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  1. The 13th Annual Legislative Summit (Washington, DC, 2010). Briefing Papers

    National Indian Education Association, 2010

    2010-01-01

    This volume contains briefing papers presented at the 13th Annual National Indian Education Association (NIEA) Legislative Summit held in Washington, DC. The following papers are included: (1) NIEA Appropriations Priorities for FY11; (2) The President's Budget Request for Fiscal Year 2011 for Native Education; (3) BIE Race to the Top; (4)…

  2. Report from the 13th Annual Western Canadian Gastrointestinal Cancer Consensus Conference; Calgary, Alberta; September 8–10, 2011

    Vickers, M.M.; Pasieka, J; Dixon, E; McEwan, S.; McKay, A; Renouf, D.; Schellenberg, D; Ruether, D.

    2012-01-01

    The 13th annual Western Canadian Gastrointestinal Cancer Consensus Conference was held in Calgary, Alberta, September 8–10, 2011. Health care professionals involved in the care of patients with gastrointestinal cancers participated in presentation and discussion sessions for the purposes of developing the recommendations presented here. This consensus statement addresses current issues in the management neuroendocrine tumours and locally advanced pancreatic cancer.

  3. The 13th Annual Meeting of the Translational Research Cancer Centers Consortium (TrC3); Immune Suppression and the Tumor Microenvironment, Columbus, Ohio; March 1–2, 2010

    Lesinski, Gregory B.; Carson, William E.; Repasky, Elizabeth A.; Wei, Weizen; Kalinski, Pawel; Lotze, Michael T; June, Carl H.; Petros, William; Muthusamy, Natarajan; Olencki, Thomas

    2010-01-01

    The Translational Research Cancer Centers Consortium (TrC3) is a cancer immunotherapy network, established to promote biologic therapeutics in the Midwestern and Northeastern regions of The United States. The 13th Annual Meeting of the TrC3 was hosted by The Ohio State University Comprehensive Cancer Center—Arthur G. James Cancer Hospital and Richard J. Solove Research Institute and took place at The Blackwell Hotel and Conference Center in Columbus, OH on March 1–2, 2010 (http://www.osuccc.o...

  4. 13th International Symposium on Experimental Robotics

    Dudek, Gregory; Khatib, Oussama; Kumar, Vijay

    2013-01-01

    The International Symposium on Experimental Robotics (ISER) is a series of bi-annual meetings which are organized in a rotating fashion around North America, Europe and Asia/Oceania. The goal of  ISER is to provide a forum for research in robotics that focuses on novelty of theoretical contributions validated by experimental results. The meetings are conceived to bring together, in a small group setting, researchers from around the world who are in the forefront of experimental robotics research. This unique reference presents the latest advances across the various fields of robotics, with ideas that are not only conceived conceptually but also explored experimentally. It collects robotics contributions on the current developments and new directions in the field of experimental robotics, which are based on the papers presented at the 13th ISER held held in Québec City, Canada, at the Fairmont Le Château Frontenac, on June 18-21, 2012. This present thirteenth edition of Experimental Robotics edited by Jayd...

  5. 13th International Conference on Information Technology

    2016-01-01

    This book collects articles presented at the 13th International Conference on Information Technology- New Generations, April, 2016, in Las Vegas, NV USA. It includes over 100 chapters on critical areas of IT including Web Technology, Communications, Security, and Data Mining.

  6. 13th International Conference on Chlamydomonas

    Silflow, Carolyn D. [University of Minnesota

    2014-03-11

    The 13th International Conference on Chlamydomonas (EMBO Workshop on the Cell and Molecular Biology of Chlamydomonas) was held May 27 to June 1, 2008 in Hyeres, France. The conference was the biennial meeting for all researchers studying the green algal systems Chlamydomonas and Volvox. The conference brought together approximately 200 investigators from around the world (North America, Asia, Europe and Australia) representing different fields and disciplines (cell biology, genetics, biochemistry, biophysics, plant physiology, genomics). It provided an opportunity for investigators from different countries to share methodologies and to discuss recent results with a focus on the Chlamydomonas experimental system.

  7. 13th International Symposium on Advanced Materials

    The 13th international symposium on advanced materials (ISAM) was held from september 23-27, 2013, at islamabad, Pakistan. The main theme of this conference was to discuss the ever increasing changes and intricacies that characterize modern industry necessitate a growing demand for technical information on advanced materials. In five day of the symposium, nearly Two hundred and twenty seven contributory and invited papers, comprising of Nineteen technical sessions and two poster sessions were presented. Renowned scientists and researchers from foreign and local institutes have shared their counter parts on the topics of common interest. This symposium provided an ideal opportunity for exchange of information amongst scientists, engineers and researchers from all over pakistan and other countries of the world. (A.B.)

  8. 13th International Conference Intelligent Autonomous Systems

    Michael, Nathan; Berns, Karsten; Yamaguchi, Hiroaki

    2016-01-01

    This book describes the latest research accomplishments, innovations, and visions in the field of robotics as presented at the 13th International Conference on Intelligent Autonomous Systems (IAS), held in Padua in July 2014, by leading researchers, engineers, and practitioners from across the world. The contents amply confirm that robots, machines, and systems are rapidly achieving intelligence and autonomy, mastering more and more capabilities such as mobility and manipulation, sensing and perception, reasoning, and decision making. A wide range of research results and applications are covered, and particular attention is paid to the emerging role of autonomous robots and intelligent systems in industrial production, which reflects their maturity and robustness. The contributions have been selected through a rigorous peer-review process and contain many exciting and visionary ideas that will further galvanize the research community, spurring novel research directions. The series of biennial IAS conferences ...

  9. The 13th Malaysian general election: Uncertainties and expectations

    Moniruzzaman, M

    2013-01-01

    Malaysia held its 13th general election on May 5, 2013 which was a contest between two coalitions, the ruling Barisan Nasional (BN) and the opposition Pakatan Rakyat (PR). The latter’s expectations for an outright win did not materialise. The election results have shown a rather status quo with minor losses and gains for the two coalitions. This study analysed the 13th election and found a number of noticeable trends. The Chinese voters have voted overwhelmingly for the opposition. The politi...

  10. The 13th CAS General Assembly Convenes in Beijing

    2006-01-01

    @@ The 13th General Assembly of CAS was held from June 5 to 8 in Beijing. Top State leaders were present at the joint opening session of the 13th CAS General Assembly and the Eighth General Assembly of the Chinese Academy of Engineering (CAE)held on the morning of June 5 at the Great Hall of the People. Chinese President HU Jintao delivered an important speech at the ceremony. State Councilor CHEN Zhili made a special report about the Framework for the National Long- and Mid-term Plan for S&T Development in China at the joint plenary session of CAS and CAE assemblies on June 6.

  11. Proceedings of the 13th symposium on advanced photon research

    The 13th Symposium on Advanced Photon Research was held at Kansai Photon Science Institute, Japan Atomic Energy Agency (JAEA-KPSI) in Kizugawa-shi, Kyoto on November 15 - 16, 2012. This report consists of invited and contributed papers presented at the oral and poster sessions in the Symposium. (author)

  12. The 13th Malaysian general election: Uncertainties and expectations

    M. Moniruzzaman

    2013-06-01

    Full Text Available Malaysia held its 13th general election on May 5, 2013 which was a contest between two coalitions, the ruling Barisan Nasional (BN and the opposition Pakatan Rakyat (PR. The latter’s expectations for an outright win did not materialise. The election results have shown a rather status quo with minor losses and gains for the two coalitions. This study analysed the 13th election and found a number of noticeable trends. The Chinese voters have voted overwhelmingly for the opposition. The political culture of Malaysia is also shifting toward more participatory type with increased social and economic mobility. The days of one-party dominance in Malaysia are apparently over. The electoral politics in Malaysia might become more polarised along ethnic lines which may require reshuffling of its coalition framework and design.

