbs) have been identified that bind to putative phospholipid and von Willebrand factor (VWF) binding epitopes and block endocytosis of fVIII by antigen presenting cells. We now demonstrate by competitive enzyme-linked immunosorbent assay and hydrogen-deuterium exchange mass spectrometry that 7 of 9 anti-human C1 mAbs tested recognize an epitope distinct from the C1 phospholipid binding site. These mAbs, designated group A, display high binding affinities for fVIII, weakly inhibit fVIII procoagulant activity, poorly inhibit fVIII binding to phospholipid, and exhibit heterogeneity with respect to blocking fVIII binding to VWF. Another mAb, designated group B, inhibits fVIII procoagulant activity, fVIII binding to VWF and phospholipid, fVIIIa incorporation into the intrinsic Xase complex, thrombin generation in plasma, and fVIII uptake by dendritic cells. Group A and B epitopes are distinct from the epitope recognized by the canonical, human-derived inhibitory anti-C1 mAb, KM33, whose epitope overlaps both groups A and B. Antibodies recognizing group A and B epitopes are present in inhibitor plasmas from patients with hemophilia A. Additionally, group A and B mAbs increase fVIII clearance and are pathogenic in a hemophilia A mouse tail snip bleeding model. Group A anti-C1 mAbs represent the first identification of pathogenic, weakly inhibitory antibodies that increase fVIII clearance. PMID:27381905

  13. Fiber-wireless transmission system of 108  Gb/sdata over 80 km fiber and 2×2multiple-input multiple-output wireless links at 100 GHz W-band frequency.

    Science.gov (United States)

    Li, Xinying; Dong, Ze; Yu, Jianjun; Chi, Nan; Shao, Yufeng; Chang, G K

    2012-12-15

    We experimentally demonstrate a seamlessly integrated fiber-wireless system that delivers a 108  Gb/s signal through 80 km fiber and 1 m wireless transport over free space at 100 GHz adopting polarization-division-multiplexing quadrature-phase-shift-keying (PDM-QPSK) modulation and heterodyning coherent detection. The X- and Y-polarization components of the optical PDM-QPSK baseband signal are simultaneously upconverted to 100 GHz wireless carrier by optical polarization-diversity heterodyne beating, and then independently transmitted and received by two pairs of transmitter and receiver antennas, which form a 2×2 multiple-input multiple-output wireless link. At the wireless receiver, two-stage downconversion is performed firstly in the analog domain based on balanced mixer and sinusoidal radio frequency signal, and then in the digital domain based on digital signal processing (DSP). Polarization demultiplexing is realized by the constant modulus algorithm in the DSP part at the receiver. The bit-error ratio for the 108  Gb/s PDM-QPSK signal is less than the pre-forward-error-correction threshold of 3.8×10(-3) after both 1 m wireless delivery at 100 GHz and 80 km single-mode fiber-28 transmission. To our knowledge, this is the first demonstration to realize 100  Gb/s signal delivery through both fiber and wireless links at 100 GHz.

  14. Immunostimulatory CpG-oligonucleotides induce functional high affinity IL-2 receptors on B-CLL cells: costimulation with IL-2 results in a highly immunogenic phenotype.

    Science.gov (United States)

    Decker, T; Schneller, F; Kronschnabl, M; Dechow, T; Lipford, G B; Wagner, H; Peschel, C

    2000-05-01

    CpG-oligodeoxynucleotides (CpG-ODN) have been shown to induce proliferation, cytokine production, and surface molecule regulation in normal and malignant human B cells. In the present study, we investigated the potential of CpG-ODN to induce functional high-affinity receptors in leukemic and normal B cells and the effects of costimulation with IL-2 on proliferation, cytokine secretion, and surface molecule regulation. Highly purified B cells from B-CLL patients and normal controls were stimulated with CpG-ODN with or without IL-2. Expression of CD25 was determined using FACS, and the presence of high-affinity IL-2 receptors was determined by scatchard analysis. Costimulatory effects of IL-2 and CpG-ODN were investigated using proliferation assays, ELISA (IL-6, TNF-alpha), and FACS analysis (CD80, CD86 expression). Reactivity of autologous and allogeneic T cells toward activated B-CLL cells was determined in mixed lymphocyte reactions and Interferon-gamma Elispot assays. The CpG-ODN DSP30 caused a significantly stronger induction of the IL-2 receptor alpha chain in malignant as compared with normal B cells (p = 0.03). This resulted in the expression of functional high-affinity IL-2 receptors in B-CLL cells, but fewer numbers of receptors with less affinity were expressed in normal B cells. Although addition of IL-2 to CpG-ODN-stimulated cells augmented proliferation in both normal B cells and B-CLL cells, no costimulatory effect on cytokine production or surface molecule expression could be observed in normal B cells. In contrast, TNF-alpha and IL-6 production was increased in B-CLL cells, and the expression of CD80 and CD86 was further enhanced when IL-2 was used as a costimulus. Autologous and allogeneic immune recognition of B-CLL cells stimulated with CpG-ODN and IL-2 was increased compared with B-CLL cells stimulated with CpG-ODN alone. Stimulation of B-CLL cells with CpG-ODN and IL-2 might be an attractive strategy for potential immunotherapies for B

  15. Human metabolites of synthetic cannabinoids JWH-018 and JWH-073 bind with high affinity and act as potent agonists at cannabinoid type-2 receptors

    International Nuclear Information System (INIS)

    Rajasekaran, Maheswari; Brents, Lisa K.; Franks, Lirit N.; Moran, Jeffery H.; Prather, Paul L.

    2013-01-01

    K2 or Spice is an emerging drug of abuse that contains synthetic cannabinoids, including JWH-018 and JWH-073. Recent reports indicate that monohydroxylated metabolites of JWH-018 and JWH-073 retain high affinity and activity at cannabinoid type-1 receptors (CB 1 Rs), potentially contributing to the enhanced toxicity of K2 compared to marijuana. Since the parent compounds also bind to cannabinoid type-2 receptors (CB 2 Rs), this study investigated the affinity and intrinsic activity of JWH-018, JWH-073 and several monohydroxylated metabolites at human CB 2 Rs (hCB 2 Rs). The affinity of cannabinoids for hCB 2 Rs was determined by competition binding studies employing CHO-hCB 2 membranes. Intrinsic activity of compounds was assessed by G-protein activation and adenylyl cyclase (AC)-inhibition in CHO-hCB 2 cells. JWH-073, JWH-018 and several of their human metabolites exhibit nanomolar affinity and act as potent agonists at hCB 2 Rs. Furthermore, a major omega hydroxyl metabolite of JWH-073 (JWH-073-M5) binds to CB 2 Rs with 10-fold less affinity than the parent molecule, but unexpectedly, is equipotent in regulating AC-activity when compared to the parent molecule. Finally, when compared to CP-55,940 and Δ 9 -tetrahydrocannabinol (Δ 9 -THC), JWH-018, JWH-018-M5 and JWH-073-M5 require significantly less CB 2 R occupancy to produce similar levels of AC-inhibition, indicating that these compounds may more efficiently couple CB 2 Rs to AC than the well characterized cannabinoid agonists examined. These results indicate that JWH-018, JWH-073 and several major human metabolites of these compounds exhibit high affinity and demonstrate distinctive signaling properties at CB 2 Rs. Therefore, future studies examining pharmacological and toxicological properties of synthetic cannabinoids present in K2 products should consider potential actions of these drugs at both CB 1 and CB 2 Rs. - Highlights: • JWH-018 and JWH-073 are synthetic cannabinoids present in abused K2

  16. Human metabolites of synthetic cannabinoids JWH-018 and JWH-073 bind with high affinity and act as potent agonists at cannabinoid type-2 receptors

    Energy Technology Data Exchange (ETDEWEB)

    Rajasekaran, Maheswari; Brents, Lisa K.; Franks, Lirit N. [Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock, AR 72205 (United States); Moran, Jeffery H. [Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock, AR 72205 (United States); Arkansas Department of Public Health, Public Health Laboratory, Little Rock, AR 72205 (United States); Prather, Paul L., E-mail: pratherpaull@uams.edu [Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock, AR 72205 (United States)

    2013-06-01

    K2 or Spice is an emerging drug of abuse that contains synthetic cannabinoids, including JWH-018 and JWH-073. Recent reports indicate that monohydroxylated metabolites of JWH-018 and JWH-073 retain high affinity and activity at cannabinoid type-1 receptors (CB{sub 1}Rs), potentially contributing to the enhanced toxicity of K2 compared to marijuana. Since the parent compounds also bind to cannabinoid type-2 receptors (CB{sub 2}Rs), this study investigated the affinity and intrinsic activity of JWH-018, JWH-073 and several monohydroxylated metabolites at human CB{sub 2}Rs (hCB{sub 2}Rs). The affinity of cannabinoids for hCB{sub 2}Rs was determined by competition binding studies employing CHO-hCB{sub 2} membranes. Intrinsic activity of compounds was assessed by G-protein activation and adenylyl cyclase (AC)-inhibition in CHO-hCB{sub 2} cells. JWH-073, JWH-018 and several of their human metabolites exhibit nanomolar affinity and act as potent agonists at hCB{sub 2}Rs. Furthermore, a major omega hydroxyl metabolite of JWH-073 (JWH-073-M5) binds to CB{sub 2}Rs with 10-fold less affinity than the parent molecule, but unexpectedly, is equipotent in regulating AC-activity when compared to the parent molecule. Finally, when compared to CP-55,940 and Δ{sup 9}-tetrahydrocannabinol (Δ{sup 9}-THC), JWH-018, JWH-018-M5 and JWH-073-M5 require significantly less CB{sub 2}R occupancy to produce similar levels of AC-inhibition, indicating that these compounds may more efficiently couple CB{sub 2}Rs to AC than the well characterized cannabinoid agonists examined. These results indicate that JWH-018, JWH-073 and several major human metabolites of these compounds exhibit high affinity and demonstrate distinctive signaling properties at CB{sub 2}Rs. Therefore, future studies examining pharmacological and toxicological properties of synthetic cannabinoids present in K2 products should consider potential actions of these drugs at both CB{sub 1} and CB{sub 2}Rs. - Highlights: • JWH-018

  17. beta-Arrestin 1 and 2 stabilize the angiotensin II type I receptor in distinct high-affinity conformations

    DEFF Research Database (Denmark)

    Sanni, S J; Hansen, J T; Bonde, M M

    2010-01-01

    The angiotensin II type 1 (AT(1)) receptor belongs to family A of 7 transmembrane (7TM) receptors. The receptor has important roles in the cardiovascular system and is commonly used as a drug target in cardiovascular diseases. Interaction of 7TM receptors with G proteins or beta-arrestins often...

  18. Mapping of barley alpha-amylases and outer subsite mutants reveals dynamic high-affinity subsites and barriers in the long substrate binding cleft

    DEFF Research Database (Denmark)

    Kandra, L.; Abou Hachem, Maher; Gyemant, G.

    2006-01-01

    Subsite affinity maps of long substrate binding clefts in barley alpha-amylases, obtained using a series of maltooligosaccharides of degree of polymerization of 3-12, revealed unfavorable binding energies at the internal subsites -3 and -5 and at subsites -8 and +3/+4 defining these subsites...... as binding barriers. Barley a-amylase I mutants Y105A and T212Y at subsite -6 and +4 resulted in release or anchoring of bound substrate, thus modifying the affinities of other high-affinity subsites (-2 and +2) and barriers. The double mutant Y105A-T212Y displayed a hybrid subsite affinity profile......, converting barriers to binding areas. These findings highlight the dynamic binding energy distribution and the versatility of long maltooligosaccharide derivatives in mapping extended binding clefts in a-amylases....

  19. Two classes of astrocytes in the adult human and pig retina in terms of their expression of high affinity NGF receptor (TrkA).

    Science.gov (United States)

    Ruiz-Ederra, Javier; Hitchcock, Peter F; Vecino, Elena

    2003-02-13

    Astrocytes have been implicated in axon guidance and synaptic regeneration in the retina and these processes involve activation of the high affinity nerve growth factor receptor, known as the tyrosine kinase A (TrkA) receptor. The purpose of the present study was to characterize the expression of TrkA in astrocytes of the adult pig and human retina. To this end, sections of human and pig retinas were immunolabeled with a combination of antibodies to glial fibrillary acidic protein (GFAP) and TrkA. Our study revealed that most of the GFAP-positive cells express TrkA, whereas a rare, novel subpopulation of astrocytes was found to be devoid of TrkA. Our results support the idea that astrocytes play an important neurotrophic role in the retina.

  20. In Vivo Neutralization of α-Cobratoxin with High-Affinity Llama Single-Domain Antibodies (VHHs) and a VHH-Fc Antibody

    Science.gov (United States)

    Richard, Gabrielle; Meyers, Ashley J.; McLean, Michael D.; Arbabi-Ghahroudi, Mehdi; MacKenzie, Roger; Hall, J. Christopher

    2013-01-01

    Small recombinant antibody fragments (e.g. scFvs and VHHs), which are highly tissue permeable, are being investigated for antivenom production as conventional antivenoms consisting of IgG or F(ab’)2 antibody fragments do not effectively neutralize venom toxins located in deep tissues. However, antivenoms composed entirely of small antibody fragments may have poor therapeutic efficacy due to their short serum half-lives. To increase serum persistence and maintain tissue penetration, we prepared low and high molecular mass antivenom antibodies. Four llama VHHs were isolated from an immune VHH-displayed phage library and were shown to have high affinity, in the low nM range, for α-cobratoxin (α–Cbtx), the most lethal component of Naja kaouthia venom. Subsequently, our highest affinity VHH (C2) was fused to a human Fc fragment to create a VHH2-Fc antibody that would offer prolonged serum persistence. After in planta (Nicotiana benthamiana) expression and purification, we show that our VHH2-Fc antibody retained high affinity binding to α–Cbtx. Mouse α–Cbtx challenge studies showed that our highest affinity VHHs (C2 and C20) and the VHH2-Fc antibody effectively neutralized lethality induced by α–Cbtx at an antibody:toxin molar ratio as low as ca. 0.75×:1. Further research towards the development of an antivenom therapeutic involving these anti-α-Cbtx VHHs and VHH2-Fc antibody molecules should involve testing them as a combination, to determine whether they maintain tissue penetration capability and low immunogenicity, and whether they exhibit improved serum persistence and therapeutic efficacy. PMID:23894495

  1. Distribution and ultrastructure of neurons in opossum piriform cortex displaying immunoreactivity to GABA and GAD and high-affinity tritiated GABA uptake

    International Nuclear Information System (INIS)

    Haberly, L.B.; Hansen, D.J.; Feig, S.L.; Presto, S.

    1987-01-01

    GABAergic neurons have been identified in the piriform cortex of the opossum at light and electron microscopic levels by immunocytochemical localization of GABA and the GABA-synthesizing enzyme glutamic acid decarboxylase and by autoradiographic visualization of high-affinity 3 H-GABA uptake. Four major neuron populations have been distinguished on the basis of soma size, shape, and segregation at specific depths and locations: large horizontal cells in layer Ia of the anterior piriform cortex, small globular cells with thin dendrites concentrated in layers Ib and II of the posterior piriform cortex, and multipolar and fusiform cells concentrated in the deep part of layer III in anterior and posterior parts of the piriform cortex and the subjacent endopiriform nucleus. All four populations were well visualized with both antisera, but the large layer Ia horizontal cells displayed only very light 3 H-GABA uptake, thus suggesting a lack of local axon collaterals or lack of high-affinity GABA uptake sites. The large, ultrastructurally distinctive somata of layer Ia horizontal cells receive a very small number of symmetrical synapses; the thin, axonlike dendrites of small globular cells are exclusively postsynaptic and receive large numbers of both symmetrical and asymmetrical synapses, in contrast to somata which receive a small number of both types; and the deep multipolar and fusiform cells receive a highly variable number of symmetrical and asymmetrical synapses on somata and proximal dendrites. Labeled puncta of axon terminal dimensions were found in large numbers in the neuropil surrounding pyramidal cell somata in layer II and in the endopiriform nucleus. Moderately large numbers of labeled puncta were found in layer I at the depth of pyramidal cell apical dendrites with greater numbers in layer Ia at the depth of distal apical segments than in layer Ib

  2. ADCC employing an NK cell line (haNK) expressing the high affinity CD16 allele with avelumab, an anti-PD-L1 antibody.

    Science.gov (United States)

    Jochems, Caroline; Hodge, James W; Fantini, Massimo; Tsang, Kwong Y; Vandeveer, Amanda J; Gulley, James L; Schlom, Jeffrey

    2017-08-01

    NK-92 cells, and their derivative, designated aNK, were obtained from a patient with non-Hodgkin lymphoma. Prior clinical studies employing adoptively transferred irradiated aNK cells have provided evidence of clinical benefit and an acceptable safety profile. aNK cells have now been engineered to express IL-2 and the high affinity (ha) CD16 allele (designated haNK). Avelumab is a human IgG1 anti-PD-L1 monoclonal antibody, which has shown evidence of clinical activity in a range of human tumors. Prior in vitro studies have shown that avelumab has the ability to mediate antibody-dependent cell-mediated cytotoxicity (ADCC) of human tumor cells when combined with NK cells. In the studies reported here, the ability of avelumab to enhance the lysis of a range of human carcinoma cells by irradiated haNK cells via the ADCC mechanism is demonstrated; this ADCC is shown to be inhibited by anti-CD16 blocking antibody and by concanamycin A, indicating the use of the granzyme/perforin pathway in tumor cell lysis. Studies also show that while NK cells have the ability to lyse aNK or haNK cells, the addition of NK cells to irradiated haNK cells does not inhibit haNK-mediated lysis of human tumor cells, with or without the addition of avelumab. Avelumab-mediated lysis of tumor cells by irradiated haNK cells is also shown to be similar to that of NK cells bearing the V/V Fc receptor high affinity allele. These studies thus provide the rationale for the clinical evaluation of the combined use of avelumab with that of irradiated adoptively transferred haNK cells. © 2017 UICC.

  3. Ferric reductase genes involved in high-affinity iron uptake are differentially regulated in yeast and hyphae of Candida albicans.

    Science.gov (United States)

    Jeeves, Rose E; Mason, Robert P; Woodacre, Alexandra; Cashmore, Annette M

    2011-09-01

    The pathogenic yeast Candida albicans possesses a reductive iron uptake system which is active in iron-restricted conditions. The sequestration of iron by this mechanism initially requires the reduction of free iron to the soluble ferrous form, which is catalysed by ferric reductase proteins. Reduced iron is then taken up into the cell by a complex of a multicopper oxidase protein and an iron transport protein. Multicopper oxidase proteins require copper to function and so reductive iron and copper uptake are inextricably linked. It has previously been established that Fre10 is the major cell surface ferric reductase in C. albicans and that transcription of FRE10 is regulated in response to iron levels. We demonstrate here that Fre10 is also a cupric reductase and that Fre7 also makes a significant contribution to cell surface ferric and cupric reductase activity. It is also shown, for the first time, that transcription of FRE10 and FRE7 is lower in hyphae compared to yeast and that this leads to a corresponding decrease in cell surface ferric, but not cupric, reductase activity. This demonstrates that the regulation of two virulence determinants, the reductive iron uptake system and the morphological form of C. albicans, are linked. Copyright © 2011 John Wiley & Sons, Ltd.

  4. Odorant-binding proteins display high affinities for behavioral attractants and repellents in the natural predator Chrysopa pallens.

    Science.gov (United States)

    Li, Zhao-Qun; Zhang, Shuai; Luo, Jun-Yu; Wang, Si-Bao; Dong, Shuang-Lin; Cui, Jin-Jie

    2015-07-01

    Chrysopa pallens is an important natural predator of various pests in many different cropping systems. Understanding the sophisticated olfactory system of insect antennae is crucial for studying the physiological bases of olfaction and could also help enhance the effectiveness of C. pallens in biological control. However, functional studies of the olfactory genes in C. pallens are still lacking. In this study, we cloned five odorant-binding protein (OBP) genes from C. pallens (CpalOBPs). Quantitative RT-PCR results indicated that the five CpalOBPs had different tissue expression profiles. Ligand-binding assays showed that farnesol, farnesene, cis-3-hexenyl hexanoate, geranylacetone, beta-ionone, octyl aldehyde, decanal, nerolidol (Kipallens. Among them, farnesene and its corresponding alcohol, farnesol, elicited remarkable repellent behavioral responses from C. pallens. Our study provides several compounds that could be selected to develop slow-release agents that attract/repel C. pallens and to improve the search for strategies to eliminate insect pests. Copyright © 2015 Elsevier Inc. All rights reserved.

  5. A binding-site barrier affects imaging efficiency of high affinity amyloid-reactive peptide radiotracers in vivo.

    Science.gov (United States)

    Wall, Jonathan S; Williams, Angela; Richey, Tina; Stuckey, Alan; Huang, Ying; Wooliver, Craig; Macy, Sallie; Heidel, Eric; Gupta, Neil; Lee, Angela; Rader, Brianna; Martin, Emily B; Kennel, Stephen J

    2013-01-01

    Amyloid is a complex pathology associated with a growing number of diseases including Alzheimer's disease, type 2 diabetes, rheumatoid arthritis, and myeloma. The distribution and extent of amyloid deposition in body organs establishes the prognosis and can define treatment options; therefore, determining the amyloid load by using non-invasive molecular imaging is clinically important. We have identified a heparin-binding peptide designated p5 that, when radioiodinated, was capable of selectively imaging systemic visceral AA amyloidosis in a murine model of the disease. The p5 peptide was posited to bind effectively to amyloid deposits, relative to similarly charged polybasic heparin-reactive peptides, because it adopted a polar α helix secondary structure. We have now synthesized a variant, p5R, in which the 8 lysine amino acids of p5 have been replaced with arginine residues predisposing the peptide toward the α helical conformation in an effort to enhance the reactivity of the peptide with the amyloid substrate. The p5R peptide had higher affinity for amyloid and visualized AA amyloid in mice by using SPECT/CT imaging; however, the microdistribution, as evidenced in micro-autoradiographs, was dramatically altered relative to the p5 peptide due to its increased affinity and a resultant "binding site barrier" effect. These data suggest that radioiodinated peptide p5R may be optimal for the in vivo detection of discreet, perivascular amyloid, as found in the brain and pancreatic vasculature, by using molecular imaging techniques; however, peptide p5, due to its increased penetration, may yield more quantitative imaging of expansive tissue amyloid deposits.

  6. Fluorinated phenmetrazine "legal highs" act as substrates for high-affinity monoamine transporters of the SLC6 family.

    Science.gov (United States)

    Mayer, Felix P; Burchardt, Nadine V; Decker, Ann M; Partilla, John S; Li, Yang; McLaughlin, Gavin; Kavanagh, Pierce V; Sandtner, Walter; Blough, Bruce E; Brandt, Simon D; Baumann, Michael H; Sitte, Harald H

    2018-05-15

    A variety of new psychoactive substances (NPS) are appearing in recreational drug markets worldwide. NPS are compounds that target various receptors and transporters in the central nervous system to achieve their psychoactive effects. Chemical modifications of existing drugs can generate NPS that are not controlled by current legislation, thereby providing legal alternatives to controlled substances such as cocaine or amphetamine. Recently, 3-fluorophenmetrazine (3-FPM), a derivative of the anorectic compound phenmetrazine, appeared on the recreational drug market and adverse clinical effects have been reported. Phenmetrazine is known to elevate extracellular monoamine concentrations by an amphetamine-like mechanism. Here we tested 3-FPM and its positional isomers, 2-FPM and 4-FPM, for their abilities to interact with plasma membrane monoamine transporters for dopamine (DAT), norepinephrine (NET) and serotonin (SERT). We found that 2-, 3- and 4-FPM inhibit uptake mediated by DAT and NET in HEK293 cells with potencies comparable to cocaine (IC 50 values 80 μM). Experiments directed at identifying transporter-mediated reverse transport revealed that FPM isomers induce efflux via DAT, NET and SERT in HEK293 cells, and this effect is augmented by the Na + /H + ionophore monensin. Each FPM evoked concentration-dependent release of monoamines from rat brain synaptosomes. Hence, this study reports for the first time the mode of action for 2-, 3- and 4-FPM and identifies these NPS as monoamine releasers with marked potency at catecholamine transporters implicated in abuse and addiction. This article is part of the Special Issue entitled 'Designer Drugs and Legal Highs.' Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  7. A binding-site barrier affects imaging efficiency of high affinity amyloid-reactive peptide radiotracers in vivo.

    Directory of Open Access Journals (Sweden)

    Jonathan S Wall

    Full Text Available Amyloid is a complex pathology associated with a growing number of diseases including Alzheimer's disease, type 2 diabetes, rheumatoid arthritis, and myeloma. The distribution and extent of amyloid deposition in body organs establishes the prognosis and can define treatment options; therefore, determining the amyloid load by using non-invasive molecular imaging is clinically important. We have identified a heparin-binding peptide designated p5 that, when radioiodinated, was capable of selectively imaging systemic visceral AA amyloidosis in a murine model of the disease. The p5 peptide was posited to bind effectively to amyloid deposits, relative to similarly charged polybasic heparin-reactive peptides, because it adopted a polar α helix secondary structure. We have now synthesized a variant, p5R, in which the 8 lysine amino acids of p5 have been replaced with arginine residues predisposing the peptide toward the α helical conformation in an effort to enhance the reactivity of the peptide with the amyloid substrate. The p5R peptide had higher affinity for amyloid and visualized AA amyloid in mice by using SPECT/CT imaging; however, the microdistribution, as evidenced in micro-autoradiographs, was dramatically altered relative to the p5 peptide due to its increased affinity and a resultant "binding site barrier" effect. These data suggest that radioiodinated peptide p5R may be optimal for the in vivo detection of discreet, perivascular amyloid, as found in the brain and pancreatic vasculature, by using molecular imaging techniques; however, peptide p5, due to its increased penetration, may yield more quantitative imaging of expansive tissue amyloid deposits.

  8. pH, Lactate, and Hypoxia: Reciprocity in Regulating High-Affinity Monocarboxylate Transporter Expression in Glioblastoma

    Directory of Open Access Journals (Sweden)

    James P. Caruso

    2017-02-01

    Full Text Available Highly malignant brain tumors harbor the aberrant propensity for aerobic glycolysis, the excessive conversion of glucose to lactic acid even in the presence of ample tissue oxygen. Lactic acid is rapidly effluxed to the tumor microenvironment via a group of plasma-membrane transporters denoted monocarboxylate transporters (MCTs to prevent “self-poisoning.” One isoform, MCT2, has the highest affinity for lactate and thus should have the ability to respond to microenvironment conditions such as hypoxia, lactate, and pH to help maintain high glycolytic flux in the tumor. Yet, MCT2 is considered to not respond to hypoxia, which is counterintuitive. Its response to tumor lactate has not been reported. In this report, we experimentally identify the transcription initiation site/s for MCT2 in astrocytes (normal and glioma (tumor. We then use a BACmid library to isolate a 4.2-kbp MCT2 promoter-exon I region and examine promoter response to glycolysis-mediated stimuli in glioma cells. Reporter analysis of nested-promoter constructs indicated response of MCT2 to hypoxia, pH, lactate, and glucose, the major physiological “players” that facilitate a tumor's growth and proliferation. Immunoblot analysis of native MCT2 expression under altered pH and hypoxia reflected the reporter data. The pH-mediated gene-regulation studies we describe are the first to record H+-based reporter studies for any mammalian system and demonstrate the exquisite response of the MCT2 gene to minute changes in tumor pH. Identical promoter usage also provides the first evidence of astrocytes harnessing the same gene regulatory regions to facilitate astrocyte-neuron lactate shuttling, a metabolic feature of normal brain.

  9. Toward 10-km mesh global climate simulations

    Science.gov (United States)

    Ohfuchi, W.; Enomoto, T.; Takaya, K.; Yoshioka, M. K.

    2002-12-01

    An atmospheric general circulation model (AGCM) that runs very efficiently on the Earth Simulator (ES) was developed. The ES is a gigantic vector-parallel computer with the peak performance of 40 Tflops. The AGCM, named AFES (AGCM for ES), was based on the version 5.4.02 of an AGCM developed jointly by the Center for Climate System Research, the University of Tokyo and the Japanese National Institute for Environmental Sciences. The AFES was, however, totally rewritten in FORTRAN90 and MPI while the original AGCM was written in FORTRAN77 and not capable of parallel computing. The AFES achieved 26 Tflops (about 65 % of the peak performance of the ES) at resolution of T1279L96 (10-km horizontal resolution and 500-m vertical resolution in middle troposphere to lower stratosphere). Some results of 10- to 20-day global simulations will be presented. At this moment, only short-term simulations are possible due to data storage limitation. As ten tera flops computing is achieved, peta byte data storage are necessary to conduct climate-type simulations at this super-high resolution global simulations. Some possibilities for future research topics in global super-high resolution climate simulations will be discussed. Some target topics are mesoscale structures and self-organization of the Baiu-Meiyu front over Japan, cyclogenecsis over the North Pacific and typhoons around the Japan area. Also improvement in local precipitation with increasing horizontal resolution will be demonstrated.

  10. An HIV-1 encoded peptide mimics the DNA binding loop of NF-κB and binds thioredoxin with high affinity

    International Nuclear Information System (INIS)

    Su Guoping; Wang Min; Taylor, Ethan Will

    2005-01-01

    Pro-fs is a human immunodeficiency virus type 1 (HIV-l)-encoded putative selenoprotein, predicted by a theoretical analysis of the viral genome; it is potentially expressed by a -1 frameshift from the protease coding region. Pro-fs has significant sequence similarity to the DNA binding loop of nuclear factor kappa B (NF-κB), which is known to bind thioredoxin (Trx). We hypothesize that the putative HIV-1 pro-fs gene product functions by mimicry of NF-κB via binding to Trx. The hypothesis was tested in vitro by co-immunoprecipitation and GST-pull down assays, using a purified mutant pro-fs protein, in which the two potential selenocysteine residues were mutated to cysteines, in order to permit expression in bacteria. Both experiments showed that pro-fs binds to human wild type Trx (Trx-wt) with high affinity. Mutation of the two conserved cysteine residues in the Trx active site redox center to serine (Ser) (Trx-CS) weakened but failed to abolish the interaction. In pro-fs-transfected 293T cells, using confocal microscopy and fluorescence resonance energy transfer (FRET), we have observed that pro-fs localizes in cell nuclei and forms oligomers. Upon stimulation by phorbol 12-myristate 13-acetate (PMA), Trx translocates into cell nuclei. Significant FRET efficiency was detected in the nuclei of PMA-stimulated 293T cells co-expressing fluorescence-tagged pro-fs and Trx-wt or Trx-CS. These results indicate that in living cells the double cysteine mutant of pro-fs binds to both Trx and Trx-CS with high affinity, suggesting that Trx-pro-fs binding is a structurally-specific interaction, involving more of the Trx molecule than just its active site cysteine residues. These results establish the capacity for functional mimicry of the Trx binding ability of the NF-κB/Rel family of transcription factors by the putative HIV-1 pro-fs protein

  11. Synthesis and characterization of [{sup 76}Br]-labeled high-affinity A{sub 3} adenosine receptor ligands for positron emission tomography

    Energy Technology Data Exchange (ETDEWEB)

    Kiesewetter, Dale O. [Positron Emission Tomography Radiochemistry Group, NIBIB, Clinical Center, National Institutes of Health, Bethesda, MD 20892 (United States)], E-mail: dk7k@nih.gov; Lang Lixin; Ma Ying; Bhattacharjee, Abesh Kumar [Positron Emission Tomography Radiochemistry Group, NIBIB, Clinical Center, National Institutes of Health, Bethesda, MD 20892 (United States); Gao, Zhan-Guo; Joshi, Bhalchandra V.; Melman, Artem; Castro, Sonia de; Jacobson, Kenneth A. [Molecular Recognition Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892 (United States)

    2009-01-15

    Introduction: Bromine-76-radiolabeled analogues of previously reported high-affinity A{sub 3} adenosine receptor (A{sub 3}AR) nucleoside ligands have been prepared as potential radiotracers for positron emission tomography. Methods: The radiosyntheses were accomplished by oxidative radiobromination on the N{sup 6}-benzyl moiety of trimethyltin precursors. Biodistribution studies of the kinetics of uptake were conducted in awake rats. Results: We prepared an agonist ligand {l_brace}[{sup 76}Br](1'S,2'R,3'S,4'R,5'S)-4'-{l_brace}2-chloro-6-[(3-bromophenylmethyl)amino] purin-9-yl{r_brace}-1'-(methylaminocarbonyl)bicyclo[3.1.0]hexane-2',3'-diol (MRS3581){r_brace} in 59% radiochemical yield with a specific activity of 19.5 GBq/{mu}mol and an antagonist ligand {l_brace}[{sup 76}Br](1'R,2'R,3'S,4'R,5'S)-4'-(6-(3-bromobenzylamino) -2-chloro-9H-purin-9-yl)bicyclo[3.1.0]hexane-2',3'-diol (MRS5147){r_brace} in 65% radiochemical yield with a specific activity of 22 GBq/{mu}mol. The resultant products exhibited the expected high affinity (K{sub i}{approx}0.6 nM) and specific binding at the human A{sub 3}AR in vitro. Biodistribution studies in the rat showed uptake in the organs of excretion and metabolism. The antagonist MRS5147 exhibited increasing uptake in testes, an organ that contains significant quantities of A{sub 3}AR, over a 2-h time course, which suggests the presence of a specific A{sub 3}AR retention mechanism. Conclusion: We were able to compare uptake of the [{sup 76}Br]-labeled antagonist MRS5147 to [{sup 76}Br]agonist MRS3581. The antagonist MRS5147 shows increasing uptake in the testes, an A{sub 3}AR-rich tissue, suggesting that this ligand may have promise as a molecular imaging agent.

  12. Sea Surface Temperature (14 KM North America)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Product shows local sea surface temperatures (degrees C). It is a composite gridded-image derived from 8-km resolution SST Observations. It is generated every 48...

  13. Climate Prediction Center IR 4km Dataset

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — CPC IR 4km dataset was created from all available individual geostationary satellite data which have been merged to form nearly seamless global (60N-60S) IR...

  14. MODIS/Aqua Clouds 5-Min L2 Swath 1km and 5km V006

    Data.gov (United States)

    National Aeronautics and Space Administration — The MODIS/Aqua Clouds 5-Min L2 Swath 1km and 5km (MYD06_L2) product consists of cloud optical and physical parameters. These parameters are derived using remotely...

  15. N- and C-terminally truncated forms of glucose-dependent insulinotropic polypeptide are high-affinity competitive antagonists of the human GIP receptor

    DEFF Research Database (Denmark)

    Hansen, L S; Sparre-Ulrich, A H; Christensen, M.

    2016-01-01

    functions and pharmacological potential. GIP(1-30)NH2 is a naturally occurring truncation of GIP(1-42). Here we characterize eight N-terminal trrncations of human GIP(1-30)NH2 : GIP(2- to 9-30)NH2 . EXPERIMENTAL APPROACH: COS-7 cells were transiently transfected with the human GIP receptor and assessed...... displayed lower affinities (Ki 2.3-347 nM) with highest affinities of GIP(3-30)NH2 and (5-30)NH2 . Agonism was only observed for GIP(1-30)NH2 with an Emax on 100% of GIP(1-42) and GIP(2-30)NH2 (Emax 20%). GIP(2- to 9-30)NH2 displayed antagonism (IC50 12-450 nM) and right-shifts of the GIP(1-42)-response......, but superior antagonist GIP(3-30)NH2 , that together with GIP(5-30)NH2 were high-affinity competitive antagonist and thus may be suitable tool compounds for basic GIP research and future pharmacological interventions....

  16. Selective cortical decrease of high-affinity choline uptake carrier in Alzheimer's disease: an autoradiographic study using 3H-hemicholinium-3

    International Nuclear Information System (INIS)

    Rodriguez-Puertas, R.; Pazos, A.; Zarranz, J.J.; Pascual, J.

    1994-01-01

    H-hemicholinium-3 (H-HC-3) binding, a marker of the presynaptic high-affinity choline uptake carrier (HACU), was measured by autoradiography in several brain regions of 17 Alzheimer's disease (AD) patients and of 11 matched controls. A significant decrease in the density of H-HC-3 binding sites was found in entorhinal cortex, hippocampus and layers I-III of the frontal cortex. By contrast, in the caudate-putamen the number of H-HC-3 binding sites in AD cases was comparable to that of control striata. These data concur with previous results using classical presynaptic markers and reflect the loss in the activity of HACU, and, hence, in the synthesis of acetylcholine, that selectively occurs in cortical areas of AD brains due to the degeneration of presynaptic cholinergic terminals arising from the basal forebrain. However, the relatively low mean reduction in HACU in cortical areas (-40 %), together with the apparent indemnity of this marker in certain severely demented AD cases, suggest that AD dementia cannot be explained simply by the loss of presynaptic terminals originating in the basal forebrain. These data seem to be a good explanation for the poor response to cholinergic replacement in AD. (author)

  17. High Affinity vs. Native Fibronectin in the Modulation of αvβ3 Integrin Conformational Dynamics: Insights from Computational Analyses and Implications for Molecular Design.

    Directory of Open Access Journals (Sweden)

    Antonella Paladino

    2017-01-01

    Full Text Available Understanding how binding events modulate functional motions of multidomain proteins is a major issue in chemical biology. We address several aspects of this problem by analyzing the differential dynamics of αvβ3 integrin bound to wild type (wtFN10, agonist or high affinity (hFN10, antagonist mutants of fibronectin. We compare the dynamics of complexes from large-scale domain motions to inter-residue coordinated fluctuations to characterize the distinctive traits of conformational evolution and shed light on the determinants of differential αvβ3 activation induced by different FN sequences. We propose an allosteric model for ligand-based integrin modulation: the conserved integrin binding pocket anchors the ligand, while different residues on the two FN10's act as the drivers that reorganize relevant interaction networks, guiding the shift towards inactive (hFN10-bound or active states (wtFN10-bound. We discuss the implications of results for the design of integrin inhibitors.

  18. The Extracellular Domain of Human High Affinity Copper Transporter (hNdCTR1), Synthesized by E. coli Cells, Chelates Silver and Copper Ions In Vivo.

    Science.gov (United States)

    Sankova, Tatiana P; Orlov, Iurii A; Saveliev, Andrey N; Kirilenko, Demid A; Babich, Polina S; Brunkov, Pavel N; Puchkova, Ludmila V

    2017-11-03

    There is much interest in effective copper chelators to correct copper dyshomeostasis in neurodegenerative and oncological diseases. In this study, a recombinant fusion protein for expression in Escherichia coli cells was constructed from glutathione-S-transferase (GST) and the N-terminal domain (ectodomain) of human high affinity copper transporter CTR1 (hNdCTR1), which has three metal-bound motifs. Several biological properties of the GST-hNdCTR1 fusion protein were assessed. It was demonstrated that in cells, the protein was prone to oligomerization, formed inclusion bodies and displayed no toxicity. Treatment of E. coli cells with copper and silver ions reduced cell viability in a dose- and time-dependent manner. Cells expressing GST-hNdCTR1 protein demonstrated resistance to the metal treatments. These cells accumulated silver ions and formed nanoparticles that contained AgCl and metallic silver. In this bacterial population, filamentous bacteria with a length of about 10 µm were often observed. The possibility for the fusion protein carrying extracellular metal binding motifs to integrate into the cell's copper metabolism and its chelating properties are discussed.

  19. The Extracellular Domain of Human High Affinity Copper Transporter (hNdCTR1, Synthesized by E. coli Cells, Chelates Silver and Copper Ions In Vivo

    Directory of Open Access Journals (Sweden)

    Tatiana P. Sankova

    2017-11-01

    Full Text Available There is much interest in effective copper chelators to correct copper dyshomeostasis in neurodegenerative and oncological diseases. In this study, a recombinant fusion protein for expression in Escherichia coli cells was constructed from glutathione-S-transferase (GST and the N-terminal domain (ectodomain of human high affinity copper transporter CTR1 (hNdCTR1, which has three metal-bound motifs. Several biological properties of the GST-hNdCTR1 fusion protein were assessed. It was demonstrated that in cells, the protein was prone to oligomerization, formed inclusion bodies and displayed no toxicity. Treatment of E. coli cells with copper and silver ions reduced cell viability in a dose- and time-dependent manner. Cells expressing GST-hNdCTR1 protein demonstrated resistance to the metal treatments. These cells accumulated silver ions and formed nanoparticles that contained AgCl and metallic silver. In this bacterial population, filamentous bacteria with a length of about 10 µm were often observed. The possibility for the fusion protein carrying extracellular metal binding motifs to integrate into the cell’s copper metabolism and its chelating properties are discussed.

  20. A viral, transporter associated with antigen processing (TAP)-independent, high affinity ligand with alternative interactions endogenously presented by the nonclassical human leukocyte antigen E class I molecule.

    Science.gov (United States)

    Lorente, Elena; Infantes, Susana; Abia, David; Barnea, Eilon; Beer, Ilan; García, Ruth; Lasala, Fátima; Jiménez, Mercedes; Mir, Carmen; Morreale, Antonio; Admon, Arie; López, Daniel

    2012-10-12

    The transporter associated with antigen processing (TAP) enables the flow of viral peptides generated in the cytosol by the proteasome and other proteases to the endoplasmic reticulum, where they complex with nascent human leukocyte antigen (HLA) class I. Later, these peptide-HLA class I complexes can be recognized by CD8(+) lymphocytes. Cancerous cells and infected cells in which TAP is blocked, as well as individuals with unusable TAP complexes, are able to present peptides on HLA class I by generating them through TAP-independent processing pathways. Here, we identify a physiologically processed HLA-E ligand derived from the D8L protein in TAP-deficient vaccinia virus-infected cells. This natural high affinity HLA-E class I ligand uses alternative interactions to the anchor motifs previously described to be presented on nonclassical HLA class I molecules. This octameric peptide was also presented on HLA-Cw1 with similar binding affinity on both classical and nonclassical class I molecules. In addition, this viral peptide inhibits HLA-E-mediated cytolysis by natural killer cells. Comparison between the amino acid sequences of the presenting HLA-E and HLA-Cw1 alleles revealed a shared structural motif in both HLA class molecules, which could be related to their observed similar cross-reactivity affinities. This motif consists of several residues located on the floor of the peptide-binding site. These data expand the role of HLA-E as an antigen-presenting molecule.

  1. High affinity radiopharmaceuticals based upon lansoprazole for PET imaging of aggregated tau in Alzheimer's disease and progressive supranuclear palsy: synthesis, preclinical evaluation, and lead selection.

    Science.gov (United States)

    Fawaz, Maria V; Brooks, Allen F; Rodnick, Melissa E; Carpenter, Garrett M; Shao, Xia; Desmond, Timothy J; Sherman, Phillip; Quesada, Carole A; Hockley, Brian G; Kilbourn, Michael R; Albin, Roger L; Frey, Kirk A; Scott, Peter J H

    2014-08-20

    Abnormally aggregated tau is the hallmark pathology of tauopathy neurodegenerative disorders and is a target for development of both diagnostic tools and therapeutic strategies across the tauopathy disease spectrum. Development of carbon-11- or fluorine-18-labeled radiotracers with appropriate affinity and specificity for tau would allow noninvasive quantification of tau burden using positron emission tomography (PET) imaging. We have synthesized [(18)F]lansoprazole, [(11)C]N-methyl lansoprazole, and [(18)F]N-methyl lansoprazole and identified them as high affinity radiotracers for tau with low to subnanomolar binding affinities. Herein, we report radiosyntheses and extensive preclinical evaluation with the aim of selecting a lead radiotracer for translation into human PET imaging trials. We demonstrate that [(18)F]N-methyl lansoprazole, on account of the favorable half-life of fluorine-18 and its rapid brain entry in nonhuman primates, favorable kinetics, low white matter binding, and selectivity for binding to tau over amyloid, is the lead compound for progression into clinical trials.

  2. High Affinity Radiopharmaceuticals Based Upon Lansoprazole for PET Imaging of Aggregated Tau in Alzheimer’s Disease and Progressive Supranuclear Palsy: Synthesis, Preclinical Evaluation, and Lead Selection

    Science.gov (United States)

    2014-01-01

    Abnormally aggregated tau is the hallmark pathology of tauopathy neurodegenerative disorders and is a target for development of both diagnostic tools and therapeutic strategies across the tauopathy disease spectrum. Development of carbon-11- or fluorine-18-labeled radiotracers with appropriate affinity and specificity for tau would allow noninvasive quantification of tau burden using positron emission tomography (PET) imaging. We have synthesized [18F]lansoprazole, [11C]N-methyl lansoprazole, and [18F]N-methyl lansoprazole and identified them as high affinity radiotracers for tau with low to subnanomolar binding affinities. Herein, we report radiosyntheses and extensive preclinical evaluation with the aim of selecting a lead radiotracer for translation into human PET imaging trials. We demonstrate that [18F]N-methyl lansoprazole, on account of the favorable half-life of fluorine-18 and its rapid brain entry in nonhuman primates, favorable kinetics, low white matter binding, and selectivity for binding to tau over amyloid, is the lead compound for progression into clinical trials. PMID:24896980

  3. Carbon-11 labelling of eticlopride in two different positions - a selective high-affinity ligand for the study of dopamine D-2 receptors using PET

    International Nuclear Information System (INIS)

    Halldin, Christer; Hall, Haakan

    1990-01-01

    A new highly selective high-affinity dopamine D-2 receptor antagonist, eticlopride ((-)-(S)-5-chloro-3-ethyl-N-(1-ethyl-2-pyrrolidinyl)methyl)-6-methoxysalicylamide), was labelled with 11 C in two different positions ([N-ethyl- 11 C]eticlopride (I) and ([methyl- 11 C]eticlopride (II)). Product I was prepared by N-alkylation of the N-desethyl compound with [ 11 C]ethyl iodide. II was prepared by O-alkylation of the diphenolic precursor with [ 11 C]methyl iodide followed by separation of the two methylated products. The radiochemical yields were 15-20% (EOB) with an overall synthesis time of 45-60 min. Both compounds were isolated by semi-preparative HPLC and the radiochemical purity was in both cases > 99%. I was injected i.v. in a Cynomolgus monkey and brain radioactivity was measured by positron emission tomography (PET). The specific activity was 70 Ci/mmol at time of injection. There was a marked accumulation of radioactivity in the basal ganglia, regions known to have a high density of dopamine D-2 receptors. (author)

  4. Selective cortical decrease of high-affinity choline uptake carrier in Alzheimer`s disease: an autoradiographic study using {sup 3}H-hemicholinium-3

    Energy Technology Data Exchange (ETDEWEB)

    Rodriguez-Puertas, R; Pazos, A [Dept. of Physiology and Pharmacology, Unit of Pharmacology, Univ. of Cantabria, Santander (Spain); Zarranz, J J [Dept. of Neuroscience, Univ. of the Basque Country, Leioa (Spain); Pascual, J [Dept. of Medicine, Service of Neurology, Univ. Hospital ` Marques de Valdecilla` , Univ. of Cantabria, Santander (Spain)

    1994-12-31

    H-hemicholinium-3 (H-HC-3) binding, a marker of the presynaptic high-affinity choline uptake carrier (HACU), was measured by autoradiography in several brain regions of 17 Alzheimer`s disease (AD) patients and of 11 matched controls. A significant decrease in the density of H-HC-3 binding sites was found in entorhinal cortex, hippocampus and layers I-III of the frontal cortex. By contrast, in the caudate-putamen the number of H-HC-3 binding sites in AD cases was comparable to that of control striata. These data concur with previous results using classical presynaptic markers and reflect the loss in the activity of HACU, and, hence, in the synthesis of acetylcholine, that selectively occurs in cortical areas of AD brains due to the degeneration of presynaptic cholinergic terminals arising from the basal forebrain. However, the relatively low mean reduction in HACU in cortical areas (-40 %), together with the apparent indemnity of this marker in certain severely demented AD cases, suggest that AD dementia cannot be explained simply by the loss of presynaptic terminals originating in the basal forebrain. These data seem to be a good explanation for the poor response to cholinergic replacement in AD. (author).

  5. PCR-identification of a Nicotiana plumbaginifolia cDNA homologous to the high-affinity nitrate transporters of the crnA family.

    Science.gov (United States)

    Quesada, A; Krapp, A; Trueman, L J; Daniel-Vedele, F; Fernández, E; Forde, B G; Caboche, M

    1997-05-01

    A family of high-affinity nitrate transporters has been identified in Aspergillus nidulans and Chlamydomonas reinhardtii, and recently homologues of this family have been cloned from a higher plant (barley). Based on six of the peptide sequences most strongly conserved between the barley and C. reinhardtii polypeptides, a set of degenerate primers was designed to permit amplification of the corresponding genes from other plant species. The utility of these primers was demonstrated by RT-PCR with cDNA made from poly(A)+ RNA from barley, C. reinhardtii and Nicotiana plumbaginifolia. A PCR fragment amplified from N. plumbaginifolia was used as probe to isolate a full-length cDNA clone which encodes a protein, NRT2;1Np, that is closely related to the previously isolated crnA homologue from barley. Genomic Southern blots indicated that there are only 1 or 2 members of the Nrt2 gene family in N. plumbaginifolia. Northern blotting showed that the Nrt2 transcripts are most strongly expressed in roots. The effects of external treatments with different N sources showed that the regulation of the Nrt2 gene(s) is very similar to that reported for nitrate reductase and nitrite reductase genes: their expression was strongly induced by nitrate but was repressed when reduced forms of N were supplied to the roots.

  6. An in vitro-identified high-affinity nucleosome-positioning signal is capable of transiently positioning a nucleosome in vivo

    Directory of Open Access Journals (Sweden)

    Gracey Lia E

    2010-07-01

    Full Text Available Abstract Background The physiological function of eukaryotic DNA occurs in the context of nucleosomal arrays that can expose or obscure defined segments of the genome. Certain DNA sequences are capable of strongly positioning a nucleosome in vitro, suggesting the possibility that favorable intrinsic signals might reproducibly structure chromatin segments. As high-throughput sequencing analyses of nucleosome coverage in vitro and in vivo have become possible, a vigorous debate has arisen over the degree to which intrinsic DNA:nucleosome affinities orchestrate the in vivo positions of nucleosomes, thereby controlling physical accessibility of specific sequences in DNA. Results We describe here the in vivo consequences of placing a synthetic high-affinity nucleosome-positioning signal, the 601 sequence, into a DNA plasmid vector in mice. Strikingly, the 601 sequence was sufficient to position nucleosomes during an early phase after introduction of the DNA into the mice (when the plasmid vector transgene was active. This positioning capability was transient, with a loss of strong positioning at a later time point when the transgenes had become silent. Conclusions These results demonstrate an ability of DNA sequences selected solely for nucleosome affinity to organize chromatin in vivo, and the ability of other mechanisms to overcome these interactions in a dynamic nuclear environment.

  7. The Mitochondrial Metallochaperone SCO1 Is Required to Sustain Expression of the High-Affinity Copper Transporter CTR1 and Preserve Copper Homeostasis

    Directory of Open Access Journals (Sweden)

    Christopher J. Hlynialuk

    2015-02-01

    Full Text Available Human SCO1 fulfills essential roles in cytochrome c oxidase (COX assembly and the regulation of copper (Cu homeostasis, yet it remains unclear why pathogenic mutations in this gene cause such clinically heterogeneous forms of disease. Here, we establish a Sco1 mouse model of human disease and show that ablation of Sco1 expression in the liver is lethal owing to severe COX and Cu deficiencies. We further demonstrate that the Cu deficiency is explained by a functional connection between SCO1 and CTR1, the high-affinity transporter that imports Cu into the cell. CTR1 is rapidly degraded in the absence of SCO1 protein, and we show that its levels are restored in Sco1−/− mouse embryonic fibroblasts upon inhibition of the proteasome. These data suggest that mitochondrial signaling through SCO1 provides a post-translational mechanism to regulate CTR1-dependent Cu import into the cell, and they further underpin the importance of mitochondria in cellular Cu homeostasis.

  8. SKF 525-A and cytochrome P-450 ligands inhibit with high affinity the binding of [3H]dextromethorphan and σligands to guinea pig brain

    International Nuclear Information System (INIS)

    Klein, M.; Canoll, P.D.; Musacchio, J.M.

    1991-01-01

    The DM 1 /σ 1 site binds dextromethorphan (DM) and σ receptor ligands. The broad binding specificity of this site and its peculiar subcellular distribution prompted us to explore the possibility that this site is a member of the cytochrome P-450 superfamily of enzymes. We tested the effects of the liver microsomal monooxygenase inhibitor SKF 525-A (Proadifen), and other P-450 substrates on the binding of [ 3 H]dextromethorphan, [ 3 H]3-(3-Hydroxyphenyl)-N-(1-propyl)piperidine and (+)-[ 3 H]1,3-Di-o-tolyl-guanidine ([ 3 H]DTG) to the guinea pig brain. SKF 525-A, l-lobeline and GBR-12909 inhibited the binding of the three labeled ligands with nM affinity. Each drug has identical nM K i values for the high-affinity site labeled by the three ligands. This indicated that they displaced the labeled ligands from the common DM 1 σ 1 site. Debrisoquine and sparteine, prototypical substrates for liver debrisoquine 4-hydroxylase, displayed K i values of 9-13 and 3-4 μM respectively against the three labeled ligands. These results, the broad specificity of the DM 1 /σ 1 binding site, and its peculiar subcellular distribution, raises the possibility that this binding site is a member of the cytochrome P-450 superfamily of isozymes, rather than a neurotransmitter receptor

  9. [Structure-functional organization of eukaryotic high-affinity copper importer CTR1 determines its ability to transport copper, silver and cisplatin].

    Science.gov (United States)

    Skvortsov, A N; Zatulovskiĭ, E A; Puchkova, L V

    2012-01-01

    It was shown recently, that high affinity Cu(I) importer eukaryotic protein CTR1 can also transport in vitro abiogenic Ag(I) ions and anticancer drug cisplatin. At present there is no rational explanation how CTR1 can transfer platinum group, which is different by coordination properties from highly similar Cu(I) and Ag(I). To understand this phenomenon we analyzed 25 sequences of chordate CTR1 proteins, and found out conserved patterns of organization of N-terminal extracellular part of CTR1 which correspond to initial metal binding. Extracellular copper-binding motifs were qualified by their coordination properties. It was shown that relative position of Met- and His-rich copper-binding motifs in CTR1 predisposes the extracellular CTR1 part to binding of copper, silver and cisplatin. Relation between tissue-specific expression of CTR1 gene, steady-state copper concentration, and silver and platinum accumulation in organs of mice in vivo was analyzed. Significant positive but incomplete correlation exists between these variables. Basing on structural and functional peculiarities of N-terminal part of CTR1 a hypothesis of coupled transport of copper and cisplatin has been suggested, which avoids the disagreement between CTR1-mediated cisplatin transport in vitro, and irreversible binding of platinum to Met-rich peptides.

  10. Establishment of a novel high-affinity IgE receptor-positive canine mast cell line with wild-type c-kit receptors

    International Nuclear Information System (INIS)

    Amagai, Yosuke; Tanaka, Akane; Ohmori, Keitaro; Matsuda, Hiroshi

    2008-01-01

    Much is known regarding participations of mast cells with innate and acquired immunity by secreting various cytokines and chemical mediators. However, details of mast cell biology still remain unclear. In this study, we successfully established a novel growth factor-independent mast cell line (MPT-1) derived from canine mast cell tumor. MPT-1 cells manifested factor-independent proliferation as floating cells containing a large amount of histamine, as well as chymase-like dog mast cell protease 3, in cytosolic granules. Particularly, MPT-1 cells expressed high-affinity IgE receptors (FcεRI) and wild-type c-kit receptors. Degranulation of MPT-1 cells was induced not only by stimulation with calcium ionophore but also by cross-linkage of the surface IgE. Given that MPT-1 is the first mast cell line with FcεRI which has no c-kit mutations, MPT-1 cells may provide great contribution for investigation of IgE-mediated activation mechanisms of mast cells, leading to development of effective treatment for allergic disorders

  11. Motif III in superfamily 2 "helicases" helps convert the binding energy of ATP into a high-affinity RNA binding site in the yeast DEAD-box protein Ded1.

    Science.gov (United States)

    Banroques, Josette; Doère, Monique; Dreyfus, Marc; Linder, Patrick; Tanner, N Kyle

    2010-03-05

    Motif III in the putative helicases of superfamily 2 is highly conserved in both its sequence and its structural context. It typically consists of the sequence alcohol-alanine-alcohol (S/T-A-S/T). Historically, it was thought to link ATPase activity with a "helicase" strand displacement activity that disrupts RNA or DNA duplexes. DEAD-box proteins constitute the largest family of superfamily 2; they are RNA-dependent ATPases and ATP-dependent RNA binding proteins that, in some cases, are able to disrupt short RNA duplexes. We made mutations of motif III (S-A-T) in the yeast DEAD-box protein Ded1 and analyzed in vivo phenotypes and in vitro properties. Moreover, we made a tertiary model of Ded1 based on the solved structure of Vasa. We used Ded1 because it has relatively high ATPase and RNA binding activities; it is able to displace moderately stable duplexes at a large excess of substrate. We find that the alanine and the threonine in the second and third positions of motif III are more important than the serine, but that mutations of all three residues have strong phenotypes. We purified the wild-type and various mutants expressed in Escherichia coli. We found that motif III mutations affect the RNA-dependent hydrolysis of ATP (k(cat)), but not the affinity for ATP (K(m)). Moreover, mutations alter and reduce the affinity for single-stranded RNA and subsequently reduce the ability to disrupt duplexes. We obtained intragenic suppressors of the S-A-C mutant that compensate for the mutation by enhancing the affinity for ATP and RNA. We conclude that motif III and the binding energy of gamma-PO(4) of ATP are used to coordinate motifs I, II, and VI and the two RecA-like domains to create a high-affinity single-stranded RNA binding site. It also may help activate the beta,gamma-phosphoanhydride bond of ATP. (c) 2009 Elsevier Ltd. All rights reserved.

  12. Design, Synthesis, and in Vitro Pharmacology of New Radiolabeled γ-Hydroxybutyric Acid Analogues Including Photolabile Analogues with Irreversible Binding to the High-Affinity γ-Hydroxybutyric Acid Binding Sites

    DEFF Research Database (Denmark)

    Sabbatini, Paola; Wellendorph, Petrine; Høg, Signe

    2010-01-01

    γ-Hydroxybutyric acid (GHB) is a psychotropic compound endogenous to the brain. Despite its potential physiological significance, the complete molecular mechanisms of action remain unexplained. To facilitate the isolation and identification of the high-affinity GHB binding site, we herein report ...

  13. Aluminium fluoride and magnesium, activators of heterotrimeric GTP-binding proteins, affect high-affinity binding of the fungal toxin fusicoccin to the fusicoccin-binding protein in oat root plasma membranes.

    NARCIS (Netherlands)

    de Boer, A.H.; Van der Molen, G.W.; Prins, H.B.A.; Korthout, H.A.A.J.; van der Hoeven, P.C.J.

    1994-01-01

    The fusicoccin-binding protein was solubilised from purified oat root plasma membranes. The solubilised protein retained full binding activity, provided that protease inhibitors were included. Sodium fluoride reduced the high-affinity [H-3]fusicoccin binding to almost zero in a

  14. Technical description of the Tansy Spectrometer, TANSY-KM5

    International Nuclear Information System (INIS)

    Grosshoeg, G.; Aronsson, D.; Hoek, M.; Rydz, R.; Norberg, L.; Urholm, L.

    1991-07-01

    The TANSY-KM5 neutron spectrometer is a system containing a lot of advanced electronic and mechanical components. The design of the system has been reported in two documents (CTH-RF--54 (INIS 17:10206) and CTH-RF--43 (INIS 14:797424)). The purpose of this documentation is to give a brief overview of the system as it is defined at the time of delivery

  15. Pulmonary Administration of GW0742, a High-Affinity Peroxisome Proliferator-Activated Receptor Agonist, Repairs Collapsed Alveoli in an Elastase-Induced Mouse Model of Emphysema.

    Science.gov (United States)

    Ozawa, Chihiro; Horiguchi, Michiko; Akita, Tomomi; Oiso, Yuki; Abe, Kaori; Motomura, Tomoki; Yamashita, Chikamasa

    2016-01-01

    Pulmonary emphysema is a disease in which lung alveoli are irreversibly damaged, thus compromising lung function. Our previous study revealed that all-trans-retinoic acid (ATRA) induces the differentiation of human lung alveolar epithelial type 2 progenitor cells and repairs the alveoli of emphysema model mice. ATRA also reportedly has the ability to activate peroxisome proliferator-activated receptor (PPAR) β/δ. A selective PPARβ/δ ligand has been reported to induce the differentiation of human keratinocytes during wound repair. Here, we demonstrate that treatment using a high-affinity PPARβ/δ agonist, GW0742, reverses the lung tissue damage induced by elastase in emphysema-model mice and improves respiratory function. Mice treated with elastase, which collapsed their alveoli, were then treated with either 10% dimethyl sulfoxide (DMSO) in saline (control group) or GW0742 (1.0 mg/kg twice a week) by pulmonary administration. Treatment with GW0742 for 2 weeks increased the in vivo expression of surfactant proteins A and D, which are known alveolar type II epithelial cell markers. GW0742 treatment also shortened the average distance between alveolar walls in the lungs of emphysema model mice, compared with a control group treated with 10% DMSO in saline. Treatment with GW0742 for 3 weeks also improved tissue elastance (cm H2O/mL), as well as the ratio of the forced expiratory volume in the first 0.05 s to the forced vital capacity (FEV 0.05/FVC). In each of these experiments, GW0742 treatment reversed the damage caused by elastase. In conclusion, PPARβ/δ agonists are potential therapeutic agents for pulmonary emphysema.

  16. Generation of high-affinity, internalizing anti-FGFR2 single-chain variable antibody fragment fused with Fc for targeting gastrointestinal cancers.

    Science.gov (United States)

    Borek, Aleksandra; Sokolowska-Wedzina, Aleksandra; Chodaczek, Grzegorz; Otlewski, Jacek

    2018-01-01

    Fibroblast growth factor receptors (FGFRs) are promising targets for antibody-based cancer therapies, as their substantial overexpression has been found in various tumor cells. Aberrant activation of FGF receptor 2 (FGFR2) signaling through overexpression of FGFR2 and/or its ligands, mutations, or receptor amplification has been reported in multiple cancer types, including gastric, colorectal, endometrial, ovarian, breast and lung cancer. In this paper, we describe application of the phage display technology to produce a panel of high affinity single chain variable antibody fragments (scFvs) against the extracellular ligand-binding domain of FGFR2 (ECD_FGFR2). The binders were selected from the human single chain variable fragment scFv phage display libraries Tomlinson I + J and showed high specificity and binding affinity towards human FGFR2 with nanomolar KD values. To improve the affinity of the best binder selected, scFvF7, we reformatted it to a bivalent diabody format, or fused it with the Fc region (scFvF7-Fc). The scFvF7-Fc antibody construct presented the highest affinity for FGFR2, with a KD of 0.76 nM, and was selectively internalized into cancer cells overexpressing FGFR2, Snu-16 and NCI-H716. Finally, we prepared a conjugate of scFvF7-Fc with the cytotoxic drug monomethyl-auristatin E (MMAE) and evaluated its cytotoxicity. The conjugate delivered MMAE selectively to FGFR2-positive tumor cells. These results indicate that scFvF7-Fc-vcMMAE is a highly potent molecule for the treatment of cancers with FGFR2 overexpression.

  17. High-affinity binding of [3H]estradiol-17 beta by an estrogen receptor in the liver of the turtle

    International Nuclear Information System (INIS)

    Ho, S.M.; Fehrer, S.; Yu, M.; Liang, L.C.; Press, D.

    1988-01-01

    Specific [3H]estradiol-17 beta ([3H]E2) binding activity (EBA) with characteristics of an estrogen receptor (ER) was demonstrated in cytosols and nuclear extracts of the female turtle, Chrysemys picta. Three different receptor assays (dextran-coated charcoal assay, hydroxylapatite batch procedure, and DNA-cellulose chromatography) were evaluated in terms of their applicability in analyzing large numbers of samples. For the measurement of cytosolic EBA, the hydroxylapatite batch procedure was found to be the most reliable assay. On the other hand, the dextran-coated charcoal assay was found to be the most appropriate method for the measurement of nuclear EBA. Turtle hepatic EBA binds [3H]E2 with high affinity (cytosolic, 17.4 +/- 2.8 X 10(9) M-1; nuclear, 17.7 +/- 1.9 X 10(9) M-1), limited capacity (cytosolic, 133.7 +/- 4.6 fmol/g tissue; nuclear, 81.1 +/- 9.0 fmol/g tissue), and strict steroid specificity. The EBA bound natural estrogens (E2, estrone, estriol) as well as the nonsteroidal estrogen, diethylstilbestrol, but exhibited little affinity for androgens, progesterone, or corticosterone. The turtle hepatic EBA resembled mammalian and avian ERs in terms of binding characteristics; however, unlike mammalian and avian ERs it was shown to be heat-labile. Incubation at 30 degrees caused rapid loss of [3H]E2 binding activity in both cytosolic and nuclear fractions. The exchange between [3H]E2 and the endogenously bound estrogen was slow at 4 and 15 degrees, but the exchange process was facilitated in the presence of the chaotropic salt, NaSCN. Establishment of quantitation methods for both cytosolic and nuclear forms of EBA will enable future investigation of the mechanism and regulation of estrogen action in the liver of this turtle species

  18. (3H)leukotriene B4 binding to the guinea pig spleen membranes: a rich tissue source for a high affinity leukotriene B4 receptor site

    International Nuclear Information System (INIS)

    Cheng, J.B.; Kohi, F.; Townley, R.G.

    1986-01-01

    To select a tissue rich for the high affinity leukotriene (LT)B 4 receptor site, they compared binding of 1 nM ( 3 H)LTB 4 (180 Ci/mmol) to the crude membrane preparations of guinea pig spleen, thymus, lung, uterus, bladder, brain, adrenal gland, small intestine, liver, kidney and heart. They found that the membrane preparations from spleen contained the highest binding activity per mg protein. They characterized the LTB 4 binding to the spleen preparation in detail. LTB 4 binding was rapid, reversible, stereoselective and saturable. The data from equilibrium experiments showed a linear Scatchard plot with a K/sub d/ of 1.6 nM and a binding site density of 259 fmol/mg prot. The rank order of agents competing for spleen ( 3 H)LTB 4 binding at 25 0 C was: LTB 4 (K/sub i/ = 2.8 nM) > 20-OH-LTB 4 (23 nM) > LTA 4 (48 nM) > LTA 4 methyl ester (0.13 μM) > 20-COOH-LTB 4 (> 6.6 μM) ≥ arachidonic acid (0.15 mM) similarly ordered FPL-55,712 (0.11 mM). At 4 0 C, LTB 4 (2.3 nM) competed at least 10x more effectively than 20-OH-LTB 4 (29 nM) and 20-COOH-LTB 4 (> 6.6 μM). HPLC analysis indicated that incubation of 84 ng LTB 4 with the spleen membrane at 25 0 C did not result in the formation of 20-OH-LTB 4 ( 3 H)LTB 4 receptor binding sites

  19. The high affinity K+ transporter AtHAK5 plays a physiological role in planta at very low K+ concentrations and provides a caesium uptake pathway in Arabidopsis.

    Science.gov (United States)

    Qi, Zhi; Hampton, Corrina R; Shin, Ryoung; Barkla, Bronwyn J; White, Philip J; Schachtman, Daniel P

    2008-01-01

    Caesium (Cs(+)) is a potentially toxic mineral element that is released into the environment and taken up by plants. Although Cs(+) is chemically similar to potassium (K(+)), and much is known about K(+) transport mechanisms, it is not clear through which K(+) transport mechanisms Cs(+) is taken up by plant roots. In this study, the role of AtHAK5 in high affinity K(+) and Cs(+) uptake was characterized. It is demonstrated that AtHAK5 is localized to the plasma membrane under conditions of K(+) deprivation, when it is expressed. Growth analysis showed that AtHAK5 plays a role during severe K(+) deprivation. Under K(+)-deficient conditions in the presence of Cs(+), Arabidopsis seedlings lacking AtHAK5 had increased inhibition of root growth and lower Cs(+) accumulation, and significantly higher leaf chlorophyll concentrations than wild type. These data indicate that, in addition to transporting K(+) in planta, AtHAK5 also transports Cs(+). Further experiments showed that AtHAK5 mediated Cs(+) uptake into yeast cells and that, although the K(+) deficiency-induced expression of AtHAK5 was inhibited by low concentrations of NH(4)(+) in planta, Cs(+) uptake by yeast was stimulated by low concentrations of NH(4)(+). Interestingly, the growth of the Arabidopsis atakt1-1 mutant was more sensitive to Cs(+) than the wild type. This may be explained, in part, by increased expression of AtHAK5 in the atakt1-1 mutant. It is concluded that AtHAK5 is a root plasma membrane uptake mechanism for K(+) and Cs(+) under conditions of low K(+) availability.

  20. Preliminary assessment of extrastriatal dopamine d-2 receptor binding in the rodent and nonhuman primate brains using the high affinity radioligand, {sup 18}F-fallypride

    Energy Technology Data Exchange (ETDEWEB)

    Mukherjee, Jogeshwar E-mail: jogeshwar-mukherjee@ketthealth.com; Yang, Z.-Y.; Brown, Terry; Lew, Robert; Wernick, Miles; Ouyang Xiaohu; Yasillo, Nicholas; Chen, C.-T.; Mintzer, Robert; Cooper, Malcolm

    1999-07-01

    We have identified the value of {sup 18}F-fallypride {l_brace}(S)-N-[(1-allyl-2-pyrrolidinyl)methyl]-5-(3-[{sup 18}F]fluoropropyl)-2,3-dim= ethoxybenzamide{r_brace}, as a dopamine D-2 receptor radiotracer for the study of striatal and extrastriatal receptors. Fallypride exhibits high affinities for D-2 and D-3 subtypes and low affinity for D-4 ({sup 3}H-spiperone IC{sub 50}s: D-2=0.05 nM [rat striata], D-3=0.30 nM [SF9 cell lines, rat recombinant], and D-4=240 nM [CHO cell lines, human recombinant]). Biodistribution in the rat brain showed localization of {sup 18}F-fallypride in striata and extrastriatal regions such as the frontal cortex, parietal cortex, amygdala, hippocampus, thalamus, and hypothalamus. In vitro autoradiographic studies in sagittal slices of the rat brain showed localization of {sup 18}F-fallypride in striatal and several extrastriatal regions, including the medulla. Positron emission tomography (PET) experiments with {sup 18}F-fallypride in male rhesus monkeys were carried out in a PET VI scanner. In several PET experiments, apart from the specific binding seen in the striatum, specific binding of {sup 18}F-fallypride was also identified in extracellular regions (in a lower brain slice, possibly the thalamus). Specific binding in the extrastriata was, however, significantly lower compared with that observed in the striata of the monkeys (extrastriata/cerebellum = 2, striata/cerebellum = 10). Postmortem analysis of the monkey brain revealed significant {sup 18}F-fallypride binding in the striata, whereas binding was also observed in extrastriatal regions such as the thalamus, cortical areas, and brain stem.

  1. SNF3 as high affinity glucose sensor and its function in supporting the viability of Candida glabrata under glucose-limited environment

    Directory of Open Access Journals (Sweden)

    Tzu Shan eNg

    2015-12-01

    Full Text Available Candida glabrata is an emerging human fungal pathogen that has efficacious nutrient sensing and responsiveness ability. It can be seen through its ability to thrive in diverse range of nutrient limited-human anatomical sites. Therefore, nutrient sensing particularly glucose sensing is thought to be crucial in contributing to the development and fitness of the pathogen. This study aimed to elucidate the role of SNF3 (Sucrose Non Fermenting 3 as a glucose sensor and its possible role in contributing to the fitness and survivability of C. glabrata in glucose-limited environment. The SNF3 knockout strain was constructed and subjected to different glucose concentrations to evaluate its growth, biofilm formation, amphotericin B susceptibility, ex vivo survivability and effects on the transcriptional profiling of the sugar receptor repressor (SRR pathway-related genes. The SNF3Δ strain showed a retarded growth in low glucose environments (0.01% and 0.1% in both fermentation and respiration-preferred conditions but grew well in high glucose concentration environments (1% and 2%. It was also found to be more susceptible to amphotericin B in low glucose environment (0.1% and macrophage engulfment but showed no difference in the biofilm formation capability. The deletion of SNF3 also resulted in the down-regulation of about half of hexose transporters genes (4 out of 9. Overall, the deletion of SNF3 causes significant reduction in the ability of C. glabrata to sense limited surrounding glucose and consequently disrupts its competency to transport and perform the uptake of this critical nutrient. This study highlighted the role of SNF3 as a high affinity glucose sensor and its role in aiding the survivability of C. glabrata particularly in glucose limited environment.

  2. A conserved motif in the linker domain of STAT1 transcription factor is required for both recognition and release from high-affinity DNA-binding sites.

    Science.gov (United States)

    Hüntelmann, Bettina; Staab, Julia; Herrmann-Lingen, Christoph; Meyer, Thomas

    2014-01-01

    Binding to specific palindromic sequences termed gamma-activated sites (GAS) is a hallmark of gene activation by members of the STAT (signal transducer and activator of transcription) family of cytokine-inducible transcription factors. However, the precise molecular mechanisms involved in the signal-dependent finding of target genes by STAT dimers have not yet been very well studied. In this study, we have characterized a sequence motif in the STAT1 linker domain which is highly conserved among the seven human STAT proteins and includes surface-exposed residues in close proximity to the bound DNA. Using site-directed mutagenesis, we have demonstrated that a lysine residue in position 567 of the full-length molecule is required for GAS recognition. The substitution of alanine for this residue completely abolished both binding to high-affinity GAS elements and transcriptional activation of endogenous target genes in cells stimulated with interferon-γ (IFNγ), while the time course of transient nuclear accumulation and tyrosine phosphorylation were virtually unchanged. In contrast, two glutamic acid residues (E559 and E563) on each monomer are important for the dissociation of dimeric STAT1 from DNA and, when mutated to alanine, result in elevated levels of tyrosine-phosphorylated STAT1 as well as prolonged IFNγ-stimulated nuclear accumulation. In conclusion, our data indicate that the kinetics of signal-dependent GAS binding is determined by an array of glutamic acid residues located at the interior surface of the STAT1 dimer. These negatively charged residues appear to align the long axis of the STAT1 dimer in a position perpendicular to the DNA, thereby facilitating the interaction between lysine 567 and the phosphodiester backbone of a bound GAS element, which is a prerequisite for transient gene induction.

  3. Dual regulation of root hydraulic conductivity and plasma membrane aquaporins by plant nitrate accumulation and high-affinity nitrate transporter NRT2.1.

    Science.gov (United States)

    Li, Guowei; Tillard, Pascal; Gojon, Alain; Maurel, Christophe

    2016-04-01

    The water status and mineral nutrition of plants critically determine their growth and development. Nitrate (NO3(-)), the primary nitrogen source of higher plants, is known to impact the water transport capacity of roots (root hydraulic conductivity, Lpr). To explore the effects and mode of action of NO3(-) on Lpr, we used an extended set of NO3(-) transport (nrt1.1, nrt1.2, nrt1.5 and nrt2.1), signaling (nrt1.1 and nrt2.1) and metabolism (nia) mutants in Arabidopsis, grown under various NO3(-) conditions. First, a strong positive relationship between Lpr and NO3(-) accumulation, in shoots rather than in roots, was revealed. Secondly, a specific 30% reduction of Lpr in nrt2.1 plants unraveled a major role for the high-affinity NO3(-) transporter NRT2.1 in increasing Lpr These results indicate that NO3(-)signaling rather than nitrogen assimilation products governs Lpr in Arabidopsis. Quantitative real-time reverse transcription-PCR and enzyme-linked immunosorbent assays (ELISAs) were used to investigate the effects of NO3(-) availability on plasma membrane aquaporin (plasma membrane intrinsic protein; PIP) expression. Whereas PIP regulation mostly occurs at the post-translational level in wild-type plants, a regulation of PIPs at both the transcriptional and translational levels was uncovered in nrt2.1 plants. In conclusion, this work reveals that control of Arabidopsis Lpr and PIP functions by NO3(-) involves novel shoot to root signaling and NRT2.1-dependent functions. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  4. Murine CMV Expressing the High Affinity NKG2D Ligand MULT-1: A Model for the Development of Cytomegalovirus-Based Vaccines

    Directory of Open Access Journals (Sweden)

    Lea Hiršl

    2018-05-01

    Full Text Available The development of a vaccine against human cytomegalovirus (CMV has been a subject of long-term medical interest. The research during recent years identified CMV as an attractive vaccine vector against infectious diseases and tumors. The immune response to CMV persists over a lifetime and its unique feature is the inflationary T cell response to certain viral epitopes. CMV encodes numerous genes involved in immunoevasion, which are non-essential for virus growth in vitro. The deletion of those genes results in virus attenuation in vivo, which enables us to dramatically manipulate its virulence and the immune response. We have previously shown that the murine CMV (MCMV expressing RAE-1γ, one of the cellular ligands for the NKG2D receptor, is highly attenuated in vivo but retains the ability to induce a strong CD8+ T cell response. Here, we demonstrate that recombinant MCMV expressing high affinity NKG2D ligand murine UL16 binding protein-like transcript (MULT-1 (MULT-1MCMV inserted in the place of its viral inhibitor is dramatically attenuated in vivo in a NK cell-dependent manner, both in immunocompetent adult mice and in immunologically immature newborns. MULT-1MCMV was more attenuated than the recombinant virus expressing RAE-1γ. Despite the drastic sensitivity to innate immune control, MULT-1MCMV induced an efficient CD8+ T cell response to viral and vectored antigens. By using in vitro assay, we showed that similar to RAE-1γMCMV, MULT-1 expressing virus provided strong priming of CD8+ T cells. Moreover, MULT-1MCMV was able to induce anti-viral antibodies, which after passing the transplacental barrier protect offspring of immunized mothers from challenge infection. Altogether, this study further supports the concept that CMV expressing NKG2D ligand possesses excellent characteristics to serve as a vaccine or vaccine vector.

  5. High-affinity interaction of hnRNP A1 with conserved RNA structural elements is required for translation and replication of enterovirus 71.

    Science.gov (United States)

    Levengood, Jeffrey D; Tolbert, Michele; Li, Mei-Ling; Tolbert, Blanton S

    2013-07-01

    Human Enterovirus 71 (EV71) is an emerging pathogen of infectious disease and a serious threat to public health. Currently, there are no antivirals or vaccines to slow down or prevent EV71 infections, thus underscoring the urgency to better understand mechanisms of host-enterovirus interactions. EV71 uses a type I internal ribosome entry site (IRES) to recruit the 40S ribosomal subunit via a pathway that requires the cytoplasmic localization of hnRNP A1, which acts as an IRES trans-activating factor. The mechanism of how hnRNP A1 trans activates EV71 RNA translation is unknown, however. Here, we report that the UP1 domain of hnRNP A1 interacts specifically with stem loop II (SLII) of the IRES, via a thermodynamically well-defined biphasic transition that involves conserved bulge 5'-AYAGY-3' and hairpin 5'-RY(U/A)CCA-3' loops. Calorimetric titrations of wild-type and mutant SLII constructs reveal these structural elements are essential to form a high-affinity UP1-SLII complex. Mutations that alter the bulge and hairpin primary or secondary structures abrogate the biphasic transition and destabilize the complex. Notably, mutations within the bulge that destabilize the complex correlate with a large reduction in IRES-dependent translational activity and impair EV71 replication. Taken together, this study shows that a conserved SLII structure is necessary to form a functional hnRNP A1-IRES complex, suggesting that small molecules that target this stem loop may have novel antiviral properties.

  6. Why to Integrate KM into everyday practices?

    International Nuclear Information System (INIS)

    Chavez Flores, A.T.

    2014-01-01

    Researchers have argued that one process to achieve a succesful KM initiative is the creation of a culture that values the transfer and creation of knowledge. Therefore, it’s neccesary for organizations to set up mechanisms by which new ideas may be shared in order to produce innovative changes

  7. 5 km de autopista al mes

    Directory of Open Access Journals (Sweden)

    Editorial, Equipo

    1972-02-01

    Full Text Available This is one of the most important civil engineering projects of its kind currently under construction in the country. It consists of 93 km of motor road, with four traffic lanes, and it includes many ancillary works, using prefabricated units, multilevel crossings and overpasses of varying lengths, some of them 600 m long. The large scale use of prefabricated elements and of very modern machinery will make it possible to complete the project several months ahead of schedule.Esta obra, que puede contarse entre las más importantes del país, comprende 93 km de autopista con cuatro circulaciones, numerosas obras de fábrica —prefabricadas—, intersecciones a distinto nivel y pasos elevados con longitudes varias, algunos de 600 metros. La prefabricación masiva y el empleo de una modernísima maquinaria permitió terminar la obra varios meses antes del plazo previsto.