  13. 13th Workshop on Radiation Monitoring for the International Space Station - Final Program

    The Workshop on Radiation Monitoring for the International Space Station (WRMISS) has been held annually since 1996. The major purpose of WRMISS is to provide a forum for discussion of technical issues concerning radiation dosimetry aboard the International Space Station. This includes discussion of new results, improved instrumentation, detector calibration, and radiation environment and transport models. The goal of WRMISS is to enhance international efforts to provide the best information on the space radiation environment in low-Earth orbit and on the exposure of astronauts and cosmonauts in order to optimize the radiation safety of the ISS crew. During the 13th Annual WRMISS, held in the Institute of Nuclear Physics (Krakow, Poland) on 8-10 September 2008, participants presented 47 lectures

  14. 13th Czechoslovak seminar on plasma physics and technology

    Of the 32 papers presented at the 13th Czechoslovak Seminar on Plasma Physics and Technology, 20 papers deal with low-temperature plasma and its applications, e.g., in thin film production, plasma chemistry and electric engineering, and with various methods of low-temperature plasma diagnostics. The investigation of hot plasma in tokamaks and stellarators is the topic of 5 papers; the latest experimental results achieved in the Czechoslovak tokamak Castor and the Soviet devices Tuman-2A, FT-1 and L-2 are reported. Five papers are devoted to high-power pulse technology applications, represented by plasma heating studies with Czechoslovak electron accelerator REBEX, the Polish plasma focus experiment PF-20, and high-current accelerators of negative ions developed in the USSR. In the only two theoretical papers, strong Langmuir turbulence and the Alfven current drive are studied. (J.U.)

  15. The Geobiology of Weathering: a 13th Hypothesis

    Schwartzman, David

    2015-01-01

    The magnitude of the biotic enhancement of weathering (BEW) has profound implications for the long-term carbon cycle. The BEW ratio is defined as how much faster the silicate weathering carbon sink is under biotic conditions than under abiotic conditions at the same atmospheric pCO2 level and surface temperature. Thus, a 13th hypothesis should be considered in addition to the 12 outlined by Brantley...(2011) regarding the geobiology of weathering: The BEW factor and its evolution over geological time can be inferred from meta-analysis of empirical and theoretical weathering studies. Estimates of the global magnitude of the BEW are presented, drawing from lab, field, watershed data and models of the long-term carbon cycle, with values ranging from one to two orders of magnitude.

  16. 13th International Conference on Distributed Computing and Artificial Intelligence

    Silvestri, Marcello; González, Sara

    2016-01-01

    The special session Decision Economics (DECON) 2016 is a scientific forum by which to share ideas, projects, researches results, models and experiences associated with the complexity of behavioral decision processes aiming at explaining socio-economic phenomena. DECON 2016 held in the University of Seville, Spain, as part of the 13th International Conference on Distributed Computing and Artificial Intelligence (DCAI) 2016. In the tradition of Herbert A. Simon’s interdisciplinary legacy, this book dedicates itself to the interdisciplinary study of decision-making in the recognition that relevant decision-making takes place in a range of critical subject areas and research fields, including economics, finance, information systems, small and international business, management, operations, and production. Decision-making issues are of crucial importance in economics. Not surprisingly, the study of decision-making has received a growing empirical research efforts in the applied economic literature over the last ...

  17. 13th International Conference on X-Ray Lasers

    Gautier, Julien; Ros, David; Zeitoun, Philippe

    2014-01-01

    These proceedings comprise of invited and contributed papers presented at the 13th International Conference on X-Ray Lasers (ICXRL 2012) which was held 11–15 June 2012 in Paris, in the famous Quartier Latin, inside the historical Center of Cordeliers. This conference is part of a continuing series dedicated to recent developments and applications of x-ray lasers and other coherent x-ray sources with attention to supporting technologies and instrumentation. New results in the generation of intense, coherent x-rays and progress towards practical devices and their applications are reported in these proceedings, including areas of research in plasma-based x-ray lasers, 4th generation accelerator-based sources and higher harmonic generation. Recent achievements related to the increase of the repetition rate up to 100 Hz and shorter wavelength collisional plasma-based soft x-ray lasers down to about 7 nm are presented. Seeding the amplifying plasma with a femtosecond high-order harmonic of infrared laser was fore...

  18. PREFACE: 13th IMEKO TC17-TC7 Joint Symposium

    Khan, Sanowar

    2010-04-01

    'Without Measurement No Science, Without Science No Measurement' The 13th IMEKO (International Measurement Confederation) TC1-TC7 Joint Symposium was held at City University London, UK from 1-3 September 2010. For the first time this Symposium also included the involvement of IMEKO Technical Committee 13 (TC13) - Measurements in Biology and Medicine. This brings an added dimension to the Symposium in London since the area of measurement science and technology in biology and medicine is an important and a fast growing one. The Symposium was organized by the City University London (www.city.ac.uk) in collaboration with the Institute of Physics (IOP), UK (www.iop.org). The work of this Symposium is reported in this volume. The scope of the Symposium included the main topics covered by the above Technical Committees - education and training in measurement and instrumentation (TC1), measurement science (TC7) and measurements in biology and medicine. These themes underpinned the strap line of the Symposium, 'Without Measurement No Science, Without Science No Measurement' with the highest number of contributions from the measurement science area. The thematic areas were led by invited presentations from each of the areas by eminent speakers. The Symposium provided a useful forum for experts working in these areas for sharing and exchanging their work and ideas. The Symposium attracted participants from many countries of the world including the United States, Japan, Russia and Ukraine. In total over sixty papers are included in the volume and they are presented under the above three key thematic areas. Each paper was independently peer-reviewed by two reviewers from a distinguished international panel. The organizers of the Symposium, City University London have pioneered the establishment of measurement and instrumentation as an academic discipline in the UK through the work of Professor Ludwik Finkelstein who was for many years Chairman of TC1 and a founding member of TC

  19. PREFACE: 13th International Conference on Liquid and Amorphous Metals

    Popel, Pjotr; Gelchinskii, Boris; Sidorov, Valeriy; Son, Leonid; Sabirzjanov, Alexandre

    2007-06-01

    The state of the art in the field of liquid and amorphous metals and alloys is regularly updated through two series of complementary international conferences, the LAM (Liquid and Amorphous Metals) and the RQ (Rapidly Quenched Materials). The first series of the conferences started as LM-1 in 1966 at Brookhaven for the basic understanding of liquid metals. The subsequent LM conferences were held in Tokyo (1972) and Bristol (1976). The conference was renewed in Grenoble (1980) as a LAM conference including amorphous metals and continued in Los Angeles (1983), Garmisch-Partenkirchen (1986), Kyoto (1989), Vienna (1992), Chicago (1995), Dortmund (1998), Yokohama (2001) and Metz (2004). The conferences are mainly devoted to liquid and amorphous metals and alloys. However, communications on some non-metallic systems such as semi conductors, quasicrystals etc, were accepted as well. The conference tradition strongly encourages the participation of junior researchers and graduate students. The 13th conference of the LAM series was organized in Ekaterinburg, Russia, by the Institute of Metallurgy of the Ural Branch of the Russian Academy of Sciences (IMet UB RAS) and Ural State Pedagogical University (USPU) and held on 8-13 July 2007 under the chairmanship of Professors Pjotr Popel (USPU) and Boris Gelchinskii (IMet UB RAS). There were 242 active and about 60 guest participants from 20 countries who attended the conference. There were no parallel sessions and all oral reports were separated into three groups: invited talks (40 min), full-scale (25 min) and brief (15 min) oral reports. The program included 10 sessions, ranging from purely theoretical subjects to technological application of molten and amorphous alloys. The following sessions took place: A) Electronic structure and transport, magnetic properties; B) Phase transitions; C) Structure; D) Atomic dynamics and transport; E) Thermodynamics; F) Modelling, simulation; G) Surface and interface; H) Mechanical properties

  20. Daegu International Poster Exhibition for Colourful Daegu & the 13th World Championships in Athletics

    McCullagh, Joseph Felix

    2008-01-01

    Design poster, exhibited for the Daegu International Poster Exhibition for Colourful Daegu and the 13th World Championships in Athletics 30th September, 2008 Daegu Culture and Arts Center, Daegu, South Korea

  1. A Discussion of the 13th Dalai Lama during the Period from 1910 to 1912

    LIANG Zhongcui

    2014-01-01

    Following the decline of the Qing dynasty and its weak foreign policy , the Tibetan people became gradually dissatisfied with: ( i ) the Qing local administration ’ s lack of progress;( ii) their succumbing to the force of Britain; and ( iii ) their wantonly selling their local benefits . Especially during the time of the 13th Dalai Lama, this kind of mood of dissatisfaction further deep-ened.In order to maintain the benefits of Tibet, the 13th Dalai Lama tried to submit reports directly to the Qing court , and was eager to get the support from the central government of the Qing dynasty . However , this effort did not reach the central gov-ernment due to the barriers created by the royally appointed minister in Tibet . The British army launched a second invasion of Tibet in 1903, and soon occupied Lhasa.To the 13th Dalai Lama’ s disappointment , Youtai, the royal minister in Ti-bet, did not consider the Tibetan ’ s strong anger about the British invasion , but tried to appease the enemy in order to safeguard “peaceful relations”. And, at the same time he framed the 13th Dalai Lama saying“his abandonment of the land became an excuse for the foreign enemy ’ s invasion”. The Qing court then , without making a detailed investigation , announced a decree that abolished Thubten Gyatso ’ s title of Dalai Lama. Since then , misunderstanding between the 13 th Dalai La-ma and the Qing court deepened further , and some upper-class Tibetans even considered leav-ing the Qing . Taking advantage of the Qing ’ s improper attitude towards the 13th Dalai Lama, Britain began officially to carry out a strategy of winning over the heart of the 13th Dalai Lama. In February, 1910, the 13th Dalai Lama leftLhasa and went into exile in India.Britain wasvery happy about the Dalai ’ s arrival in India.The British man with whom the 13th Dalai Lamahad the most contact during his stay in British Indi -a was Charles Alfred Bell, a political officer ofSikkim.Bell noted the