  8. Long-term, on-site borehole monitoring of gases released from an “active” fault system at 3.6km depth, TauTona Gold Mine, South Africa

    Science.gov (United States)

    Lippmann-Pipke, J.; Erzinger, J.; Zimmer, M.; Kujawa, C.; Boettcher, M. S.; Moller, H.; van Heerden, E.; Bester, A.; Reches, Z.

    2009-12-01

    Fluid transport and seismicity are interrelated. Fluids can trigger earthquakes and seismic activity can release fluids from rock formations. The study of this relationship requires direct and near-field observations at focal depth. The international DAFSAM-NELSAM*-projects focus on building the earthquake laboratory in deep gold mines in South Africa. Our DAFGAS-project (Drilling Active Faults - Gas Analytical System) aims to quantify the gases released during seismic events. One motivation for the project is to investigate the hypothesis that released fluids might be a nutrient supply for microbial ecosystems in active fault zones. Extensive underground activities started in 2004 with establishing a 25m2 cubby within the Pretorius fault zone at 3.6 km depth for the save installation of comprehensive technical equipment. For DAFGAS, subsequently two different gas analytical units were installed to measure gases collected in a 40 m long borehole crossing the fault. The DAFGAS Team and our collaborators overcame numerous technical problems. Since 2007 a dedicated air-conditioned box protects a mass spectrometer, pumps, a PC, a radon detector and electronics from the harsh underground environment. Since 2009 gas sensitive sensors and a data logger replace the spectrometer and the PC. In parallel the NELSAM project has installed 9 seismometers in a narrow network surrounding the gas collection system. The accelerometers and geophones record mining activities (e.g. drilling and ore-production blasts) as well as tens of mining-induced earthquakes (magnitude ≥ -4) on and around the Pretorius Fault each day. Data from three years is presented: Borehole temperature at 40m increased by about 0.8 °C/year to 52.3 °C; different scales of pressure variations on surface (869±5) mbar (three-week mean, maximal and minimal daily mean) and below surface (1130±15) mbar are explained by the barometric formula. The major gas concentrations are constant and air-like with 78 % N2

  9. PdeH, a high-affinity cAMP phosphodiesterase, is a key regulator of asexual and pathogenic differentiation in Magnaporthe oryzae.

    Directory of Open Access Journals (Sweden)

    Ravikrishna Ramanujam

    2010-05-01

    Full Text Available Cyclic AMP-dependent pathways mediate the communication between external stimuli and the intracellular signaling machinery, thereby influencing important aspects of cellular growth, morphogenesis and differentiation. Crucial to proper function and robustness of these signaling cascades is the strict regulation and maintenance of intracellular levels of cAMP through a fine balance between biosynthesis (by adenylate cyclases and hydrolysis (by cAMP phosphodiesterases. We functionally characterized gene-deletion mutants of a high-affinity (PdeH and a low-affinity (PdeL cAMP phosphodiesterase in order to gain insights into the spatial and temporal regulation of cAMP signaling in the rice-blast fungus Magnaporthe oryzae. In contrast to the expendable PdeL function, the PdeH activity was found to be a key regulator of asexual and pathogenic development in M. oryzae. Loss of PdeH led to increased accumulation of intracellular cAMP during vegetative and infectious growth. Furthermore, the pdeHDelta showed enhanced conidiation (2-3 fold, precocious appressorial development, loss of surface dependency during pathogenesis, and highly reduced in planta growth and host colonization. A pdeHDelta pdeLDelta mutant showed reduced conidiation, exhibited dramatically increased (approximately 10 fold cAMP levels relative to the wild type, and was completely defective in virulence. Exogenous addition of 8-Br-cAMP to the wild type simulated the pdeHDelta defects in conidiation as well as in planta growth and development. While a fully functional GFP-PdeH was cytosolic but associated dynamically with the plasma membrane and vesicular compartments, the GFP-PdeL localized predominantly to the nucleus. Based on data from cAMP measurements and Real-Time RTPCR, we uncover a PdeH-dependent biphasic regulation of cAMP levels during early and late stages of appressorial development in M. oryzae. We propose that PdeH-mediated sustenance and dynamic regulation of cAMP signaling

  10. Preclinical evaluation of multistep targeting of diasialoganglioside GD2 using a IgG-scFv bispecific antibody with high affinity for GD2 and DOTA metal complex

    Science.gov (United States)

    Cheal, Sarah M.; Xu, Hong; Guo, Hong-fen; Zanzonico, Pat B.; Larson, Steven M.; Cheung, Nai-Kong

    2014-01-01

    Bispecific antibodies (BsAb) have proven to be useful targeting vectors for pretargeted radioimmunotherapy (PRIT). We sought to overcome key PRIT limitations such as high renal radiation exposure and immunogenicity (e.g. of streptavidin-antibody fusions), to advance clinical translation of this PRIT strategy for diasialoganglioside GD2-positive (GD2(+)) tumors. For this purpose, a IgG-scFv BsAb was engineered using the sequences for the anti-GD2 humanized monoclonal antibody hu3F8 (1) and C825, a murine scFv antibody with high affinity for the chelator 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) complexed with beta-particle emitting radiometals such as 177Lu and 90Y (2, 3). A three-step regimen including hu3F8-C825, a dextran-based clearing agent, and p-aminobenzyl-DOTA radiolabeled with 177Lu (as 177Lu-DOTA-Bn; t1/2 = 6.71 days (d)) was optimized in immunocompromised mice carrying subcutaneous (s.c.) human GD2(+) neuroblastoma (NB) xenografts. Absorbed doses for tumor and normal tissues were ∼85 cGy/MBq and ≤3.7 cGy/MBq, respectively, with therapeutic indicies (TI) of 142 for blood and 23 for kidney. A therapy study (n = 5 per group; tumor volume: 240 ± 160 mm3) with three successive PRIT cycles (total 177Lu: ∼33 MBq; tumor dose ∼3400 cGy), revealed complete tumor response in 5/5 animals, with no recurrence up to 28 d post-treatment. Tumor ablation was confirmed histologically in 4/5 mice, and normal organs showed minimal overall toxicities. All non-treated mice required sacrifice within 12 d (>1.0 cm3 tumor volume). We conclude that this novel anti-GD2 PRIT approach has sufficient TI to successfully ablate s.c. GD2(+)–NB in mice while sparing kidney and bone marrow. PMID:24944121

  11. Labeling by ( sup 3 H)1,3-di(2-tolyl)guanidine of two high affinity binding sites in guinea pig brain: Evidence for allosteric regulation by calcium channel antagonists and pseudoallosteric modulation by sigma ligands

    Energy Technology Data Exchange (ETDEWEB)

    Rothman, R.B.; Reid, A.; Mahboubi, A.; Kim, C.H.; De Costa, B.R.; Jacobson, A.E.; Rice, K.C. (National Institute of Mental Health, Bethesda, MD (USA))

    1991-02-01

    Equilibrium binding studies with the sigma receptor ligand ({sup 3}H)1,3-di(2-tolyl)guanidine (({sup 3}H)DTG) demonstrated two high affinity binding sites in membranes prepared from guinea pig brain. The apparent Kd values of DTG for sites 1 and 2 were 11.9 and 37.6 nM, respectively. The corresponding Bmax values were 1045 and 1423 fmol/mg of protein. Site 1 had high affinity for (+)-pentazocine, haloperidol, (R)-(+)-PPP, carbepentane, and other sigma ligands, suggesting a similarity with the dextromethorphan/sigma 1 binding site described by Musacchio et al. (Life Sci. 45:1721-1732 (1989)). Site 2 had high affinity for DTG and haloperidol (Ki = 36.1 nM) and low affinity for most other sigma ligands. Kinetic experiments demonstrated that ({sup 3}H)DTG dissociated in a biphasic manner from both site 1 and site 2. DTG and haloperidol increased the dissociation rate of ({sup 3}H)DTG from site 1 and site 2, demonstrating the presence of pseudoallosteric interactions. Inorganic calcium channel blockers such as Cd2+ selectively increased the dissociation rate of ({sup 3}H)DTG from site 2, suggesting an association of this binding site with calcium channels.

  12. Labeling by [3H]1,3-di(2-tolyl)guanidine of two high affinity binding sites in guinea pig brain: Evidence for allosteric regulation by calcium channel antagonists and pseudoallosteric modulation by sigma ligands

    International Nuclear Information System (INIS)

    Rothman, R.B.; Reid, A.; Mahboubi, A.; Kim, C.H.; De Costa, B.R.; Jacobson, A.E.; Rice, K.C.

    1991-01-01

    Equilibrium binding studies with the sigma receptor ligand [ 3 H]1,3-di(2-tolyl)guanidine ([ 3 H]DTG) demonstrated two high affinity binding sites in membranes prepared from guinea pig brain. The apparent Kd values of DTG for sites 1 and 2 were 11.9 and 37.6 nM, respectively. The corresponding Bmax values were 1045 and 1423 fmol/mg of protein. Site 1 had high affinity for (+)-pentazocine, haloperidol, (R)-(+)-PPP, carbepentane, and other sigma ligands, suggesting a similarity with the dextromethorphan/sigma 1 binding site described by Musacchio et al. [Life Sci. 45:1721-1732 (1989)]. Site 2 had high affinity for DTG and haloperidol (Ki = 36.1 nM) and low affinity for most other sigma ligands. Kinetic experiments demonstrated that [ 3 H]DTG dissociated in a biphasic manner from both site 1 and site 2. DTG and haloperidol increased the dissociation rate of [ 3 H]DTG from site 1 and site 2, demonstrating the presence of pseudoallosteric interactions. Inorganic calcium channel blockers such as Cd2+ selectively increased the dissociation rate of [ 3 H]DTG from site 2, suggesting an association of this binding site with calcium channels

  13. Interactions of dopaminergic agonists and antagonists with dopaminergic D3 binding sites in rat striatum. Evidence that [3H]dopamine can label a high affinity agonist-binding state of the D1 dopamine receptor

    International Nuclear Information System (INIS)

    Leff, S.E.; Creese, I.

    1985-01-01

    The interactions of dopaminergic agonists and antagonists with 3 H-agonist labeled D3 dopaminergic binding sites of rat striatum have been characterized by radioligand-binding techniques. When the binding of [ 3 H]dopamine and [ 3 H]apomorphine to D2 dopamine receptors is blocked by the inclusion of D2 selective concentrations of unlabeled spiroperidol or domperidone, these ligands appear to label selectively the previously termed D3 binding site. Antagonist/[ 3 H]dopamine competition curves are of uniformly steep slope (nH . 1.0), suggesting the presence of a single D3 binding site. The relative potencies of antagonists to inhibit D3 specific [ 3 H]dopamine binding are significantly correlated with their potencies to block D1 dopamine receptors as measured by the inhibition of both dopamine-stimulated adenylate cyclase and [ 3 H]flupentixol-binding activities. The affinities of agonists to inhibit D3 specific [ 3 H]dopamine binding are also correlated with estimates of these agonists affinities for the high affinity binding component of agonist/[ 3 H]flupentixol competition curves. Both D3 specific [ 3 H] dopamine binding and the high affinity agonist-binding component of dopamine/[ 3 H]flupentixol competition curves show a similar sensitivity to guanine nucleotides. Taken together, these data strongly suggest that the D3 binding site is related to a high affinity agonist-binding state of the D1 dopamine receptor

  14. Geometry and Width of the Ringwoodite - Bridgmanite+Ferropericlase Binary Loop in the System (Mg,Fe)2SiO4: Interpretation of the Sharpness of the 660-km Discontinuity

    Science.gov (United States)

    Katsura, T.; Ishii, T.; Huang, R.; Maeda, F.; Yuan, L.; Bhat, S.; Farla, R. J.; Kawazoe, T.; Tsujino, N.; Liu, Z.; Fei, H.; Wang, L.; Druzhbin, D.; Yamamoto, T.; Kulik, E.; Koemets, I.; Higo, Y.; Tange, Y.

    2017-12-01

    Short wave-length seismic reflections from the 660 km discontinuity (D660) indicated an extreme sharp, namely less than 2 km thick boundary. To explain this observation, Ito and Takahashi [1989] estimated that the pressure interval of the dissociation of ringwoodite (Rw) to bridgmanite (Brg) plus ferropericlase (fPc) is work to estimate the Rw-Brg-fPc pressure interval at 1700 K with uncertainty in the pressure estimation as low as 0.05 GPa. The majority of our efforts have been made to determine difference in transition pressure between Fo100 and Fo70 compositions. For this purpose, we prepared fine-grained mixtures of olivine (Ol), enstatite (En) and fPc with these two compositions and loaded in a MAP cell together with an MgO pressure marker and a WRe thermocouple. The cell was compressed to ca. 6 MN, heated to 1100 K to convert Ol and En to Rw and akimotoite (Ak), respectively, and further compressed until the sample pressure reached 22 24 GPa. Then, the sample was heated to 1700 K, and the temperature was maintained for 1-2 hours. Simultaneously, after several trials, the press load was increased at the right rate at the temperature to maintain nearly constant pressure within 0.2 GPa. X-ray diffractions of the samples and pressure maker were taken to determine pressure and phases present. An important finding is that the transition pressure in Fo70 is 0.1 GPa higher than that in Fo100. Namely, the hypothetical transition pressure in Fe2SiO4 is 0.3 GPa higher than the transition pressure in Mg2SiO4. This geometry is contrary to the previous understanding. The obtained pressure difference and the partition coefficients of Fe and Mg among Brg, Rw and fPc suggest that the pressure interval is almost zero, which explains the sharpness of the D660.

  15. Characterization of a novel variant of amino acid transport system asc in erythrocytes from Przewalski's horse (Equus przewalskii).

    Science.gov (United States)

    Fincham, D A; Ellory, J C; Young, J D

    1992-08-01

    In thoroughbred horses, red blood cell amino acid transport activity is Na(+)-independent and controlled by three codominant genetic alleles (h, l, s), coding for high-affinity system asc1 (L-alanine apparent Km for influx at 37 degrees C congruent to 0.35 mM), low-affinity system asc2 (L-alanine Km congruent to 14 mM), and transport deficiency, respectively. The present study investigated amino acid transport mechanisms in red cells from four wild species: Przewalski's horse (Equus przewalskii), Hartmann's zebra (Zebra hartmannae), Grevy's zebra (Zebra grevyi), and onager (Equus hemonius). Red blood cell samples from different Przewalski's horses exhibited uniformly high rates of L-alanine uptake, mediated by a high-affinity asc1-type transport system. Mean apparent Km and Vmax values (+/- SE) for L-alanine influx at 37 degrees C in red cells from 10 individual animals were 0.373 +/- 0.068 mM and 2.27 +/- 0.11 mmol (L cells.h), respectively. As in thoroughbreds, the Przewalski's horse transporter interacted with dibasic as well as neutral amino acids. However, the Przewalski asc1 isoform transported L-lysine with a substantially (6.4-fold) higher apparent affinity than its thoroughbred counterpart (Km for influx 1.4 mM at 37 degrees C) and was also less prone to trans-stimulation effects. The novel high apparent affinity of the Przewalski's horse transporter for L-lysine provides additional key evidence of functional and possible structural similarities between asc and the classical Na(+)-dependent system ASC and between these systems and the Na(+)-independent dibasic amino acid transport system y+. Unlike Przewalski's horse, zebra red cells were polymorphic with respect to L-alanine transport activity, showing high-affinity or low-affinity saturable mechanisms of L-alanine uptake. Onager red cells transported this amino acid with intermediate affinity (apparent Km for influx 3.0 mM at 37 degrees C). Radiation inactivation analysis was used to estimate the target

  16. 320 Gb/s Single polarization OTDM Transmission over 80 km Standard Transmission Fiber

    DEFF Research Database (Denmark)

    Siahlo, Andrei; Seoane, Jorge; Clausen, Anders

    2005-01-01

    Optical time-division multiplexing (OTDM) is an attractive technique for increasing the capacity of optical transmission systems. 320 Gbit/s single-channel and single-polarization error-free transmission over continuous spans of either 80 km SMF or 77 km NZDSF are realized.......Optical time-division multiplexing (OTDM) is an attractive technique for increasing the capacity of optical transmission systems. 320 Gbit/s single-channel and single-polarization error-free transmission over continuous spans of either 80 km SMF or 77 km NZDSF are realized....

  17. Teleportation of entanglement over 143 km.

    Science.gov (United States)

    Herbst, Thomas; Scheidl, Thomas; Fink, Matthias; Handsteiner, Johannes; Wittmann, Bernhard; Ursin, Rupert; Zeilinger, Anton

    2015-11-17

    As a direct consequence of the no-cloning theorem, the deterministic amplification as in classical communication is impossible for unknown quantum states. This calls for more advanced techniques in a future global quantum network, e.g., for cloud quantum computing. A unique solution is the teleportation of an entangled state, i.e., entanglement swapping, representing the central resource to relay entanglement between distant nodes. Together with entanglement purification and a quantum memory it constitutes a so-called quantum repeater. Since the aforementioned building blocks have been individually demonstrated in laboratory setups only, the applicability of the required technology in real-world scenarios remained to be proven. Here we present a free-space entanglement-swapping experiment between the Canary Islands of La Palma and Tenerife, verifying the presence of quantum entanglement between two previously independent photons separated by 143 km. We obtained an expectation value for the entanglement-witness operator, more than 6 SDs beyond the classical limit. By consecutive generation of the two required photon pairs and space-like separation of the relevant measurement events, we also showed the feasibility of the swapping protocol in a long-distance scenario, where the independence of the nodes is highly demanded. Because our results already allow for efficient implementation of entanglement purification, we anticipate our research to lay the ground for a fully fledged quantum repeater over a realistic high-loss and even turbulent quantum channel.

  18. The diffusion of radioactive gases in the meso-scale (20 km-400 km)

    International Nuclear Information System (INIS)

    Wippermann, F.

    1974-01-01

    The term ''Mesoscale'' refers to distances between 20 km and 400 km from the source; in defining this range, the structure of atmospheric turbulence is taken into account. To arrive at an evaluation of diffusion in the mesoscale, quantitative methods from the microscale (source distance 400 km) are extrapolated into the mesoscale. In the first case a table is given to read off the minimum factor by which the concentration is reduced in the mesoscale as the source distance increases to obtain the diffusion for the worst possible case, the existence of a mixing-layer topped by a temperature inversion, was assumed. For this it was essential, first of all, to determine the source distance xsub(D) beyond which the diffusing gases are completely mixed within the mixing-layer of thickness D. To make allowance for all possible thicknesses of this mixing-layer, a measurement carried out at ground level at only 10 km from the source can be used to calculate the correct concentrations in the mixing-layer; the dilution factors will then be related to this value. Possible ways of an improved incorporation of certain factors in the diffusion estimate, such as the topography of the earth's surface, the roughness of terrain, the vertical profiles of wind and exchange coefficients and the effects of non-stability are given in the last section

  19. Venous hemogasometry of equines finalists in 90 km endurance races

    Directory of Open Access Journals (Sweden)

    Cinthia B.S. Dumont

    2014-06-01

    Full Text Available Front of exercise, the organic systems may suffer water-electrolyte and acid-base imbalances, particularly in the case of blood gases, demonstrating variations from different causes, whether respiratory and/or metabolic. Understanding the physiological adaptations to exercise is essential in the search for the optimum performance. In this way, this study measured the venous blood gases (pO2, pCO2, as well as the oxygen saturation (SatO2 in healthy equines, Arabian horses finalists in 90km endurance races. A total of fourteen Arabian horses were evaluated, nine males and five females, between six and 12 years old, finalists in 90km endurance races. There was a significant reduction in pO2, pCO2 and SatO2 after the exercise, however, the values remained within the normality range, and did not change the athletic performance of the animals, indicating a temporary alteration, assuming thus a character of physiological response to the exercise performed. The equines, finalists in 90 Km endurance races, demonstrated efficient ventilatory process, without any alterations in the athletic performance, being adapted to the type of exercise imposed.

  20. KARAKTERISTIK KM. ZAISAN STAR AKIBAT PERUBAHAN MUATAN

    Directory of Open Access Journals (Sweden)

    Samuel Samuel

    2014-02-01

    Full Text Available KM. Zaisan Star yang semula merupakan kapal general cargo dimodifikasi menjadi kapal pengangkut kendaraan (vehicle carrier dengan penambahan geladak pada ruang muat dan diatas geladak utama. Penelitian ini bertujuan mengetahui nilai stabilitas dan kekuatan memanjang kapal dari 32 simulasi kondisi karena pengaruh pengisian geladak muat dan kondisi pelayaran kapal. Perhitungan dan analisa pada penelitian ini dilakukan dengan metode pendekatan rumus stabilitas dan kekuatan memanjang kapal yang terintegrasi pada perangkat lunak pekapalan yang mengacu standar IMO dan Rules BKI. Hasil analisa stabilitas menunjukkan nilai GZ terendah pada kondisi XXXI dengan 1,103 m sedangkan kriteria minimumnya 0,200 m. Nilai GM terendah pada kondisi XXXII dengan 1,160 m, sedangkan nilai minimumnya 0,150 m. Pada analisa kekuatan memanjang diperoleh nilai tegangan geladak kondisi air tenang 0,009 N/mm2, sagging 0,013 N/mm2 dan hogging 3,40 N/mm2 serta tegangan alas kondisi air tenang 0,020 N/mm2, sagging 0,029 N/mm2 dan hogging 7,825 N/mm2, nilai tersebut tidak melebihi nilai tegangan ijin kapal 188,815 N/mm2. Perhitungan modulus penampang menunjukkan nilai modulus penampang geladak 831,990 m3 dan alas 1913,974 m3, nilai tersebut memenuhi nilai modulus minimum kapal 0,1824 m3. Perhitungan momen inersia menunjukkan nilai momen inersia sebesar 2899,540 m4, nilai ini memenuhi nilai minimum momen inersia kapal 0,4103 m4.

  1. Exploring the Mediation Between KM Infrastructure Capabilities and Organisational Performance: The Penetration of Learning by KM Practices

    OpenAIRE

    Meng-Lin Shih; Shu-Hui Chuang; Chechen Liao

    2009-01-01

    Previous studies have examined the relationship between knowledge management (KM) infrastructure capabilities and organisational performance. However, most studies neglect the mediating effect of organisational learning by KM practices (OLKMP) in the relationship between KM infrastructure capabilities and organisational performance. This study uses the survey method to discuss the relationships governing KM infrastructure capabilities, OLKMP and organisational performance. Results of the anal...

  2. Molecular cloning of a second subunit of the receptor for human granulocyte - macrophage colony-stimulating factor (GM-CSF): Reconstitution of a high-affinity GM-CSF receptor

    International Nuclear Information System (INIS)

    Hayashida, Kazuhiro; Kitamura, Toshio; Gorman, D.M.; Miyajima, Atsushi; Arai, Kenichi; Yokota, Takashi

    1990-01-01

    Using the mouse interleukin 3 (IL-3) receptor cDNA as a probe, the authors obtained a monologous cDNA (KH97) from a cDNA library of a human hemopoietic cell line, TF-1. The protein encoded by the KH97 cDNA has 56% amino acid sequence identity with the mouse IL-3 receptor and retains features common to the family of cytokine receptors. Fibroblasts transfected with the KH97 cDNA expressed a protein of 120 kDa but did not bind any human cytokines, including IL-3 and granulocyte - macrophage colony-stimulating factor (GM-CSF). Interestingly, cotransfection of cDNAs for KH97 and the low-affinity human GM-CSF receptor in fibroblasts resulted in formation of a high-affinity receptor for GM-CSF. The dissociation rate of GM-CSF from the reconstituted high-affinity receptor was slower than that from the low-affinity site, whereas the association rate was unchanged. Cross-linking of 125 I-labeled GM-CSF to fibroblasts cotransfected with both cDNAs revealed the same cross-linking patterns as in TF-1 cells - i.e., two major proteins of 80 and 120 kDa which correspond to the low-affinity GM-CSF receptor and the KH97 protein, respectively. These results indicate that the high-affinity GM-CSF receptor is composed of at least two components in a manner analogous to the IL-2 receptor. They therefore propose to designate the low-affinity GM-CSF receptor and the KH97 protein as the α and β subunits of the GM-CSF receptor, respectively

  3. Quality control considerations for the KM3NeT Very Large Volume Neutrino Telescope

    Energy Technology Data Exchange (ETDEWEB)

    Sollima, C., E-mail: c.sollima@ing.unipi.it

    2013-10-11

    Within the KM3NeT project a quality management system was proposed that included a qualification process and a database to store information on the design. This paper highlights quality control procedures applicable to KM3NeT and describes the database.

  4. GHRSST Level 4 Australian Bureau of Meteorology GAMSSA_28km Global SST:1

    Data.gov (United States)

    National Aeronautics and Space Administration — The GAMSSA v1.0 system blends NAVOCEANO's GAC 9.9 km x 4.4 km resolution AVHRR L2P SST1m data (NOAA-17, NOAA-18 and METOP-A), European Space Agency's 0.17 AATSR skin...

  5. The landscape basis of the complex monitoring of 30-km zone of NPP

    International Nuclear Information System (INIS)

    Malisheva, L.L.; Romanchuk, S.P.; Schur, U.V.; Rybalko, S.I.; Proskura, N.I.; Lury, D.I.

    1992-01-01

    This publication reviews works on the problem of creation the landscape basis of complex monitoring in 30 km zone of NPP(nuclear power plants). The objectives, subject and designation of monitoring system are considered. The monitoring network and organization of systematical observations are justified. The structure of radiation monitoring in NPP 30 km zone is considered in details. (author)

  6. The C-terminal SH2 domain of p85 accounts for the high affinity and specificity of the binding of phosphatidylinositol 3-kinase to phosphorylated platelet-derived growth factor beta receptor.

    Science.gov (United States)

    Klippel, A; Escobedo, J A; Fantl, W J; Williams, L T

    1992-01-01

    Upon stimulation by its ligand, the platelet-derived growth factor (PDGF) receptor associates with the 85-kDa subunit of phosphatidylinositol (PI) 3-kinase. The 85-kDa protein (p85) contains two Src homology 2 (SH2) domains and one SH3 domain. To define the part of p85 that interacts with the PDGF receptor, a series of truncated p85 mutants was analyzed for association with immobilized PDGF receptor in vitro. We found that a fragment of p85 that contains a single Src homology domain, the C-terminal SH2 domain (SH2-C), was sufficient for directing the high-affinity interaction with the receptor. Half-maximal binding of SH2-C to the receptor was observed at an SH2-C concentration of 0.06 nM. SH2-C, like full-length p85, was able to distinguish between wild-type PDGF receptor and a mutant receptor lacking the PI 3-kinase binding site. An excess of SH2-C blocked binding of full-length p85 and PI 3-kinase to the receptor but did not interfere with the binding of two other SH2-containing proteins, phospholipase C-gamma and GTPase-activating protein. These results demonstrate that a region of p85 containing a single SH2 domain accounts both for the high affinity and specificity of binding of PI 3-kinase to the PDGF receptor. Images PMID:1312663

  7. Evaluation of the Plant-Craig stochastic convection scheme (v2.0) in the ensemble forecasting system MOGREPS-R (24 km) based on the Unified Model (v7.3)

    Science.gov (United States)

    Keane, Richard J.; Plant, Robert S.; Tennant, Warren J.

    2016-05-01

    The Plant-Craig stochastic convection parameterization (version 2.0) is implemented in the Met Office Regional Ensemble Prediction System (MOGREPS-R) and is assessed in comparison with the standard convection scheme with a simple stochastic scheme only, from random parameter variation. A set of 34 ensemble forecasts, each with 24 members, is considered, over the month of July 2009. Deterministic and probabilistic measures of the precipitation forecasts are assessed. The Plant-Craig parameterization is found to improve probabilistic forecast measures, particularly the results for lower precipitation thresholds. The impact on deterministic forecasts at the grid scale is neutral, although the Plant-Craig scheme does deliver improvements when forecasts are made over larger areas. The improvements found are greater in conditions of relatively weak synoptic forcing, for which convective precipitation is likely to be less predictable.

  8. MODIS 3km Aerosol Product: Algorithm and Global Perspective

    Science.gov (United States)

    Remer, L. A.; Mattoo, S.; Levy, R. C.; Munchak, L.

    2013-01-01

    After more than a decade of producing a nominal 10 km aerosol product based on the dark target method, the MODIS aerosol team will be releasing a nominal 3 km product as part of their Collection 6 release. The new product differs from the original 10 km product only in the manner in which reflectance pixels are ingested, organized and selected by the aerosol algorithm. Overall, the 3 km product closely mirrors the 10 km product. However, the finer resolution product is able to retrieve over ocean closer to islands and coastlines, and is better able to resolve fine aerosol features such as smoke plumes over both ocean and land. In some situations, it provides retrievals over entire regions that the 10 km product barely samples. In situations traditionally difficult for the dark target algorithm, such as over bright or urban surfaces the 3 km product introduces isolated spikes of artificially high aerosol optical depth (AOD) that the 10 km algorithm avoids. Over land, globally, the 3 km product appears to be 0.01 to 0.02 higher than the 10 km product, while over ocean, the 3 km algorithm is retrieving a proportionally greater number of very low aerosol loading situations. Based on collocations with ground-based observations for only six months, expected errors associated with the 3 km land product are determined to be greater than for the 10 km product: 0.05 0.25 AOD. Over ocean, the suggestion is for expected errors to be the same as the 10 km product: 0.03 0.05 AOD. The advantage of the product is on the local scale, which will require continued evaluation not addressed here. Nevertheless, the new 3 km product is expected to provide important information complementary to existing satellite-derived products and become an important tool for the aerosol community.

  9. Modeling a complex system of multipurpose reservoirs under prospective scenarios (hydrology, water uses, water management): the case of the Durance River basin (South Eastern France, 12 800 km2)

    Science.gov (United States)

    Monteil, Céline; Hendrickx, Frédéric; Samie, René; Sauquet, Eric

    2015-04-01

    but also evapotranspiration process. However changes in total precipitation are highly uncertain. The six tested rainfall-runoff models project reduced flows, especially in the spring and summer seasons. Depending on the socio-economic scenarios and the area, the downstream total water needs could decrease or remain stable. Considering the present day constraints, these changes would lead to a decrease in energy production (mainly due to reduced annual inflows) and to less flexibility for hydropower management during winter peak energy demand. Results of the R²D² 2050 project suggest also that the downscaling methods still fail to reproduce some crucial aspects of the climate at regional scale. Unexpected biases are propagated along the chain of models. The key issue to simulate accurately reservoir operations under present and future climate conditions is the filling curves that depict the balance between water supply and demand. Probabilistic filling curves were calibrated here to meet the constraint on water level in summer objective nine years over ten. A large proportion of regional climates generated over the baseline period lead to unrealistic curves, pointing out higher levels of requirement in models output to assess global change impacts on water management systems like on the Durance River basin.

  10. Synthesis, in vitro validation and in vivo pharmacokinetics of [{sup 125}I]N-[2-(4-iodophenyl)ethyl]-N-methyl-2-(1-piperidinyl) ethylamine: A high-affinity ligand for imaging sigma receptor positive tumors

    Energy Technology Data Exchange (ETDEWEB)

    John, Christy S; Gulden, Mary E; Vilner, Bertold J; Bowen, Wayne D

    1996-08-01

    N-[2-(4-iodophenyl)ethyl]-N-methyl-2-(1-piperidinyl)ethylamine, IPEMP, and the corresponding bromo derivative, BrPEMP, have been synthesized and characterized. Both BrPEMP and IPEMP were evaluated for sigma-1 and sigma-2 subtype receptor affinities and found to possess very high affinities for both receptor subtypes. The precursor for radioiodination n-tributylstannylphenylethylpiperidinylethylamine was prepared from its bromo derivative by palladium-catalyzed stannylation reaction. Radioiodinated 4-[{sup 125}I]PEMP was readily prepared in high yields and high specific activity by oxidative iododestannylation reaction using chloramine-T as oxidizing agent. Sites labeled by 4-[{sup 125}I]PEMP in guinea pig brain membranes showed high affinity for BD1008, haloperidol, and (+)-pentazocine (Ki = 5.06 {+-} 0.40, 32.6 {+-} 2.75, and 48.1 {+-} 8.60 nM, respectively), which is consistent with sigma receptor pharmacology. Competition binding studies of 4-[{sup 125}I]PEMP in melanoma (A375) and MCF-7 breast cancer cells showed a high affinity, dose-dependent inhibition of binding with known sigma ligand N-[2-(3,4-dichlorophenyl)ethyl]-N-methyl-2-(1-pyrrolidinyl) ethylamine, BD1008 (Ki = 5, 11 nM, respectively), supporting the labeling of sigma sites in these cells. Haloperidol, however showed a weaker (Ki 100-200 nM) affinity for the sites labeled by 4-[{sup 125}I]PEMP in these cells. Biodistribution studies of 4-[{sup 125}I]PEMP in rats showed a fast clearance of this radiopharmaceutical from blood, liver, lung, and other organs. A co-injection of 4-IPEMP with 4-[{sup 125}I]PEMP resulted in 37%, 69%, and 35% decrease in activity in liver, kidney, and brain (organs possessing sigma receptors), respectively at 1-h postinjection. These results suggest that 4-[{sup 125}I]PEMP is a promising radiopharmaceutical for pursuing further studies in animal models with tumors.

  11. The TorR High-Affinity Binding Site Plays a Key Role in Both torR Autoregulation and torCAD Operon Expression in Escherichia coli

    OpenAIRE

    Ansaldi, Mireille; Simon, Gwénola; Lepelletier, Michèle; Méjean, Vincent

    2000-01-01

    In the presence of trimethylamine N-oxide (TMAO), the TorS-TorR two-component regulatory system induces the torCAD operon, which encodes the TMAO respiratory system of Escherichia coli. The sensor protein TorS detects TMAO and transphosphorylates the response regulator TorR which, in turn, activates transcription of torCAD. The torR gene and the torCAD operon are divergently transcribed, and the short torR-torC intergenic region contains four direct repeats (the tor boxes) which proved to be ...

  12. Analysis of the HLA-DR peptidome from human dendritic cells reveals high affinity repertoires and nonconventional pathways of peptide generation

    NARCIS (Netherlands)

    Ciudad, M Teresa; Sorvillo, Nicoletta; van Alphen, Floris P.J.; Catalán, Diego; Meijer, Sander; Voorberg, Jan; Jaraquemada, Dolores

    Dendritic cells (DCs) are the major professional APCs of the immune system; however, their MHC-II-associated peptide repertoires have been hard to analyze, mostly because of their scarce presence in blood and tissues. In vitro matured human monocyte-derived DCs (MoDCs) are widely used as

  13. Rainforest metropolis casts 1,000-km defaunation shadow.

    Science.gov (United States)

    Tregidgo, Daniel J; Barlow, Jos; Pompeu, Paulo S; de Almeida Rocha, Mayana; Parry, Luke

    2017-08-08

    Tropical rainforest regions are urbanizing rapidly, yet the role of emerging metropolises in driving wildlife overharvesting in forests and inland waters is unknown. We present evidence of a large defaunation shadow around a rainforest metropolis. Using interviews with 392 rural fishers, we show that fishing has severely depleted a large-bodied keystone fish species, tambaqui ( Colossoma macropomum ), with an impact extending over 1,000 km from the rainforest city of Manaus (population 2.1 million). There was strong evidence of defaunation within this area, including a 50% reduction in body size and catch rate (catch per unit effort). Our findings link these declines to city-based boats that provide rural fishers with reliable access to fish buyers and ice and likely impact rural fisher livelihoods and flooded forest biodiversity. This empirical evidence that urban markets can defaunate deep into rainforest wilderness has implications for other urbanizing socioecological systems.

  14. Calibration methods and tools for KM3NeT

    Directory of Open Access Journals (Sweden)

    Kulikovskiy Vladimir

    2016-01-01

    Full Text Available The KM3NeT detectors, ARCA and ORCA, composed of several thousands digital optical modules, are in the process of their realization in the Mediterranean Sea. Each optical module contains 31 3-inch photomultipliers. Readout of the optical modules and other detector components is synchronized at the level of sub-nanoseconds. The position of the module is measured by acoustic piezo detectors inside the module and external acoustic emitters installed on the bottom of the sea. The orientation of the module is obtained with an internal attitude and heading reference system chip. Detector calibration, i.e. timing, positioning and sea-water properties, is overviewed in this talk and discussed in detail in this conference. Results of the procedure applied to the first detector unit ready for installation in the deep sea will be shown.

  15. Affinities and densities of high-affinity [3H]muscimol (GABA-A) binding sites and of central benzodiazepine receptors are unchanged in autopsied brain tissue from cirrhotic patients with hepatic encephalopathy

    International Nuclear Information System (INIS)

    Butterworth, R.F.; Lavoie, J.; Giguere, J.F.; Pomier-Layrargues, G.

    1988-01-01

    The integrity of GABA-A receptors and of central benzodiazepine receptors was evaluated in membrane preparations from prefrontal cortex and caudate nuclei obtained at autopsy from nine cirrhotic patients who died in hepatic coma and an equal number of age-matched control subjects. Histopathological studies revealed Alzheimer Type II astrocytosis in all cases in the cirrhotic group; controls were free from neurological, psychiatric or hepatic diseases. Binding to GABA-A receptors was studied using [ 3 H]muscimol as radioligand. The integrity of central benzodiazepine receptors was evaluated using [ 3 H]flunitrazepam and [ 3 H]Ro15-1788. Data from saturation binding assays was analyzed by Scatchard plot. No modifications of either affinities (Kd) or densities (Bmax) of [ 3 H]muscimol of central benzodiazepine binding sites were observed. These findings do not support recent suggestions that alterations of either high-affinity GABA or benzodiazepine receptors play a significant role in the pathogenesis of hepatic encephalopathy

  16. Identification of the ligand-binding subunit of the human 5-hydroxytryptamine1A receptor with N-(p-azido-m-[125I] iodophenethyl)spiperone, a high affinity radioiodinated photoaffinity probe

    International Nuclear Information System (INIS)

    Raymond, J.R.; Fargin, A.; Lohse, M.J.; Regan, J.W.; Senogles, S.E.; Lefkowitz, R.J.; Caron, M.G.

    1989-01-01

    The ligand-binding subunit of the human 5-hydroxytryptamine1A (5-HT1A) receptor transiently expressed in COS-7 cells and of the native human 5-HT1A receptor derived from hippocampus and frontal cortex were identified by photoaffinity labeling with N-(p-azido-m-[125I]iodophenethyl)spiperone [( 125I]N3-NAPS), previously characterized as a high affinity radioiodinated D2-dopamine receptor probe. The identity of the ligand-binding subunit was confirmed by immunoprecipitation with an antipeptide rabbit antiserum, JWR21, raised against a synthetic peptide derived from the predicted amino acid sequence of the putative third intracellular loop of the human 5-HT1A receptor. In transiently transfected COS-7 cells expressing 14 +/- 3 pmol/mg of protein human 5-HT1A receptors, a single broad 75-kDa band was photoaffinity labeled by [125I]N3-NAPS. This band displayed the expected pharmacology of the 5-HT1A receptor, as evidenced by the ability of a series of competing ligands to block [125I]N3-NAPS photoincorporation. Moreover, antiserum JWR21 specifically and quantitatively immunoprecipitated the 75-kDa photoaffinity-labeled band from a soluble extract of the transfected COS-7 cell membranes, further confirming its identity. Finally, utilizing a combination of photoaffinity labeling and immunoprecipitation, the native ligand-binding subunit of 62-64 kDa was identified in human hippocampus and frontal cortex. The availability of the high specific activity, high affinity, photoaffinity ligand [125I]N3-NAPS and of a potent immunoprecipitating antiserum (JWR21) should greatly facilitate the biochemical characterization of the human 5-HT1A receptor

  17. CmRBP50 protein phosphorylation is essential for assembly of a stable phloem-mobile high-affinity ribonucleoprotein complex.

    Science.gov (United States)

    Li, Pingfang; Ham, Byung-Kook; Lucas, William J

    2011-07-01

    RNA-binding proteins (RBPs) form ribonucleoprotein (RNP) complexes that play crucial roles in RNA processing for gene regulation. The angiosperm sieve tube system contains a unique population of transcripts, some of which function as long-distance signaling agents involved in regulating organ development. These phloem-mobile mRNAs are translocated as RNP complexes. One such complex is based on a phloem RBP named Cucurbita maxima RNA-binding protein 50 (CmRBP50), a member of the polypyrimidine track binding protein family. The core of this RNP complex contains six additional phloem proteins. Here, requirements for assembly of this CmRBP50 RNP complex are reported. Phosphorylation sites on CmRBP50 were mapped, and then coimmunoprecipitation and protein overlay studies established that the phosphoserine residues, located at the C terminus of CmRBP50, are critical for RNP complex assembly. In vitro pull-down experiments revealed that three phloem proteins, C. maxima phloem protein 16, C. maxima GTP-binding protein, and C. maxima phosphoinositide-specific phospholipase-like protein, bind directly with CmRBP50. This interaction required CmRBP50 phosphorylation. Gel mobility-shift assays demonstrated that assembly of the CmRBP50-based protein complex results in a system having enhanced binding affinity for phloem-mobile mRNAs carrying polypyrimidine track binding motifs. This property would be essential for effective long-distance translocation of bound mRNA to the target tissues.

  18. Silver Nanoparticle-Directed Mast Cell Degranulation Is Mediated through Calcium and PI3K Signaling Independent of the High Affinity IgE Receptor.

    Directory of Open Access Journals (Sweden)

    Nasser B Alsaleh

    Full Text Available Engineered nanomaterial (ENM-mediated toxicity often involves triggering immune responses. Mast cells can regulate both innate and adaptive immune responses and are key effectors in allergic diseases and inflammation. Silver nanoparticles (AgNPs are one of the most prevalent nanomaterials used in consumer products due to their antimicrobial properties. We have previously shown that AgNPs induce mast cell degranulation that was dependent on nanoparticle physicochemical properties. Furthermore, we identified a role for scavenger receptor B1 (SR-B1 in AgNP-mediated mast cell degranulation. However, it is completely unknown how SR-B1 mediates mast cell degranulation and the intracellular signaling pathways involved. In the current study, we hypothesized that SR-B1 interaction with AgNPs directs mast cell degranulation through activation of signal transduction pathways that culminate in an increase in intracellular calcium signal leading to mast cell degranulation. For these studies, we utilized bone marrow-derived mast cells (BMMC isolated from C57Bl/6 mice and RBL-2H3 cells (rat basophilic leukemia cell line. Our data support our hypothesis and show that AgNP-directed mast cell degranulation involves activation of PI3K, PLCγ and an increase in intracellular calcium levels. Moreover, we found that influx of extracellular calcium is required for the cells to degranulate in response to AgNP exposure and is mediated at least partially via the CRAC channels. Taken together, our results provide new insights into AgNP-induced mast cell activation that are key for designing novel ENMs that are devoid of immune system activation.

  19. Synthesis, biodistribution and in vitro evaluation of brain permeable high affinity type 2 cannabinoid receptor agonists [11C]MA2 and [18F]MA3

    Directory of Open Access Journals (Sweden)

    Muneer Ahamed

    2016-09-01

    Full Text Available Abstract The type 2 cannabinoid receptor (CB2 is a member of the endocannabinoid system and is known for its important role in (neuroinflammation. A PET-imaging agent that allows in vivo visualization of CB2 expression may thus allow quantification of neuroinflammation. In this paper, we report the synthesis, radiosynthesis, biodistribution and in vitro evaluation of a carbon-11 ([11C]MA2 and a fluorine-18 ([18F]MA3 labeled analogue of a highly potent N-arylamide oxadiazole CB2 agonist (EC50 = 0.015 nM. MA2 and MA3 behaved as potent CB2 agonist (EC50: 3 nM and 0.1 nM, respectively and their in vitro binding affinity for hCB2 was found to be 87 nM and 0.8 nM, respectively. Also MA3 (substituted with a fluoro ethyl group was found to have higher binding affinity and EC50 values when compared to the originally reported trifluoromethyl analogue 12. [11C]MA2 and [18F]MA3 were successfully synthesized with good radiochemical yield, high radiochemical purity and high specific activity. In mice, both tracers were efficiently cleared from blood and all major organs by the hepatobiliary pathway and importantly these compounds showed high brain uptake. In conclusion, [11C]MA2 and [18F]MA3 are shown to be high potent CB2 agonists with good brain uptake, these favorable characteristics makes them potential PET probes for in vivo imaging of brain CB2 receptors. However in view of its higher affinity and selectivity, further detailed evaluation of MA3 as a PET tracer for CB2 is warranted.

  20. Synthesis, Biodistribution and In vitro Evaluation of Brain Permeable High Affinity Type 2 Cannabinoid Receptor Agonists [11C]MA2 and [18F]MA3.

    Science.gov (United States)

    Ahamed, Muneer; van Veghel, Daisy; Ullmer, Christoph; Van Laere, Koen; Verbruggen, Alfons; Bormans, Guy M

    2016-01-01

    The type 2 cannabinoid receptor (CB2) is a member of the endocannabinoid system and is known for its important role in (neuro)inflammation. A PET-imaging agent that allows in vivo visualization of CB2 expression may thus allow quantification of neuroinflammation. In this paper, we report the synthesis, radiosynthesis, biodistribution and in vitro evaluation of a carbon-11 ([ 11 C]MA2) and a fluorine-18 ([ 18 F]MA3) labeled analog of a highly potent N -arylamide oxadiazole CB2 agonist (EC 50 = 0.015 nM). MA2 and MA3 behaved as potent CB2 agonist (EC 50 : 3 nM and 0.1 nM, respectively) and their in vitro binding affinity for h CB2 was found to be 87 nM and 0.8 nM, respectively. Also MA3 (substituted with a fluoro ethyl group) was found to have higher binding affinity and EC 50 values when compared to the originally reported trifluoromethyl analog 12 . [ 11 C]MA2 and [ 18 F]MA3 were successfully synthesized with good radiochemical yield, high radiochemical purity and high specific activity. In mice, both tracers were efficiently cleared from blood and all major organs by the hepatobiliary pathway and importantly these compounds showed high brain uptake. In conclusion, [ 11 C]MA2 and [ 18 F]MA3 are shown to be high potent CB2 agonists with good brain uptake, these favorable characteristics makes them potential PET probes for in vivo imaging of brain CB2 receptors. However, in view of its higher affinity and selectivity, further detailed evaluation of MA3 as a PET tracer for CB2 is warranted.

  1. Linear and Nonlinear Infrasound Propagation to 1000 km

    Science.gov (United States)

    2015-12-15

    AFRL-RV-PS- AFRL-RV-PS- TR-2016-0017 TR-2016-0017 LINEAR AND NONLINEAR INFRASOUND PROPAGATION TO 1000 KM Catherine de Groot-Hedlin Scripps...Nonlinear Infrasound Propagation to 1000 km 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 62601F 6. AUTHOR(S) Catherine de Groot

  2. 7 CFR 29.1035 - Mixed color (KM).

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Mixed color (KM). 29.1035 Section 29.1035 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing... Type 92) § 29.1035 Mixed color (KM). Distinctly different colors of the type mingled together. (See...

  3. Status of the KM3NeT project

    International Nuclear Information System (INIS)

    Katz, U.F.

    2009-01-01

    KM3NeT is a future research infrastructure in the Mediterranean Sea, hosting a cubic-kilometre scale neutrino telescope and nodes for associated sciences such as marine biology, oceanology and geophysics. The status of the KM3NeT project and the progress made in the EU-funded Design Study is reviewed. Some physics studies indicating the sensitivity of the KM3NeT neutrino telescope are highlighted and selected major technical design options to be further pursued are described. Finally, the remaining steps towards construction of KM3NeT will be discussed. This document reflects the status of the KM3NeT Conceptual Design Report (CDR), which has been presented to the public for the first time at the VLVnT08 Workshop.

  4. Knowledge Management (KM): a new management paradigm in MINT

    International Nuclear Information System (INIS)

    Amy Hamijah Ab Hamid

    2006-01-01

    Knowledge management (KM) is one of the leading approaches to improve organizational benefits by introducing into an organization a range of specific processes and practices for identifying and capturing knowledge, know-how, expertise and other intellectual properties, and for making such knowledge assets available for transfer and reuse across the organization. This paper identified and compared some of the KM theories, methodologies and best practices from various literatures. It also pointed out the KM Life Cycle and KM Transformation through Organization as a prior case study. This forms the guidelines for MINT, local agencies and overseas conglomerates experiences through some challenges and opportunities identification. As MINT is still in the early stages in doing so, it is important to start small but continuously doing incremental improvement overtime. Finally, an improvement towards KM culture in MINT is proposed referring to the ideas and facts stated above

  5. Evaluation of NLDAS 12-km and downscaled 1-km temperature products in New York State for potential use in health exposure response studies

    Science.gov (United States)

    Estes, M. G., Jr.; Insaf, T.; Crosson, W. L.; Al-Hamdan, M. Z.

    2017-12-01

    Heat exposure metrics (maximum and minimum daily temperatures,) have a close relationship with human health. While meteorological station data provide a good source of point measurements, temporal and spatially consistent temperature data are needed for health studies. Reanalysis data such as the North American Land Data Assimilation System's (NLDAS) 12-km gridded product are an effort to resolve spatio-temporal environmental data issues; the resolution may be too coarse to accurately capture the effects of elevation, mixed land/water areas, and urbanization. As part of this NASA Applied Sciences Program funded project, the NLDAS 12-km air temperature product has been downscaled to 1-km using MODIS Land Surface Temperature patterns. Limited validation of the native 12-km NLDAS reanalysis data has been undertaken. Our objective is to evaluate the accuracy of both the 12-km and 1-km downscaled products using the US Historical Climatology Network station data geographically dispersed across New York State. Statistical methods including correlation, scatterplots, time series and summary statistics were used to determine the accuracy of the remotely-sensed maximum and minimum temperature products. The specific effects of elevation and slope on remotely-sensed temperature product accuracy were determined with 10-m digital elevation data that were used to calculate percent slope and link with the temperature products at multiple scales. Preliminary results indicate the downscaled temperature product improves accuracy over the native 12-km temperature product with average correlation improvements from 0.81 to 0.85 for minimum and 0.71 to 0.79 for maximum temperatures in 2009. However, the benefits vary temporally and geographically. Our results will inform health studies using remotely-sensed temperature products to determine health risk from excessive heat by providing a more robust assessment of the accuracy of the 12-km NLDAS product and additional accuracy gained from

  6. Evaluation of the 7-km GEOS-5 Nature Run

    Science.gov (United States)

    Gelaro, Ronald; Putman, William M.; Pawson, Steven; Draper, Clara; Molod, Andrea; Norris, Peter M.; Ott, Lesley; Prive, Nikki; Reale, Oreste; Achuthavarier, Deepthi; hide

    2015-01-01

    This report documents an evaluation by the Global Modeling and Assimilation Office (GMAO) of a two-year 7-km-resolution non-hydrostatic global mesoscale simulation produced with the Goddard Earth Observing System (GEOS-5) atmospheric general circulation model. The simulation was produced as a Nature Run for conducting observing system simulation experiments (OSSEs). Generation of the GEOS-5 Nature Run (G5NR) was motivated in part by the desire of the OSSE community for an improved high-resolution sequel to an existing Nature Run produced by the European Centre for Medium-Range Weather Forecasts (ECMWF), which has served the community for several years. The intended use of the G5NR in this context is for generating simulated observations to test proposed observing system designs regarding new instruments and their deployments. Because NASA's interest in OSSEs extends beyond traditional weather forecasting applications, the G5NR includes, in addition to standard meteorological components, a suite of aerosol types and several trace gas concentrations, with emissions downscaled to 10 km using ancillary information such as power plant location, population density and night-light information. The evaluation exercise described here involved more than twenty-five GMAO scientists investigating various aspects of the G5NR performance, including time mean temperature and wind fields, energy spectra, precipitation and the hydrological cycle, the representation of waves, tropical cyclones and midlatitude storms, land and ocean surface characteristics, the representation and forcing effects of clouds and radiation, dynamics of the stratosphere and mesosphere, and the representation of aerosols and trace gases. Comparisons are made with observational data sets when possible, as well as with reanalyses and other long model simulations. The evaluation is broad in scope, as it is meant to assess the overall realism of basic aspects of the G5NR deemed relevant to the conduct of OSSEs

  7. KM3NeT tower data acquisition and data transport electronics

    Science.gov (United States)

    Nicolau, C. A.; Ameli, F.; Biagioni, A.; Capone, A.; Frezza, O.; Lonardo, A.; Masullo, R.; Mollo, C. M.; Orlando, A.; Simeone, F.; Vicini, P.

    2016-04-01

    In the framework of the KM3Net European project, the production stage of a large volume underwater neutrino telescope has started. The forthcoming installation includes 8 towers and 24 strings, that will be installed 100 km off-shore Capo Passero (Italy) at 3500 m depth. The KM3NeT tower, whose layout is strongly based on the NEMO Phase-2 prototype tower deployed in March 2013, has been re-engineered and partially re-designed in order to optimize production costs, power consumption, and usability. This contribution gives a description of the main electronics, including front-end, data transport and clock distribution system, of the KM3NeT tower detection unit.

  8. KM3NeT tower data acquisition and data transport electronics

    Directory of Open Access Journals (Sweden)

    Nicolau C.A.

    2016-01-01

    Full Text Available In the framework of the KM3Net European project, the production stage of a large volume underwater neutrino telescope has started. The forthcoming installation includes 8 towers and 24 strings, that will be installed 100 km off-shore Capo Passero (Italy at 3500 m depth. The KM3NeT tower, whose layout is strongly based on the NEMO Phase-2 prototype tower deployed in March 2013, has been re-engineered and partially re-designed in order to optimize production costs, power consumption, and usability. This contribution gives a description of the main electronics, including front-end, data transport and clock distribution system, of the KM3NeT tower detection unit.

  9. New model for the lunar interior to 250 km

    International Nuclear Information System (INIS)

    Piwinskii, A.J.; Duba, A.G.

    1975-01-01

    A new model for the structure of the lunar interior to about a 250-km depth is proposed. It is suggested that this region is composed of plagioclase-bearing rocks, and that the 65-km seismic discontinuity represents the appearance of garnet. A variety of rock types mainly composed of plagioclase, pyroxene, olivine and garnet is envisioned, with at least half of the outer 250 km of the moon made of plagioclase, which dominates the electrical conductivity. This model agrees with recent petrological and electrical conductivity results and does not violate velocity-depth profiles obtained from elastic-wave studies of lunar and terrestrial materials. (U.S.)

  10. Global 1-km Sea Surface Temperature (G1SST)

    Data.gov (United States)

    National Aeronautics and Space Administration — JPL OurOcean Portal: A daily, global Sea Surface Temperature (SST) data set is produced at 1-km (also known as ultra-high resolution) by the JPL ROMS (Regional Ocean...

  11. Status of the KM3NeT project

    International Nuclear Information System (INIS)

    Margiotta, A

    2014-01-01

    KM3NeT is a deep-sea research infrastructure being constructed in the Mediterranean Sea. It will be installed at three sites: KM3NeT-Fr, offshore Toulon, France, KM3NeT-It, offshore Portopalo di Capo Passero, Sicily (Italy) and KM3NeT-Gr, offshore Pylos, Peloponnese, Greece. It will host the next generation Cherenkov neutrino telescope and nodes for a deep sea multidisciplinary observatory, providing oceanographers, marine biologists, and geophysicists with real time measurements. The neutrino telescope will search for Galactic and extra-Galactic sources of neutrinos, complementing IceCube in its field of view. The detector will have a modular structure and consists of six building blocks, each including about one hundred Detection Units (DUs). Each DU will be equipped with 18 multi-PMT digital optical modules. The first phase of construction has started and shore and deep-sea infrastructures hosting the future KM3NeT detector are being prepared in France near Toulon and in Italy, near Capo Passero in Sicily. The technological solutions for KM3NeT and the expected performance of the detector are presented and discussed

  12. Magma reservoir at Mt. Vesuvius: Deeper than 10 km

    International Nuclear Information System (INIS)

    Natale, M.; Luongo, G.; Nunziata, C.; Panza, G.F.

    2005-07-01

    One- and two-dimensional Vp models were obtained by TomoVes experiment, all characterized by low Vp in the uppermost 500 m and a sharp discontinuity at about 2-3 km beneath the volcano. Large amplitude late arrivals were identified as P- to S-phases converted at the top, between 8 and 10 km deep, of a low velocity layer with a dramatic drop of Vs, from approximately 3.6 km/s to less than 1.0 km/s. Here we synthesize the interpretation of Rayleigh wave dispersion measurements, made by several authors, to delineate the extent of such anomalous layer of hot, partially molten, crust material. Our non-linear inversion of broad-band dispersion measurements, gives a thickness not greater than 0.35 km, if we assume Vs equal to 1.0 km/s. The volume occupied by this very low velocity layer, sill shaped, is compatible with the size of Mt. Vesuvius cone, but it develops above a much larger hot mass which could be the parental source as the erupted products are only few percent of magma chamber. (author)

  13. Monastrol, a 3,4-dihydropyrimidin-2(1H)-thione, as structural scaffold for the development of modulators for GHB high-affinity binding sites and α1β2δ GABAA receptors

    DEFF Research Database (Denmark)

    Damgaard, Maria; Al-Khawaja, Anas; Nittegaard-Nielsen, Mia

    2017-01-01

    -affinity binding and is furthermore reported as an allosteric modulator selective for the α1β2δ GABAARs. Therefore, structural determinants for selectivity at the two targets were investigated. 39 structural diverse monastrol analogues were synthesized by employing the Biginelli cyclocondensation and examined......-affinity binding. However, three analogues of monastrol (11, 12 and 24) enhanced the maximal binding of [(3)H]NCS-382 to a higher maximal level than seen for monastrol itself. Selected compounds were further characterized as modulators at α1β2δ, α1β2γ2s and α1β2 GABAARs. Most of these modulators were shown to have...... δ-specific GABA-potentiating effects. The dual effect shown for monastrol to modulate the GHB high-affinity binding and α1β2δ GABAAR activity was also shown for the compounds 11, 18 and 24. Compound 29 displayed minimal modulatory effect on GABAARs and therefore appears to be a GHB high...

  14. HAMS: High-Affinity Mass Spectrometry Screening. A High-Throughput Screening Method for Identifying the Tightest-Binding Lead Compounds for Target Proteins with No False Positive Identifications.

    Science.gov (United States)

    Imaduwage, Kasun P; Go, Eden P; Zhu, Zhikai; Desaire, Heather

    2016-11-01

    A major challenge in drug discovery is the identification of high affinity lead compounds that bind a particular target protein; these leads are typically identified by high throughput screens. Mass spectrometry has become a detection method of choice in drug screening assays because the target and the ligand need not be modified. Label-free assays are advantageous because they can be developed more rapidly than assays requiring labels, and they eliminate the risk of the label interfering with the binding event. However, in commonly used MS-based screening methods, detection of false positives is a major challenge. Here, we describe a detection strategy designed to eliminate false positives. In this approach, the protein and the ligands are incubated together, and the non-binders are separated for detection. Hits (protein binders) are not detectable by MS after incubation with the protein, but readily identifiable by MS when the target protein is not present in the incubation media. The assay was demonstrated using three different proteins and hundreds of non-inhibitors; no false positive hits were identified in any experiment. The assay can be tuned to select for ligands of a particular binding affinity by varying the quantity of protein used and the immobilization method. As examples, the method selectively detected inhibitors that have K i values of 0.2 μM, 50 pM, and 700 pM. These findings demonstrate that the approach described here compares favorably with traditional MS-based screening methods. Graphical Abstract ᅟ.

  15. Similarities and Differences in Pacing Patterns in a 161-km and 101-km Ultra-Distance Road Race.

    Science.gov (United States)

    Tan, Philip L S; Tan, Frankie H Y; Bosch, Andrew N

    2016-08-01

    Tan, PLS, Tan, FHY, and Bosch, AN. Similarities and differences in pacing patterns in a 161-km and 101-km ultra-distance road race. J Strength Cond Res 30(8): 2145-2155, 2016-The purpose of this study was to establish and compare the pacing patterns of fast and slow finishers in a tropical ultra-marathon. Data were collected from the Craze Ultra-marathon held on the 22nd and 21st of September in 2012 and 2013, respectively. Finishers of the 161-km (N = 47) and 101-km (N = 120) categories of the race were divided into thirds (groups A-C) by merit of finishing time. Altogether, 17 and 11 split times were recorded for the 161-km and 101-km finishers, respectively, and used to calculate the mean running speed for each distance segment. Running speed for the first segment was normalized to 100, with all subsequent splits adjusted accordingly. Running speed during the last 5 km was calculated against the mean race pace to establish the existence of an end spurt. A reverse J-shaped pacing profile was demonstrated in all groups for both distance categories and only 38% of the finishers executed an end spurt. In the 101-km category, in comparison with groups B and C, group A maintained a significantly more even pace (p = 0.013 and 0.001, respectively) and completed the race at a significantly higher percent of initial starting speed (p = 0.001 and 0.001, respectively). Descriptive data also revealed that the top 5 finishers displayed a "herd-behavior" by staying close to the lead runner in the initial portion of the race. These findings demonstrate that to achieve a more even pace, recreational ultra-runners should adopt a patient sustainable starting speed, with less competitive runners setting realistic performance goals whereas competitive runners with a specific time goal to consider running in packs of similar pace.

  16. Coherent Detection for 1550 nm, 5 Gbit/s VCSEL Based 40 km Bidirectional PON Transmission

    DEFF Research Database (Denmark)

    Jensen, Jesper Bevensee; Rodes Lopez, Roberto; Zibar, Darko

    2011-01-01

    Coherent detection of directly modulated 1550nm VCSELs in 5Gbit/s bidirectional 40km SSMF PON-links is presented. Receiver sensitivity of –37.3dBm after transmission is achieved with 30dB system margin, corresponding to 1:1024 passive powersplitting.......Coherent detection of directly modulated 1550nm VCSELs in 5Gbit/s bidirectional 40km SSMF PON-links is presented. Receiver sensitivity of –37.3dBm after transmission is achieved with 30dB system margin, corresponding to 1:1024 passive powersplitting....

  17. Acoustic calibration for the KM3NeT pre-production module

    Energy Technology Data Exchange (ETDEWEB)

    Enzenhöfer, A., E-mail: alexander.enzenhoefer@physik.uni-erlangen.de [Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen Centre for Astroparticle Physics, Erwin-Rommel-Str. 1, D-91058 Erlangen (Germany)

    2013-10-11

    The proposed large scale Cherenkov neutrino telescope KM3NeT will carry photo-sensors on flexible structures, the detection units. The Mediterranean Sea, where KM3NeT will be installed, constitutes a highly dynamic environment in which the detection units are constantly in motion. Thus it is necessary to monitor the exact sensor positions continuously to achieve the desired resolution for the neutrino telescope. A common way to perform this monitoring is the use of acoustic positioning systems with emitters and receivers based on the piezoelectric effect. The acoustic receivers are attached to detection units whereas the emitters are located at known positions on the sea floor. There are complete commercial systems for this application with sufficient precision. But these systems are limited in the use of their data and inefficient as they were designed to perform only this single task. Several working groups in the KM3NeT consortium are cooperating to custom-design a positioning system for the specific requirements of KM3NeT. Most of the studied solutions hold the possibility to extend the application area from positioning to additional tasks like acoustic particle detection or monitoring of the deep-sea acoustic environment. The KM3NeT Pre-Production Module (PPM) is a test system to verify the correct operation and interoperability of the major involved hardware and software components developed for KM3NeT. In the context of the PPM, alternative designs of acoustic sensors including small piezoelectric elements equipped with preamplifiers inside the same housing as the optical sensors will be tested. These will be described in this article.

  18. A high resolution (1 km) groundwater model for Indonesia

    Science.gov (United States)

    Sutanudjaja, Edwin; Verkaik, Jarno; de Graaf, Inge; van Beek, Rens; Erkens, Gilles; Bierkens, Marc

    2015-04-01

    Groundwater is important in many parts of Indonesia. It serves as a primary source of drinking water and industrial activities. During times of drought, it sustains water flows in streams, rivers, lakes and wetlands, and thus support ecosystem habitat and biodiversity as well as preventing hazardous forest fire. Besides its importance, groundwater is known as a vulnerable resource as unsustainable groundwater exploitation and management occurs in many areas of the country. Therefore, in order to ensure sustainable management of groundwater resources, monitoring and predicting groundwater changes in Indonesia are imperative. However, large extent groundwater models to assess these changes on a regional scale are almost non-existent and are hampered by the strong topographical and lithological transitions that characterize Indonesia. In this study, we built an 1 km resolution groundwater model for the entire Indonesian archipelago (total inland area: about 2 million km2). We adopted the approaches of Sutanudjaja et al. (2011, 2014a) and de Graaf et al. (2014) in order to make a MODFLOW (Harbaugh et al., 2000) groundwater model by using only global datasets. Aquifer schematization and properties of the groundwater model were developed from available global lithological maps (e.g. Dürr et al., 2005; Gleeson et al., 2011; Hartmann & Moorsdorf, 2012; Gleeson et al., 2014). We forced the groundwater model with the recent output of global hydrological model PCR-GLOBWB version 2.0 (Sutanudjaja et al., 2014b; van Beek et al., 2011), specifically the long term average of groundwater recharge and average surface water levels derived from channel discharge. Simulation results were promising. The MODFLOW model converged with realistic aquifer properties (i.e. transmissivities) and produced reasonable groundwater head spatial distribution reflecting the positions of major groundwater bodies and surface water bodies in the country. In Vienna, we aim to show and demonstrate these

  19. Knowledge management (KM) processes in organizations theoretical foundations and practice

    CERN Document Server

    McInerney, Claire R

    2011-01-01

    Knowledge Management (KM) is an effort to increase useful knowledge in the organization. It is a natural outgrowth of late twentieth century movements to make organizational management and operations more effective, of higher quality, and more responsive to constituents in a rapidly changing global environment. This document traces the evolution of KM in organizations, summarizing the most influential research and literature in the field. It also presents an overview of selected common and current practices in knowledge management, including the relationship between knowledge management and de

  20. Evaluation of triggering schemes for KM3NeT

    Energy Technology Data Exchange (ETDEWEB)

    Seitz, T., E-mail: Thomas.Seitz@physik.uni-erlangen.de [Erlangen Centre for Astroparticle Physics, Erwin-Rommel-Str. 1, 91058 Erlangen (Germany); Herold, B., E-mail: Bjoern.Herold@physik.uni-erlangen.de [Erlangen Centre for Astroparticle Physics, Erwin-Rommel-Str. 1, 91058 Erlangen (Germany); Shanidze, R., E-mail: shanidze@physik.uni-erlangen.de [Erlangen Centre for Astroparticle Physics, Erwin-Rommel-Str. 1, 91058 Erlangen (Germany)

    2013-10-11

    The future neutrino telescope KM3NeT, to be built in the Mediterranean Sea, will be the largest of its kind. It will include nearly two hundred thousand photomultiplier tubes (PMT) mounted in multi-PMT digital optical modules (DOM). The dominant source of the PMT signals is decays of {sup 40}K and marine fauna bioluminescence. Selection of neutrino and muon events from this continuous optical background signals requires the implementation of fast and efficient triggers. Various schemes for the filtering of background data and the selection of neutrino and muon events were evaluated for the KM3NeT telescope using Monte Carlo simulations.

  1. High-Affinity Low-Capacity and Low-Affinity High-Capacity N-Acetyl-2-Aminofluorene (AAF) Macromolecular Binding Sites Are Revealed During the Growth Cycle of Adult Rat Hepatocytes in Primary Culture.

    Science.gov (United States)

    Koch, Katherine S; Moran, Tom; Shier, W Thomas; Leffert, Hyam L

    2018-05-01

    Long-term cultures of primary adult rat hepatocytes were used to study the effects of N-acetyl-2-aminofluorene (AAF) on hepatocyte proliferation during the growth cycle; on the initiation of hepatocyte DNA synthesis in quiescent cultures; and, on hepatocyte DNA replication following the initiation of DNA synthesis. Scatchard analyses were used to identify the pharmacologic properties of radiolabeled AAF metabolite binding to hepatocyte macromolecules. Two classes of growth cycle-dependent AAF metabolite binding sites-a high-affinity low-capacity site (designated Site I) and a low-affinity high-capacity site (designated Site II)-associated with two spatially distinct classes of macromolecular targets, were revealed. Based upon radiolabeled AAF metabolite binding to purified hepatocyte genomic DNA or to DNA, RNA, proteins, and lipids from isolated nuclei, Site IDAY 4 targets (KD[APPARENT] ≈ 2-4×10-6 M and BMAX[APPARENT] ≈ 6 pmol/106 cells/24 h) were consistent with genomic DNA; and with AAF metabolized by a nuclear cytochrome P450. Based upon radiolabeled AAF binding to total cellular lysates, Site IIDAY 4 targets (KD[APPARENT] ≈ 1.5×10-3 M and BMAX[APPARENT] ≈ 350 pmol/106 cells/24 h) were consistent with cytoplasmic proteins; and with AAF metabolized by cytoplasmic cytochrome P450s. DNA synthesis was not inhibited by concentrations of AAF that saturated DNA binding in the neighborhood of the Site I KD. Instead, hepatocyte DNA synthesis inhibition required higher concentrations of AAF approaching the Site II KD. These observations raise the possibility that carcinogenic DNA adducts derived from AAF metabolites form below concentrations of AAF that inhibit replicative and repair DNA synthesis.

  2. In Vitro and In Vivo Evaluation of Alexa Fluor 680-Bombesin[7–14]NH2 Peptide Conjugate, a High-Affinity Fluorescent Probe with High Selectivity for the Gastrin-Releasing Peptide Receptor

    Directory of Open Access Journals (Sweden)

    Lixin Ma

    2007-05-01

    Full Text Available Gastrin-releasing peptide (GRP receptors are overexpressed on several types of human cancer cells, including breast, prostate, small cell lung, and pancreatic cancers. Bombesin (BBN, a 14–amino acid peptide that is an analogue of human GRP, binds to GRP receptors with very high affinity and specificity. The aim of this study was to develop a new fluorescent probe based on BBN having high tumor uptake and optimal pharmacokinetics for specific targeting and optical imaging of human breast cancer tissue. In this study, solid-phase peptide synthesis was used to produce H2N-glycylglycylglycine-BBN[7–14]NH2 peptide with the following general sequence: H2N-G-G-G-Q-W-A-V-G-H-L-M-(NH2. This conjugate was purified by reversed-phase high-performance liquid chromatography and characterized by electrospray-ionization mass spectra. The fluorescent probe Alexa Fluor 680-G-G-G-BBN[7–14]NH2 conjugate was prepared by reaction of Alexa Fluor 680 succinimidyl ester to H2N-G-G-G-BBN[7–14]NH2 in dimethylformamide (DMF. In vitro competitive binding assays, using 125I-Tyr4-BBN as the radiolabeling gold standard, demonstrated an inhibitory concentration 50% value of 7.7 ± 1.4 nM in human T-47D breast cancer cells. Confocal fluorescence microscopy images of Alexa Fluor 680-G-G-G-BBN[7–14]NH2 in human T-47D breast cancer cells indicated specific uptake, internalization, and receptor blocking of the fluorescent bioprobe in vitro. In vivo investigations in SCID mice bearing xenografted T-47D breast cancer lesions demonstrated the ability of this new conjugate to specifically target tumor tissue with high selectivity and affinity.

  3. Allocating emissions to 4 km and 1 km horizontal spatial resolutions and its impact on simulated NOx and O3 in Houston, TX

    Science.gov (United States)

    Pan, Shuai; Choi, Yunsoo; Roy, Anirban; Jeon, Wonbae

    2017-09-01

    A WRF-SMOKE-CMAQ air quality modeling system was used to investigate the impact of horizontal spatial resolution on simulated nitrogen oxides (NOx) and ozone (O3) in the Greater Houston area (a non-attainment area for O3). We employed an approach recommended by the United States Environmental Protection Agency to allocate county-based emissions to model grid cells in 1 km and 4 km horizontal grid resolutions. The CMAQ Integrated Process Rate analyses showed a substantial difference in emissions contributions between 1 and 4 km grids but similar NOx and O3 concentrations over urban and industrial locations. For example, the peak NOx emissions at an industrial and urban site differed by a factor of 20 for the 1 km and 8 for the 4 km grid, but simulated NOx concentrations changed only by a factor of 1.2 in both cases. Hence, due to the interplay of the atmospheric processes, we cannot expect a similar level of reduction of the gas-phase air pollutants as the reduction of emissions. Both simulations reproduced the variability of NASA P-3B aircraft measurements of NOy and O3 in the lower atmosphere (from 90 m to 4.5 km). Both simulations provided similar reasonable predictions at surface, while 1 km case depicted more detailed features of emissions and concentrations in heavily polluted areas, such as highways, airports, and industrial regions, which are useful in understanding the major causes of O3 pollution in such regions, and to quantify transport of O3 to populated communities in urban areas. The Integrated Reaction Rate analyses indicated a distinctive difference of chemistry processes between the model surface layer and upper layers, implying that correcting the meteorological conditions at the surface may not help to enhance the O3 predictions. The model-observation O3 bias in our studies (e.g., large over-prediction during the nighttime or along Gulf of Mexico coastline), were due to uncertainties in meteorology, chemistry or other processes. Horizontal grid

  4. The 27-km circular path of the LHC tunnel

    CERN Multimedia

    AC-DI-MM

    1994-01-01

    This aerial view of the CERN site shows the path of the 27-km circumference tunnel that housed the LEP accelerator and now contains the accelerator for CERN's new flagship project, the LHC. The ring stretches from Geneva airport, which can be seen on the lower left, to the French countryside.

  5. 40km tunnel will try to unravel secrets of universe

    CERN Multimedia

    Connor, Steve

    2004-01-01

    An international panel of particle physicists has decided that the high-nergy linera collider will use revolutionary superconducting technology to shed light on the origin and nature of the universe. The collider will be housed in a dead-straight tunnel up to 40km long (1 page)

  6. Gravity wave vertical energy flux at 95 km

    Science.gov (United States)

    Jacob, P. G.; Jacka, F.

    1985-01-01

    A three-field photometer (3FP) located at Mt. Torrens near Adelaide, is capable of monitoring different airglow emissions from three spaced fields in the sky. A wheel containing up to six different narrow bandpass interference filters can be rotated, allowing each of the filters to be sequentially placed into each of the three fields. The airglow emission of interest is the 557.7 nm line which has an intensity maximum at 95 km. Each circular field of view is located at the apexes of an equilateral triangle centered on zenith with diameters of 5 km and field separations of 13 km when projected to the 95-km level. The sampling period was 30 seconds and typical data lengths were between 7 and 8 hours. The analysis and results from the interaction of gravity waves on the 557.7 nm emission layer are derived using an atmospheric model similar to that proposed by Hines (1960) where the atmosphere is assumed isothermal and perturbations caused by gravity waves are small and adiabatic, therefore, resulting in linearized equations of motion. In the absence of waves, the atmosphere is also considered stationary. Thirteen nights of quality data from January 1983 to October 1984, covering all seasons, are used in this analysis.

  7. Magical properties of a 2540 km baseline superbeam experiment

    International Nuclear Information System (INIS)

    Raut, Sushant K.; Singh, Ravi Shanker; Uma Sankar, S.

    2011-01-01

    Lack of any information on the CP violating phase δ CP weakens our ability to determine neutrino mass hierarchy. Magic baseline of 7500 km was proposed to overcome this problem. However, to obtain large enough fluxes, at this very long baseline, one needs new techniques of generating high intensity neutrino beams. In this Letter, we highlight the magical properties of a 2540 km baseline. At such a baseline, using a narrow band neutrino superbeam whose no oscillation event rate peaks around the energy 3.5 GeV, we can determine neutrino mass hierarchy independently of the CP phase. For sin 2 2θ 13 ≥0.05, a very modest exposure of 10 Kiloton-years is sufficient to determine the hierarchy. For 0.02≤sin 2 2θ 13 ≤0.05, an exposure of about 100 Kiloton-years is needed.

  8. Hypervelocity launch capabilities to over 10 km/s

    International Nuclear Information System (INIS)

    Chhabildas, L.C.

    1991-01-01

    Very high pressure and acceleration is necessary to launch flier plates to hypervelocities. In addition, the high pressure loading must be uniform, structured, and shockless, i.e., time-dependent to prevent the flier plate from either fracturing or melting. In this paper, a novel technique is described which allows the use of megabar level loading pressures, and 10 9 g acceleration to launch intact flier plates to velocities of 12.2 km/s. 32 refs., 2 figs

  9. Model evaluation for travel distances 30 to 140 km

    International Nuclear Information System (INIS)

    Pendergast, M.M.

    1978-01-01

    The assessment of environmental effects from industrial pollution for travel distances over 50 km has been made largely without verification of the models used. Recently the Savannah River Laboratory, in cooperation with the Air Resources Laboratory of NOAA, has compiled a data base capable of providing this important verification. The data consists of (1) hourly release rates of 85 Kr from 62 m stacks near the center of the Savannah River Plant (SRP), Aiken, SC; (2) turbulence quality meteorological data from seven 62 m towers at SRP and the 335 m WJBF-TV tower at Beech Island, SC, located 25 km from the center of the SRP; (3) National Weather Service surface and upper air observations including Bush Field Airport, Augusta, Ga., about 30 km from the center of SRP; (4) hourly estimates of the mixing depth obtained with an acoustic sounder located on the SRP; and (5) weekly and 10-hour averaged 85 Kr air concentrations at 13 sites surrounding the SRP at distances ranging between 30 and 143 km. An earlier report has shown that annual averaged air concentrations for 1975 agree with observed values at the 13 sites within a factor of two (Pendergast, 1977). This report presents more detailed results based upon 10-hour averaged air concentrations. The models evaluated were variations of the stability wind-rose model and a segmented plume model. The meteorological models depend upon several key input variables: (1) stability category, (2) sigma/sub y/ and sigma/sub z/ curves, (3) wind velocity, and (4) mixing depth. Each of these key variables can be estimated by a variety of methods averaging processes. Several of the more commonly used methods for estimating the four key variables were evaluated using calculated and measured 85 Kr air concentrations. Estimates of error were obtained for monthly and 10-hour sampling times

  10. Activation of molybdenite concentrate of KM-4 index mark

    International Nuclear Information System (INIS)

    Bogomolov, A.M.; Mitskevich, A.A.; Shuvaev, A.P.