  2. 13th International Workshop on Condensed Matter Theories

    1990-01-01

    This volume gathers the invited talks of the XIII International Work­ shop on Condensed Matter Theories which took place in Campos do Jordao near Sao Paulo, Brazil, August 6-12, 1989. It contains contributions in a wide variety of fields including neutral quantum and classical fluids, electronic systems, composite materials, plasmas, atoms, molecules and nuclei, and as this year's workshop reflected the natural preoccupation in materials science with its spectacular prospect for mankind, room tempera­ ture super-conductivity. All topics are treated from a common viewpoint: that of many-body physics, whether theoretical or simu1ational. Since the very first workshop, held at the prestigious Instituto de Fisica Teorica in Sao Paulo, and organized by the same organizer of the 1989 workshop, Professor Valdir Casaca Aguilera-Navarro, the meeting has taken place annually six times in Latin America, four in Europe and three in the United States. Its principal objective has been to innitiate and nurture collaborati...

  3. 13th EU-US Transport Task Force Workshop on transport in fusion plasmas

    Connor, J.W.; Fasoli, A.; Hidalgo, C.;

    2009-01-01

    This report summarizes the contributions presented at the 13th EU-US Transport Task Force Workshop on transport in fusion plasmas, held in Copenhagen, Denmark, 1-4 September 2008. There were sessions on core heat and particle transport; core and edge momentum transport; edge and scrape-off-layer ......This report summarizes the contributions presented at the 13th EU-US Transport Task Force Workshop on transport in fusion plasmas, held in Copenhagen, Denmark, 1-4 September 2008. There were sessions on core heat and particle transport; core and edge momentum transport; edge and scrape...

  4. Feasibility of 13th lymph nodes as sentinels for periampullary carcinoma

    白晓枫

    2013-01-01

    Objective To study the feasibility and clinical value of 13th lymph nodes in predicting general lymph nodes metastases for periampullary carcinoma.Methods A total of 77 patients with pathologically confirmed periampullary carcinoma were recruited.And 26 (18 males and 8

  5. Abstracts of 13th International Congress on Plasma Physics (ICPP 2006). Published in 2 volumes

    This report contains the presentation on the 13-th International Congress on Plasma Physics (ICPP 2006). Five main topics are covered: fundamental problems of plasma physics; fusion plasmas; plasmas in astrophysics and space physics; plasmas in applications and technologies; complex plasmas

  6. Proceedings of the 13th annual meeting on upper atmosphere studies by optical methods

    A total of 41 papers were presented under the following session topics: Atmospheric emissions; auroral features and dynamics; auroral pulsations; airglow and atmospheric parameters; atmospheric constituents; instrumentation and data handling; and observation programs. The report presents the full text or abstracts of 31 of the lectures given at the meeting. 25 of the papers are seperate input to the data base from this report

  7. The 13 th Annual Intelligent Ground Vehicle Competition: intelligent ground vehicles created by intelligent teams

    Theisen, Bernard L.

    2005-10-01

    The Intelligent Ground Vehicle Competition (IGVC) is one of three, unmanned systems, student competitions that were founded by the Association for Unmanned Vehicle Systems International (AUVSI) in the 1990s. The IGVC is a multidisciplinary exercise in product realization that challenges college engineering student teams to integrate advanced control theory, machine vision, vehicular electronics, and mobile platform fundamentals to design and build an unmanned system. Teams from around the world focus on developing a suite of dual-use technologies to equip ground vehicles of the future with intelligent driving capabilities. Over the past 13 years, the competition has challenged undergraduate, graduate and Ph.D. students with real world applications in intelligent transportation systems, the military and manufacturing automation. To date, teams from over 50 universities and colleges have participated. This paper describes some of the applications of the technologies required by this competition and discusses the educational benefits. The primary goal of the IGVC is to advance engineering education in intelligent vehicles and related technologies. The employment and professional networking opportunities created for students and industrial sponsors through a series of technical events over the three-day competition are highlighted. Finally, an assessment of the competition based on participant feedback is presented.

  8. Proceedings of the 13th annual meeting of Japanese Society of Radiation Safety Management 2014 Tokushima

    This is the entitled program and proceedings held from December 3rd through 5th of 2014. Focusing on the issues of Fukushima Accident, the sessions including poster, invited/special speeches etc. are education on radiation, radiation measurement, risk communication, dose level evaluation, radioactivity in environment, contamination inspection, decontamination, management of radiation emission equipment and its effective use. (J.P.N)

  9. Females do not have more injury road accidents on Friday the 13th

    Summala Heikki

    2004-11-01

    Full Text Available Abstract Background This study reinvestigated the recent finding that females – but not males – die in traffic accidents on Friday the 13th more often than on other Fridays (Näyhä S: Traffic deaths and superstition on Friday the 13th. Am J Psychiatry 2002, 159: 2110–2111. The current study used matched setting and injury accident data base that is more numerous than fatality data. If such an effect would be caused by impaired psychic and psychomotor functioning due to more frequent anxiety among women, it should also appear in injury crashes. Methods We used the national Finnish road accident database for 1989–2002. To control seasonal variation, 21 Fridays the 13th were compared in a matched design to previous and following Fridays, excluding all holidays, on number of accidents, male/female responsibility for accidents, and the number of dead, injured and overall number of active participants (drivers, pedestrians and bicyclists as a consequence of the accident. Results There were no significant differences in any examined aspect of road injury accidents among the three Fridays, either in females or males. Women were not overrepresented in crashes that occurred on Fridays 13th. Conclusion There is no consistent evidence for females having more road traffic crashes on Fridays the 13th, based on deaths or road accident statistics. However, this does not imply a non-existent effect of superstition related anxiety on accident risk as no exposure-to-risk data are available. People who are anxious of "Black Friday" may stay home, or at least avoid driving a car.

  10. Field Trials of the Prototype Pounder Rig, Uganda, 20th August - 13th November 1999.

    Ball, Peter; Danert, Kerstin

    1999-01-01

    A prototype low-cost drilling rig was imported to Uganda in August 1999 and handed over to Mpigi District Government. Field trials of this machine were undertaken between 20 th August and 13 th November 1999. This work was enabled through a tripartite Memorandum of Understanding between the Cranfield University/DFID Low Cost well Drilling Project, the Directorate of Water Development (DWD) of the Government of Uganda, and the local Government of Mpigi District. A total of fo...

  11. Irradiation experiments of the 13th-15th OGL-1 fuel assemblies

    The Oarai Gas Loop-1, OGL-1, which had been installed in the Japan Materials Testing Reactor (JMTR) of the Oarai Research Establishment of the Japan Atomic Energy Research Institute (JAERI), was an in-pile helium loop for irradiation of high-temperature gas-cooled reactor fuels at high pressure and temperature. This report describes results of fabrication, irradiation and post-irradiation examinations (PIEs) of the 13th-15th OGL-1 fuel assemblies. The 13th and 15th fuel assemblies employed the first-charge fuel of the High Temperature Engineering Test Reactor (HTTR). The 13th assembly was loaded with a high quality fuel, whose as-produced failure fraction had been drastically decreased, compared with that for fuels before that time. The 15th assembly was loaded with a fuel, which had been produced by the same apparatus that was used afterwards for the first charge fuel of the HTTR. Both of these fuel assemblies gave good results in PIEs as well as in the fission-gas release rates during irradiation. The 14th fuel assembly used a trial product of an advanced fuel for high burnup utilization, which employed coated fuel particles (CFPs) with thicker coating layers than those for the first charge fuel. This fuel assembly indicated a spike release of fission gas during irradiation at 1500degC after a transient temperature increase up to this value. As a whole, all of the 13th - 15th assemblies demonstrated good performance of the loaded fuels, giving significantly lower values in fission-gas release rates during irradiation and in failure fractions of CFPs after irradiation, than the corresponding design limit values for the first charge fuel of the HTTR. (author)

  12. Irradiation experiments of the 13th-15th OGL-1 fuel assemblies

    Hayashi, Kimio; Sawa, Kazuhiro; Kitajima, Toshio [Japan Atomic Energy Research Inst., Oarai, Ibaraki (Japan). Oarai Research Establishment; Shiratori, Tetsuo; Kikuchi, Hironobu; Fukuda, Kousaku; Itoh, Tadaharu; Waragai, Heita [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment

    2000-01-01

    The Oarai Gas Loop-1, OGL-1, which had been installed in the Japan Materials Testing Reactor (JMTR) of the Oarai Research Establishment of the Japan Atomic Energy Research Institute (JAERI), was an in-pile helium loop for irradiation of high-temperature gas-cooled reactor fuels at high pressure and temperature. This report describes results of fabrication, irradiation and post-irradiation examinations (PIEs) of the 13th-15th OGL-1 fuel assemblies. The 13th and 15th fuel assemblies employed the first-charge fuel of the High Temperature Engineering Test Reactor (HTTR). The 13th assembly was loaded with a high quality fuel, whose as-produced failure fraction had been drastically decreased, compared with that for fuels before that time. The 15th assembly was loaded with a fuel, which had been produced by the same apparatus that was used afterwards for the first charge fuel of the HTTR. Both of these fuel assemblies gave good results in PIEs as well as in the fission-gas release rates during irradiation. The 14th fuel assembly used a trial product of an advanced fuel for high burnup utilization, which employed coated fuel particles (CFPs) with thicker coating layers than those for the first charge fuel. This fuel assembly indicated a spike release of fission gas during irradiation at 1500degC after a transient temperature increase up to this value. As a whole, all of the 13th - 15th assemblies demonstrated good performance of the loaded fuels, giving significantly lower values in fission-gas release rates during irradiation and in failure fractions of CFPs after irradiation, than the corresponding design limit values for the first charge fuel of the HTTR. (author)

  13. Color-coordinate system from a 13th-century account of rainbows.

    Smithson, Hannah E; Anderson, Philip S; Dinkova-Bruun, Greti; Fosbury, Robert A E; Gasper, Giles E M; Laven, Philip; McLeish, Tom C B; Panti, Cecilia; Tanner, Brian K

    2014-04-01

    We present a new analysis of Robert Grosseteste's account of color in his treatise De iride (On the Rainbow), dating from the early 13th century. The work explores color within the 3D framework set out in Grosseteste's De colore [see J. Opt. Soc. Am. A29, A346 (2012)], but now links the axes of variation to observable properties of rainbows. We combine a modern understanding of the physics of rainbows and of human color perception to resolve the linguistic ambiguities of the medieval text and to interpret Grosseteste's key terms. PMID:24695192

  14. Report of the 13th IEA workshop on radiation effects in ceramic insulators

    The 13th IEA Workshop on Radiation Effects in Ceramic Insulators, based on Annex II: Experimentation on Radiation Damage in Fusion Materials, to the IEA Implementing Agreement for a Programme of Research and Development on Radiation Damage in Fusion Materials, was held on the 9th, December, 2003, at Kyoto International Conference Center, in Kyoto, Japan, in conjunction with the 11th International Conference on Fusion Reactor Materials (ICFRM-11). 44 participants from 10 countries (26 from Japan, 5 from Spain, 3 from Belgium, 3 from USA, 2 from RF, each 1 from Austria, Greece, Italy, Romania, and UK) gathered together and discussed following issues extensively, with the newest experimental results, after the welcome remarks by one of the organizer and chairpersons, Dr. E.R. Hodgson of CIEMAT. Effects of electric field on radiation induced microstructural evolution, parasitic electrical current and voltage induced in cables and wires by radiation effects, optical materials, IFMIF related issues and fundamental aspects were discussed. Significant results such as an observation of γ-alumina and aluminum colloid formation for the Radiation Induced Electrical Degradation mechanism are obtained. This report is workshop summary, abstracts and documents of the 13th IEA Workshop on Radiation Effects in Ceramic Insulators. (author)

  15. The ultrasonographic diagnosis of fetal encephalocele at 13th gestational week

    Šorak Marija

    2010-01-01

    Full Text Available Background. Encephalocele presents a rare anomaly of central nervous system, developed as a consequence of neural tube closing defect during early embrional development, and it is described by a baggy formation which prolaborates through the pores of the scull, filled with brain tissue, cerebrospinal liquor and entwined with meninges. According to literature search, until this day, the earliest it can be ultrasonically detected is the 13th gestation week, with the appliance of three-dimensional ultrasound. Case report. We presented 25 years old patient, ultrasonically diagnosed with occipital fetal encephalocela at the 13th gestation week. A gestation sack was located in the right uteral corn of the two-corned uterus with one cervix. The diagnosis was confirmed also by trippled value of alpha-fetoprotein in maternal serum: 75,98 IU/mL. Conclusion. Ultrasonic examination is the method of choice for prenatal detection of a fetal anomaly. It is possible to diagnose encephalocele if it prominates above the limits of the scull.

  16. The 13th International ICIT Conference Progress in Cryogenics and Isotopes Separation. Proceedings

    The proceedings of the 13th International ICIT Conference Progress in Cryogenics and Isotopes Separation, held in Calimanesti-Caciulata, Romania on 7-9 November 2007 contain plenary lectures (15 lectures) and 8 sections addressing the following subject matters: Section 1. Stable Isotopes Physics. Technologies and Applications (5 papers); Section 2. Cryogenics Technologies and Equipment (9 papers); Section 3. Science and Materials Engineering (17 papers); Section 4. Nuclear Power (22 papers); Section 5. Hydrogen and Fuel Cells (7 papers); Section 6. Environmental Protection. Industrial Risk (24 papers); Section 7. Laboratory Analysis Methods (10 papers); Section 8. Business and Technological Incubator BTI - ICIT RM VALCEA. The proceedings are completed by an Addendum consisting of 5 papers. A number of 45 papers are available in abstract form only and accordingly full text entered the corresponding records

  17. City state institutions under municipal or family rule in 13th-century Alessandria

    Alberto Luongo

    2011-11-01

    Full Text Available In the 13th century many attempts were made to set up personal rule over Alessandria, with varying degrees of success. The first was Manfredi II Lancia, viceroy of Emperor Frederick II. This was followed by Guglielmo VII of Monferrato, whose plans were opposed initially by Oberto Pelavicino and subsequently by Carlo of Angiò. Dominion over Alessandria actively involved the population and the dignitaries of the city (divided by loyalty to the rival Lanzavecchia and Del Pozzo families. They were able to establish or end the rule of one family or another. Hence, the political élite of Alessandria took advantage of the aspirations of foreign rulers to solve internal problems, by according them with different powers as the situation demanded and as the loyalties of the rival factions changed.

  18. 13th International Symposium on Distributed Computing and Artificial Intelligence 2016

    Semalat, Ali; Bocewicz, Grzegorz; Sitek, Paweł; Nielsen, Izabela; García, Julián; Bajo, Javier

    2016-01-01

    The 13th International Symposium on Distributed Computing and Artificial Intelligence 2016 (DCAI 2016) is a forum to present applications of innovative techniques for studying and solving complex problems. The exchange of ideas between scientists and technicians from both the academic and industrial sector is essential to facilitate the development of systems that can meet the ever-increasing demands of today’s society. The present edition brings together past experience, current work and promising future trends associated with distributed computing, artificial intelligence and their application in order to provide efficient solutions to real problems. This symposium is organized by the University of Sevilla (Spain), Osaka Institute of Technology (Japan), and the Universiti Teknologi Malaysia (Malaysia).

  19. Certification of GMP is imperative necessity on radiopharmaceutical manufacturers

    The general circumstances on the conception, quality of GMP and Chinese GMP for radiopharmaceutical manufacturers are introduced briefly. It explains that certification of GMP is imperative necessity on radiopharmaceutical manufacturers

  20. Do-Fluoride "Cryolite By- product Carbon White" Awarded the 13th China Excellent Patent Award%Do-Fluoride "Cryolite By- product Carbon White" Awarded the 13th China Excellent Patent Award

    2011-01-01

    On November 4, the results of the 13th China Patent Awards were publicized by the State Intellectual Property Office of the People's Republic of China. The patent of "production method of cryolite by-product carbon white" declared by Henan Province Jiaozuo Do-Fluoride Company was awarded China Excellent Patent Award.