    1991-01-01

    A study was made on the effect of mechanical activation in DU-73 disintegrator on structural characteristics, dissolution rate and leaching degree of molybdenite in KM-4 concentrated. The following facts were established in experiments: high amorphization degree of basal planes (plate surfaces) ∼ 80%, increase of oxidation degree by 35% on the average. All this testify to the fact, that oxidation reaction in activated molybdenite proceeds both on plate edges and on broken basal planes

  11. Radiosynthesis and in vitro evaluation of 2-(N-alkyl-N-1'-11C-propyl)amino-5-hydroxytetralin analogs as high affinity agonists for dopamine D-2 receptors

    International Nuclear Information System (INIS)

    Shi Bingzhi; Narayanan, Tanjore K.; Yang, Z.-Y.; Christian, Bradley T.; Mukherjee, Jogeshwar

    1999-01-01

    We have developed radiotracers based on agonists that may potentially allow the in vivo assessment of the high affinity (HA) state of the dopamine D-2 receptors. The population of HA state, which is likely the functional state of the receptor, may be altered in certain diseases. We carried out radiosyntheses and evaluated the binding affinities, lipophilicity, and in vitro autoradiographic binding characteristics of three dopamine D-2 receptor agonists: (±)-2-(N,N-dipropyl)amino-5-hydroxytetralin (5-OH-DPAT), (±)-2-(N-phenethyl-N-propyl)amino-5-hydroxytetralin (PPHT), and (±)-2-(N-cyclohexylethyl-N-propyl)amino-5-hydroxytetralin (ZYY-339). In 3 H-spiperone assays using rat striata, ZYY-339 exhibited subnanomolar affinity for D-2 receptor sites ( IC 50 =0.010 nM), PPHT was somewhat weaker ( IC 50 =0.65 nM), and 5-OH-DPAT exhibited the weakest affinity ( IC 50 =2.5 nM) of the three compounds. Radiosynthesis of these derivatives, 2-(N-propyl-N-1'- 11 C-propyl)amino-5-hydroxytetralin ( 11 C-5-OH-DPAT), 2-(N-phenethyl-N-1'- 11 C-propyl)amino-5-hydroxytetralin ( 11 C-PPHT), and 2-(N-cyclohexylethyl-N-1'- 11 C-propyl)amino-5-hydroxytetralin ( 11 C-ZYY-339) was achieved by first synthesizing 11 C-1-propionyl chloride and subsequent coupling with the appropriate secondary amine precursor to form the respective amide, which was then reduced to provide the desired tertiary amine products. The final products were obtained by reverse-phase high performance liquid chromatography (HPLC) purification in radiochemical yields of 5-10% after 60-75 min from the end of 11 CO 2 trapping and with specific activities in the range of 250-1,000 Ci/mmol. In vitro autoradiographs in rat brain slices with 11 C-5-OH-DPAT, 11 C-PPHT, and 11 C-ZYY-339 revealed selective binding of the three radiotracers to the dopamine D-2 receptors in the striata

  12. (D-Pen2,4 prime -125I-Phe4,D-Pen5)enkephalin: A selective high affinity radioligand for delta opioid receptors with exceptional specific activity

    Energy Technology Data Exchange (ETDEWEB)

    Knapp, R.J.; Sharma, S.D.; Toth, G.; Duong, M.T.; Fang, L.; Bogert, C.L.; Weber, S.J.; Hunt, M.; Davis, T.P.; Wamsley, J.K. (Department of Pharmacology, University of Arizona, College of Medicine, Tucson (United States))

    1991-09-01

    (D-Pen2,4{prime}-125I-Phe4,D-Pen5)enkephalin ((125I)DPDPE) is a highly selective radioligand for the delta opioid receptor with a specific activity (2200 Ci/mmol) that is over 50-fold greater than that of tritium-labeled DPDPE analogs. (125I)DPDPE binds to a single site in rat brain membranes with an equilibrium dissociation constant (Kd) value of 421 {plus minus} 67 pM and a receptor density (Bmax) value of 36.4 {plus minus} 2.7 fmol/mg protein. The high affinity of this site for delta opioid receptor ligands and its low affinity for mu or kappa receptor-selective ligands are consistent with its being a delta opioid receptor. The distribution of these sites in rat brain, observed by receptor autoradiography, is also consistent with that of delta opioid receptors. Association and dissociation binding kinetics of 1.0 nM (125I) DPDPE are monophasic at 25 degrees C. The association rate (k + 1 = 5.80 {plus minus} 0.88 {times} 10(7) M-1 min-1) is about 20- and 7-fold greater than that measured for 1.0 nM (3H) DPDPE and 0.8 nM (3H) (D-Pen2,4{prime}-Cl-Phe4, D-Pen5)enkephalin, respectively. The dissociation rate of (125I)DPDPE (0.917 {plus minus} 0.117 {times} 10(-2) min-1) measured at 1.0 nM is about 3-fold faster than is observed for either of the other DPDPE analogs. The rapid binding kinetics of (125I)DPDPE is advantageous because binding equilibrium is achieved with much shorter incubation times than are required for other cyclic enkephalin analogs. This, in addition to its much higher specific activity, makes (125I)DPDPE a valuable new radioligand for studies of delta opioid receptors.

  13. Dimerization Is Not a Determining Factor for Functional High Affinity Human Plasminogen Binding by the Group A Streptococcal Virulence Factor PAM and Is Mediated by Specific Residues within the PAM a1a2 Domain*

    Science.gov (United States)

    Bhattacharya, Sarbani; Liang, Zhong; Quek, Adam J.; Ploplis, Victoria A.; Law, Ruby; Castellino, Francis J.

    2014-01-01

    A emm53 subclass of Group A Streptococcus pyogenes (GAS) interacts tightly with human plasma plasminogen (hPg) and plasmin (hPm) via the kringle 2 (K2hPg) domain of hPg/hPm and the N-terminal a1a2 regions of a GAS coiled-coil M-like protein (PAM). Previous studies have shown that a monomeric PAM fragment, VEK30 (residues 97–125 + Tyr), interacted specifically with isolated K2hPg. However, the binding strength of VEK30 (KD = 56 nm) was ∼60-fold weaker than that of full-length dimeric PAM (KD = 1 nm). To assess whether this attenuated binding was due to the inability of VEK30 to dimerize, we defined the minimal length of PAM required to dimerize using a series of peptides with additional PAM residues placed at the NH2 and COOH termini of VEK30. VEK64 (PAM residues 83–145 + Tyr) was found to be the smallest peptide that adopted an α-helical dimer, and was bound to K2hPg with nearly the same affinity as PAM (KD = 1–2 nm). However, addition of two PAM residues (Arg126-His127) to the COOH terminus of VEK30 (VEK32) maintained a monomeric peptidic structure, but exhibited similar K2hPg binding affinity as full-length dimeric PAM. We identified five residues in a1a2 (Arg113, His114, Glu116, Arg126, His127), mutation of which reduced PAM binding affinity for K2hPg by ∼1000-fold. Replacement of these critical residues by Ala in the GAS genome resulted in reduced virulence, similar to the effects of inactivating the PAM gene entirely. We conclude that rather than dimerization of PAM, the five key residues in the binding domain of PAM are essential to mediate the high affinity interaction with hPg, leading to increased GAS virulence. PMID:24962580

  14. Dimerization is not a determining factor for functional high affinity human plasminogen binding by the group A streptococcal virulence factor PAM and is mediated by specific residues within the PAM a1a2 domain.

    Science.gov (United States)

    Bhattacharya, Sarbani; Liang, Zhong; Quek, Adam J; Ploplis, Victoria A; Law, Ruby; Castellino, Francis J

    2014-08-01

    A emm53 subclass of Group A Streptococcus pyogenes (GAS) interacts tightly with human plasma plasminogen (hPg) and plasmin (hPm) via the kringle 2 (K2hPg) domain of hPg/hPm and the N-terminal a1a2 regions of a GAS coiled-coil M-like protein (PAM). Previous studies have shown that a monomeric PAM fragment, VEK30 (residues 97-125 + Tyr), interacted specifically with isolated K2hPg. However, the binding strength of VEK30 (KD = 56 nm) was ∼60-fold weaker than that of full-length dimeric PAM (KD = 1 nm). To assess whether this attenuated binding was due to the inability of VEK30 to dimerize, we defined the minimal length of PAM required to dimerize using a series of peptides with additional PAM residues placed at the NH2 and COOH termini of VEK30. VEK64 (PAM residues 83-145 + Tyr) was found to be the smallest peptide that adopted an α-helical dimer, and was bound to K2hPg with nearly the same affinity as PAM (KD = 1-2 nm). However, addition of two PAM residues (Arg(126)-His(127)) to the COOH terminus of VEK30 (VEK32) maintained a monomeric peptidic structure, but exhibited similar K2hPg binding affinity as full-length dimeric PAM. We identified five residues in a1a2 (Arg(113), His(114), Glu(116), Arg(126), His(127)), mutation of which reduced PAM binding affinity for K2hPg by ∼ 1000-fold. Replacement of these critical residues by Ala in the GAS genome resulted in reduced virulence, similar to the effects of inactivating the PAM gene entirely. We conclude that rather than dimerization of PAM, the five key residues in the binding domain of PAM are essential to mediate the high affinity interaction with hPg, leading to increased GAS virulence. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  15. The 1 km resolution global data set: needs of the International Geosphere Biosphere Programme

    Science.gov (United States)

    Townshend, J.R.G.; Justice, C.O.; Skole, D.; Malingreau, J.-P.; Cihlar, J.; Teillet, P.; Sadowski, F.; Ruttenberg, S.

    1994-01-01

    Examination of the scientific priorities for the International Geosphere Biosphere Programme (IGBP) reveals a requirement for global land data sets in several of its Core Projects. These data sets need to be at several space and time scales. Requirements are demonstrated for the regular acquisition of data at spatial resolutions of 1 km and finer and at high temporal frequencies. Global daily data at a resolution of approximately 1 km are sensed by the Advanced Very High Resolution Radiometer (AVHRR), but they have not been available in a single archive. It is proposed, that a global data set of the land surface is created from remotely sensed data from the AVHRR to support a number of IGBP's projects. This data set should have a spatial resolution of 1 km and should be generated at least once every 10 days for the entire globe. The minimum length of record should be a year, and ideally a system should be put in place which leads to the continuous acquisition of 1 km data to provide a base line data set prior to the Earth Observing System (EOS) towards the end of the decade. Because of the high cloud cover in many parts of the world, it is necessary to plan for the collection of data from every orbit. Substantial effort will be required in the preprocessing of the data set involving radiometric calibration, atmospheric correction, geometric correction and temporal compositing, to make it suitable for the extraction of information.

  16. Three-dimensional spatial structures of solar wind turbulence from 10 000-km to 100-km scales

    Directory of Open Access Journals (Sweden)

    Y. Narita

    2011-10-01

    Full Text Available Using the four Cluster spacecraft, we have determined the three-dimensional wave-vector spectra of fluctuating magnetic fields in the solar wind. Three different solar wind intervals of Cluster data are investigated for this purpose, representing three different spatial scales: 10 000 km, 1000 km, and 100 km. The spectra are determined using the wave telescope technique (k-filtering technique without assuming the validity of Taylor's frozen-in-flow hypothesis nor are any assumptions made as to the symmetry properties of the fluctuations. We find that the spectra are anisotropic on all the three scales and the power is extended primarily in the directions perpendicular to the mean magnetic field, as might be expected of two-dimensional turbulence, however, the analyzed fluctuations are not axisymmetric. The lack of axisymmetry invalidates some earlier techniques using single spacecraft observations that were used to estimate the percentage of magnetic energy residing in quasi-two-dimensional power. However, the dominance of two-dimensional turbulence is consistent with the relatively long mean free paths of cosmic rays in observed in the heliosphere. On the other hand, the spectra also exhibit secondary extended structures oblique from the mean magnetic field direction. We discuss possible origins of anisotropy and asymmetry of solar wind turbulence spectra.

  17. Mapping the global land surface using 1 km AVHRR data

    Science.gov (United States)

    Lauer, D.T.; Eidenshink, J.C.

    1998-01-01

    The scientific requirements for mapping the global land surface using 1 km advanced very high resolution radiometer (AVHRR) data have been set forth by the U.S. Global Change Research Program; the International Geosphere Biosphere Programme (IGBP); The United Nations; the National Oceanic and Atmospheric Administration (NOAA); the Committee on Earth Observations Satellites; and the National Aeronautics and Space Administration (NASA) mission to planet Earth (MTPE) program. Mapping the global land surface using 1 km AVHRR data is an international effort to acquire, archive, process, and distribute 1 km AVHRR data to meet the needs of the international science community. A network of AVHRR receiving stations, along with data recorded by NOAA, has been acquiring daily global land coverage since April 1, 1992. A data set of over 70,000 AVHRR images is archived and distributed by the United States Geological Survey (USGS) EROS Data Center, and the European Space Agency. Under the guidance of the IGBP, processing standards have been developed for calibration, atmospheric correction, geometric registration, and the production of global 10-day maximum normalized difference vegetation index (NDVI) composites. The major uses of the composites are for the study of surface vegetation condition, mapping land cover, and deriving biophysical characteristics of terrestrial ecosystems. A time-series of 54 10-day global vegetation index composites for the period of April 1, 1992 through September 1993 has been produced. The production of a time-series of 33 10-day global vegetation index composites using NOAA-14 data for the period of February 1, 1995 through December 31, 1995 is underway. The data products are available from the USGS, in cooperation with NASA's MTPE program and other international organizations.

  18. Nanobeacon: A low cost time calibration instrument for the KM3NeT neutrino telescope

    Energy Technology Data Exchange (ETDEWEB)

    Calvo, David [IFIC. Instituto de Física Corpuscular, CSIC-Universidad de Valencia, C/Catedrático José Beltrán, 2. 46980 Paterna (Spain); Collaboration: KM3NeT Collaboration

    2014-11-18

    The KM3NeT collaboration aims at the construction of a multi-km3 high-energy neutrino telescope in the Mediterranean Sea consisting of a matrix of pressure resistant glass spheres holding each one a set (31) of small area photomultipliers. The main goal of the telescope is to observe cosmic neutrinos through the Cherenkov light induced in sea water by charged particles produced in neutrino interactions with the surrounding medium. A relative time calibration between photomultipliers of the order of 1 ns is required to achieve an optimal performance. Due to the high volume to be covered by KM3NeT, a cost reduction of the different systems is a priority. To this end a very low price calibration device, the so called Nanobeacon, has been designed and developed. At present one of such devices has already been integrated successfully at the KM3NeT telescope and eight of them in the Nemo Tower Phase II. In this article the main properties and operation of this device are described.

  19. Projected Population Proximity Indices (30km for 2005, 2030 & 2050

    Directory of Open Access Journals (Sweden)

    Neil S. Alexander

    2013-07-01

    Full Text Available This data package includes nine population proximity index layers for 2005, 2030 and 2050, for rural, urban and total populations.  The layers are distributed as 1km GeoTIFFs and GeoJPGss at 1km. The aim of these layers is to describe the population which may be likely to visit a specific locality where access is determined by Euclidean distance. By using the layers alongside other geographic datasets relating to disease risk it may help identify where people may come into contact with a disease.  Human population layers are often used in models to identify risk areas where humans and viruses interact, however most pathogens are not restricted to areas of human habitation: many are found in lesser populated areas such as forests.  This dataset will help identify less populated areas that may well still receive high visitor numbers. The layers have been projected to 2030 and 2050 to enable projections of human/disease interfaces in the medium-term which are required to inform policy makers at country and continental level. Urban and rural populations have been separated into individual layers as in some cases it is useful to distinguish between the behaviour and associated risks attributed to the different population segments.  There may be a different risk of contacting diseases in rural habitats for rural workers than for than urban visitors.

  20. Synthesis and pre-clinical evaluation of a new class of high-affinity {sup 18}F-labeled PSMA ligands for detection of prostate cancer by PET imaging

    Energy Technology Data Exchange (ETDEWEB)

    Kelly, James; Amor-Coarasa, Alejandro; Williams, Clarence; Ponnala, Shashikanth [Weill Cornell Medicine, Division of Radiopharmaceutical Sciences and Molecular Imaging Innovations Institute, Department of Radiology, New York, NY (United States); Nikolopoulou, Anastasia [Weill Cornell Medicine, Division of Radiopharmaceutical Sciences and Molecular Imaging Innovations Institute, Department of Radiology, New York, NY (United States); Weill Cornell Medicine, Citigroup Biomedical Imaging Center, New York, NY (United States); Kim, Dohyun [Weill Cornell Medicine, Citigroup Biomedical Imaging Center, New York, NY (United States); Babich, John W. [Weill Cornell Medicine, Division of Radiopharmaceutical Sciences and Molecular Imaging Innovations Institute, Department of Radiology, New York, NY (United States); Weill Cornell Medicine, Citigroup Biomedical Imaging Center, New York, NY (United States); Weill Cornell Medicine, Meyer Cancer Center, New York, NY (United States)

    2017-04-15

    Current clinical imaging of PSMA-positive prostate cancer by positron emission tomography (PET) mainly features {sup 68}Ga-labeled tracers, notably [{sup 68}Ga]Ga-PSMA-HBED-CC. The longer half-life of fluorine-18 offers significant advantages over Ga-68, clinically and logistically. We aimed to develop high-affinity PSMA inhibitors labeled with fluorine-18 as alternative tracers for prostate cancer. Six triazolylphenyl ureas and their alkyne precursors were synthesized from the Glu-urea-Lys PSMA binding moiety. PSMA affinity was determined in a competitive binding assay using LNCaP cells. The [{sup 18}F]triazoles were isolated following a Cu(I)-catalyzed click reaction between the alkynes and [{sup 18}F]fluoroethylazide. The {sup 18}F-labeled compounds were evaluated in nude mice bearing LNCaP tumors and compared to [{sup 68}Ga]Ga-PSMA-HBED-CC and [{sup 18}F]DCFPyL. Biodistribution studies of the two tracers with the highest imaged-derived tumor uptake and highest PSMA affinity were undertaken at 1 h, 2 h and 4 h post-injection (p.i.), and co-administration of PMPA was used to determine whether uptake was PSMA-specific. F-18-labeled triazolylphenyl ureas were prepared with a decay-corrected RCY of 20-40 %, >98 % radiochemical and chemical purity, and specific activity of up to 391 GBq/μmol. PSMA binding (IC{sub 50}) ranged from 3-36 nM. The position of the triazole influenced tumor uptake (3 > 4 > 2), and direct conjugation of the triazole with the phenylurea moiety was preferred to insertion of a spacer group. Image-derived tumor uptake ranged from 6-14 %ID/g at 2 h p.i., the time of maximum tumor uptake; uptake of [{sup 68}Ga]Ga-PSMA-HBED-CC and [{sup 18}F]DCFPyL was 5-6 %ID/g at 1-3 h p.i., the time of maximum tumor uptake. Biodistribution studies of the two most promising compounds gave maximum tumor uptakes of 10.9 ± 1.0 % and 14.3 ± 2.5 %ID/g, respectively, as compared to 6.27 ± 1.44 %ID/g for [{sup 68}Ga]Ga-PSMA-HBED-CC. Six [{sup 18}F

  1. Synthesis and pre-clinical evaluation of a new class of high-affinity "1"8F-labeled PSMA ligands for detection of prostate cancer by PET imaging

    International Nuclear Information System (INIS)

    Kelly, James; Amor-Coarasa, Alejandro; Williams, Clarence; Ponnala, Shashikanth; Nikolopoulou, Anastasia; Kim, Dohyun; Babich, John W.

    2017-01-01

    Current clinical imaging of PSMA-positive prostate cancer by positron emission tomography (PET) mainly features "6"8Ga-labeled tracers, notably ["6"8Ga]Ga-PSMA-HBED-CC. The longer half-life of fluorine-18 offers significant advantages over Ga-68, clinically and logistically. We aimed to develop high-affinity PSMA inhibitors labeled with fluorine-18 as alternative tracers for prostate cancer. Six triazolylphenyl ureas and their alkyne precursors were synthesized from the Glu-urea-Lys PSMA binding moiety. PSMA affinity was determined in a competitive binding assay using LNCaP cells. The ["1"8F]triazoles were isolated following a Cu(I)-catalyzed click reaction between the alkynes and ["1"8F]fluoroethylazide. The "1"8F-labeled compounds were evaluated in nude mice bearing LNCaP tumors and compared to ["6"8Ga]Ga-PSMA-HBED-CC and ["1"8F]DCFPyL. Biodistribution studies of the two tracers with the highest imaged-derived tumor uptake and highest PSMA affinity were undertaken at 1 h, 2 h and 4 h post-injection (p.i.), and co-administration of PMPA was used to determine whether uptake was PSMA-specific. F-18-labeled triazolylphenyl ureas were prepared with a decay-corrected RCY of 20-40 %, >98 % radiochemical and chemical purity, and specific activity of up to 391 GBq/μmol. PSMA binding (IC_5_0) ranged from 3-36 nM. The position of the triazole influenced tumor uptake (3 > 4 > 2), and direct conjugation of the triazole with the phenylurea moiety was preferred to insertion of a spacer group. Image-derived tumor uptake ranged from 6-14 %ID/g at 2 h p.i., the time of maximum tumor uptake; uptake of ["6"8Ga]Ga-PSMA-HBED-CC and ["1"8F]DCFPyL was 5-6 %ID/g at 1-3 h p.i., the time of maximum tumor uptake. Biodistribution studies of the two most promising compounds gave maximum tumor uptakes of 10.9 ± 1.0 % and 14.3 ± 2.5 %ID/g, respectively, as compared to 6.27 ± 1.44 %ID/g for ["6"8Ga]Ga-PSMA-HBED-CC. Six ["1"8F]triazolylphenyl ureas were prepared in good radiochemical yield

  2. A Case Analysis to Increase Awareness of Current USMC Knowledge Management (KM) Practices

    Science.gov (United States)

    2013-09-01

    Knowledge Management System KPI Key Performance Indicator KS Knowledge Superiority KVA Knowledge Value Added MAGTF Marine Air Ground Task Force...quantitative KPIs [key performance indicators] developed to monitor efficiency and some qualitative KPIs to assess effectiveness in the implementation...integration,” and the level of control is “qualitative and quantitative KPIs in place to monitor the implementation of an effective and efficient KM

  3. Ambient Seismic Imaging of Hydraulically Active Fractures at km Depths

    Science.gov (United States)

    Malin, P. E.; Sicking, C.

    2017-12-01

    Streaming Depth Images of ambient seismic signals using numerous, densely-distributed, receivers have revealed their connection to hydraulically active fractures at 0.5 to 5 km depths. Key for this type of imaging is very high-fold stacking over both multiple receives and periods of a few hours. Also important is suppression of waveforms from fixed, repeating sources such as pumps, generators, and traffic. A typical surface-based ambient SDI survey would use a 3D seismic receiver grid. It would have 1,000 to 4,000 uniformly distributed receivers at a density of 50/km2over the target. If acquired by borehole receivers buried 100 m deep, the density can be dropped by an order of magnitude. We show examples of the acquisition and signal processing scenarios used to produce the ambient images. (Sicking et al., SEG Interpretation, Nov 2017.) While the fracture-fluid source connection of SDI has been verified by drilling and various types of hydraulic tests, the precise nature of the signal's origin is not clear. At the current level of observation, the signals do not have identifiable phases, but can be focused using P wave velocities. Suggested sources are resonances of pressures fluctuations in the fractures, or small, continuous, slips on fractures surfaces. In either case, it appears that the driving mechanism is tectonic strain in an inherently unstable crust. Solid earth tides may enhance these strains. We illustrate the value of the ambient SDI method in its industrial application by showing case histories from energy industry and carbon-capture-sequestration projects. These include ambient images taken before, during, and after hydraulic treatments in un-conventional reservoirs. The results show not only locations of active fractures, but also their time responses to stimulation and production. Time-lapse ambient imaging can forecast and track events such as well interferences and production changes that can result from nearby treatments.

  4. Exposure of Postnatal Rats to a Static Magnetic Field of 0.14 T Influences Functional Laterality of the Hippocampal High-Affinity Choline Uptake System in Adulthood; In Vitro Test With Magnetic Nanoparticles

    Czech Academy of Sciences Publication Activity Database

    Krištofíková, Z.; Čermák, M.; Benešová, O.; Klaschka, Jan; Zach, P.

    2005-01-01

    Roč. 30, č. 2 (2005), s. 253-262 ISSN 0364-3190 R&D Projects: GA MZd NF7576 Keywords : magnetic nanoparticles * choline transport * cholinergic * functional impairment * hippocampus * laterality * magnetoreception * static magnetic field Subject RIV: BB - Applied Statistics, Operational Research Impact factor: 2.187, year: 2005

  5. Progress toward a Km-scale neutrino detector in the deep ocean

    Energy Technology Data Exchange (ETDEWEB)

    Stokstad, R.G.

    1997-11-01

    The best particles for observing distant objects are photons and neutrinos. Because of the neutrino`s weak interaction cross section, detectors suitable for astronomy must be very large and well shielded from cosmic rays. Eventually, a detector with the order of a square km of effective area will be needed for systematic observations of distant point sources such as active galactic nuclei. Prototype detectors are currently being developed at several sites in the ocean, at Lake Baikal, and in Antarctica. This talk summarizes the status of the projects that use the deep ocean for the detector medium and shielding: DUMAND, NESTOR and ANTARES. Technical developments will be needed for a future km-scale detector; progress on one of these, a digital electronic system, is also described.

  6. Progress toward a Km-scale neutrino detector in the deep ocean

    International Nuclear Information System (INIS)

    Stokstad, R.G.

    1997-11-01

    The best particles for observing distant objects are photons and neutrinos. Because of the neutrino's weak interaction cross section, detectors suitable for astronomy must be very large and well shielded from cosmic rays. Eventually, a detector with the order of a square km of effective area will be needed for systematic observations of distant point sources such as active galactic nuclei. Prototype detectors are currently being developed at several sites in the ocean, at Lake Baikal, and in Antarctica. This talk summarizes the status of the projects that use the deep ocean for the detector medium and shielding: DUMAND, NESTOR and ANTARES. Technical developments will be needed for a future km-scale detector; progress on one of these, a digital electronic system, is also described

  7. Ultra-high velocity impacts: Cratering studies of microscopic impacts from 3 km/s to 30 km/s

    International Nuclear Information System (INIS)

    Stradling, G.L.; Idzorek, G.C.; Shafer, B.P.; Curling, H.L. Jr.; Collopy, M.T.; Hopkins Blossom, A.A.; Fuerstenau, S.

    1992-01-01

    Cratering experiments performed under carefully controlled conditions at impact velocities ranging from 3 km/s to 30 km/s into a wide variety of target materials are presented. These impact experiments use the 6 MV vertical Van de Graaff accelerator of the Ion Beam Facility at the Los Alamos National Laboratory to electrostatically accelerate highly charged iron microspheres. The sub-micron spheres, from a random size distribution, are shocklessly accelerated along an 8 m flight path. Ultra-sensitive charge detectors monitor the passage of the projectiles at a rate of up to 100 projectiles/second. An online computer records and displays in real time the charge, velocity and mass of the projectiles and provides cross correlation between the events observed by the several in-flight charge detectors and impact detectors. Real-time logic controls an electrostatic kicker which deflects projectiles of selected charge and velocity onto the target. Thus each experiment consists of an ensemble of 10 to 40 impacts onto a single target within a narrow window of the projectile parameter space, providing excellent statistical resolution of each data point. The target materials used include single crystal copper and single crystal aluminum, gold, and quartz as well as pyrolytic graphic and epoxy used in composite materials of interest to space applications. We also conducted impact experiments onto thin Mylar and nickel foils. This paper presents these experiments and summarizes the cratering characterization performed to date. Emphasis is placed on cratering results in several materials over a range of impact velocities

  8. 1km Global Terrestrial Carbon Flux: Estimations and Evaluations

    Science.gov (United States)

    Murakami, K.; Sasai, T.; Kato, S.; Saito, M.; Matsunaga, T.; Hiraki, K.; Maksyutov, S. S.

    2017-12-01

    Estimating global scale of the terrestrial carbon flux change with high accuracy and high resolution is important to understand global environmental changes. Furthermore the estimations of the global spatiotemporal distribution may contribute to the political and social activities such as REDD+. In order to reveal the current state of terrestrial carbon fluxes covering all over the world and a decadal scale. The satellite-based diagnostic biosphere model is suitable for achieving this purpose owing to observing on the present global land surface condition uniformly at some time interval. In this study, we estimated the global terrestrial carbon fluxes with 1km grids by using the terrestrial biosphere model (BEAMS). And we evaluated our new carbon flux estimations on various spatial scales and showed the transition of forest carbon stocks in some regions. Because BEAMS required high resolution meteorological data and satellite data as input data, we made 1km interpolated data using a kriging method. The data used in this study were JRA-55, GPCP, GOSAT L4B atmospheric CO2 data as meteorological data, and MODIS land product as land surface satellite data. Interpolating process was performed on the meteorological data because of insufficient resolution, but not on MODIS data. We evaluated our new carbon flux estimations using the flux tower measurement (FLUXNET2015 Datasets) in a point scale. We used 166 sites data for evaluating our model results. These flux sites are classified following vegetation type (DBF, EBF, ENF, mixed forests, grass lands, croplands, shrub lands, Savannas, wetlands). In global scale, the BEAMS estimations was underestimated compared to the flux measurements in the case of carbon uptake and release. The monthly variations of NEP showed relatively high correlations in DBF and mixed forests, but the correlation coefficients of EBF, ENF, and grass lands were less than 0.5. In the meteorological factors, air temperature and solar radiation showed

  9. Hydrogeological monitoring (0-15 km of the Chernobyl' station affected zone)

    International Nuclear Information System (INIS)

    Sitnikov, A.B.; Dzheko, S.P.

    1992-01-01

    Hydrogeological monitoring (HM) is aimed at finding out the regularities of formation of subsurface water, as well as unfavorable phenomena and parameters used for forecasts. To the main objects of HM belong: surface storm run-off; soil moisture of the aeration zone; underground water; water-bearing horizons in Buchak and Senomazh depositions. Criteria for grounding the system of monitoring and control are presented. The elements of the system for tracking local HM in 10-15 km zone are given. Potential centres of unfavorable phenomena, as well as the objects to be protected are noted. 3 figs

  10. CHARACTERIZATION AND MODELING OF SOLITON TRANSMISSION AT 2.5 GB/S OVER 200 KM

    Directory of Open Access Journals (Sweden)

    KHALID A. S. AL-KHATEEB

    2010-09-01

    Full Text Available Soliton characteristics and soliton transmission have been simulated using a VPI simulator. Simulation was also used to construct and study a soliton communication system. Near soliton pulses emitted by an actively mode-locked laser is then compressed in a dispersion-compensating fiber (DCF to produce solitons. The effects of non-linearity and active pre-chirping of mode-locked laser diode sources were also investigated. Assessment on a modeled system using real data shows that propagation over 250 km at 2.5 Gb/s in standard fibers with 20 ps pulse widths is possible in the 1550 nm wavelength range.

  11. Full 40 km crustal reflection seismic datasets in several Indonesian basins

    Science.gov (United States)

    Dinkelman, M. G.; Granath, J. W.; Christ, J. M.; Emmet, P. A.; Bird, D. E.

    2010-12-01

    sedimentary and burial metamorphosed sedimentary rock that we divide into two packages on the basis of seismic character. The upper is 8-15 km of undeformed late Precambrian sediments the top of which ties Eocambrian rocks in wells in offshore New Guinea. This package appears to correlate to the Wessel Group in northern Australia. The lower package is composed of 10-15 km of strongly bedded, presumably burial metamorphosed rocks that make up the bulk of the lower crust. These may equate to any of a number of northern Australian Mesoproterozoic basins. This lower package offlaps ‘pods’ of seismically transparent basement (?Paleoproterozoic or Archean) that make up at most the lowermost 15 km of the 40 km PSDM line. Both Precambrian packages appear to be craton-margin sedimentary wedges, the younger overlapping the older. The SE extent of the lowermost package is deformed in a thrust system which may mark the event that detached it from its original underlying oceanic or transitional crust during cratonization. The SPAN programs are important new data sets to clarify and in some cases solve outstanding problems in basin architecture and tectonic evolution.

  12. ASIC design in the KM3NeT detector

    International Nuclear Information System (INIS)

    Gajanana, D; Gromov, V; Timmer, P

    2013-01-01

    In the KM3NeT project [1], Cherenkov light from the muon interactions with transparent matter around the detector, is used to detect neutrinos. Photo multiplier tubes (PMT) used as photon sensor, are housed in a glass sphere (aka Optical Module) to detect single photons from the Cherenkov light. The PMT needs high operational voltage ( ∼ 1.5 kV) and is generated by a Cockroft-Walton (CW) multiplier circuit. The electronics required to control the PMT's and collect the signals is integrated in two ASIC's namely: 1) a front-end mixed signal ASIC (PROMiS) for the readout of the PMT and 2) an analog ASIC (CoCo) to generate pulses for charging the CW circuit and to control the feedback of the CW circuit. In this article, we discuss the two integrated circuits and test results of the complete setup. PROMiS amplifies the input charge, converts it to a pulse width and delivers the information via LVDS signals. These LVDS signals carry accurate information on the Time of arrival ( 2 C bus. This unique combination of the ASIC's results in a very cost and power efficient PMT base design.

  13. Stratospheric microbiology at 20 km over the Pacific Ocean

    Science.gov (United States)

    Smith, David J.; Griffin, Dale W.; Schuerger, Andrew C.

    2010-01-01

    An aerobiology sampling flight at 20 km was conducted on 28 April 2008 over the Pacific Ocean (36.5° N, 118–149° W), a period of time that coincided with the movement of Asian dust across the ocean. The aim of this study was to confirm the presence of viable bacteria and fungi within a transoceanic, atmospheric bridge and to improve the resolution of flight hardware processing techniques. Isolates of the microbial strains recovered were analyzed with ribosomal ribonucleic acid (rRNA) sequencing to identify bacterial species Bacillus sp., Bacillus subtilis, Bacillus endophyticus, and the fungal genus Penicillium. Satellite imagery and ground-based radiosonde observations were used to measure dust movement and characterize the high-altitude environment at the time of collection. Considering the atmospheric residency time (7–10 days), the extreme temperature regime of the environment (-75°C), and the absence of a mechanism that could sustain particulates at high altitude, it is unlikely that our samples indicate a permanent, stratospheric ecosystem. However, the presence of viable fungi and bacteria in transoceanic stratosphere remains relevant to understanding the distribution and extent of microbial life on Earth.

  14. Production of Biodiesel from Waste Vegetable Oil via KM Micromixer

    Directory of Open Access Journals (Sweden)

    M. F. Elkady

    2015-01-01

    Full Text Available The production of biodiesel from waste vegetable oils through its pretreatment followed by transesterification process in presence of methanol was investigated using a KM micromixer reactor. The parameters affecting biodiesel production process such as alcohol to oil molar ratio, catalyst concentration, the presence of tetrahydrofuran (THF as a cosolvent, and the volumetric flow rates of inlet fluids were optimized. The properties of the produced biodiesel were compared with its parent waste oil through different characterization techniques. The presence of methyl ester groups at the produced biodiesel was confirmed using both the gas chromatography-mass spectrometry (GC-MS and the infrared spectroscopy (FT-IR. Moreover, the thermal analysis of the produced biodiesel and the comparable waste oil indicated that the product after the transesterification process began to vaporize at 120°C which makes it lighter than its parent oil which started to vaporize at around 300°C. The maximum biodiesel production yield of 97% was recorded using 12 : 1 methanol to oil molar ratio in presence of both 1% NaOH and THF/methanol volume ratio 0.3 at 60 mL/h flow rate.

  15. SoilGrids1km — Global Soil Information Based on Automated Mapping

    Science.gov (United States)

    Hengl, Tomislav; de Jesus, Jorge Mendes; MacMillan, Robert A.; Batjes, Niels H.; Heuvelink, Gerard B. M.; Ribeiro, Eloi; Samuel-Rosa, Alessandro; Kempen, Bas; Leenaars, Johan G. B.; Walsh, Markus G.; Gonzalez, Maria Ruiperez

    2014-01-01

    Background Soils are widely recognized as a non-renewable natural resource and as biophysical carbon sinks. As such, there is a growing requirement for global soil information. Although several global soil information systems already exist, these tend to suffer from inconsistencies and limited spatial detail. Methodology/Principal Findings We present SoilGrids1km — a global 3D soil information system at 1 km resolution — containing spatial predictions for a selection of soil properties (at six standard depths): soil organic carbon (g kg−1), soil pH, sand, silt and clay fractions (%), bulk density (kg m−3), cation-exchange capacity (cmol+/kg), coarse fragments (%), soil organic carbon stock (t ha−1), depth to bedrock (cm), World Reference Base soil groups, and USDA Soil Taxonomy suborders. Our predictions are based on global spatial prediction models which we fitted, per soil variable, using a compilation of major international soil profile databases (ca. 110,000 soil profiles), and a selection of ca. 75 global environmental covariates representing soil forming factors. Results of regression modeling indicate that the most useful covariates for modeling soils at the global scale are climatic and biomass indices (based on MODIS images), lithology, and taxonomic mapping units derived from conventional soil survey (Harmonized World Soil Database). Prediction accuracies assessed using 5–fold cross-validation were between 23–51%. Conclusions/Significance SoilGrids1km provide an initial set of examples of soil spatial data for input into global models at a resolution and consistency not previously available. Some of the main limitations of the current version of SoilGrids1km are: (1) weak relationships between soil properties/classes and explanatory variables due to scale mismatches, (2) difficulty to obtain covariates that capture soil forming factors, (3) low sampling density and spatial clustering of soil profile locations. However, as the Soil

  16. SoilGrids1km--global soil information based on automated mapping.

    Directory of Open Access Journals (Sweden)

    Tomislav Hengl

    Full Text Available BACKGROUND: Soils are widely recognized as a non-renewable natural resource and as biophysical carbon sinks. As such, there is a growing requirement for global soil information. Although several global soil information systems already exist, these tend to suffer from inconsistencies and limited spatial detail. METHODOLOGY/PRINCIPAL FINDINGS: We present SoilGrids1km--a global 3D soil information system at 1 km resolution--containing spatial predictions for a selection of soil properties (at six standard depths: soil organic carbon (g kg-1, soil pH, sand, silt and clay fractions (%, bulk density (kg m-3, cation-exchange capacity (cmol+/kg, coarse fragments (%, soil organic carbon stock (t ha-1, depth to bedrock (cm, World Reference Base soil groups, and USDA Soil Taxonomy suborders. Our predictions are based on global spatial prediction models which we fitted, per soil variable, using a compilation of major international soil profile databases (ca. 110,000 soil profiles, and a selection of ca. 75 global environmental covariates representing soil forming factors. Results of regression modeling indicate that the most useful covariates for modeling soils at the global scale are climatic and biomass indices (based on MODIS images, lithology, and taxonomic mapping units derived from conventional soil survey (Harmonized World Soil Database. Prediction accuracies assessed using 5-fold cross-validation were between 23-51%. CONCLUSIONS/SIGNIFICANCE: SoilGrids1km provide an initial set of examples of soil spatial data for input into global models at a resolution and consistency not previously available. Some of the main limitations of the current version of SoilGrids1km are: (1 weak relationships between soil properties/classes and explanatory variables due to scale mismatches, (2 difficulty to obtain covariates that capture soil forming factors, (3 low sampling density and spatial clustering of soil profile locations. However, as the SoilGrids system is

  17. File Specification for the 7-km GEOS-5 Nature Run, Ganymed Release Non-Hydrostatic 7-km Global Mesoscale Simulation

    Science.gov (United States)

    da Silva, Arlindo M.; Putman, William; Nattala, J.

    2014-01-01

    This document describes the gridded output files produced by a two-year global, non-hydrostatic mesoscale simulation for the period 2005-2006 produced with the non-hydrostatic version of GEOS-5 Atmospheric Global Climate Model (AGCM). In addition to standard meteorological parameters (wind, temperature, moisture, surface pressure), this simulation includes 15 aerosol tracers (dust, sea-salt, sulfate, black and organic carbon), O3, CO and CO2. This model simulation is driven by prescribed sea-surface temperature and sea-ice, daily volcanic and biomass burning emissions, as well as high-resolution inventories of anthropogenic sources. A description of the GEOS-5 model configuration used for this simulation can be found in Putman et al. (2014). The simulation is performed at a horizontal resolution of 7 km using a cubed-sphere horizontal grid with 72 vertical levels, extending up to to 0.01 hPa (approximately 80 km). For user convenience, all data products are generated on two logically rectangular longitude-latitude grids: a full-resolution 0.0625 deg grid that approximately matches the native cubed-sphere resolution, and another 0.5 deg reduced-resolution grid. The majority of the full-resolution data products are instantaneous with some fields being time-averaged. The reduced-resolution datasets are mostly time-averaged, with some fields being instantaneous. Hourly data intervals are used for the reduced-resolution datasets, while 30-minute intervals are used for the full-resolution products. All full-resolution output is on the model's native 72-layer hybrid sigma-pressure vertical grid, while the reduced-resolution output is given on native vertical levels and on 48 pressure surfaces extending up to 0.02 hPa. Section 4 presents additional details on horizontal and vertical grids. Information of the model surface representation can be found in Appendix B. The GEOS-5 product is organized into file collections that are described in detail in Appendix C. Additional

  18. Control and evaluation programs for TANSY-KM5

    International Nuclear Information System (INIS)

    Grosshoeg, G.; Aronsson, D.; Hoek, M.; Rydz, R.; Norberg, L.; Urholm, L.

    1991-01-01

    Our first task was to purchase a computer and to create a basic software library for the preliminary test of the components. Our choice was a Starburst system from CES. The computer was a PDP-11/70 on a chip. It is the same computer as the ones used by the auxiliary CAMAC crate controllers in the TANSY system. However, the host has a 256 kbyte memory instead of the 64 kbyte memory used in the crate controllers. The operation system is RSX11-M. It is a real time system with properties suitable for laboratory works. However, the computer works in 64 kbyte partitions which limits the size of the programs. Four program systems were written. The two first were one-parameter systems for amplitude and time measurements using the ADCs and TDCs which later should be included in the TANSY system. These systems were supplemented with two two-parameter systems, one for time and amplitude measurements and one for time measurements on 16 detectors simultaneously. The display capabilities of these program systems have been used extensively by the TANSY programs. The programs needed for the manipulation of the system give the user a possibility to manually run the system without any intervention from the normal CODAS control system. The programs have been used for calibration measurements as well as test of the TANSY system. The evaluation programs use the output file from a normal measurement. The data file may be tested for errors. The true events can be sorted into different configurations. The final one is the neutron energy sort program, which gives the neutron energy distribution

  19. A 80 km reach fully passive WDM-PON based on reflective ONUs

    DEFF Research Database (Denmark)

    Presi, Marco; Proietti, Roberto; Prince, Kamau

    2008-01-01

    We propose a novel line coding combination (Inverse RZ coding in downlink and RZ in uplink) that extends the reach of WDM Passive Optical Networks based on Reflective SOAs with no in-line amplification. We achieved full downstream remodulation even when feeding the reflective SOA with power level...... as low as -35dBm, thus increasing the system power budget. We experimentally assessed this scheme for a fully passive, full-duplex and symmetrical 1.25Gb/s WDM-PON over a 80km G.652 feeder....