  1. Proceedings of the 13th meeting for tandem accelerators and related techniques

    NONE

    2001-02-01

    The 13th Meeting for Tandem Accelerators and Related Techniques was held on June 8-9, 2000 at Mutsu, Aomori Prefecture, under the auspices of Marine Research Laboratory, Mutsu Establishment, the Japan Atomic Energy Research Institute. About 60 people participated in the meeting from 27 universities and organizations, many of which have a tandem accelerator or an electrostatic accelerator. The objectives of the meeting are to exchange information on tandem accelerates or electrostatic accelerators by making a presentation on related research and topics, and also to enhance mutual friendship of participants, resulting in the contribution to associated R and D. The meeting mainly consisted of presentations by participants and corresponding Q and A; the present situation on facilities of accelerator mass spectrometers (AMS), operations of and research on tandem accelerators or electrostatic accelerators, applications of AMS, related techniques and applications using accelerators, and so on. This report summarizes the contents of presentations made in the meeting. The 30 of the presented papers are indexed individually. (J.P.N.)

  2. The 13th international workshop on targetry and target chemistry proceedings

    Haroun, S. (SFU, TRIUMF (Canada)); Givskov, A.; Jensen, Mikael (Technical Univ. of Denmark. Risoe National Laboratory for Sustainable Energy, Radiation Research Division, Roskilde (Denmark))

    2011-06-15

    This report contains the complete proceedings of the 13th International Workshop on Targetry and Target Chemistry. The Workshop was held at Risoe National Laboratory for Sustainable Energy on July 26-28 2010. The workshop deals with the development of methods and systems for efficient production of radioactive isotopes with accelerators. The WTTC series of workshops was initiated for the purpose of exchanging information about the problems and solutions associated with the production of radioisotopes for biomedical research and their applications to the diagnosis and treatment of disease. The goal of the WTTC is to advance the science associated with radioisotope production targetry. The Workshops are designed to bring experienced targetry scientists together with newcomers to the field, both from industry and academia, to discuss issues of targetry and target chemistry and approaches to exploring in situ target chemistry and the engineering required to optimize production yields. In the workshop, experience, ideas and information are freely and openly shared; learning and collaborations are fostered, with active participation by all attendees. This participation includes both formal and informal sessions. The present proceedings captures both submitted abstracts and the actual presentations showed during the very successful workshop meeting number 13 in the row, the WTTC13. (Author)

  3. The Serbo-Bulgarian relations at the end of the 13th century

    Mišić Siniša

    2009-01-01

    Full Text Available Relations between the Serbs and the Bulgarians at the end of the 13th century has been a part of global international relations in the Balkans and in the Danube region. The Tatars (Mongols as the powerful warring people represent external factor that highly influenced those relations. They also made powerful impact on the Serbian and Bulgarian states of the time. The relations between the Serbs and the Bulgarians had not been at the time determined by their attitude toward the Byzantine Empire which was of the Serbian state, while Bulgaria went through tough period of disintegration of the central power. The internal affaires of those two states influenced their mutual relations. During the war with the Byzantine Empire, king Milutin tried to keep Bulgaria on his side, which is the reason why he became related by the marriage to the Bulgarian imperial family. However, strong involvement of Nogay made this alliance non-useful. After 1299 Serbia lost interest for Bulgaria, and behavior toward the queen Ana made two states enter the 14th century as the enemies. The close relations would be maintained with Vidin. In the frame of those relations Milutin's marriage and release of the Bulgarian princess Ana should be regarded, while Stefan's marriage had been motivated differently and happened around 1305/1306.

  4. The 13th international workshop on targetry and target chemistry proceedings

    This report contains the complete proceedings of the 13th International Workshop on Targetry and Target Chemistry. The Workshop was held at Risoe National Laboratory for Sustainable Energy on July 26-28 2010. The workshop deals with the development of methods and systems for efficient production of radioactive isotopes with accelerators. The WTTC series of workshops was initiated for the purpose of exchanging information about the problems and solutions associated with the production of radioisotopes for biomedical research and their applications to the diagnosis and treatment of disease. The goal of the WTTC is to advance the science associated with radioisotope production targetry. The Workshops are designed to bring experienced targetry scientists together with newcomers to the field, both from industry and academia, to discuss issues of targetry and target chemistry and approaches to exploring in situ target chemistry and the engineering required to optimize production yields. In the workshop, experience, ideas and information are freely and openly shared; learning and collaborations are fostered, with active participation by all attendees. This participation includes both formal and informal sessions. The present proceedings captures both submitted abstracts and the actual presentations showed during the very successful workshop meeting number 13 in the row, the WTTC13. (Author)

  5. Proceedings of the 13th meeting for tandem accelerators and related techniques

    The 13th Meeting for Tandem Accelerators and Related Techniques was held on June 8-9, 2000 at Mutsu, Aomori Prefecture, under the auspices of Marine Research Laboratory, Mutsu Establishment, the Japan Atomic Energy Research Institute. About 60 people participated in the meeting from 27 universities and organizations, many of which have a tandem accelerator or an electrostatic accelerator. The objectives of the meeting are to exchange information on tandem accelerates or electrostatic accelerators by making a presentation on related research and topics, and also to enhance mutual friendship of participants, resulting in the contribution to associated R and D. The meeting mainly consisted of presentations by participants and corresponding Q and A; the present situation on facilities of accelerator mass spectrometers (AMS), operations of and research on tandem accelerators or electrostatic accelerators, applications of AMS, related techniques and applications using accelerators, and so on. This report summarizes the contents of presentations made in the meeting. The 30 of the presented papers are indexed individually. (J.P.N.)

  6. IAEA technical meeting: 13th meeting of the IFRC Subcommittee on Atomic and Molecular Data for Fusion. Summary report

    This report briefly describes the proceedings, conclusions and recommendations of the 13th Meeting of the Subcommittee on Atomic and Molecular Data for Fusion of the International Fusion Research Council held on 24-25 June, 2002 at the IAEA Headquarters in Vienna Austria. The report includes an Executive Summary of the Subcommittee from this Meeting. (author)

  7. Residual risks of the 13th amendment to the German Atomic Energy Act

    The 13th amendment to the German Atomic Energy Act, which was adopted by the German federal parliament on June 30 and entered into force on August 6, 2011, must be judged in the light of its genesis. Federal Chancellor Merkel, in her government declaration of June 9, 2011, had mentioned topics such as residual risk, safety standards, and risk assumptions, on which the federal government, in the week after the event of March 11, 2011, had commissioned the Advisory Committee on Reactor Safeguards (RSK) to conduct a comprehensive safety review of all German nuclear power plants, and appointed an Ethics Committee to write an opinion on safe energy supply. On the basis of quickly drafted reports, the federal cabinet, without any international harmonization (EU, IEA), adopted a draft opt out law on June 6, 2011. How should the declarations by the Federal Chancellor on June 9, 2011 be classified in terms of atomic energy law? In her words, it all revolved around the residual risk. The debate, which has been shifted to the realm of constitutional law, is open to considerations and steps to attack the new opt out law on grounds of material unconstitutionality (violation of the property guarantee under Sec. 14 or the principle of equality under Art.3 of the Basic Law). As far as final storage is concerned, the amendment to the German Atomic Energy Act announced still for this year, also for transposition of the EURATOM Directive of July 19, 2011 about nuclear waste management, the ''re-assessment of the residual risk'' is not likely to play a role. All these events are reminiscent of a sentence by former Federal Chancellor Schmidt: ''The history of the NATO dual-track decision remains a textbook case showing that even in a democracy emotions using ethical arguments, mixed with demagogy, can become strong enough to cast aside balanced reason.'' There is also a distinction by Max Weber between ''ethics of ideology'' and ''ethics of responsibility''. (orig.)

  8. PREFACE: 13th International Workshop on Slow Positron Beam Techniques and Applications (SLOPOS13)

    2014-04-01

    These proceedings originate from the 13th International Workshop on Slow Positron Beam Techniques and Applications SLOPOS13 which was held at the campus of the Technische Universität München in Garching between 15th-20th September, 2013. This event is part of a series of triennial SLOPOS conferences. In total 123 delegates from 21 countries participated in the SLOPOS13. The excellent scientific program comprised 50 talks and 58 posters presented during two poster sessions. It was very impressive to learn about novel technical developments on positron beam facilities and the wide range of their applications all over the world. The workshop reflected the large variety of positron beam experiments covering fundamental studies, e.g., for efficient production of anti-hydrogen as well as applied research on defects in bulk materials, thin films, surfaces, and interfaces. The topics comprised: . Positron transport and beam technology . Pulsed beams and positron traps . Defect profiling in bulk and layered structures . Nanostructures, porous materials, thin films . Surfaces and interfaces . Positronium formation and emission . Positron interactions with atoms and molecules . Many positrons and anti-hydrogen . Novel experimental techniques The international advisory committee of SLOPOS awarded student prizes for the best presented scientific contributions to a team of students from Finland, France, and the NEPOMUC team at TUM. The conference was overshadowed by the sudden death of Professor Klaus Schreckenbach immediately before the workshop. In commemoration of him as a spiritus rectus of the neutron induced positron source a minutes' silence was hold. We are most grateful for the hard work of the Local Organising Committee, the help of the International Advisory Committee, and all the students for their friendly and efficient support during the meeting. The workshop could not have occurred without the generous support of the Heinz Maier-Leibnitz Zentrum (MLZ), Deutsche

  9. PREFACE: 13th High-Tech Plasma Processes Conference (HTPP-2014)

    2014-11-01

    The High-Tech Plasma Processes - 13th European Plasma Conference (HTPP-2014) was held in Toulouse (France) on 22-27 June 2014. The conference series started in 1990 as a thermal plasma conference and has gradually expanded to include other related topics. Now the High-Tech Plasma Processes - European Plasma Conference (HTPP) is an international conference organised in Europe every two years with topics encompassing the whole field of plasma processing science. The aim of the conference is to bring different scientific communities together, to facilitate contacts between science, technology and industry and to provide a platform for the exploration of both the fundamental topics and new applications of plasmas. For this edition of HTPP, as was the case for the last, we have acheived a well balanced participation from the communities of both thermal and non-thermal plasma researchers. 142 people from 17 countries attended the conference with the total number of contributions being 155, consisting of 8 plenary and 8 invited talks plus 51 oral and 88 poster contributions. We have received numerous papers corresponding to the contributions of HTPP-2014 that have been submitted for publication in this volume of Journal of Physics: Conference Series. Each submitted contribution has been peer reviewed (60 referees with at least two reviewing each paper) and the Editors are very grateful to the referees for their careful support in improving the original manuscripts. In total, 52 manuscripts have been accepted for publication covering a range of topics of plasma processing science from plasma fundamentals to process applications through to experiments, diagnostics and modelling. We have grouped the papers into the following 5 topics: - Arc-Materials Interaction and Metallurgy - Plasma Torches and Spraying - Synthesis of Powders and Nanomaterials - Deposition and Surface Treatment - Non-Equilibrium Plasmas We deeply thank the authors for their enthusiastic and high

  10. Ancient earthen constructions and antiseismic dwellings: Boni's project after the earthquake of 13th January 1915 (Avezzano, Italy)

    E. M. Beranger

    1995-01-01

    After the earthquake of 13th January 1915, the archaeologist Giacomo Boni (1859-1925), inspired by ancient building techniques. proposed the reconstruction of Marsica and the Middle Liri Valley with earthen dwellings. With the cooperation of the Italian Ministry of Agriculture, he organized an exhibition where he collected samples of vegetal trellises which were an essential support to implement these techniques. Furthermore, he rebuilt two huts on the Palatine (Orti Farnesiani): ...

  11. Notarial acts «ad pedes consulum». About some Milanese trial records (12th-13th Century

    Marta Luigina Mangini

    2014-06-01

    Full Text Available Milan’s documentary heritage created between the 12th and the 13th century is to be found by searching the records of the administrative offices and religious institutions that regularly had dealings with it. The aim of this paper is to examine some of these records anew, particularly evaluating the evolution of the constants and variables that have strongly influenced their creation and preservation over time.

  12. Computational Models of Cognitive Processes: Proceedings of the 13th Neural Computation and Psychology Workshop (Ncpw13)

    Mayor, Julien; Gomez, Pablo

    2013-01-01

    Computational Models of Cognitive Processes collects refereed versions of papers presented at the 13th Neural Computation and Psychology Workshop (NCPW13) that took place in July 2012, in San Sebastian (Spain). This workshop series is a well-established and unique forum that brings together researchers from such diverse disciplines as artificial intelligence, cognitive science, computer science, neurobiology, philosophy and psychology to discuss their latest work on models of cognitive proces...

  13. 13th FINA World Championship finals: stroke kinematical and race times according to performance, gender and event

    Jesus, S.; Costa, M. J.; Marinho, D.A.; Garrido, N.; Silva, A. J.; Barbosa, Tiago M.

    2011-01-01

    The aim of this work was to compare the stroke kinematics and race times of the freestyle final races at the 13th FINA World Championships between: (i) the three medalists versus the last three finalists; (ii) males versus female swimmers; (iii) all events in each gender. Data was collected from the champioships official web site. There were no significant differences in the stroke kinematics neither in the race times between medallists and non-medallists. There were significant effects in th...

  14. ARC Code TI: Geometry Manipulation Protocol (GMP)

    National Aeronautics and Space Administration — The Geometry Manipulation Protocol (GMP) is a library which serializes datatypes between XML and ANSI C data structures to support CFD applications. This library...

  15. EDITORIAL: Invited review and topical lectures from the 13th International Congress on Plasma Physics

    Zagorodny, A.; Kocherga, O.

    2007-05-01

    The 13th International Congress on Plasma Physics (ICPP 2006) was organized, on behalf of the International Advisory Committee of the ICPP series, by the National Academy of Sciences of Ukraine and the Bogolyubov Institute for Theoretical Physics (BITP) and held in Kiev, Ukraine, 22 26 May 2006. The Congress Program included the topics: fundamental problems of plasma physics; fusion plasmas; plasmas in astrophysics and space physics; plasmas in applications and technologies; complex plasmas. A total of 305 delegates from 30 countries took part in the Congress. The program included 9 invited review lectures, 32 invited topical and 313 contributed papers (60 of which were selected for oral presentation). The Congress Program was the responsibility of the International Program Committee: Anatoly Zagorodny (Chairman) Bogolyubov Institute for Theoretical Physics, Ukraine Olha Kocherga (Scientific Secretary) Bogolyubov Institute for Theoretical Physics, Ukraine Boris Breizman The University of Texas at Austin, USA Iver Cairns School of Physics, University of Sydney, Australia Tatiana Davydova Institute for Nuclear Research, Ukraine Tony Donne FOM-Institute for Plasma Physics, Rijnhuizen, The Netherlands Nikolai S Erokhin Space Research Institute of RAS, Russia Xavier Garbet CEA, France Valery Godyak OSRAM SYLVANIA, USA Katsumi Ida National Institute for Fusion Science, Japan Alexander Kingsep Russian Research Centre `Kurchatov Institute', Russia E P Kruglyakov Budker Institute of Nuclear Physics, Russia Gregor Morfill Max-Planck-Institut für extraterrestrische Physik, Germany Osamu Motojima National Institute for Fusion Science, Japan Jef Ongena ERM-KMS, Brussels and EFDA-JET, UK Konstantyn Shamrai Institute for Nuclear Research, Ukraine Raghvendra Singh Institute for Plasma Research, India Konstantyn Stepanov Kharkiv Institute of Physics and Technology, Ukraine Masayoshi Tanaka National Institute for Fusion Science, Japan Nodar Tsintsadze Physics Institute, Georgia The

  16. PREFACE: EMAS 2013 Workshop: 13th European Workshop on Modern Developments and Applications in Microbeam Analysis

    Llovet, Xavier, Dr; Matthews, Mr Michael B.; Brisset, François, Dr; Guimarães, Fernanda, Dr; Vieira, Professor Joaquim M., Dr

    2014-03-01

    This volume of the IOP Conference Series: Materials Science and Engineering contains papers from the 13th Workshop of the European Microbeam Analysis Society (EMAS) on Modern Developments and Applications in Microbeam Analysis which took place from the 12th to the 16th of May 2013 in the Centro de Congressos do Alfândega, Porto, Portugal. The primary aim of this series of workshops is to assess the state-of-the-art and reliability of microbeam analysis techniques. The workshops also provide a forum where students and young scientists starting out on a career in microbeam analysis can meet and discuss with the established experts. The workshops have a very specific format comprising invited plenary lectures by internationally recognized experts, poster presentations by the participants and round table discussions on the key topics led by specialists in the field. This workshop was organized in collaboration with LNEG - Laboratório Nacional de Energia e Geologia and SPMICROS - Sociedade Portuguesa de Microscopia. The technical programme included the following topics: electron probe microanalysis, future technologies, electron backscatter diffraction (EBSD), particle analysis, and applications. As at previous workshops there was also a special oral session for young scientists. The best presentation by a young scientist was awarded with an invitation to attend the 2014 Microscopy and Microanalysis meeting at Hartford, Connecticut. The prize went to Shirin Kaboli, of the Department of Metals and Materials Engineering of McGill University (Montréal, Canada), for her talk entitled ''Plastic deformation studies with electron channelling contrast imaging and electron backscattered diffraction''. The continuing relevance of the EMAS workshops and the high regard in which they are held internationally can be seen from the fact that 74 posters from 21 countries were on display at the meeting and that the participants came from as far away as Japan, Canada and the USA. A

  17. PREFACE: 13th General Conference of the Condensed Matter Division of the European Physical Society

    Hoffmann, H.; Klein, R.; Schwoerer, M.