  20. ANALISA KEANDALAN SISTEM BAHAN BAKAR MOTOR INDUK PADA KM. LEUSER

    Directory of Open Access Journals (Sweden)

    Eko Sasmito Hadi

    2012-04-01

    Full Text Available Fuel oil system is one of the vital main engine supporting systems of MV. Leuser because this system is directly influence the main engine performance. The failure of fuel system should be the main engine shut down. The bigger affect was the ship could not be operated so the financial lost would be occurred. The aim of the research is an analysis the fuel system by approximation on reliability based. An analysis of the system was using qualitative and quantitative approximation. Qualitative analysis could be achieved by the Failure Mode and Effect Analysis (FMEA and the Fault Tree Analysis (FTA. The quantitative analysis could be achieved by the Monte Carlo simulation. The resulted of the research, severity components such as filter, separator, transfer pump and booster pump was obtained by the severity classification and failure probability. The simulation would be achieved the value availability at 0.97 and the value of MTTFF (Mean Time to First Failure is 2423.8363 hours. The simulation used some scenarios, the system availability would be more decrease if there were more stand by components occur the failure in the one of the components at 0,893.

  1. The Effect of Creatin and Carnitine Supplementation on 5 kmClassic and 10 km Free Styles Race Performance of Cross Country Skiers

    OpenAIRE

    Ebru ÇETİN

    2004-01-01

    This İnvestigation examined the effect of creatin and carnitine supplementation on 5 km classic and 10 km free styles race performance of competitive cross country skiers.Eighteen highly trained (12 male and 6 female) cross country skiers aged 13-16 years seperated into 3 equal groups. All groups participated in the 5 km classic and 10 km free races styles in Erciyes at 2200m altitude ski center before the carnitine and creatine loading. After the race subjects were seperated into carnitine, ...

  2. Data collection program for TANSY-KM5

    International Nuclear Information System (INIS)

    Grosshoeg, G.

    1991-01-01

    The purpose of the TANSY data collection program is dual. During normal measurement it fetches the incoming data, compresses them in a suitable shape and places the result in the CAMAC memory modules. The data should afterwards be taken care of by the JET Codas system. It may also be used as a stand-alone system. Calibrations can be done controlled by a terminal connected to the auxiliary CAMAC controller or by the host computer system. The program is written for the Starburst, the CES auxiliary CAMAC controller ACC 2180. It is essentially a PDP 11/70 computer on a microchip offering several advanced computer options. The program is written in assembler code in order to achieve shortest possible collection times. Some examples of user programs for the host are presented. They are written for RSX11/M operating system in a host Starburst. (au)

  3. Geology and petrography in basaltic rocks (Arapey formation) cropping out in road 4 between Arapey river (92 km) and Artigas city (200 Km)

    International Nuclear Information System (INIS)

    Oyhantcabal, P.; Pineiro, G.

    2007-01-01

    This contribution presents a geological map of the basaltic flows of Arapey formation (Mezosoic) cropping out in Road 4 between the Arapey river (92 Km) and Artigas city (200 Km) together with the description of the petrographic features of the different portions of the 13 recognized flow units. (author)

  4. Future changes in the East Asian rain band projected by global atmospheric models with 20-km and 60-km grid size

    Energy Technology Data Exchange (ETDEWEB)

    Kusunoki, Shoji; Mizuta, Ryo [Meteorological Research Institute, Climate Research Department, Tsukuba, Ibaraki (Japan); Matsueda, Mio [Japan Agency for Marine-Earth Science and Technology (JAMSTEC), Tsukuba, Ibaraki (Japan)

    2011-12-15

    Global warming projection experiments were conducted using a 20-km mesh global atmospheric model, focusing on the change in the rain band of East Asian summer monsoon. To assess the uncertainty of climate change projections, we performed ensemble simulations with the 60-km resolution model combining four different SSTs and three atmospheric initial conditions. In the present-day climate simulations, the 20-km model reproduces the rain band of East Asian summer monsoon better than lower resolution models in terms of geographical distribution and seasonal march. In the future climate simulation by the 20-km model, precipitation increases over the Yangtze River valley in May through July, Korean peninsula in May, and Japan in July. The termination of rainy season over Japan tends to be delayed until August. Ensemble simulations by the 60-km model show that precipitation in the future climate for July increases over the Yangtze River valley, the East China Sea and Japan. These changes in precipitation are partly consistent with those projected by the 20-km model. Simulations by the 20-km and 60-km models consistently show that in the future climate the termination of rainy season over Japan tends to be delayed until August. The changes in the vertically integrated water vapor flux show the intensification of clockwise moisture transport over the western Pacific subtropical high. Most precipitation changes over the East Asia can be interpreted as the moisture convergence resulting from change in the horizontal transport of water vapor. (orig.)

  5. Geology and petrography of the basaltic rocks (Arapey formation) cropping out in toad 4 between Arapey river (92 km) and Artigas (200 Km)

    International Nuclear Information System (INIS)

    Oyhantcabal, P.; Pineiro, G.

    2007-01-01

    This contribution presents a geological map of the basaltic flows of Arapey formation (Mezosoic) cropping out in Road 4 between the Arapey river (92 Km) and Artigas city (200 Km) together with the description of the petrographic features of the different portions of the 13 recognized flow units. (author)

  6. Geology and petrography in basaltic rocks (Arapey formation) cropping out in road 30 between the Bella Union round point (27 km) and Penas cuesta (225 Km)

    International Nuclear Information System (INIS)

    Oyhantcabal, P.; Pineiro, G.

    2007-01-01

    This contribution presents a geological map of the basaltic flows of Arapey formation (Mezosoic) cropping out in Road 30 between the Bella Union round point (27 Km) and Pena s cuesta (225 Km) together with the description of the petrographic features of the different portions of the 20 recognized flow units

  7. Photonics-oriented data transmission network for the KM3NeT prototype detector

    International Nuclear Information System (INIS)

    Hoek, M. van der; Mos, S.; Schmelling, J.W.; Hogenbirk, J.; Heine, E.; Jansweijer, P.; Kieft, G.; Peek, H.; Timmer, P.; Wolf, E. de; Zwart, A.

    2013-01-01

    The design of the readout and data acquisition system of the future KM3NeT neutrino telescope employs 10 Gbps photonic technologies for data transmission to shore. The photonic architecture can handle standard transmission protocols. The generic scheme is based on DWDM technology using lasers on shore and optical modulators in each of the 12,800 Digital Optical Modules arranged on several hundred vertical detection units anchored to the seabed. Each module will house 31 photomultipliers together with auxiliary instrumentation and readout electronics. A 100 km electro-optical fibre cable will connect the optical modules to the store. The readout system will guarantee an individual optical connection between each optical module and the shore. A small-scale prototype of a detection unit with four optical modules is in a realization phase and will allow for in situ testing of the data transmission network. We will present results of laboratory tests of the photonics-oriented transmission layer of the network that have been realized for the prototype detection unit

  8. MODIS/Aqua Clouds 5-Min L2 Swath 1km and 5km V5.1

    Data.gov (United States)

    National Aeronautics and Space Administration — MODIS was launched aboard the Aqua satellite on May 04, 2002 (1:30 pm equator crossing time) as part of NASA's Earth Observing System (EOS) mission. MODIS with its...

  9. KM Tools and Technologies that Share Distribute Knowledge

    Directory of Open Access Journals (Sweden)

    Ileana Hauer

    2008-10-01

    Full Text Available The exponential increase in information, primarily due to the electronic capture of data and its storage in vast data warehouses, has created a demand for analyzing the large amount of data generated by today’s organizations so that enterprise can respond quickly to fast changing markets. There are various tools and technologies that can be used to share and distribute knowledge, include e-mail, groupware, data mining ,expert systems and others. The paper outlines these technologies, which dominate the technical tools for sharing knowledge from an organizations data assets and finally. The case study, an Expert System, use expert knowledge to attain highlevel decision performance in a narrow domain.

  10. Gate-keeper module for TANSY-KM5

    International Nuclear Information System (INIS)

    Rydz, R.; Norberg, L.; Urholm, L.; Grosshoeg, G.

    1991-01-01

    The purpose of the Gate-keeper is the control of the RDCs, the ADCs, and the constant fraction discriminator. The Gate-keeper synchronizes the units and ensures that the data taking is clean and not intermixed with other events. There are six Gate-Keepers in the system, one for each proton detector. All input circuits are designed to accept TTL as well as negative NIM signals. The output is 50 ohm TTL or negative NIM as defined by internal jumpers

  11. The prototype detection unit of the KM3NeT detector

    NARCIS (Netherlands)

    Adrian-Martinez, S.; Ageron, M.; Aharonian, F.; Aiello, S.; Albert, A.; Ameli, F.; Anassontzis, E. G.; Androulakis, G. C.; Anghinolfi, M.; Anton, G.; Anvar, S.; Ardid, M.; Avgitas, T.; Balasi, K.; Band, H.; Barbarino, G.; Barbarito, E.; Barbato, F.; Baret, B.; Baron, S.; Barrios, J.; Belias, A.; Berbee, E.; van den Berg, A. M.; Berkien, A.; Bertin, V.; Beurthey, S.; van Beveren, V.; Beverini, N.; Biagi, S.; Biagioni, A.; Bianucci, S.; Billault, M.; Birbas, A.; Rookhuizen, H. Boer; Bormuth, R.; Bouche, V.; Bouhadef, B.; Bourlis, G.; Boutonnet, C.; Bouwhuis, M.; Bozza, C.; Bruijn, R.; Brunner, J.; Cacopardo, G.; Dorosti-Hasankiadeh, Q.; Hevinga, M. A.; Kavatsyuk, O.; Löhner, H.; van Wooning, R. H. L.

    2016-01-01

    A prototype detection unit of the KM3NeT deep-sea neutrino telescope has been installed at 3500m depth 80 km offshore the Italian coast. KM3NeT in its final configuration will contain several hundreds of detection units. Each detection unit is a mechanical structure anchored to the sea floor, held

  12. Plasmid (pKM101)-mediated enhancement of repair and mutagenesis: dependence on chromosomal genes in 'Escherichia coli' K-12

    International Nuclear Information System (INIS)

    Walker, G.C.

    1977-01-01

    The drug resistance plasmid pKM101 plays a major role in the Ames Salmonella/microsome carcinogen detecting system by enhancing chemical mutagenesis. It is shown that in Escherichia coli K-12 the plasmid pKM101 enhances both spontaneous and methyl methanesulfonate-caused reversion of an ochre mutation, bacterial survival after ultaviolet irradiation, and reactivation of ultraviolet-irradiated lambda in unirradiated cells. All these effects are shown to be dependent on the recA + lexA + genotype but not on the recB + recC + or recF + genotypes. The recA lexA-dependence of the plasmid-mediated repair and mutagenesis suggests an interaction with the cell's inducible error-prone repair system. The presence of pKM101 is shown to cause an additional increase in methyl methanesulfonate mutagenesis in a tif mutant beyond that caused by growth at 42 0 . The presence of the plasmid raises the level of the Weigle-reactivation curve for the reactivation of ultraviolet-irradiated lambda in E. coli and causes a shift of the maximum to a higher UV fluence. These observations suggest that pKM101 does not exert its effects by altering the regulation of the cell's error-prone repair system but rather by supplying a mechanistic component or components. (orig.) [de

  13. The competitive advantage of a dual-transporter system.

    Science.gov (United States)

    Levy, Sagi; Kafri, Moshe; Carmi, Miri; Barkai, Naama

    2011-12-09

    Cells use transporters of different affinities to regulate nutrient influx. When nutrients are depleted, low-affinity transporters are replaced by high-affinity ones. High-affinity transporters are helpful when concentrations of nutrients are low, but the advantage of reducing their abundance when nutrients are abundant is less clear. When we eliminated such reduced production of the Saccharomyces cerevisiae high-affinity transporters for phosphate and zinc, the elapsed time from the initiation of the starvation program until the lack of nutrients limited growth was shortened, and recovery from starvation was delayed. The latter phenotype was rescued by constitutive activation of the starvation program. Dual-transporter systems appear to prolong preparation for starvation and to facilitate subsequent recovery, which may optimize sensing of nutrient depletion by integrating internal and external information about nutrient availability.

  14. Estimation of Global 1km-grid Terrestrial Carbon Exchange Part I: Developing Inputs and Modelling

    Science.gov (United States)

    Sasai, T.; Murakami, K.; Kato, S.; Matsunaga, T.; Saigusa, N.; Hiraki, K.

    2015-12-01

    Global terrestrial carbon cycle largely depends on a spatial pattern in land cover type, which is heterogeneously-distributed over regional and global scales. However, most studies, which aimed at the estimation of carbon exchanges between ecosystem and atmosphere, remained within several tens of kilometers grid spatial resolution, and the results have not been enough to understand the detailed pattern of carbon exchanges based on ecological community. Improving the sophistication of spatial resolution is obviously necessary to enhance the accuracy of carbon exchanges. Moreover, the improvement may contribute to global warming awareness, policy makers and other social activities. In this study, we show global terrestrial carbon exchanges (net ecosystem production, net primary production, and gross primary production) with 1km-grid resolution. As methodology for computing the exchanges, we 1) developed a global 1km-grid climate and satellite dataset based on the approach in Setoyama and Sasai (2013); 2) used the satellite-driven biosphere model (Biosphere model integrating Eco-physiological And Mechanistic approaches using Satellite data: BEAMS) (Sasai et al., 2005, 2007, 2011); 3) simulated the carbon exchanges by using the new dataset and BEAMS by the use of a supercomputer that includes 1280 CPU and 320 GPGPU cores (GOSAT RCF of NIES). As a result, we could develop a global uniform system for realistically estimating terrestrial carbon exchange, and evaluate net ecosystem production in each community level; leading to obtain highly detailed understanding of terrestrial carbon exchanges.

  15. An overview of knowledge management (KM) issues for implementation in consultant firms in Malaysian construction industry

    Science.gov (United States)

    Othman, Azlan; Ismail, Syuhaida; Yahya, Khairulzan

    2017-12-01

    In the past few years, there has been a growing interest in treating knowledge as a significant organisational resource. Thus, effective development and implementation of KM requires a foundation in several rich literatures. As a preparation for the competitive industrial nation, KM is an important countenance that should be the point of convergence for the industry players. This paper wishes to draw the attention on the current situation of KM practice, focusing on consultant firms in Malaysian construction industry. Questionnaires were distributed to about 200 respondents working in the industry, with the objective of appraising the KM implementation amongst consultant firms working in construction industry in Malaysia. This paper also gives the overview on KM definition, process, understanding and challenges in construction industry, besides the critical success factor of KM implementation. The literature is restricted on the recent KM study of 17 years research from 2000 to 2017. Finally, this paper proposes the conceptual ideas of relationship between KM process, KM understanding and KM challenges with critical success factor of KM implementation.

  16. [An oral chemical vaccine from the hypertoxigenic strains of the causative agent of cholera KM-76 Inaba and KM-68 Ogawa].

    Science.gov (United States)

    Dzhaparidze, M N; Naumov, A V; Nikitina, G P; Meleshchenko, M V; Dobrova, G V; Zavorotnykh, V I; Gracheva, V P; Zakharova, T L

    1991-04-01

    The material on the development of chemical vaccine, prepared from two newly formed strains (KM-76 Inaba and KM-68 Ogawa) and intended for oral administration, is presented. The conditions for the submerged cultivation of these strains have been established, which makes it possible to increase the production of choleragen 8- to 10-fold and O-antigen 3- to 4-fold in comparison with V. cholerae natural strain 569B. The maximum accumulation of neuraminidase, protease, phospholipase, along with choleragen, has been registered in the logarithmic phase and that of O-antigen, in the stationary phase of growth. The use of strains KM-76 and KM-68 has led to the fourfold increase of the specific activity of the main immunogens, thus permitting the respective increase of the yield of the oral vaccine without changes in its high capacity for the formation of specific antibodies and its low residual toxigenicity.

  17. Fc receptor-γ subunits with both polar or non-polar amino acids at position of T22 are capable of restoring surface expression of the high-affinity IgE receptor and degranulation in γ subunit-deficient rat basophilic leukemia cells

    Czech Academy of Sciences Publication Activity Database

    Rashid, A.; Housden, J.E.; Helm, B.A.; Dráber, Petr

    2013-01-01

    Roč. 53, č. 3 (2013), s. 270-273 ISSN 0161-5890 R&D Projects: GA MŠk LD12073; GA ČR GA301/09/1826; GA ČR GAP302/10/1759; GA ČR(CZ) GBP302/12/G101; GA MŠk 1M0506 Grant - others:European Cooperation in Science and Technology(XE) BM1007; AV ČR(CZ) M200520901 Institutional research plan: CEZ:AV0Z50520514 Keywords : allergy * high-affinity IgE receptor * plasma membrane * transmembrane signaling * 3-helix assembly model Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.003, year: 2013

  18. [(3)H]8-Ethyl-4-methyl-2-phenyl-(8R)-4,5,7,8-tetrahydro-1H-imidazo[2,1-i]-purin-5-one ([(3)H]PSB-11), a novel high-affinity antagonist radioligand for human A(3) adenosine receptors.

    Science.gov (United States)

    Müller, Christa E; Diekmann, Martina; Thorand, Mark; Ozola, Vita

    2002-02-11

    This study describes the preparation and binding properties of [(3)H]PSB-11, a novel, potent, and selective antagonist radioligand for human A(3) adenosine receptors (ARs). [(3)H]PSB-11 binding to membranes of Chinese hamster ovary (CHO) cells expressing the human A(3) AR was saturable and reversible. Saturation experiments showed that [(3)H]PSB-11 labeled a single class of binding sites with high affinity (K(D)=4.9 nM) and limited capacity (B(max)=3500 fmol/mg of protein). PSB-11 is highly selective versus the other adenosine receptor subtypes. The new radioligand shows an extraordinarily low degree of non-specific binding rendering it a very useful tool for studying the (patho)physiological roles of A(3 )ARs.

  19. Lower thermosphere (80-100 km) dynamics response to solar and geomagnetic activity: Overview

    International Nuclear Information System (INIS)

    Kazimirovsky, E.S.

    1989-01-01

    The variations of solar and geomagnetic activity may affect the thermosphere circulation via plasma heating and electric fields, especially at high latitudes. The possibility exists that the energy involved in auroral and magnetic storms can produce significant changes of mesosphere and lower thermosphere wind systems. A study of global radar measurements of winds at 80 to 100 km region revealed the short term effects (correlation between wind field and geomagnetic storms) and long term variations over a solar cycle. It seems likely that the correlation results from a modification of planetary waves and tides propagated from below, thus altering the dynamical regime of the thermosphere. Sometimes the long term behavior points rather to a climatic variation with the internal atmospheric cause than to a direct solar control

  20. Radionuclide distributions in phytocenoses elements of the Chernobyl' NPP 30-km zone

    International Nuclear Information System (INIS)

    Fedotov, I.S.; Mishenkov, N.N.; Arkhipov, N.P.

    1989-01-01

    In order to estimate the radioecological situation in phytocenoses of the 30-km zone consisting in the main of conifer and conifer-foliage forests, the studies, which give, an opportunity to divide the pine forests into five zones according to degrees of radiation injury character revealing, are made. These zones are characterized by total death, strong injury, intermediate injury, weak injury and stimulation. Radionuclides redistribution in the system including wood fier, forest litter and soil, their accumulation in organs and elements if each cenosis component are studied. The characteristics of experimental sections are given. The data on radionuclide distributions in soil profile of forest tracts, radionuclide concentrations in pine organs, radionuclide contents in mushrooms (conifers), contamination distribution (%) in pines under different levels of soil contamination are given. 6 tabs

  1. Seismic Observations Indicating That the 2015 Ogasawara (Bonin) Earthquake Ruptured Beneath the 660 km Discontinuity

    Science.gov (United States)

    Kuge, Keiko

    2017-11-01

    The termination of deep earthquakes at a depth of 700 km is a key feature for understanding the physical mechanism of deep earthquakes. The 680 km deep 30 May 2015, Ogasawara (Bonin) earthquake (Mw 7.9) and its aftershocks were recorded by seismic stations at distances from 7° to 19°. Synthetic seismograms indicate that the P waveforms depend on whether the earthquake is located above or below the 660 km discontinuity. In this study, I show that broadband recordings indicate that the 2015 earthquake may have occurred below the 660 km velocity discontinuity. Recordings of the P wave from the strongest aftershock lack evidence for wave triplication expected when a subhorizontal discontinuity underlies the hypocenter. Theoretical waveforms computed with a 660 km discontinuity above the aftershock and mainshock match the observed waveforms more accurately. These observations may indicate earthquake ruptures due to mantle minerals other than olivine or strong deformation of the 660 km phase transition.

  2. Developing a Maturity Model for Knowledge Management (KM) in the Digital Age

    OpenAIRE

    Thornley, Clare; Carcary, Marian; Connolly, Niall; O'Duffy, Michael; Pierce, John

    2016-01-01

    This paper arises from a work-in-progress academia/industry collaborative research project to develop a knowledge management (KM) maturity model as a component (critical capability) of the IT Capability Maturity Framework (IT-CMF). The aim of the project is to develop a knowledge management (KM) maturity model that is ‘fit for purpose’ for organisations in the digital age. In undertaking this work it became clear that, outside of the fundamental challenges of KM, new significant challenges an...

  3. Protective effect of kombucha mushroom (KM) tea on phenol-induced cytotoxicity in albino mice.

    Science.gov (United States)

    Yapar, Kursad; Cavusoglu, Kultigin; Oruc, Ertan; Yalcin, Emine

    2010-09-01

    The present study was carried out to evaluate the protective role of kombucha mushroom (KM) tea on cytotoxicity induced by phenol (PHE) in mice. We used weight gain and micronucleus (MN) frequency as indicators of cytotoxicity and supported these parameters with pathological findings. The animals were randomly divided into seven groups: (Group I) only tap water (Group II) 1000 microl kg(-1) b. wt KM-tea, (Group III) 35 mg kg(-1) body wt. PHE (Group IV) 35 mg kg(-1) body wt. PHE + 250 microl kg(-1) b. wt KM-tea (Group V) 35 mg kg(-1) b. wt PHE + 500 microl kg(-1) b. wt KM-tea (Group VI) 35 mg kg(-1) b. wt PHE + 750 microl kg(-1) b. wt KM-tea, (Group VII) 35 mg kg(-1) b. wt PHE + 1000 microl kg(-1) b. wt KM-tea, for 20 consecutive days by oral gavage. The results indicated that all KM-tea supplemented mice showed a lower MN frequency than erythrocytes in only PHE-treated group. There was an observable regression on account of lesions in tissues of mice supplemented with different doses of KM-tea in histopathological observations. In conclusion, the KM-tea supplementation decreases cytotoxicity induced by PHE and its protective role is dose-dependent.

  4. Experimental Demonstration of 32 Gbaud 4-PAM for Data Center Interconnections of up to 320 km

    DEFF Research Database (Denmark)

    Madsen, Peter; Suhr, Lau Frejstrup; Clausen, Anders

    2017-01-01

    This paper presents experimental results demonstrating a 64 Gbps 4-PAM transmission over a 320 km SSMF span employing standard 80 km fiber spans for metro links. The receiver consists of a LPF and a DFE utilizing the DD-LMS algorithm.......This paper presents experimental results demonstrating a 64 Gbps 4-PAM transmission over a 320 km SSMF span employing standard 80 km fiber spans for metro links. The receiver consists of a LPF and a DFE utilizing the DD-LMS algorithm....

  5. Early changes of the anemia phenomenon in male 100-km ultramarathoners

    Directory of Open Access Journals (Sweden)

    Yu-Hui Chiu

    2015-02-01

    Conclusion: Running a 100-km ultramarathon will induce substantial sports anemia, and oxidative stress response, hemolysis, hematuria, and gastrointestinal bleeding are typical factors that contribute to its onset.

  6. Identification of high-affinity anti-IL-1 α autoantibodies in normal human serum as an interfering substance in a sensitive enzyme-linked immunosorbent assay for IL-1 α

    International Nuclear Information System (INIS)

    Mae, N.; Liberato, D.J.; Chizzonite, R.; Satoh, H.

    1991-01-01

    A highly reproducible, sensitive, and specific sandwich enzyme-linked immunosorbent assay (ELISA) for recombinant human IL-1 α (rhIL-1 alpha) has been developed. Results from this ELISA have demonstrated that the concentration of rhIL-1 α added to normal human serum (NHS) decreased by 16.3% after 3 h and 24.9% after 6 h at room temperature. Molecular exclusion column chromatography with Sephacryl S-300 HR revealed that 125I-labeled IL-1 α added to normal human serum rapidly formed higher molecular weight complexes without indication of proteolytic degradation. The observed reduction in immunoreactivity was correlated with this protein complex formation and accounted for the apparent instability of rhIL-1 α in NHS. Immunoblot analysis indicated that the molecular weight of the binding protein was 150-160K, and the IL-1 α binding activity was removed and recovered from NHS by Protein-G affinity chromatography; indicating that the binding protein was IL-1 α-specific IgG. The binding of 125I-labeled IL-1 α to the serum binding proteins could be inhibited by unlabeled IL-1 alpha (IC50 = 7.4 x 10(-11) M) but not by unlabeled IL-1 β. Kinetic analysis with 125I-labeled IL-1 alpha revealed that the average binding affinity of these IL-1 α-specific IgGs was 4.7 x 10(10) M-1. These results suggest that these autoantibodies may interfere with the detection of IL-1 α in human serum by various assay systems and also could be a regulator of circulating IL-1 α

  7. High-Mannose Specific Lectin and Its Recombinants from a Carrageenophyta Kappaphycus alvarezii Represent a Potent Anti-HIV Activity Through High-Affinity Binding to the Viral Envelope Glycoprotein gp120.

    Science.gov (United States)

    Hirayama, Makoto; Shibata, Hiromi; Imamura, Koji; Sakaguchi, Takemasa; Hori, Kanji

    2016-02-01

    We previously reported that a high-mannose binding lectin KAA-2 from the red alga Kappaphycus alvarezii, which is an economically important species and widely cultivated as a source of carrageenans, had a potent anti-influenza virus activity. In this study, the full-length sequences of two KAA isoforms, KAA-1 and KAA-2, were elucidated by a combination of peptide mapping and complementary DNA (cDNA) cloning. They consisted of four internal tandem-repeated domains, which are conserved in high-mannose specific lectins from lower organisms, including a cyanobacterium Oscillatoria agardhii and a red alga Eucheuma serra. Using an Escherichia coli expression system, an active recombinant form of KAA-1 (His-tagged rKAA-1) was successfully generated in the yield of 115 mg per liter of culture. In a detailed oligosaccharide binding analysis by a centrifugal ultrafiltration-HPLC method with 27 pyridylaminated oligosaccharides, His-tagged rKAA-1 and rKAA-1 specifically bound to high-mannose N-glycans with an exposed α1-3 mannose in the D2 arm as the native lectin did. Predicted from oligosaccharide binding specificity, a surface plasmon resonance analysis revealed that the recombinants exhibit strong interaction with gp120, a heavily glycosylated envelope glycoprotein of HIV with high association constants (1.48 - 1.61 × 10(9) M(-1)). Native KAAs and the recombinants inhibited the HIV-1 entry at IC50s of low nanomolar levels (7.3-12.9 nM). Thus, the recombinant proteins would be useful as antiviral reagents targeting the viral surface glycoproteins with high-mannose N-glycans, and the cultivated alga K. alvarezii could also be a good source of not only carrageenans but also this functional lectin(s).

  8. Position calibration for the future KM3NeT detector

    International Nuclear Information System (INIS)

    Motz, Holger

    2010-01-01

    Deep-sea neutrino telescopes consist of an array of photomultipliers to detect Cherenkov light emitted by neutrino-induced muons and particle showers in the surrounding sea water, allowing for reconstruction of the neutrino direction from position and timing of the Cherenkov photons. Since the photomultipliers are in most cases mounted on flexible structures, e.g. lines, and move with the sea current, a positioning system is required to determine the precise location of each sensor. The positioning system of the ANTARES neutrino telescope is based on acoustic triangulation using hydrophones mounted along the lines in combination with tiltmeters and compasses and provides centimetre precision alignment. For the future KM3NeT detector an Optical Module with integrated Piezo sensors for position calibration is proposed as a cost-effective solution. The performance of this system is tested with several sensors of the AMADEUS project, which is integrated in ANTARES to study the background for acoustic detection of highest energy neutrinos.

  9. Resting sympatho-vagal balance is related to 10 km running performance in master endurance athletes

    Directory of Open Access Journals (Sweden)

    Angelo Cataldo

    2018-02-01

    Full Text Available Relationships between heart rate recovery after exercise (HRR, baseline heart rate variability measures (HRV, and time to perform a 10Km running trial (t10Km were evaluated in "master" athletes of endurance to assess whether the measured indexes may be useful for monitoring the training status of the athletes. Ten “master” athletes of endurance, aged 40-60 years, were recruited. After baseline measures of HRV, the athletes performed a graded maximal test on treadmill and HRR was measured at 1 and 2 minutes from recovery. Subsequently they performed a 10Km running trial and t10Km was related to HRV and HRR indexes. The time to perform a 10Km running trial was significantly correlated with baseline HRV indexes. No correlation was found between t10Km and HRR. Baseline HRV measures, but not HRR, were significantly correlated with the time of performance on 10km running in “master” athletes. The enhanced parasympathetic function at rest appears to be a condition to a better performance on 10km running. HRV can be simple and useful measurements for monitoring the training stratus of athletes and their physical condition in proximity of a competition.

  10. MODIS/Terra Calibrated Radiances 5-Min L1B Swath 1km V006

    Data.gov (United States)

    National Aeronautics and Space Administration — The MODIS/Terra Calibrated Radiances 5-Min L1B Swath 1km (MOD021KM) contains calibrated and geolocated at-aperture radiances for 36 discrete bands located in the 0.4...

  11. 10 km running performance predicted by a multiple linear regression model with allometrically adjusted variables.

    Science.gov (United States)

    Abad, Cesar C C; Barros, Ronaldo V; Bertuzzi, Romulo; Gagliardi, João F L; Lima-Silva, Adriano E; Lambert, Mike I; Pires, Flavio O

    2016-06-01

    The aim of this study was to verify the power of VO 2max , peak treadmill running velocity (PTV), and running economy (RE), unadjusted or allometrically adjusted, in predicting 10 km running performance. Eighteen male endurance runners performed: 1) an incremental test to exhaustion to determine VO 2max and PTV; 2) a constant submaximal run at 12 km·h -1 on an outdoor track for RE determination; and 3) a 10 km running race. Unadjusted (VO 2max , PTV and RE) and adjusted variables (VO 2max 0.72 , PTV 0.72 and RE 0.60 ) were investigated through independent multiple regression models to predict 10 km running race time. There were no significant correlations between 10 km running time and either the adjusted or unadjusted VO 2max . Significant correlations (p 0.84 and power > 0.88. The allometrically adjusted predictive model was composed of PTV 0.72 and RE 0.60 and explained 83% of the variance in 10 km running time with a standard error of the estimate (SEE) of 1.5 min. The unadjusted model composed of a single PVT accounted for 72% of the variance in 10 km running time (SEE of 1.9 min). Both regression models provided powerful estimates of 10 km running time; however, the unadjusted PTV may provide an uncomplicated estimation.

  12. A Co-Creation Blended KM Model for Cultivating Critical-Thinking Skills

    Science.gov (United States)

    Yeh, Yu-chu

    2012-01-01

    Both critical thinking (CT) and knowledge management (KM) skills are necessary elements for a university student's success. Therefore, this study developed a co-creation blended KM model to cultivate university students' CT skills and to explore the underlying mechanisms for achieving success. Thirty-one university students participated in this…

  13. Ad-libitum drinking and performance during a 40-km cycling time trial in the heat

    NARCIS (Netherlands)

    Berkulo, M.A.R.; Bol, S.; Levels, K.; Lamberts, R.P.; Daanen, H.A.M.; Noakes, T.D.

    2015-01-01

    The aim of this study was to investigate if drinking ad-libitum can counteract potential negative effects of a hypohydrated start caused by fluid restriction during a 40-km time trial (TT) in the heat. Twelve trained males performed one 40-km cycling TT euhydrated (EU: no water during the TT) and

  14. Ad-libitum drinking and performance during a 40-km cycling time trial in the heat

    NARCIS (Netherlands)

    Berkulo, M.A.R.; Bol, S.; Levels, K.; Lamberts, R.P.; Daanen, H.A.M.; Noakes, T.D.

    2016-01-01

    The aim of this study was to investigate if drinking ad-libitum can counteract potential negative effects of a hypohydrated start caused by fluid restriction during a 40-km time trial (TT) in the heat. Twelve trained males performed one 40-km cycling TT euhydrated (EU: no water during the TT) and

  15. Assessment of the 2,4 km run as a predictor of aerobic capacity ...

    African Journals Online (AJOL)

    Since the 2,4 km run time test is routinely used in military training programmes as an indicator of aerobic capacity and its possible improvement, an attempt was made to: (i) establish a regression equation of V02max V. 2,4 km run time in a group of 20 young military volunteers; and (ii) determine whether this equation could ...

  16. MODIS/Terra Calibrated Radiances 5-Min L1B Swath 1km Subsetted V005

    Data.gov (United States)

    National Aeronautics and Space Administration — This data type (MOD02SSH) is a subsample from the MODIS Level 1B 1-km data. Every fifth pixel is taken from the MOD021KM product and written out to MOD02SSH. The...

  17. Towards an integrated methodology to develop KM solutions with the support of agents

    NARCIS (Netherlands)

    Guizzardi-Silva Souza, R.; Dignum, V.; Perini, A.; Wagner, G.; Thompson, C.; Hexmoor, H.

    Activities related to Knowledge Management (KM) processes require changes within the organizational processes in order to accommodate new goals and tasks, besides changing the way people view and do their work. In this paper, we argue that agents are suitable for modeling KM contexts, due to their

  18. Characteristic of lipid metabolism and state of free-radical processes in workers of 30-km alienation zone at Chernobyl Atomic Power Plant

    International Nuclear Information System (INIS)

    Chayalo, P.P.; Chobot'ko, G.M.; Palamar, L.A.; Kolesnik, L.L.; Kuznetsov, G.P.

    1997-01-01

    The study involved 54 men aged 35-50 working in 30 km alienation zone at the Chernobyl Atomic Power Plant. Blood serum and erythrocyte lipid peroxidation indices were estimated. Investigation of peroxidation processes in the erythrocytes allowed to reveal changes in glutation system, they being characterized by its amount elevation against the background of glutation transferase activity increase both in the persons, working in the 30 km zone, and in those from 'Ukryttia' Establishment

  19. Design and mass production of the optical modules for KM3NeT-Italia project

    Directory of Open Access Journals (Sweden)

    Leonora Emanuele

    2016-01-01

    Full Text Available The KM3NeT European project aims at constructing a km3 underwater neutrino telescope in the depths of the Mediterranean Sea. The first phase that is under construction will comprise eight tower-like detection structures (KM3NeT-Italia, which will form the internal core of a km3-scale detector. The detection element of KM3NeT-Italia, the optical module, is made of a 13-inch pressure-resistant glass-vessel that contains a single 10-inch photomultiplier and the relative electronics. The design of the whole optical module, the main results obtained from the massive photomultipliers measurements, and the foremost phases of the mass production procedure performed at the production site of Catania are also presented.

  20. Pure Rotational Raman Lidar for Temperature Measurements from 5-40 Km Over Wuhan, China

    Directory of Open Access Journals (Sweden)

    Li Yajuan

    2016-01-01

    Full Text Available In this paper a pure rotational Raman lidar (PRR was established for the atmospheric temperature measurements from 5 km to 40 km over Wuhan, China (30.5°N, 114.5°E. To extract the expected PRR signals and simultaneously suppress the elastically backscattered light, a high-spectral resolution polychromator for light splitting and filtering was designed. Observational results revealed that the temperature difference measured by PRR lidar and the local radiosonde below 30 km was less than 3.0 K. The good agreement validated the reliability of the PRR lidar. With the 1-h integration and 150-m spatial resolution, the statistical temperature error for PRR lidar increases from 0.4 K at 10 km up to 4 K at altitudes of about 30 km. In addition, the whole night temperature profiles were obtained for study of the long-term observation of atmospheric fluctuations.

  1. Dynamic behaviour of the contact line for 350 km/h on the new line Wuhan - Guangzhou; Dynamisches Verhalten der Oberleitung fuer 350 km/h auf der neuen Strecke Wuhan - Guangzhou

    Energy Technology Data Exchange (ETDEWEB)

    Zimmert, Gerhard [Balfour Beatty Rail, Beijing (China)

    2010-04-15

    Only after four and a half years of construction activities regular services started on the approximately 1 000 km long new high-speed line Wuhan - Guangzhou in the People's Republic of China. Achieving 320 to 330 km/h commercial speed, this line is the fastest railway connection in the world. The dynamic interaction between contact line and pantograph determines the maximally possible speed to an increasing extent. Worldwide, there is only low experience on the contact line dynamic behavior at this speed level. Balfour Beatty Rail designed the contact line for this installation, participated essentially in the implementation and, therefore, was in charge of proving the contact line quality. A series of test runs concerning the geometrical requirements and the contact force behaviour eventually proved the suitability of the system. (orig.)

  2. Design and stabilization works of the km 767 slope of Bolivia-Brazil gas pipeline; Projeto e obra de estabilizacao do talude do km 767 do gasoduto Bolivia-Brasil

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Hudson R.; Vasconcellos, Carlos Renato Aragonez de [TBG - Transportadora Brasileira Gasoduto Bolivia Brasil S.A., Rio de Janeiro, RJ (Brazil)

    2005-07-01

    The Bolivia-Brazil Natural Gas Pipeline starts at Santa Cruz de La Sierra city, in Bolivia, and goes until Canoas City (RS) in Brazil, with a total extent of 3,150 km. The pipeline crosses in the 2,593 km established in Brazilian soil, the most diverse types of geology and geomorphology. Along the line, the right-of-way (ROW) also crosses a lot of roads, railways, rivers and lakes. During a routine inspection (foot patrol), signs of instability were detected at an embankment slope of a highway of the Santa Catarina state, at the pipeline crossing. An eventual failure of this slope could put the pipeline at risk. The aim of this paper is to present the aspects of the stabilizations phases, since field investigation, design, works, instrumentation, until monitoring. Emphasis is given to the design criteria to pipeline safety. The solution adopted is composite by soil nailing, a changing of slope inclination and superficial drainage system. (author)

  3. Effects of training and anthropometric factors on marathon and 100 km ultramarathon race performance

    Directory of Open Access Journals (Sweden)

    Tanda G

    2015-04-01

    Full Text Available Giovanni Tanda,1 Beat Knechtle2,3 1Polytechnic School, University of Genoa, Genoa, Italy; 2Gesundheitszentrum St Gallen, St Gallen, 3Institute of Primary Care, University of Zurich, Zurich, Switzerland Background: Marathon (42 km and 100 km ultramarathon races are increasing in popularity. The aim of the present study was to investigate the potential associations of anthropometric and training variables with performance in these long-distance running competitions. Methods: Training and anthropometric data from a large cohort of marathoners and 100 km ultramarathoners provided the basis of this work. Correlations between training and anthropometric indices of subjects and race performance were assessed using bivariate and multiple regression analyses. Results: A combination of volume and intensity in training was found to be suitable for prediction of marathon and 100 km ultramarathon race pace. The relative role played by these two variables was different, in that training volume was more important than training pace for the prediction of 100 km ultramarathon performance, while the opposite was found for marathon performance. Anthropometric characteristics in terms of body fat percentage negatively affected 42 km and 100 km race performance. However, when this factor was relatively low (ie, less than 15% body fat, the performance of 42 km and 100 km races could be predicted solely on the basis of training indices. Conclusion: Mean weekly training distance run and mean training pace were key predictor variables for both marathon and 100 km ultramarathon race performance. Predictive correlations for race performance are provided for runners with a relatively low body fat percentage. Keywords: running, performance, training indices, body fat, sports training

  4. Evaluation of the Impacts of Assimilating the TAMDAR Data on 12/4 km Grid WRF-Based RTFDDA Simulations over the CONUS

    Directory of Open Access Journals (Sweden)

    Yongxin Zhang

    2016-01-01

    Full Text Available An analysis of the impacts of assimilating the Tropospheric Airborne Meteorological Data Report (TAMDAR data with the Weather Research and Forecasting- (WRF- real-time four-dimensional data assimilation (RTFDDA and forecasting system over the Contiguous US (CONUS is presented. The impacts of the horizontal resolution increase from 12 km to 4 km on the WRF-RTFDDA simulations are also examined in conjunction with the TAMDAR data impacts. The assimilation of the TAMDAR data reduces the root mean squared error of the moisture field predictions and increases the correlation between the predictions and the observations for both domains with 12 km and 4 km grid spacings. The TAMDAR data reduce the model dry biases in the middle and lower levels by adding moisture at those levels. Assimilating the TAMDAR data improves temperature predictions at middle to high levels and wind speed predictions at all levels especially for the 12 km domain. Increasing the horizontal resolution from 12 km to 4 km results in significantly larger impacts on surface variables than assimilating the TAMDAR data.

  5. Photoaffinity labelling of high affinity dopamine binding proteins

    International Nuclear Information System (INIS)

    Ross, G.M.; McCarry, B.E.; Mishra, R.K.

    1986-01-01

    A photoactive analogue of the dopamine agonist 2-amino-6,7-dihydroxy-1,2,3,4-tetrahydronapthalene (ADTN) has been synthesized and used to photoaffinity label dopamine binding proteins prepared from bovine caudate nucleus. N-(3-]N'-4-azidobenzamidol]-aminopropyl)-aminopropyl)-ADTN (AzB-AP-ADTN) was incubated with caudate membranes and irradiated with UV light. Membranes were then repeatedly washed by centrifugation to remove excess photolabel. A binding assay, using ( 3 H)-SCH 23390 (a D 1 specific antagonist), was then performed to evaluate the loss of receptor density in the photolyzed preparation. AzB-AP-ADTN irreversibly blocked ( 3 H)-SCH 23390 binding in a dose-dependent manner. Scatchard analysis revealed a decrease in the B/sub max/, with no significant change in the K/sub d/, of ( 3 H)-SCH 23390 binding. Compounds which compete for D 1 receptor binding (such as dopamine, SKF 38393 or apomorphine), proteted the SCH 23390 binding site from inactivation. This data would suggest that the novel photoaffinity ligand, AzB-AP-ADTN, can covalently label the D 1 (adenylate cyclase linked) dopamine receptor

  6. Global Investigation of the Mg Atom and ion Layers using SCIAMACHY/Envisat Observations between 70 km and 150 km Altitude and WACCM-MG Model Results

    Science.gov (United States)

    Langowski, M.; vonSavigny, C.; Burrows, J. P.; Feng, W.; Plane, J. M. C.; Marsh, D. R.; Janches, Diego; Sinnhuber, M.; Aikin, A. C.

    2014-01-01

    Mg and Mg+ concentration fields in the upper mesosphere/lower thermosphere (UMLT) region are retrieved from SCIAMACHY/Envisat limb measurements of Mg and Mg+ dayglow emissions using a 2-D tomographic retrieval approach. The time series of monthly means of Mg and Mg+ for number density as well as vertical column density in different latitudinal regions are shown. Data from the limb mesosphere-thermosphere mode of SCIAMACHY/Envisat are used, which covers the 50 km to 150 km altitude region with a vertical sampling of 3.3 km and a highest latitude of 82 deg. The high latitudes are not covered in the winter months, because there is no dayglow emission during polar night. The measurements were performed every 14 days from mid-2008 until April 2012. Mg profiles show a peak at around 90 km altitude with a density between 750 cm(exp-3) and 2000 cm(exp-3). Mg does not show strong seasonal variation at mid-latitudes. The Mg+ peak occurs 5-15 km above the neutral Mg peak at 95-105 km. Furthermore, the ions show a significant seasonal cycle with a summer maximum in both hemispheres at mid- and high-latitudes. The strongest seasonal variations of the ions are observed at mid-latitudes between 20-40 deg and densities at the peak altitude range from 500 cm(exp-3) to 6000 cm(exp-3). The peak altitude of the ions shows a latitudinal dependence with a maximum at mid-latitudes that is up to 10 km higher than the peak altitude at the equator. The SCIAMACHY measurements are compared to other measurements and WACCM model results. In contrast to the SCIAMACHY results, the WACCM results show a strong seasonal variability for Mg with a winter maximum, which is not observable by SCIAMACHY, and globally higher peak densities. Although the peak densities do not agree the vertical column densities agree, since SCIAMACHY results show a wider vertical profile. The agreement of SCIAMACHY and WACCM results is much better for Mg+, showing the same seasonality and similar peak densities. However

  7. Uptake of [N-Me-3H]-choline by synaptosomes from the central nervous system of Locusta migratoria

    International Nuclear Information System (INIS)

    Breer, H.

    1982-01-01

    The accumulation of 3H-choline by isolated synaptosomes from the central nervous system of locust was studied at concentrations varying from 0.05 to 40 microM. Kinetic analysis of the saturable process revealed a high-affinity and a low-affinity system. The high-affinity uptake was competitively inhibited by hemicholinium-3 and was absolutely dependent on external sodium. Elevated potassium concentrations inhibited choline uptake. The choline uptake by insect synaptosomes was found to be remarkably resistant to a variety of metabolic inhibitors. The reduced choline uptake under depolarizing conditions (high potassium concentration or veratridine) in the absence of calcium implies that electrochemical gradients are important for high-affinity choline uptake. Depolarization of preloaded synaptosomes under appropriate conditions resulted in a significant release of newly accumulated choline radioactivity

  8. A radiolabeled peptide ligand of the hERG channel, [125I]-BeKm-1

    DEFF Research Database (Denmark)

    Angelo, Kamilla; Korolkova, Yuliya V; Grunnet, Morten

    2003-01-01

    The wild-type scorpion toxin BeKm-1, which selectively blocks human ether-a-go-go related (hERG) channels, was radiolabeled with iodine at tyrosine 11. Both the mono- and di-iodinated derivatives were found to be biologically active. In electrophysiological patch-clamp recordings mono-[127I]-BeKm-1...... had a concentration of half-maximal inhibition (IC50 value) of 27 nM, while wild-type BeKm-1 inhibited hERG channels with an IC50 value of 7 nM. Mono-[125I]-BeKm-1 was found to bind in a concentration-dependent manner and with picomolar affinity to hERG channel protein in purified membrane vesicles...... of [125I]-BeKm-1 to the hERG channel to an IC50 of 7 nM. In autoradiographic studies on rat hearts, binding of [125I]-BeKm-1 was dose-dependent and could partially be displaced by the addition of excess amounts of non-radioactive BeKm-1. The density of the radioactive signal was equally distributed...

  9. Effect of β-alanine supplementation on 20 km cycling time trial performance

    Directory of Open Access Journals (Sweden)

    Ruth Margaret JAMES

    2014-09-01

    Full Text Available The effects of β-alanine supplementation on high-intensity cycling performance and capacity have been evaluated, although the effects on longer duration cycling performance are unclear. Nineteen UK category 1 male cyclists completed four 20 km cycling time trials, two before and two after supplementation with either 6.4 g•d-1 β-alanine (n = 10; BA or a matched placebo (n = 9; P. Performance time for the 20 km time trial and 1 km split times were recorded. There was no significant effect of β-alanine supplementation on 20 km time trial performance (BA-pre 1943 ± 129 s; BA-post 1950 ± 147 s; P-pre 1989 ± 106 s; P-post 1986 ± 115 s or on the performance of each 1 km split. The effect of β-alanine on 20 km time trial performance was deemed unclear as determined by magnitude based inferences. Supplementation with 6.4 g•d-1 of β-alanine for 4 weeks did not affect 20 km cycling time trial performance in well trained male cyclists.

  10. Drift velocities of 150-km Field-Aligned Irregularities observed by the Equatorial Atmosphere Radar

    Directory of Open Access Journals (Sweden)

    Yuichi Otsuka

    2013-11-01

    Full Text Available Between 130 and 170 km altitude in the daytime ionosphere, the so-called 150-km field-aligned irregularities (FAIs have been observed since the 1960s at equatorial regions with several very high frequency (VHF radars. We report statistical results of 150-km FAI drift velocities on a plane perpendicular to the geomagnetic field, acquired by analyzing the Doppler velocities of 150-km FAIs observed with the Equatorial Atmosphere Radar (EAR at Kototabang, Indonesia during the period from Aug. 2007 to Oct. 2009. We found that the southward/upward perpendicular drift velocity of the 150-km FAIs tends to decrease in the afternoon and that this feature is consistent with that of F-region plasma drift velocities over the magnetic equator. The zonal component of the 150-km FAI drift velocity is westward and decreases with time, whereas the F-region plasma drift velocity observed with the incoherent scatter radar at Jicamarca, Peru, which is westward, reaches a maximum at about noon. The southward/upward and zonal drift velocities of the 150-km FAIs are smaller than that of the F-region plasma drift velocity by approximately 3 m/s and 25 m/s, respectively, on average. The large difference between the 150-km FAI and F-region plasma drift velocities may not arise from a difference in the magnetic latitudes at which their electric fields are generated. Electric fields generated at the altitude at which the 150-km FAIs occur may not be negligible.

  11. Digital optical modules for the KM3NeT neutrino telescope

    Energy Technology Data Exchange (ETDEWEB)

    Kalekin, Oleg [Universitaet Erlangen, ECAP (Germany); Collaboration: ANTARES-KM3NeT-Erlangen-Collaboration

    2015-07-01

    KM3NeT is multi-cubic-kilometer neutrino telescope under construction in the Mediterranean Sea. In the currently running Phase 1 of the project, almost 30 detection units - 700 m tall vertical structures holding 18 Digital Optical Modules (DOMs) each - will be produced and deployed. A KM3NeT DOM consists of a pressure resistant glass sphere encapsulating 31 photomultiplier tubes of 80 mm diameter, readout electronics and additional instrumentation for calibration and monitoring. The Erlangen Centre for Astroparticle Physics is one of the DOM integration sites of the project. This contribution describes the design, functionality and integration procedure of the KM3NeT DOM.

  12. KM3NeT/ARCA sensitivity and discovery potential for neutrino point-like sources

    Directory of Open Access Journals (Sweden)

    Trovato A.

    2016-01-01

    Full Text Available KM3NeT is a large research infrastructure with a network of deep-sea neutrino telescopes in the abyss of the Mediterranean Sea. Of these, the KM3NeT/ARCA detector, installed in the KM3NeT-It node of the network, is optimised for studying high-energy neutrinos of cosmic origin. Sensitivities to galactic sources such as the supernova remnant RXJ1713.7-3946 and the pulsar wind nebula Vela X are presented as well as sensitivities to a generic point source with an E−2 spectrum which represents an approximation for the spectrum of extragalactic candidate neutrino sources.

  13. Flooding Causes Analysis in The Engine Room of KM. Nusantara Akbar

    Directory of Open Access Journals (Sweden)

    Trika Pitana

    2017-03-01

    Full Text Available The study analyzes about the cause of the leak in KM Nusantara Akbar’s engine room. However, this research is focus on  leakage caused by shaft deflectio From the field data provided by the KNKT, know the cause of the leak is packing driven by a shaft deflection and therefore can not hold the rate of water. Analysis was done by a review of the technical and non-technical factors, because the scene of the ship accident was not purely due to technical factors but also non-technical factors. The first step is analyzing the ship document, such as the certificate of ship from classification, incident reports, docking report, ship crew certificates and other ship document. Then on the next step, the evidence that has been obtained from the analysis of the documents related to the  ship sail is use to make the analysis using 5 whys method to looking for the root cause. According to the analysis that has been done, the technical cause of the leak vessel caused by system shafting system is reconditioned flange bolts that have been damaged and the addition of flax on the bearing shaft has to cope without straightening axle deflection itself. While the Autodesk Inventor 3D modeling software obtained bolt broken because the act force is 10782.31 N with the shear stress 2.230 MPa, while the maximum force in the normal load is 9.434,531.N with shear force 1,951 Mpa. Then from the cause from non-technical factors are equipment / spare part to overcome the failure of shafting system is very less, the workplace is dirty and uncomfortable and happened miss communication between the crew in the engine room to overcome the leakage.

  14. Patterns of Mass Mortality among Rocky Shore Invertebrates across 100 km of Northeastern Pacific Coastline.

    Directory of Open Access Journals (Sweden)

    Laura J Jurgens

    Full Text Available Mass mortalities in natural populations, particularly those that leave few survivors over large spatial areas, may cause long-term ecological perturbations. Yet mass mortalities may remain undocumented or poorly described due to challenges in responding rapidly to unforeseen events, scarcity of baseline data, and difficulties in quantifying rare or patchily distributed species, especially in remote or marine systems. Better chronicling the geographic pattern and intensity of mass mortalities is especially critical in the face of global changes predicted to alter regional disturbance regimes. Here, we couple replicated post-mortality surveys with preceding long-term surveys and historical data to describe a rapid and severe mass mortality of rocky shore invertebrates along the north-central California coast of the northeastern Pacific Ocean. In late August 2011, formerly abundant intertidal populations of the purple sea urchin (Strongylocentrotus purpuratus, a well-known ecosystem engineer, and the predatory six-armed sea star (Leptasterias sp. were functionally extirpated from ~100 km of coastline. Other invertebrates, including the gumboot chiton (Cryptochiton stelleri the ochre sea star (Pisaster ochraceus, and subtidal populations of purple sea urchins also exhibited elevated mortality. The pattern and extent of mortality suggest the potential for long-term population, community, and ecosystem consequences, recovery from which may depend on the different dispersal abilities of the affected species.

  15. Combined Opto-Acoustical sensor modules for KM3NeT

    International Nuclear Information System (INIS)

    Enzenhöfer, A.

    2013-01-01

    KM3NeT is a future multi-cubic-kilometre water Cherenkov neutrino telescope currently entering a first construction phase. It will be located in the Mediterranean Sea and comprise about 600 vertical structures called detection units. Each of these detection units has a length of several hundred metres and is anchored to the sea bed on one side and held taut by a buoy on the other side. The detection units are thus subject to permanent movement due to sea currents. Modules holding photosensors and additional equipment are equally distributed along the detection units. The relative positions of the photosensors has to be known with an uncertainty below 20 cm in order to achieve the necessary precision for neutrino astronomy. These positions can be determined with an acoustic positioning system: dedicated acoustic emitters located at known positions and acoustic receivers along each detection unit. This article describes the approach to combine an acoustic receiver with the photosensors inside one detection module using a common power supply and data readout. The advantage of this approach lies in a reduction of underwater connectors and module configurations as well as in the compactification of the detection units integrating the auxiliary devices necessary for their successful operation.

  16. MODIS/Terra Geolocation Fields 5-Min L1A Swath 1km V005

    Data.gov (United States)

    National Aeronautics and Space Administration — The geolocation fields are calculated for each 1 km MODIS Instantaneous Field of Views (IFOV) for all orbits daily. The locations and ancillary information...

  17. LBA-ECO CD-04 Dendrometry, km 83 Tower Site, Tapajos National Forest, Brazil

    Data.gov (United States)

    National Aeronautics and Space Administration — A dendrometry study was conducted at the logged forest tower site, km 83 site, Tapajos National Forest, Para, Brazil over a period of 4 years following the...

  18. 14 km Sea Surface Temperature for North America, 1986 - present (NODC Accession 0099042)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This product presents local sea surface temperatures (degrees C). It is a composite gridded-image derived from 8-km resolution SST observations collected by Advanced...

  19. LBA-ECO CD-04 Meteorological and Flux Data, km 83 Tower Site, Tapajos National Forest

    Data.gov (United States)

    National Aeronautics and Space Administration — Tower flux measurements of carbon dioxide,water vapor, heat, and meteorological variables were obtained at the Tapajos National Forest, km 83 site, Santarem, Para,...

  20. Gridded 5km GHCN-Daily Temperature and Precipitation Dataset, Version 1

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The Gridded 5km GHCN-Daily Temperature and Precipitation Dataset (nClimGrid) consists of four climate variables derived from the GHCN-D dataset: maximum temperature,...

  1. SAFARI 2000 Land Cover from AVHRR, 8-km, 1984 (DeFries et al.)

    Data.gov (United States)

    National Aeronautics and Space Administration — This data set consists of a southern African subset of the University of Maryland (UMD) 8-km Global Land Cover product in ASCII GRID and binary image files formats.

  2. LBA Regional Land Cover from AVHRR, 8-km, 1984 (DeFries et al.)

    Data.gov (United States)

    National Aeronautics and Space Administration — This data set is a subset of an 8-km global land cover product (DeFries et al. 1998). This subset was created for the study area of the Large Scale...

  3. LBA Regional Tree Cover from AVHRR, 1-km, 1992-1993 (DeFries et al.)

    Data.gov (United States)

    National Aeronautics and Space Administration — This data set is a subset of the 1-km global tree cover data set (DeFries et al. 1999) developed at the Laboratory for Global Remote Sensing Studies (LGRSS) at the...

  4. LBA Regional Tree Cover from AVHRR, 1-km, 1992-1993 (DeFries et al.)

    Data.gov (United States)

    National Aeronautics and Space Administration — ABSTRACT: This data set is a subset of the 1-km global tree cover data set (DeFries et al. 1999) developed at the Laboratory for Global Remote Sensing Studies...

  5. SAFARI 2000 Tree Cover from AVHRR, 1-km, 1992-1993 (DeFries et al.)

    Data.gov (United States)

    National Aeronautics and Space Administration — This data set consists of a southern African subset of the 1-km Global Tree Cover Data Set developed at the Laboratory for Global Remote Sensing Studies (LGRSS) at...

  6. SAFARI 2000 Land Cover from AVHRR, 8-km, 1984 (DeFries et al.)

    Data.gov (United States)

    National Aeronautics and Space Administration — ABSTRACT: This data set consists of a southern African subset of the University of Maryland (UMD) 8-km Global Land Cover product in ASCII GRID and binary image files...

  7. SAFARI 2000 Tree Cover from AVHRR, 1-km, 1992-1993 (DeFries et al.)

    Data.gov (United States)

    National Aeronautics and Space Administration — ABSTRACT: This data set consists of a southern African subset of the 1-km Global Tree Cover Data Set developed at the Laboratory for Global Remote Sensing Studies...

  8. LBA Regional Land Cover from AVHRR, 8-km, 1984 (DeFries et al.)

    Data.gov (United States)

    National Aeronautics and Space Administration — ABSTRACT: This data set is a subset of an 8-km global land cover product (DeFries et al. 1998). This subset was created for the study area of the Large Scale...

  9. LBA-ECO CD-04 Leaf Litter Data, km 83 Tower Site, Tapajos National Forest, Brazil

    Data.gov (United States)

    National Aeronautics and Space Administration — Above-ground litter productivity was measured in a 18 ha plot adjacent to the eddy flux tower at the logged forest tower site, km 83, Tapajos National Forest, Para,...

  10. LBA-ECO CD-10 Forest Litter Data for km 67 Tower Site, Tapajos National Forest

    Data.gov (United States)

    National Aeronautics and Space Administration — ABSTRACT: This data set contains a single text file which reports litter type and mass in the old-growth upland forest at the Para Western (Santarem) - km 67,...

  11. LBA-ECO CD-04 Leaf Area Index, km 83 Tower Site, Tapajos National Forest, Brazil

    Data.gov (United States)

    National Aeronautics and Space Administration — ABSTRACT: Leaf area index was estimated in an 18 ha plot at the logged forest tower site, km 83, Tapajos National Forest, Para, Brazil. The plot was adjacent to the...

  12. LBA-ECO CD-04 Leaf Litter Data, km 83 Tower Site, Tapajos National Forest, Brazil

    Data.gov (United States)

    National Aeronautics and Space Administration — ABSTRACT: Above-ground litter productivity was measured in a 18 ha plot adjacent to the eddy flux tower at the logged forest tower site, km 83, Tapajos National...

  13. LBA-ECO CD-04 Leaf Area Index, km 83 Tower Site, Tapajos National Forest, Brazil

    Data.gov (United States)

    National Aeronautics and Space Administration — Leaf area index was estimated in an 18 ha plot at the logged forest tower site, km 83, Tapajos National Forest, Para, Brazil. The plot was adjacent to the eddy flux...

  14. Spetsialistid soovitavad kesklinnas piirata sõidukite kiirust 40 km/h-ni / Askur Alas

    Index Scriptorium Estoniae

    Alas, Askur, 1973-

    2004-01-01

    Liiklusspetsialistid soovitavad inimvigastustega õnnetuste vähendamiseks piirata Tallinna kesklinnas kiirust 40 km/h-ni. Linna liiklusteenistus ei pea seda põhjendatuks. Lisa: Helsingi positiivne kogemus

  15. NACP North American 8-km Net Ecosystem Exchange and Component Fluxes, 2004

    Data.gov (United States)

    National Aeronautics and Space Administration — ABSTRACT: This data set provides modeled carbon flux estimates at 8-km spatial resolution over North America for the year 2004 of (1) net ecosystem exchange (NEE) of...

  16. NACP North American 8-km Net Ecosystem Exchange and Component Fluxes, 2004

    Data.gov (United States)

    National Aeronautics and Space Administration — This data set provides modeled carbon flux estimates at 8-km spatial resolution over North America for the year 2004 of (1) net ecosystem exchange (NEE) of carbon...

  17. Internal γ Decay and the Superallowed Branching Ratio for the β+ Emitter Km38

    Science.gov (United States)

    Leach, K. G.; Svensson, C. E.; Ball, G. C.; Leslie, J. R.; Austin, R. A. E.; Bandyopadhyay, D.; Barton, C.; Bassiachvilli, E.; Ettenauer, S.; Finlay, P.; Garrett, P. E.; Grinyer, G. F.; Hackman, G.; Melconian, D.; Morton, A. C.; Mythili, S.; Newman, O.; Pearson, C. J.; Pearson, M. R.; Phillips, A. A.; Savajols, H.; Schumaker, M. A.; Wong, J.

    2008-05-01

    The branching ratio for the superallowed β+ decay of Km38 was measured at TRIUMF’s ISAC radioactive ion beam facility. The M3 internal transition between the isomer and the ground state of Km38 was observed with a branching ratio of 330(43) ppm. A search for the nonanalogue β-decay branch to the first excited 0+ state in Ar38 was also performed and yielded an upper limit of ≤12ppm at 90% C.L. These measurements lead to a revised superallowed branching ratio for Km38 of 99.967(4)%, and increase the Km38 ft value by its entire quoted uncertainty to ft=3052.1(10)s. Implications for tests of the nuclear-structure dependent corrections in superallowed β decays and the extraction of the Cabibbo-Kobayashi-Maskawa matrix element Vud are discussed.

  18. MODIS/Aqua Aerosol 5-Min L2 Swath 10km V006

    Data.gov (United States)

    National Aeronautics and Space Administration — The MODIS/Aqua Aerosol 5-Min L2 Swath 10km (MYD04_L2) product continues to provide full global coverage of aerosol properties from the Dark Target (DT) and Deep Blue...

  19. LBA-ECO CD-10 Temperature Profiles at km 67 Tower Site, Tapajos National Forest

    Data.gov (United States)

    National Aeronautics and Space Administration — This data set contains a single text file which reports temperature measurements at the Para Western (Santarem) - km 67, Primary Forest Tower Site. This site is in...

  20. LBA-ECO CD-10 Temperature Profiles at km 67 Tower Site, Tapajos National Forest

    Data.gov (United States)

    National Aeronautics and Space Administration — ABSTRACT: This data set contains a single text file which reports temperature measurements at the Para Western (Santarem) - km 67, Primary Forest Tower Site. This...

  1. LBA-ECO CD-10 Forest Litter Data for km 67 Tower Site, Tapajos National Forest

    Data.gov (United States)

    National Aeronautics and Space Administration — This data set contains a single text file which reports litter type and mass in the old-growth upland forest at the Para Western (Santarem) - km 67, Primary Forest...

  2. LBA-ECO ND-02 Soil Gas Flux, Rainfall Exclusion, km 67, Tapajos National Forest

    Data.gov (United States)

    National Aeronautics and Space Administration — ABSTRACT: This data set reports the results of a rainfall exclusion experiment in the Tapajos National Forest (Flona-Tapajos) at km 67 along the Santarem-Cuiaba...

  3. LBA-ECO ND-02 Soil Gas Flux, Rainfall Exclusion, km 67, Tapajos National Forest

    Data.gov (United States)

    National Aeronautics and Space Administration — This data set reports the results of a rainfall exclusion experiment in the Tapajos National Forest (Flona-Tapajos) at km 67 along the Santarem-Cuiaba BR-163...

  4. Foot Temperatures and Toe Blood Flow during a 12 km Winter Hike and Guard Duty

    National Research Council Canada - National Science Library

    Mekjavic, Igor B; Kocjan, Nina; Vrhovec, Miro; Golja, Petra; House, Carol; Eiken, Ola

    2005-01-01

    .... During the 3-week study, the trails were covered with snow. Peripheral vasodilatation, presumably as a result of the elevated core temperature, maintained average skin temperature constant during the 12 km hike, and increased toe temperature...

  5. Depth and sharpness variations of 410-km and 660-km discontinuities in North China Craton from dense array ambient noise interferometry

    Science.gov (United States)

    Yao, H.; Feng, J.; Poli, P.; Fang, L.; Wu, Y.

    2017-12-01

    Recent studies have demonstrated that body waves between pairs of stations can be successfully retrieved from ambient noise cross-correlation at both regional and global scales, although surface waves are the dominant signals. However, it is still difficult to use these retrieved body wave signals to map lateral depth variations of main structural discontinuities or velocity contrasts because of its low signal to noise ratio (SNR). In this research, based on a dense seismic array in eastern North China Craton, reflected P-wave signals from 410-km and 660-km discontinuities can be successfully recovered from ambient noise cross-correlation. To improve SNR of the reflected phases, the cross-correlations are stacked within each bin with the phase-weighted stack (PWS) method. In addition, there exist apparent spatial variations of the relative amplitude ratios between the reflected P410P and P660P phases. The retrieved P410P and P660P phases from stacked correlations reveal lateral variations of both depths and sharpness of the 410-km and 660-km discontinuities along two profiles, which may be related with hot material upwelling and the effect of stagnant Pacific plate in the transition zone beneath North China Craton. The imaging results are generally consistent with the results from teleseismic receiver functions, which demonstrate the possibility of mapping high-resolution topography and sharpness of deep internal discontinuities without earthquake-station geometric limitations. Future joint imaging with both interferometric and passive signals will be better for understanding interface architectures and related dynamic processes of the Earth.

  6. Exploration of the dopamine transporter: in vitro and in vivo characterization of a high-affinity and high-specificity iodinated tropane derivative (E)-N-(3-iodoprop-2-enyl)-2β-carbomethoxy- 3β-(4'-methylphenyl)nortropane (PE2I)

    International Nuclear Information System (INIS)

    Guilloteau, Denis; Emond, Patrick; Baulieu, Jean-Louis; Garreau, Lucette; Frangin, Yves; Pourcelot, Leandre; Mauclaire, Laurent; Besnard, Jean-Claude; Chalon, Sylvie

    1998-01-01

    For the diagnosis and follow-up of neurodegenerative diseases, many cocaine derivatives have been proposed as radioligands to explore the dopamine transporter. As none of them have all the criteria of specificity and kinetics for human use, we have developed a new derivative, (E)-N-(3-iodoprop-2-enyl)-2β-carbomethoxy-3β-(4'-methylphenyl)nortropane (PE2I), which displays promising properties. We report the characterization of PE2I in vitro on rat striatal membranes and in vivo in rats and in monkeys. PE2I had a high affinity (Kd=0.09±0.01 nM) and high specificity for the dopamine transporter. In rats we observed a high accumulation in the striatum; by contrast, a very low fixation was measured in the cortex. Moreover, a preinjection of a saturating dose of GBR 12909 prevented the striatal accumulation of PE2I by 74%. These results confirmed the specificity of PE2I for the dopamine transporter. In vivo in monkeys, SPECT studies showed a high accumulation in striatum. Moreover, an equilibrium state was obtained 1 h after injection. PE2I seemed to be the most promising ligand for the dopamine transporter exploration by SPECT using a single-day protocol.

  7. Projection of the change in future extremes over Japan using a cloud-resolving model: (2) Precipitation Extremes and the results of the NHM-1km experiments

    Science.gov (United States)

    Kanada, S.; Nakano, M.; Nakamura, M.; Hayashi, S.; Kato, T.; Kurihara, K.; Sasaki, H.; Uchiyama, T.; Aranami, K.; Honda, Y.; Kitoh, A.

    2008-12-01

    In order to study changes in the regional climate in the vicinity of Japan during the summer rainy season due to global warming, experiments by a semi-cloud resolving non-hydrostatic model with a horizontal resolution of 5km (NHM-5km) have been conducted from June to October by nesting within the results of the 10-year time-integrated experiments using a hydrostatic atmospheric general circulation model with a horizontal grid of 20 km (AGCM-20km: TL959L60) for the present and future up to the year 2100. A non-hydrostatic model developed by the Japan Meteorological Agency (JMA) (JMA-NHM; Saito et al. 2001, 2006) was adopted. Detailed descriptions of the NHM-5km are shown by the poster of Nakano et al. Our results show that rainy days over most of the Japanese Islands will decrease in June and July and increase in August and September in the future climate. Especially, remarkable increases in intense precipitations such as larger than 150 - 300 mm/day are projected from the present to future climate. The 90th percentiles of regional largest values among maximum daily precipitations (R-MDPs) grow 156 to 207 mm/day in the present and future climates, respectively. It is well-known that the horizontal distribution of precipitation, especially the heavy rainfall in the vicinity of Japan, much depends on the topography. Therefore, higher resolution experiments by a cloud-resolving model with a horizontal resolution of 1km (NHM-1km) are one-way nested within the results of NHM-5km. The basic frame and design of the NHM-1km is the same as those of the NHM-5km, but the topography is finer and no cumulus parameterization is used in the NHM-1km experiments. The NHM-1km, which treats the convection and cloud microphysics explicitly, can represent not only horizontal distributions of rainfall in detail but also the 3-dimensional structures of meso-beta-scale convective systems (MCSs). Because of the limitation of computation resources, only heavy rainfall events that rank in top

  8. Full-duplex transmission of IEEE 802.11ac-compliant MIMO WLAN signals over a 2-km 7-core fiber

    NARCIS (Netherlands)

    Fan, Yuting; Li, Jianqiang; Lei, Yi; Tang, Ming; Yin, Feifei; Dai, Yitang; Xu, Kun

    2017-01-01

    In this Letter, we experimentally demonstrate a full-duplex transmission system of IEEE 802.11ac-compliant multiple-input multiple-output (MIMO) signals over a 2-km 7-core fiber for in-building wireless local-area network (WLAN) distributed antenna systems. For full-duplex 3 � 3 MIMO

  9. Future changes in Asian summer monsoon precipitation extremes as inferred from 20-km AGCM simulations

    Science.gov (United States)

    Lui, Yuk Sing; Tam, Chi-Yung; Lau, Ngar-Cheung

    2018-04-01

    This study examines the impacts of climate change on precipitation extremes in the Asian monsoon region during boreal summer, based on simulations from the 20-km Meteorological Research Institute atmospheric general circulation model. The model can capture the summertime monsoon rainfall, with characteristics similar to those from Tropical Rainfall Measuring Mission and Asian Precipitation-Highly-Resolved Observational Data Integration Towards Evaluation. By comparing the 2075-2099 with the present-day climate simulations, there is a robust increase of the mean rainfall in many locations due to a warmer climate. Over southeastern China, the Baiu rainband, Bay of Bengal and central India, extreme precipitation rates are also enhanced in the future, which can be inferred from increases of the 95th percentile of daily precipitation, the maximum accumulated precipitation in 5 consecutive days, the simple daily precipitation intensity index, and the scale parameter of the fitted gamma distribution. In these regions, with the exception of the Baiu rainband, most of these metrics give a fractional change of extreme rainfall per degree increase of the lower-tropospheric temperature of 5 to 8.5% K-1, roughly consistent with the Clausius-Clapeyron relation. However, over the Baiu area extreme precipitation change scales as 3.5% K-1 only. We have also stratified the rainfall data into those associated with tropical cyclones (TC) and those with other weather systems. The AGCM gives an increase of the accumulated TC rainfall over southeastern China, and a decrease in southern Japan in the future climate. The latter can be attributed to suppressed TC occurrence in southern Japan, whereas increased accumulated rainfall over southeastern China is due to more intense TC rain rate under global warming. Overall, non-TC weather systems are the main contributor to enhanced precipitation extremes in various locations. In the future, TC activities over southeastern China tend to further

  10. Results of the first detection units of KM3NeT

    Science.gov (United States)

    Biagi, Simone; KM3NeT Collaboration

    2017-12-01

    The KM3NeT collaboration is building a km3-scale neutrino telescope in the Mediterranean Sea. The current phase of construction comprises the deep-sea and onshore infrastructures at two installation sites and the installation of the first detection units for the "ARCA" (Astroparticle Research with Cosmics in the Abyss) and "ORCA" (Oscillation Research with Cosmics in the Abyss) detector. At the KM3NeT-It site, 80 km offshore Capo Passero, Italy, the first 32 detection units for the ARCA detector are being installed and at the KM3Net-Fr site, 40 km offshore Toulon, France, 7 detection units for the ORCA detector will be deployed. The second phase of KM3NeT foresees the completion of ARCA for neutrino astronomy at energies above TeV and ORCA for neutrino mass hierarchy studies at energies in the GeV range. The basic element of the KM3NeT detector is the detection unit. In the ARCA geometry, the detection unit is a 700 m long vertical structure hosting 18 optical modules. Each optical module comprises 31 3 in photomultiplier tubes, instruments to monitor environmental parameters, and the electronic boards for the digitisation of the PMT signals and the management of data acquisition. In their final configuration, both ARCA and ORCA will be composed of about 200 detection units. The first detection unit was installed at the KM3NeT-It site in December 2015. It is active and taking data since its connection to the subsea network. The time of arrival and the duration of photon hits on each of the photomultipliers is measured with a time resolution of 1 ns and transferred onshore where the measurements are processed, triggered and stored on disk. A time calibration procedure, based on data recorded with flashing LED beacons during dedicated periods, allows for time synchronisation of the signals from the optical modules at the nanosecond level. In May 2016, an additional detection unit was installed at the KM3NeT-It site. The first results with two active detection units

  11. Effects of Iron and Aluminum on Phase Boundaries at 600-800 km Depths

    Science.gov (United States)

    Shim, Sang-Heon; Ye, Yu; Prakapenka, Vitali; Meng, Yue

    2014-05-01

    High-resolution seismic studies have reported complex discontinuity structures at 600-800 km depths. However, the origin of the structures have not been well understood. In order to understand compositional effects, we have measured the post-spinel, post-garnet, and post-ilmenite phase boundaries in MgO-Al2O3-SiO2 (iron free) and CaO-MgO-Al2O3-SiO2-FeO (iron bearing) systems with pyrolitic oxide ratios. In-situ X-ray diffraction measurements were performed at 20-30 GPa and 1500-2300 K in the laser-heated diamond-anvil cell at the GSECARS and HPCAT sectors of the Advanced Photon Source. We use the Pt and Au pressure scales for the iron-free and iron-bearing compositions, respectively. The Pt and Au scales were calibrated with respect to each other in separate experiments. In most experiments, Ar was cryogenically loaded in the sample chamber as a thermal insulation and pressure transmitting medium, except for a few experiments where a KCl medium was used. At temperatures above 1900 K, the post-garnet transition occurs at higher pressures than the post-spinel transition in both the iron-free and iron-bearing systems. At lower temperatures, while the post-ilmenite transition occurs at nearly same pressures as the post-spinel transition in the iron-bearing system, the post-ilmenite transition occurs at slightly higher pressure (1 GPa) than the post-spinel transitions in the iron-free system. In the iron-free system, akimotoite is stable to much higher temperature (2300 K) than previously thought. In the iron-bearing system, the stability of akimotoite is limited to 2050 K. Our data indicate that Al partitions more into akimotoite than garnet in the iron-free system, which is the opposite to what has been found in iron-bearing systems. The high Al content in akimotoite seems to be responsible for the high-temperature stability of akimotoite in the iron-free system. The Clapeyron slope of the post-garnet boundary is greater by a factor of 2.5 in the iron-bearing system

  12. The isolated 678-km deep 30 May 2015 MW 7.9 Ogasawara (Bonin) Islands earthquake

    Science.gov (United States)

    Ye, L.; Lay, T.; Zhan, Z.; Kanamori, H.; Hao, J.

    2015-12-01

    Deep-focus earthquakes, located 300 to 700 km below the Earth's surface within sinking slabs of relatively cold oceanic lithosphere, are mysterious phenomena. Seismic radiation from deep events is essentially indistinguishable from that for shallow stick-slip frictional-sliding earthquakes, but the confining pressure and temperature are so high for deep-focus events that a distinct process is likely needed to account for their abrupt energy release. The largest recorded deep-focus earthquake (MW 7.9) in the Izu-Bonin slab struck on 30 May 2015 beneath the Ogasawara (Bonin) Islands, isolated from prior seismicity by over 100 km in depth, and followed by only 2 small aftershocks. Globally, this is the deepest (678 km) major (MW > 7) earthquake in the seismological record. Seismicity indicates along-strike contortion of the Izu-Bonin slab, with horizontal flattening near a depth of 550 km in the Izu region and progressive steepening to near-vertical toward the south above the location of the 2015 event. Analyses of a large global data set of P, SH and pP seismic phases using short-period back-projection, subevent directivity, and broadband finite-fault inversion indicate that the mainshock ruptured a shallowly-dipping fault plane with patchy slip that spread over a distance of ~40 km with variable expansion rate (~5 km/s down-dip initially, ~3 km/s up-dip later). During the 17 s rupture duration the radiated energy was ~3.3 x 1016 J and the stress drop was ~38 MPa. The radiation efficiency is moderate (0.34), intermediate to that of the 1994 Bolivia and 2013 Sea of Okhotsk MW 8.3 earthquakes, indicating a continuum of processes. The isolated occurrence of the event suggests that localized stress concentration associated with the pronounced deformation of the Izu-Bonin slab likely played a role in generating this major earthquake.

  13. Temperature lidar measurements from 1 to 105 km altitude using resonance, Rayleigh, and Rotational Raman scattering

    Directory of Open Access Journals (Sweden)

    M. Alpers

    2004-01-01

    Full Text Available For the first time, three different temperature lidar methods are combined to obtain time-resolved complete temperature profiles with high altitude resolution over an altitude range from the planetary boundary layer up to the lower thermosphere (about 1–105 km. The Leibniz-Institute of Atmospheric Physics (IAP at Kühlungsborn, Germany (54° N, 12° E operates two lidar instruments, using three different temperature measurement methods, optimized for three altitude ranges: (1 Probing the spectral Doppler broadening of the potassium D1 resonance lines with a tunable narrow-band laser allows atmospheric temperature profiles to be determined at metal layer altitudes (80–105 km. (2 Between about 20 and 90 km, temperatures were calculated from Rayleigh backscattering by air molecules, where the upper start values for the calculation algorithm were taken from the potassium lidar results. Correction methods have been applied to account for, e.g. Rayleigh extinction or Mie scattering of aerosols below about 32 km. (3 At altitudes below about 25 km, backscattering in the Rotational Raman lines is strong enough to obtain temperatures by measuring the temperature dependent spectral shape of the Rotational Raman spectrum. This method works well down to about 1 km. The instrumental configurations of the IAP lidars were optimized for a 3–6 km overlap of the temperature profiles at the method transition altitudes. We present two night-long measurements with clear wave structures propagating from the lower stratosphere up to the lower thermosphere.

  14. 1.28 Tbit/s/channel single-polarization DQPSK transmission over 525 km using ultrafast time-domain optical Fourier transformation

    DEFF Research Database (Denmark)

    Guan, P.; Mulvad, Hans Christian Hansen; Tomiyama, Y.

    2010-01-01

    A single-channel 1.28 Tbit/s transmission over 525 km is demonstrated for the first time with a single-polarization DQPSK signal. Ultrafast time-domain optical Fourier transformation is successfully applied to DQPSK signals and results in improved performance and increased system margin.......A single-channel 1.28 Tbit/s transmission over 525 km is demonstrated for the first time with a single-polarization DQPSK signal. Ultrafast time-domain optical Fourier transformation is successfully applied to DQPSK signals and results in improved performance and increased system margin....