    1993-01-01

    The 13th General Conference of the Condensed Matter Division of the European Physical Society was held in conjunction with the Frühjahrstagung des Arbeitskreises Festkörperphysik der Deutschen Physikalischen Gesellschaft from March 29 till April 2, 1993, in Regensburg. The programme comprised 3,134 contributions : 8 Plenary Talks, 171 Invited Talks, 1,480 Contributed Talks, 1,441 Poster Presentations, 1 Public Evening Talk and 33 Exhibitors Reports. The abstracts have been published as Europhysics Conference Abstracts, Volume 17A/Verhandlungen der Deutschen Physikalischen Gesellschaft 5/1993. The table (see PDF file) shows the distribution of the Plenary and Invited Speakers as well as of the participants according to countries within and outside of Europe. The conference was the largest meeting of physicists held in Germany to date. It was a manifestation of the enormous scientific activity in both basic and applied research in the fields of Condensed Matter Physics in Europe. Most of the research work, which was presented at the conference, was done by young physicists. They represent a large human capital in Europe. Most of the senior physicists and many of our young colleagues maintain scientific cooperations, and also personal friendships, which are and which have been almost independent of national barriers over the past three decades. The latter is to a large extent due to the European Physical Society which always cultivated these contacts, especially between the eastern and western parts of Europe. We would like to express our sincere thanks to the members of the Programme Committee. By their intensive work, which was free from national interests, a scientific programme was prepared, which covered the entire field of Condensed Matter Physics. About 70% of the Plenary and Invited Speakers came from 20 different foreign countries and about 30% from Germany. The meeting therefore has been a truly European Conference. For the young physicists, the number of

  18. Proceedings, 13th International Symposium on Nuclei in the Cosmos (NIC XIII)

    Elekes, Zoltán

    2015-01-01

    Nuclei in the Cosmos is the foremost bi-annual conference of nuclear physicists, astrophysicists, cosmochemists, and others to survey the recent achievements in Nuclear Astrophysics. As an interdisciplinary meeting it promotes mutual understanding and collaboration over fields fundamental to solve a range of open questions, from the origin of the elements to stellar evolution. Inherent part of the conference is a school devoted to students and young scientists where prominent scientists introduce the field of nuclear astrophysics to the participants.

  19. Proceedings of RSEEM 2006 : 13th Research Symposium on Emerging Electronic Markets

    2006-01-01

    Electronic markets have been a prominent topic of research for the past decade. Moreover, we have seen the rise but also the disappearance of many electronic marketplaces in practice. Today, electronic markets are a firm component of inter-organisational exchanges and can be observed in many branches. The Research Symposium on Emerging Electronic Markets is an annual conference bringing together researchers working on various topics concerning electronic markets in research and practice. T...

  20. Direct activation of PDE5 by cGMP

    Mullershausen, Florian; Friebe, Andreas; Feil, Robert; Thompson, W. Joseph; Hofmann, Franz; Koesling, Doris

    2003-01-01

    In platelets, the nitric oxide (NO)–induced cGMP response is indicative of a highly regulated interplay of cGMP formation and cGMP degradation. Recently, we showed that within the NO-induced cGMP response in human platelets, activation and phosphorylation of phosphodiesterase type 5 (PDE5) occurred. Here, we identify cyclic GMP-dependent protein kinase I as the kinase responsible for the NO-induced PDE5 phosphorylation. However, we demonstrate that cGMP can directly activate PDE5 without phos...

  1. PREFACE: 13th International Conference on Electrorheological Fluids and Magnetorheological Suspensions (ERMR2012)

    Unal, Halil Ibrahim

    2013-02-01

    Conference photograph The 13th International Conference on Electrorheological Fluids and Magnetorheological Suspensions (ERMR2012) was held in Ankara, Turkey at Gazi University in the Architect Kemaleddin historical hall on 2-6 July 2012. The first International Conference on Electrorheological Fluids and Magnetorheological Suspensions took place nearly 25 years ago and this conference continued the same tradition by providing an arena for researchers around the world to present their new research findings in these fields, and gave them the opportunity to learn about the latest research and technology and to renew their acquaintances. The meeting brought together scientists and engineers in multidisciplinary areas such as chemical engineering, mechanical engineering, materials science and engineering, physics, chemistry and polymer science and technology, to explore the state-of-art technology, identify key areas to be focused on and discuss their problems/issues. All oral presentations were held in a single session to enhance the interactions between the scientists and engineers. The ERMR2012 Conference included plenary lectures given by prominent leaders in their respective fields. About 130 participants from more than 50 organizations attended the conference and 15 plenary speeches, 64 oral presentations and 57 poster presentations took place in the following areas: (i) synthesis, characterization and processing of novel ER/MR materials, (ii) dynamics, chain and structure formation of ER/MR materials, (iii) ER/MR elastomers, ferrogels and their characterizations, (iv) rheological techniques and measurements of ER/MR materials, (v) modeling and simulations of ER/MR materials, (vi) device development and control techniques and (vii) applications of ER/MR materials. The ERMR2012 International Conference began with Turkish classical music performed by the musicians of the Ministry of Culture and Tourism. Rector Professor Dr R Ayhan welcomed the participants and the

  2. 15th International Congress on Plasma Physics & 13th Latin American Workshop on Plasma Physics

    Soto, Leopoldo

    2014-05-01

    The International Advisory Committee of the 15th International Congress on Plasma Physics (ICPP 2010) and the International Advisory Committee of the 13th Latin American Workshop on Plasma Physics (LAWPP 2010), together agreed to carry out this combined meeting ICPP-LAWPP-2010 in Santiago de Chile, 8-13 August 2010, on occasion of the Bicentennial of Chilean Independence. The ICPP-LAWPP-2010 was organized by the Thermonuclear Plasma Department of the Chilean Nuclear Energy Commission (CCHEN) as part of the official program within the framework of the Chilean Bicentennial. The event was also a scientific and academic activity of the project ''Center for Research and Applications in Plasma Physics and Pulsed Power, P4'', supported by National Scientific and Technological Commission, CONICYT-Chile, under grant ACT-26. The International Congress on Plasma Physics was first held in Nagoya, in 1980, and followed by the Congresses: Gothenburg (1982), Lausanne (1984), Kiev (1987), New Delhi (1989), Innsbruck (1992), Foz do Iguacu (1994), Nagoya (1996), Prague (1998), Quebec City (2000), Sydney (2002), Nice (2004), Kiev (2006), and Fukuoka (2008). The purpose of the Congress is to discuss the recent progress and future views in plasma science, including fundamental plasma physics, fusion plasmas, astrophysical plasmas, and plasma applications, and so forth. The Latin American Workshop on Plasma Physics was first held in 1982 in Cambuquira, Brazil, followed by the Workshops: Medellín (1985), Santiago (1988), Buenos Aires (1990), Mexico City (1992), Foz do Iguacu (1994, also combined with ICPP), Caracas (1997), Tandil (1998), La Serena (2000), Sao Pedro (2003), Mexico City (2005), and Caracas (2007). The Latin American Workshop on Plasma Physics is a communication forum of the achievements of the plasma-physics regional community, fostering collaboration between plasma scientists within the region and elsewhere. The program of the ICPP-LAWPP-2010 included the topics

  3. A cracked mirror? - forming the ideal ruler in Epirus and Nicaea in the first half of the 13th century

    Vukašinović Milan

    2015-01-01

    Full Text Available During the first half of the 13th century both Byzantine Empire and the image of its ideal ruler had to undergo a transformation. By applying mostly the narratological analysis to the parenetic texts written in the two successor states of the Empire, the paper sheds light on the dynamic ‘negotiations’ within the Roman elites of the place that the Emperor should have inside the symbolic order, and suggests a possible model of approach to other Byzantine texts and periods. [Projekat Ministarstva nauke Republike Srbije, br. 177015

  4. Proceedings of the 13th China symposium on computer and network applications in modern science and technology

    The 13th China symposium on Computer and Network Applications in Modern Science and Technology was held by Nuclear Electronics and Nuclear Detecting Technology Branch Society on 4-9 November, 2007 in Shenzhen of Guangdong province. 79 articles are collected in the proceedings. The contents include calculation method and software technology, data processing and analysis technique, data acquisition and automatization, network and safety, grid computing technology, information system and database application, system design and development as well as operation maintenance, computer and microprocessor application system and so on.