  15. Comparison between reflectivity statistics at heights of 3 and 6 km and rain rate statistics at ground level

    Science.gov (United States)

    Crane, R. K.

    1975-01-01

    An experiment was conducted to study the relations between the empirical distribution functions of reflectivity at specified locations above the surface and the corresponding functions at the surface. A bistatic radar system was used to measure continuously the scattering cross section per unit volume at heights of 3 and 6 km. A frequency of 3.7 GHz was used in the tests. It was found that the distribution functions for reflectivity may significantly change with height at heights below the level of the melting layer.

  16. Repair promoted by plasmid pKM101 is different from SOS repair

    International Nuclear Information System (INIS)

    Goze, A.; Devoret, R.

    1979-01-01

    In E. coli K12 bacteria carrying plasmid pKM101, prophage lambda was induced at UV doses higher than in plasmid-less parental bacteria. UV-induced reactivation per se was less effective. Bacteria with pKM101 showed no alteration in their division cycle. Plasmid PKM101 coded for a constitutive error-prone repair different from the inducible error-prone repair called SOS repair. Plasmid pKM101 protected E. coli bacteria from UV damage but slightly sensitized them to X-ray lesions. Protection against UV damage was effective in mutant bacteria deficient in DNA excision-repair provided that the recA, lexA and uvrE genes were functional. Survival of phages lambda and S13 after UV irradiation was enhanced in bacteria carrying plasmid pKM101; phage lambda mutagenesis was also increased. Plasmid pKM101 repaired potentially lethal DNA lesions, although Wild-type DNA sequences may not necessarily be restored; hence the mutations observed are the traces of the original DNA lesions. (Auth.)

  17. Zonal asymmetry of daytime 150-km echoes observed by Equatorial Atmosphere Radar in Indonesia

    Directory of Open Access Journals (Sweden)

    T. Yokoyama

    2009-03-01

    Full Text Available Multi-beam observations of the daytime ionospheric E-region irregularities and the so-called 150-km echoes with the 47-MHz Equatorial Atmosphere Radar (EAR in West Sumatra, Indonesia (0.20° S, 100.32° E, 10.36° S dip latitude are presented. 150-km echoes have been frequently observed by the EAR, and their characteristics are basically the same as the equatorial ones, except for an intriguing zonal asymmetry; stronger echoes in lower altitudes in the east directions, and weaker echoes in higher altitudes in the west. The highest occurrence is seen at 5.7° east with respect to the magnetic meridian, and the altitude gradually increases as viewing from the east to west. Arc structures which return backscatter echoes are proposed to explain the asymmetry. While the strength of radar echoes below 105 km is uniform within the wide coverage of azimuthal directions, the upper E-region (105–120 km echoes also show a different type of zonal asymmetry, which should be generated by an essentially different mechanism from the lower E-region and 150-km echoes.

  18. KAJIAN REPARASI PENGECATAN PADA LAMBUNG KAPAL (STUDI KASUS KM. KIRANA 3

    Directory of Open Access Journals (Sweden)

    Zulfaidah Ariany

    2014-10-01

    Full Text Available [Title: Study on Painting Repair of Hull Ship (Case Study of KM. Kirana 3] Regular and scheduled maintenance are important things to make sure any ships could work and sail properly. So the main purpose to carry peoples or deliver the goods can be run smoothly. Painting is one of that items. It can protect the body or their parts from rust attack, in common word, corrosion. Estimating the paints consumption upon maintenances are very useful for ship owners, project owners neither shipyards. Upon ship KM. Kirana’s hull reparation, an analysis process can calculate the estimation of her paints consumption: For hull paint reparation of KM Kirana 3, it needed estimated 8 cane (20 liter for bottom painting, 13 canes for bottom top, 42 canes for topside and 6 canes for weather and upper deck.

  19. Naturally enhanced ion-line spectra around the equatorial 150-km region

    Directory of Open Access Journals (Sweden)

    J. L. Chau

    2009-03-01

    Full Text Available For many years strong radar echoes coming from 140–170 km altitudes at low latitudes have been associated to the existence of field-aligned irregularities (FAIs (the so called 150-km echoes. In this work, we present frequency spectra as well as angular distribution of 150-km echoes. When the 150-km region is observed with beams perpendicular to the magnetic field (B the observed radar spectra are very narrow with spectral widths between 3–12 m/s. On the other hand, when few-degrees off-perpendicular beams are used, the radar spectra are wide with spectral widths comparable to those expected from ion-acoustic waves at these altitudes (>1000 m/s. Moreover the off-perpendicular spectral width increases with increasing altitude. The strength of the received echoes is one to two orders of magnitude stronger than the expected level of waves in thermal equilibrium at these altitudes. Such enhancement is not due to an increase in electron density. Except for the enhancement in power, the spectra characteristics of off-perpendicular and perpendicular echoes are in reasonable agreement with expected incoherent scatter spectra at these angles and altitudes. 150-km echoes are usually observed in narrow layers (2 to 5. Bistatic common volume observations as well as observations made few kilometers apart show that, for most of the layers, there is very high correlation on power fluctuations without a noticeable time separation between simultaneous echoes observed with Off-perpendicular and Perpendicular beams. However, in one of the central layers, the echoes are the strongest in the perpendicular beam and absent or very weak in the off-perpendicular beams, suggesting that they are generated by a plasma instability. Our results indicate that most echoes around 150-km region are not as aspect sensitive as originally thought, and they come from waves that have been enhanced above waves in thermal equilibrium.

  20. Naturally enhanced ion-line spectra around the equatorial 150-km region

    Directory of Open Access Journals (Sweden)

    J. L. Chau

    2009-03-01

    Full Text Available For many years strong radar echoes coming from 140–170 km altitudes at low latitudes have been associated to the existence of field-aligned irregularities (FAIs (the so called 150-km echoes. In this work, we present frequency spectra as well as angular distribution of 150-km echoes. When the 150-km region is observed with beams perpendicular to the magnetic field (B the observed radar spectra are very narrow with spectral widths between 3–12 m/s. On the other hand, when few-degrees off-perpendicular beams are used, the radar spectra are wide with spectral widths comparable to those expected from ion-acoustic waves at these altitudes (>1000 m/s. Moreover the off-perpendicular spectral width increases with increasing altitude. The strength of the received echoes is one to two orders of magnitude stronger than the expected level of waves in thermal equilibrium at these altitudes. Such enhancement is not due to an increase in electron density. Except for the enhancement in power, the spectra characteristics of off-perpendicular and perpendicular echoes are in reasonable agreement with expected incoherent scatter spectra at these angles and altitudes. 150-km echoes are usually observed in narrow layers (2 to 5. Bistatic common volume observations as well as observations made few kilometers apart show that, for most of the layers, there is very high correlation on power fluctuations without a noticeable time separation between simultaneous echoes observed with Off-perpendicular and Perpendicular beams. However, in one of the central layers, the echoes are the strongest in the perpendicular beam and absent or very weak in the off-perpendicular beams, suggesting that they are generated by a plasma instability. Our results indicate that most echoes around 150-km region are not as aspect sensitive as originally thought, and they come from waves that have been enhanced above waves in thermal equilibrium.

  1. Probability density functions for radial anisotropy: implications for the upper 1200 km of the mantle

    Science.gov (United States)

    Beghein, Caroline; Trampert, Jeannot

    2004-01-01

    The presence of radial anisotropy in the upper mantle, transition zone and top of the lower mantle is investigated by applying a model space search technique to Rayleigh and Love wave phase velocity models. Probability density functions are obtained independently for S-wave anisotropy, P-wave anisotropy, intermediate parameter η, Vp, Vs and density anomalies. The likelihoods for P-wave and S-wave anisotropy beneath continents cannot be explained by a dry olivine-rich upper mantle at depths larger than 220 km. Indeed, while shear-wave anisotropy tends to disappear below 220 km depth in continental areas, P-wave anisotropy is still present but its sign changes compared to the uppermost mantle. This could be due to an increase with depth of the amount of pyroxene relative to olivine in these regions, although the presence of water, partial melt or a change in the deformation mechanism cannot be ruled out as yet. A similar observation is made for old oceans, but not for young ones where VSH> VSV appears likely down to 670 km depth and VPH> VPV down to 400 km depth. The change of sign in P-wave anisotropy seems to be qualitatively correlated with the presence of the Lehmann discontinuity, generally observed beneath continents and some oceans but not beneath ridges. Parameter η shows a similar age-related depth pattern as shear-wave anisotropy in the uppermost mantle and it undergoes the same change of sign as P-wave anisotropy at 220 km depth. The ratio between dln Vs and dln Vp suggests that a chemical component is needed to explain the anomalies in most places at depths greater than 220 km. More tests are needed to infer the robustness of the results for density, but they do not affect the results for anisotropy.

  2. Off-fault seismicity suggests creep below 10 km on the northern San Jacinto Fault

    Science.gov (United States)

    Cooke, M. L.; Beyer, J. L.

    2017-12-01

    Within the San Bernardino basin, CA, south of the juncture of the San Jacinto (SJF) and San Andreas faults (SAF), focal mechanisms show normal slip events that are inconsistent with the interseismic strike-slip loading of the region. High-quality (nodal plane uncertainty faults [Anderson et al., 2004]. However, the loading of these normal slip events remains enigmatic because the region is expected to have dextral loading between large earthquake events. These enigmatic normal slip events may be loaded by deep (> 10 km depth) spatially creep along the northern SJF. Steady state models show that over many earthquake cycles, the dextral slip rate on the northern SJF increases southward, placing the San Bernardino basin in extension. In the absence of recent large seismic events that could produce off-fault normal focal mechanisms in the San Bernardino basin, non-uniform deep aseismic slip on the SJF could account for this seismicity. We develop interseismic models that incorporate spatially non-uniform creep below 10 km on the SJF based on steady-state slip distribution. These model results match the pattern of deep normal slip events within the San Bernardino basin. Such deep creep on the SJF may not be detectable from the geodetic signal due to the close proximity of the SAF, whose lack of seismicity suggests that it is locked to 20 km. Interseismic models with 15 km locking depth on both faults are indistinguishable from models with 10 km locking depth on the SJF and 20 km locking depth on the SAF. This analysis suggests that the microseismicity in our multi-decadal catalog may record both the interseismic dextral loading of the region as well as off-fault deformation associated with deep aseismic creep on the northern SJF. If the enigmatic normal slip events of the San Bernardino basin are included in stress inversions from the seismic catalog used to assess seismic hazard, the results may provide inaccurate information about fault loading in this region.

  3. Acute Impact of Inhaled Short Acting B-Agonists on 5 Km Running Performance

    Directory of Open Access Journals (Sweden)

    John Dickinson

    2014-06-01

    Full Text Available Whilst there appears to be no ergogenic effect from inhaled salbutamol no study has investigated the impact of the acute inhalation of 1600 µg, the World Anti-Doping Agency (WADA daily upper limit, on endurance running performance. To investigate the ergogenic effect of an acute inhalation of short acting β2-agonists at doses up to 1600 µg on 5 km time trial performance and resultant urine concentration. Seven male non-asthmatic runners (mean ± SD; age 22.4 ± 4.3 years; height 1.80 ± 0.07 m; body mass 76.6 ± 8.6 kg provided written informed consent. Participants completed six 5 km time-trials on separate days (three at 18 °C and three at 30 °C. Fifteen minutes prior to the initiation of each 5 km time-trial participants inhaled: placebo (PLA, 800 µg salbutamol (SAL800 or 1600 µg salbutamol (SAL1600. During each 5 km time-trial HR, VO2, VCO2, VE, RPE and blood lactate were measured. Urine samples (90 ml were collected between 30-180 minutes post 5 km time-trial and analysed for salbutamol concentration. There was no significant difference in total 5 km time between treatments (PLA 1714.7 ± 186.2 s; SAL800 1683.3 ± 179.7 s; SAL1600 1683.6 ± 190.7 s. Post 5 km time-trial salbutamol urine concentration between SAL800 (122.96 ± 69.22 ug·ml-1 and SAL1600 (574.06 ± 448.17 ug·ml-1 were not significantly different. There was no improvement in 5 km time-trial performance following the inhalation of up to 1600 µg of salbutamol in non-asthmatic athletes. This would suggest that the current WADA guidelines, which allow athletes to inhale up to 1600 µg per day, is sufficient to avoid pharmaceutical induced performance enhancement.

  4. Study of data filtering algorithms for the KM3NeT neutrino telescope

    Energy Technology Data Exchange (ETDEWEB)

    Herold, B., E-mail: Bjoern.Herold@physik.uni-erlangen.d [Erlangen Centre for Astroparticle Physics, Erwin-Rommel-Str. 1, 91058 Erlangen (Germany); Seitz, T., E-mail: Thomas.Seitz@physik.uni-erlangen.d [Erlangen Centre for Astroparticle Physics, Erwin-Rommel-Str. 1, 91058 Erlangen (Germany); Shanidze, R., E-mail: shanidze@physik.uni-erlangen.d [Erlangen Centre for Astroparticle Physics, Erwin-Rommel-Str. 1, 91058 Erlangen (Germany)

    2011-01-21

    The photomultiplier signals above a defined threshold (hits) are the main data collected from the KM3NeT neutrino telescope. The neutrino and muon events will be reconstructed from these signals. However, in the deep sea the dominant source of hits are the decays of {sup 40}K isotope and marine fauna bioluminescence. The selection of neutrino and muon events requires the implementation of fast and efficient data filtering algorithms for the reduction of accidental background event rates. A possible data filtering scheme for the KM3NeT neutrino telescope is discussed in the paper.

  5. A New Mission for Schools in Hong Kong in the 21st Century: Promoting Effective KM

    Directory of Open Access Journals (Sweden)

    Esther Chan

    2005-10-01

    Full Text Available Knowledge Management, though a hot topic in business, is quite a new concept in schools and in the education sector in Hong Kong. KM involves identifying, preserving, sharing and making the knowledge assets grow. Proper management of the knowledge assets will make the schools or educational organizations more responsive to changes and to operate successfully in the information age. Thus, there is a need to promote effective knowledge management in schools. The first section of the paper aims to define KM and the relationship of knowledge, information and data. The second section discusses some practical ways that lead to effective handling of knowledge in schools.

  6. Performances and long-term stability of the LHAASO-KM2A prototype array

    International Nuclear Information System (INIS)

    Liu Jia; Sheng XiangDong; He Huihai; Zhao Jing; Chang Jinfan; Gu Minhao; Hou Chao; Lu Hongkui

    2014-01-01

    A prototype array for the LHAASO-KM2A, which consists of 42 detector units and fully overlaps the ARGO-YBJ experiment, was set up at the Yangbajing cosmic ray observatory and has been in stable operation since October 2010. The resulting performances of the KM2A electromagnetic particle detector prototypes fully meet the design requirements. Through hybrid observation of cosmic ray showers with the ARGO-YBJ experiment, the performances and long-term stability of the prototype array are tested and the results are consistent with expectation. The cosmic ray moon shadow observed by the prototype array is also presented. (authors)

  7. Configuration Management (CM) Support for KM Processes at NASA/Johnson Space Center (JSC)

    Science.gov (United States)

    Cioletti, Louis

    2010-01-01

    Collection and processing of information are critical aspects of every business activity from raw data to information to an executable decision. Configuration Management (CM) supports KM practices through its automated business practices and its integrated operations within the organization. This presentation delivers an overview of JSC/Space Life Sciences Directorate (SLSD) and its methods to encourage innovation through collaboration and participation. Specifically, this presentation will illustrate how SLSD CM creates an embedded KM activity with an established IT platform to control and update baselines, requirements, documents, schedules, budgets, while tracking changes essentially managing critical knowledge elements.

  8. One kilometer (1 km) electric solar wind sail tether produced automatically.

    Science.gov (United States)

    Seppänen, Henri; Rauhala, Timo; Kiprich, Sergiy; Ukkonen, Jukka; Simonsson, Martin; Kurppa, Risto; Janhunen, Pekka; Hæggström, Edward

    2013-09-01

    We produced a 1 km continuous piece of multifilament electric solar wind sail tether of μm-diameter aluminum wires using a custom made automatic tether factory. The tether comprising 90,704 bonds between 25 and 50 μm diameter wires is reeled onto a metal reel. The total mass of 1 km tether is 10 g. We reached a production rate of 70 m/24 h and a quality level of 1‰ loose bonds and 2‰ rebonded ones. We thus demonstrated that production of long electric solar wind sail tethers is possible and practical.

  9. Range Reference Atmosphere 0-70 Km Altitude. Kwajalein Missile Range, Kwajalein, Marshall Islands

    Science.gov (United States)

    1982-01-01

    DOCUMENT 360-82 KWAJALEIN MISSILE RANGE KWAJALEIN, MARSHALL ISLANDS RANGE REFERENCE ATMOSPHERE 0-70 KM ALTITUDE, C00 L’’I METEOROLOGY GROUP .RANGE...34Reference Atmosphere (Part 1), Kwajale 4n Missile Range, Kwajalein, Marshall Islands ," ADA002664. * 19. KEY WORDS (Continue on revorsae d. If necoeewy...CLASSIFICATION OF TIlS PAGE (Whe~n Data EnterecD -v DOCUMENT 360-82 Vo- KWAJALEIN MISSILE RANGE KWAJALEIN, MARSHALL ISLANDS RANGE REFERENCE ATMOSPHERE 0-70 km

  10. Inter-data center 28 Gbaud 4-PAM transmission over 240 km standard single mode fiber

    DEFF Research Database (Denmark)

    Madsen, Peter; Suhr, Lau Frejstrup; Tafur Monroy, Idelfonso

    2018-01-01

    We report on achieving 28 Gbaud 4-PAM transmission with post-equalization over a 240 km SSMF link without re-engineering the transmission link design. The results demonstrate the prospect of re-using conventional links for inter data center connections.......We report on achieving 28 Gbaud 4-PAM transmission with post-equalization over a 240 km SSMF link without re-engineering the transmission link design. The results demonstrate the prospect of re-using conventional links for inter data center connections....

  11. AMSR-E/Aqua surface soil moisture (LPRM) L3 1 day 25 km x 25 km ascending V002 (LPRM_AMSRE_A_SOILM3) at GES DISC

    Data.gov (United States)

    National Aeronautics and Space Administration — AMSR-E/Aqua surface soil moisture (LPRM) L3 1 day 25 km x 25 km ascending V002 is a Level 3 (gridded) data set. Its land surface parameters, surface soil moisture,...

  12. Production and qualification of 40 km of Al-stabilized NbTi cable for the ATLAS experiment at CERN

    CERN Document Server

    Baccaglioni, G; Cartegni, G C; Horváth, I L; Neuenschwander, J; Pedrini, D; Rossi, L; Volpini, G

    2002-01-01

    The production of the conductor for the superconducting toroids of the ATLAS experiment at LHC (CERN) is now in progress. The toroid system, composed of one barrel toroid (BT) and two end cap toroids (ECTs), exploits aluminum-clad Rutherford-type NbTi conductors of large size (57 * 12 mm for BT, 42 * 12 mm for ECTs) and high critical current (Ic) (58 kA for BT and 60 kA for ECTs @ 4.2 K, 5 T). Some 55 km of conductor are required for the BT and 26 km for the ECTs, respectively. An Italian-Swiss (ETH Zurich and INFN) consortium is in charge of the delivery of half of the whole amount. This paper describes the results of this production with particular emphasis to the quality control system developed to monitor the production with both on-line controls and the post-production quality assessment protocols. The main result is the confirmation that the technologies selected and the whole process are reliable and reproducible over large production quantities. The overall degradation due to the cabling and the co-ex...

  13. AMSR-E/Aqua Daily L3 6.25 km 89 GHz Brightness Temperature (Tb) Polar Grids V002

    Data.gov (United States)

    National Aeronautics and Space Administration — The AMSR-E/Aqua Level-3 6.25 km daily sea ice product includes 89.0 GHz brightness temperature averages (daily, ascending, and descending) on a 6.25 km polar...

  14. Characterisation of the Hamamatsu photomultipliers for the KM3NeT Neutrino Telescope

    NARCIS (Netherlands)

    Aiello, S.; Akrame, S.E.; Amélineau, F.; Anassontzis, E.G.; Andre, M.; Androulakis, G.; Anghinolfi; Anton, G.; Ardid, M.; Aublin, J.; Avgitas, T.; Baars, M.; Bagatelas, C.; Barbarino, G.; Baret, B.; Barrios-Martí, J.; Belias, A.; Berbee, E.; van den Berg, A.; Bertin, V.; Biagi, S.; Biagioni, A.; Biernoth, C.; Bormuth, R.; Boumaaza, J.; Bourret, S.; Bouwhuis, M.; Bozza, C.; Brânzas, H.; Briukhanova, N.; Bruijn, R.; Brunner, J.; Buis, E.; Buompane, R,; Busto, J.; Calvo, D.; Capone, A.; Caramete, L.; Celli, S.; Chabab, M.; Cherubini, S.; Chiarella, V.; Chiarusi, T.; Circella, M.; Cocimano, R.; Coelho, J.A.B.; Coleiro, A.; Molla, M.C.; Coniglione, R.; Coyle, P.; Creusot, A.; Cuttone, G.; D’Onofrio, A.; Dallier, R.; De Sio, C.; Di Palma, I.; Díaz, A.F.; Distefano, C.; Domi, A.; Donà, R.; Donzaud, C.; Dornic, D.; Dörr, M.; Durocher, M.; Eberl, T.; Van Eijk, D.; El Bojaddaini, I.; Elsaesser, D.; Enzenhöfer, A.; Ferrara, G.; Fusco, L.A.; Gal, T.; Garufi, F.; Gauchery, S.; Geißelsöder, S.; Gialanella, L.; Giorgio, E.; Giuliante, A.; Gozzini, S.R.; Gracia-Ruiz, R.; Graf, K.; Grasso, D.; Grégoire, T.; Grella, G.; Hallmann, S.; van Haren, H.; Heid, T.; Heijboer, A.; Hekalo, A.; Hernandez-Rey, J.J.; Hofestädt, J.; Illuminati, G.; James, C.W.; Jongen, M.; Jongewaard, B.; de Jong, M.; de Jong, P.; Kadler, M.; Kalaczynski, P.; Kalekin, O.; Katz, U.F.; Khan Chowdhury, N.R.; Kieft, G.; Kießling, D.; Koffeman, E.N.; Kooijman, P.; Kouchner, A.; Kreter, M.; Kulikovskiy, V.; Lahmann, R.; Le Breton, A.; Leone, F.; Leonora, E.; Levi, G.; Lincetto, M.; Lonardo, A.; Longhitano, F.; Lotze, M.; Loucatos, S.; Maggi, G.; Manczak, J.; Mannheim, K.; Margiotta, A.; Marinelli, A.; Markou, C.; Martin, L.; Martínez-Mora, J.A.; Martini, A.; Marzaioli, F.; Mele, R.; Melis, K.W.; Migliozzi, P.; Migneco, E.; Mijakowski, P.; Mollo, C.M.; Morganti, M.; Moser, M.; Moussa, A.; Muller, R.; Musumeci, M.; Nauta, L.; Navas, S.; Nicolau, C.A.; Nielsen, C.; Organokov, M.; Orlando, A.; Panagopoulos, V.; Papalashvili, G.; Papaleo, R.; Pavalas, G.E.; Pellegrini, G.; Pellegrino, C.; Pérez Romero, J.; Perrin-Terrin, M.; Piattelli, P.; Pikounis, K.; Pisanti, O.; Poirè, C.; Polydefki, G.; Poma, G.E.; Popa, V.; Post, M.; Pradier, T.; Pühlhofer, G.; Pulvirenti, S.; Quinn, L.; Raffaelli, F.; Randazzo, N.; Razzaque, S.; Real, D.; Resvanis, L.; Reubelt, J.; Riccobene, G.; Richer, M.; Rovelli, A.; Salvadori, I.; Samtleben, D.F.E.; Sánchez-Losa, A.; Sanguineti, M.; Santangelo, A.; Sapienza, P.; Schermer, B.; Sciacca, V.; Seneca, J.; Sgura, I.; Shanidze, R.; Sharma, A.; Simeone, F.; Sinopoulou, A.; Spisso, B.; Spurio, M.; Stavropoulos, D.; Steijger, J.; Stellacci, S.M.; Strandberg, B.; Stransky, D.; Stüven, T.; Taiuti, M.; Tatone, F.; Tayalati, Y.; Tenllado, E.; Thakore, T.; Timmer, P.; Trovato, A.; Tsagkli, S.; Tzamariudaki, E.; Tzanetatos, D.; Valieri, C.; Vallage, B.; Van Elewyck, V.; Versari, F.; Viola, S.; Vivolo, D.; Volkert, M.; de Waardt, L.; Wilms, J.; de Wolf, E.; Zaborov, D.; Zornoza, J.D.; Zúñiga, J.

    2018-01-01

    The Hamamatsu R12199-02 3-inch photomultiplier tube is the photodetector chosen for the first phase of the KM3NeT neutrino telescope. About 7000 photomultipliers have been characterised for dark count rate, timing spread and spurious pulses. The quantum efficiency, the gain and the peak-to-valley

  15. A method to stabilise the performance of negatively fed KM3NeT photomultipliers

    NARCIS (Netherlands)

    Adrián-Martínez, S.; Ageron, M.; Aiello, S.; Albert, A.; Ameli, F.; Anassontzis, E.G.; Andre, M.; Androulakis, G.; Anghinolfi, M.; Anton, G.; Ardid, M.; Avgitas, T.; Barbarino, G.; Barbarito, E.; Baret, B.; Barrios-Martí, J.; Belias, A.; Berbee, E.; Berg, A. van den; Bertin, V.; Beurthey, S.; Beveren, V. van; Beverini, N.; Biagi, S.; Biagioni, A.; Billault, M.; Bondì, M.; Bormuth, R.; Bouhadef, B.; Bourlis, G.; Bourret, S.; Boutonnet, C.; Bouwhuis, M.; Bozza, C.; Bruijn, R.; Brunner, J.; Buis, E.J.; Buompane, R.; Busto, J.; Cacopardo, G.; Caillat, L.; Calamai, M.; Calvo, D.; Capone, A.; Caramete, L.; Cecchini, S.; Celli, S.; Champion, C.; Cherubini, S.; Chiarella, V.; Chiarelli, L.; Chiarusi, T.; Circella, M.; Classen, L.; Cobas, D.; Cocimano, R.; Coelho, J.A.B.; Coleiro, A.; Colonges, S.; Coniglione, R.; Cordelli, M.; Cosquer, A.; Coyle, P.; Creusot, A.; Cuttone, G.; D'Amato, C.; D'Amico, A.; D'Onofrio, A.; De Bonis, G.; De Sio, C.; Di Capua, F.; Di Palma, I.; Distefano, C.; Donzaud, C.; Dornic, D.; Dorosti-Hasankiadeh, Q.; Drakopoulou, E.; Drouhin, D.; Durocher, M.; Eberl, T.; Eichie, S.; Van Eijk, D.; El Bojaddaini, I.; Elsaesser, D.; Enzenhöfer, A.; Favaro, M.; Fermani, P.; Ferrara, G.; Frascadore, G.; Furini, M.; Fusco, L.A.; Gal, T.; Galatà, S.; Garufi, F.; Gay, P.; Gebyehu, M.; Giacomini, F.; Gialanella, L.; Giordano, V.; Gizani, N.; Gracia, R.; Graf, K.; Grégoire, T.; Grella, G.; Grmek, A.; Guerzoni, M.; Habel, R.; Hallmann, S.; Haren, H. van; Harissopulos, S.; Heid, T.; Heijboer, A.; Heine, E.; Henry, S.; Hernández-Rey, J.J.; Hevinga, M.; Hofestädt, J.; Hugon, C.M.F.; Illuminati, G.; James, C.W.; Jansweijer, P.; Jongen, M.; Jong, M. de; Kadler, M.; Kalekin, O.; Kappes, A.; Katz, U.F.; Keller, P.; Kieft, G.; Kießling, D.; Koffeman, E.N.; Kooijman, P.; Kouchner, A.; Kreter, M.; Kulikovskiy, V.; Lahmann, R.; Lamare, P.; Leisos, A.; Leonora, E.; Clark, M.L.; Liolios, A.; Alvarez, C.D.L.; Lo Presti, D.; Löhner, H.; Lonardo, A.; Lotze, M.; Loucatos, S.; Maccioni, E.; Mannheim, K.; Manzali, M.; Margiotta, A.; Margotti, A.; Marinelli, A.; Mariš, O.; Markou, C.; Martínez-Mora, J.A.; Martini, A.; Marzaioli, F.; Mele, R.; Melis, K.W.; Michael, T.; Migliozzi, P.; Migneco, E.; Mijakowski, P.; Miraglia, A.; Mollo, C.M.; Mongelli, M.; Morganti, M.; Moussa, A.; Musico, P.; Musumeci, M.; Nicolau, C.A.; Olcina, I.; Olivetto, C.; Orlando, A.; Orzelli, A.; Pancaldi, G.; Paolucci, A.; Papaikonomou, A.; Papaleo, R.; Pǎvǎlaš, G.E.; Peek, H.; Pellegrini, G.; Pellegrino, C.; Perrina, C.; Pfutzner, M.; Piattelli, P.; Pikounis, K.; Poma, G.E.; Popa, V.; Pradier, T.; Pratolongo, F.; Pühlhofer, G.; Pulvirenti, S.; Quinn, L.; Racca, C.; Raffaelli, F.; Randazzo, N.; Real, D.; Resvanis, L.; Reubelt, J.; Riccobene, G.; Rossi, C.; Rovelli, A.; Saldaña, M.; Salvadori, I.; Samtleben, D.F.E.; Sánchez García, A.; Sánchez Losa, A.; Sanguineti, M.; Santangelo, A.; Santonocito, D.; Sapienza, P.; Schimmel, F.; Schmelling, J.; Schnabel, J.; Sciacca, V.; Sedita, M.; Seitz, T.; Sgura, I.; Simeone, F.; Sipala, V.; Spisso, B.; Spurio, M.; Stavropoulos, G.; Steijger, J.; Stellacci, S.M.; Stransky, D.; Taiuti, M.; Tayalati, Y.; Terrasi, F.; Tézier, D.; Theraube, S.; Timmer, P.; Tönnis, C.; Trasatti, L.; Travaglini, R.; Trovato, A.; Tsirigotis, A.; Tzamarias, S.; Tzamariudaki, E.; Vallage, B.; Van Elewyck, V.; Vermeulen, J.; Versari, F.; Vicini, P.; Viola, S.; Vivolo, D.; Volkert, M.; Wiggers, L.; Wilms, J.; Wolf, E. de; Zachariadou, K.; Zani, S.; Zornoza, J.D.; Zúñiga, J.

    2016-01-01

    The KM3NeT research infrastructure, currently under construction in the Mediterranean Sea, will host neutrino telescopes for the identification of neutrino sources in the Universe and for studies of the neutrino mass hierarchy. These telescopes will house hundreds of thousands of photomultiplier

  16. Letter of intent for KM3NeT 2.0

    NARCIS (Netherlands)

    Adrián-Martínez, S.; Ageron, M.; Aharonian, F.; Aiello, S.; Albert, A.; Ameli, F.; Anassontzis, E.; Andre, M.; Androulakis, G.; Anghinolfi, M.; Anton, G.; Ardid, M.; Avgitas, T.; Barbarino, G.; Barbarito, E.; Baret, B.; Barrios-Martí, J.; Belhorma, B.; Belias, A.; Berbee, E.; Berg, A. van den; Bertin, V.; Beurthey, S.; Beveren, V. van; Beverini, N.; Biagi, S.; Biagioni, A.; Billault, M.; Bondì, M.; Bormuth, R.; Bouhadef, B.; Bourlis, G.; Bourret, S.; Boutonnet, C.; Bouwhuis, M.; Bozza, C.; Bruijn, R.; Brunner, J.; Buis, E.J.; Busto, J.; Cacopardo, G.; Caillat, L.; Calamai, M.; Calvo, D.; Capone, A.; Caramete, L.; Cecchini, S.; Celli, S.; Champion, C.; El Moursli, R.C.; Cherubini, S.; Chiarusi, T.; Circella, M.; Classen, L.; Cocimano, R.; Coelho, J.A.B.; Coleiro, A.; Colonges, S.; Coniglione, R.; Cordelli, M.; Cosquer, A.; Coyle, P.; Creusot, A.; Cuttone, G.; D'Amico, A.; De Bonis, G.; De Rosa, G.; De Sio, C.; Di Capua, F.; Di Palma, I.; Díaz García, A.F.; Distefano, C.; Donzaud, C.; Dornic, D.; Dorosti-Hasankiadeh, Q.; Drakopoulou, E.; Drouhin, D.; Drury, L.; Durocher, M.; Eberl, T.; Eichie, S.; Eijk, D. van; El Bojaddaini, I.; El Khayati, N.; Elsaesser, D.; Enzenhöfer, A.; Fassi, F.; Favali, P.; Fermani, P.; Ferrara, G.; Filippidis, C.; Frascadore, G.; Fusco, L.A.; Gal, T.; Galatà, S.; Garufi, F.; Gay, P.; Gebyehu, M.; Giordano, V.; Gizani, N.; Gracia, R.; Graf, K.; Grégoire, T.; Grella, G.; Habel, R.; Hallmann, S.; Haren, H. van; Harissopulos, S.; Heid, T.; Heijboer, A.; Heine, E.; Henry, S.; Hernández-Rey, J.J.; Hevinga, M.; Hofestädt, J.; Hugon, C.M.F.; Illuminati, G.; James, C.W.; Jansweijer, P.; Jongen, M.; Jong, M. de; Kadler, M.; Kalekin, O.; Kappes, A.; Katz, U.F.; Keller, P.; Kieft, G.; Kießling, D.; Koffeman, E.N.; Kooijman, P.; Kouchner, A.; Kulikovskiy, V.; Lahmann, R.; Lamare, P.; Leisos, A.; Leonora, E.; Clark, M.L.; Liolios, A.; Alvarez, C.D.L.; Lo Presti, D.; Löhner, H.; Lonardo, A.; Lotze, M.; Loucatos, S.; MacCioni, E.; Mannheim, K.; Margiotta, A.; Marinelli, A.; Maris, O.; Markou, C.; Martínez-Mora, J.A.; Martini, A.; Mele, R.; Melis, K.W.; Michael, T.; Migliozzi, P.; Migneco, E.; Mijakowski, P.; Miraglia, A.; Mollo, C.M.; Mongelli, M.; Morganti, M.; Moussa, A.; Musico, P.; Musumeci, M.; Navas, S.; Nicolau, C.A.; Olcina, I.; Olivetto, C.; Orlando, A.; Papaikonomou, A.; Papaleo, R.; Pavalas, G.E.; Peek, H.; Pellegrino, C.; Perrina, C.; Pfutzner, M.; Piattelli, P.; Pikounis, K.; Poma, G.E.; Popa, V.; Pradier, T.; Pratolongo, F.; Pühlhofer, G.; Pulvirenti, S.; Quinn, L.; Racca, C.; Raffaelli, F.; Randazzo, N.; Rapidis, P.; Razis, P.; Real, D.; Resvanis, L.; Reubelt, J.; Riccobene, G.; Rossi, C.; Rovelli, A.; Saldaña, M.; Salvadori, I.; Samtleben, D.F.E.; García, A.S.; Losa, A.S.; Sanguineti, M.; Santangelo, A.; Santonocito, D.; Sapienza, P.; Schimmel, F.; Schmelling, J.; Sciacca, V.; Sedita, M.; Seitz, T.; Sgura, I.; Simeone, F.; Siotis, I.; Sipala, V.; Spisso, B.; Spurio, M.; Stavropoulos, G.; Steijger, J.; Stellacci, S.M.; Stransky, D.; Taiuti, M.; Tayalati, Y.; Tézier, D.; Theraube, S.; Thompson, L.; Timmer, P.; Tönnis, C.; Trasatti, L.; Trovato, A.; Tsirigotis, A.; Tzamarias, S.; Tzamariudaki, E.; Vallage, B.; Elewyck, V. van; Vermeulen, J.; Vicini, P.; Viola, S.; Vivolo, D.; Volkert, M.; Voulgaris, G.; Wiggers, L.; Wilms, J.; De Wolf, E.; Zachariadou, K.; Zornoza, J.D.; Zúñiga, J.

    2016-01-01

    The main objectives of the KM3NeT Collaboration are (i) the discovery and subsequent observation of high-energy neutrino sources in the Universe and (ii) the determination of the mass hierarchy of neutrinos. These objectives are strongly motivated by two recent important discoveries, namely: (1) the

  17. Time-resolved wave-profile measurements at impact velocities of 10 km/s

    Energy Technology Data Exchange (ETDEWEB)

    Chhabildas, L.C.; Furnish, M.D.; Reinhart, W.D.

    1998-06-01

    Development of well-controlled hypervelocity launch capabilities is the first step to understand material behavior at extreme pressures and temperatures not available using conventional gun technology. In this paper, techniques used to extend both the launch capabilities of a two-stage light-gas gun to 10 km/s and their use to determine material properties at pressures and temperature states higher than those ever obtained in the laboratory are summarized. Time-resolved interferometric techniques have been used to determine shock loading and release characteristics of materials impacted by titanium and aluminum fliers launched by the only developed three-stage light-gas gun at 10 km/s. In particular, the Sandia three stage light gas gun, also referred to as the hypervelocity launcher, HVL, which is capable of launching 0.5 mm to 1.0 mm thick by 6 mm to 19 mm diameter plates to velocities approaching 16 km/s has been used to obtain the necessary impact velocities. The VISAR, interferometric particle-velocity techniques has been used to determine shock loading and release profiles in aluminum and titanium at impact velocities of 10 km/s.

  18. Immunoglobulin GM and KM genes and measles vaccine-induced humoral immunity.

    Science.gov (United States)

    Ovsyannikova, Inna G; Larrabee, Beth R; Schaid, Daniel J; Poland, Gregory A

    2017-10-04

    Identifying genetic polymorphisms that explain variations in humoral immunity to live measles virus vaccine is of great interest. Immunoglobulin GM (heavy chain) and KM (light chain) allotypes are genetic markers known to be associated with susceptibility to several infectious diseases. We assessed associations between GM and KM genotypes and measles vaccine humoral immunity (neutralizing antibody titers) in a combined cohort (n=1796) of racially diverse healthy individuals (age 18-41years). We did not discover any significant associations between GM and/or KM genotypes and measles vaccine-induced neutralizing antibody titers. African-American subjects had higher neutralizing antibody titers than Caucasians (1260mIU/mL vs. 740mIU/mL, p=7.10×10 -13 ), and those titers remained statistically significant (p=1.68×10 -09 ) after adjusting for age at enrollment and time since last vaccination. There were no statistically significant sex-specific differences in measles-induced neutralizing antibody titers in our study (p=0.375). Our data indicate a surprising lack of evidence for an association between GM and KM genotypes and measles-specific neutralizing antibody titers, despite the importance of these immune response genes. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Radiation state of environment in the 30-km zone of Chernobyl NPP, water objects

    International Nuclear Information System (INIS)

    Sukhoruchkin, A.K.

    1994-01-01

    The level of radioactivity in the zone of Chernobyl NPP before the accident is shown. The extremely high level of radioactive contamination produced by the release from the ChNPP at the accident declined as short-lived nuclides decayed. The aerial γ surveys of the 5 km zone of the ChNPP in October, 1986 and September, 1987 are shown. The radiation hygienic parameters of long-lived nuclides at the moment of release are presented. The main contribution to the dose in inhalation is expected from transuranium nuclides, and in ingestion, 90 Sr. The radiation situation in the 30 km zone is at present determined by 137 Cs, 90 Sr and transuranium nuclides, and such nuclide mixture is very unfavorable from the radiobiological point of view. The dosimetric monitoring of them is discussed. The soil contamination density in the 5 km zone is shown. The circumstances of the rich water resources in the region are explained, and the state of contamination is shown. The observation of radioactive contamination of the surface water of Pripyat River is described, and the results are shown. Much concern in the 30 km zone was aroused by the radiation state of groundwater at the places of the forced temporary burial of solid radioactive waste. The state of the cooling water pond in relation to Pripyat River is reported. (K.I.)

  20. Surface to 90 km winds for Kennedy Space Center, Florida, and Vandenberg AFB, California

    Science.gov (United States)

    Johnson, D. L.; Brown, S. C.

    1979-01-01

    Bivariate normal wind statistics for a 90 degree flight azimuth, from 0 through 90 km altitude, for Kennedy Space Center, Florida, and Vandenberg AFB, California are presented. Wind probability distributions and statistics for any rotation of axes can be computed from the five given parameters.