  5. The Successive CME on 13th; 14th and 15th February 2011 and Forbush decrease on 18 February 2011

    Aims. We analyze the kinematics of three interplanetary coronal mass ejections (ICMEs) that occurred on 13th, 14th and 15th February 2011 in the active region AR 11155 and have shown that they appeared at the Earth orbit on February, 18th and caused Forbush decrease (FD). Methods. The solar coordinates of flares are (S19W03), (S20W14) and (S21W18). The kinematic curves were obtained using STEREO (A and B) data. Additionally, we explore the possibility of the CME-CME interaction for these three events. We compare obtained estimates of ICME arrival with the in-situ measurements from WIND satellite at L1 point and with ground-based cosmic ray data obtained from SEVAN network. Results. The acceleration of each CME is highly correlated with the associated SXR flares energy release. CMEs that erupted at 13 and 14 Feb 2011 are not associated with prominence eruption; maximum velocity was vmax approc. 550 ± 50 km/s and vmax approx. 400 ± 50 km/s, respectively. However, 15 Feb 2011 CME is connected with much more violent eruption associated with a prominence, with maximum velocity of vmax approx. 1400 ± 50 km/s. The last overtakes 13th and 14th Feb CMEs at distances of 32 and 160 R☉, respectively.

  6. Overseas Voter Mobilisation in Singapore: Implications from Malaysia’s 13th General Election

    James Gomez

    2013-01-01

    Full Text Available This paper discusses voter mobilisation and other election-related activities of Malaysian voters living, studying and working in Singapore in the context of Malaysia’s 13th general election (GE13. According to the World Bank, nearly 400,000 Malaysians reside in the city-state. Thus these figures represent a significant Malaysian voter pool based in Singapore. Efforts to mobilise these voters for general elections or other causes have political implications for both countries, which became apparent following Singapore-based Malaysians’ activities to encourage Malaysians to return home to cast their votes during the 13th general election. Singapore’s strict public assembly laws led to several legal issues related to the voter mobilisation and election campaign activities undertaken by Malaysians in the city-state. These legal issues became a source of friction between the two countries during the elections as government leaders and authorities on both sides of the causeway accused the other of interfering in domestic political matters. Given the growing number of Malaysians in Singapore and the likely repeat of Malaysian voter mobilisation activities in Singapore in the run-up to the fourteenth general election (GE14 in 2018, issues related to the election activities of Malaysian voters in Singapore stand to be another set of factors that will shape the health of bilateral relations between these two countries.

  7. Atmospheric CO2 during the 13th century AD: reconciliation of data from ice core measurements and stomatal frequency analysis

    Atmospheric CO2 reconstructions are currently available from direct measurements of air enclosures in Antarctic ice and, alternatively, from stomatal frequency analysis performed on fossil leaves. A period where both methods consistently provide evidence for natural CO2 changes is during the 13th century ad. The results of the two independent methods differ significantly in the amplitude of the estimated CO2 changes (10 ppmv ice versus 34 ppmv stomatal frequency). Here, we compare the stomatal frequency and ice core results by using a firn diffusion model in order to assess the potential influence of smoothing during enclosure on the temporal resolution as well as the amplitude of the CO2 changes. The seemingly large discrepancies between the amplitudes estimated by the contrasting methods diminish when the raw stomatal data are smoothed in an analogous way to the natural smoothing which occurs in the firn

  8. 13th IUPAC International Congress of Pesticide Chemistry: Crop, Environment, and Public Health Protection, Technologies for a Changing World.

    McConnell, Laura L; Racke, Kenneth D; Hapeman, Cathleen J; Seiber, James N

    2016-01-13

    This introductory paper provides an overview of Perspectives papers written by plenary speakers from the 13th IUPAC International Congress of Pesticide Chemistry held in San Francisco, CA, USA, in August 2014. This group of papers emphasizes some of the emerging issues and challenges at the forefront of agricultural research: sustainability; agriculture's response to climate change and population growth; pollinator health and risk assessment; and global food production and food security. In addition, as part of the Congress, a workshop on "Developing Global Leaders for Research, Regulation, and Stewardship of Crop Protection Chemistry in the 21st Century" identified specific recommendations to attract the best scientists to agricultural science, to provide opportunities to study and conduct research on crop protection chemistry topics, and to improve science communication skills. PMID:26709728

  9. 2015 Special Sessions of the 13th International Conference on Practical Applications of Agents and Multi-Agent Systems

    Hernández, Josefa; Mathieu, Philippe; Campbell, Andrew; Fernández-Caballero, Antonio; Moreno, María; Julián, Vicente; Alonso-Betanzos, Amparo; Jiménez-López, María; Botti, Vicente; Trends in Practical Applications of Agents, Multi-Agent Systems and Sustainability : the PAAMS Collection

    2015-01-01

    This volume presents the papers that have been accepted for the 2015 special sessions of the 13th International Conference on Practical Applications of Agents and Multi-Agent Systems, held at University of Salamanca, Spain, at 3rd-5th June, 2015: Agents Behaviours and Artificial Markets (ABAM); Agents and Mobile Devices (AM); Multi-Agent Systems and Ambient Intelligence (MASMAI); Web Mining and Recommender systems (WebMiRes); Learning, Agents and Formal Languages (LAFLang); Agent-based Modeling of Sustainable Behavior and Green Economies (AMSBGE); Emotional Software Agents (SSESA) and Intelligent Educational Systems (SSIES). The volume also includes the paper accepted for the Doctoral Consortium in PAAMS 2015. PAAMS, the International Conference on Practical Applications of Agents and Multi-Agent Systems is an evolution of the International Workshop on Practical Applications of Agents and Multi-Agent Systems. PAAMS is an international yearly tribune to present, to discuss, and to disseminate the latest develo...

  10. Cyclic GMP pathways in hepatic encephalopathy. Neurological and therapeutic implications.

    Montoliu, Carmina; Rodrigo, Regina; Monfort, Pilar; Llansola, Marta; Cauli, Omar; Boix, Jordi; Elmlili, Nisrin; Agusti, Ana; Felipo, Vicente

    2010-03-01

    Cyclic GMP (cGMP) modulates important cerebral processes including some forms of learning and memory. cGMP pathways are strongly altered in hyperammonemia and hepatic encephalopathy (HE). Patients with liver cirrhosis show reduced intracellular cGMP in lymphocytes, increased cGMP in plasma and increased activation of soluble guanylate cyclase by nitric oxide (NO) in lymphocytes, which correlates with minimal HE assessed by psychometric tests. Activation of soluble guanylate cyclase by NO is also increased in cerebral cortex, but reduced in cerebellum, from patients who died with HE. This opposite alteration is reproduced in vivo in rats with chronic hyperammonemia or HE. A main pathway modulating cGMP levels in brain is the glutamate-NO-cGMP pathway. The function of this pathway is impaired both in cerebellum and cortex of rats with hyperammonemia or HE. Impairment of this pathway is responsible for reduced ability to learn some types of tasks. Restoring the pathway and cGMP levels in brain restores learning ability. This may be achieved by administering phosphodiesterase inhibitors (zaprinast, sildenafil), cGMP, anti-inflammatories (ibuprofen) or antagonists of GABAA receptors (bicuculline). These data support that increasing cGMP by safe pharmacological means may be a new therapeutic approach to improve cognitive function in patients with minimal or clinical HE. PMID:20195723

  11. Microbiological criteria for good manufacturing practice (GMP)

    Good manufacturing practice (GMP) consist of an effective manufacturing operation and an effective application of food control. GMP is best supported by the Hazard Analysis Critical Control Point system (HACCP) of the preventive quality assurance, which requires that food irradiation as any food processing technology should be used only with foods of an acceptable quality and adequate handling and storage procedures should precede and follow the processing. The paper concentrates on the first element of the HACCP system for an irradiation plant: the incoming product control, i.e. whether GMP of foods to be irradiated can be assessed by establishing microbiological criteria for their previous good manufacturing practice. In this regard, it summarizes considerations and findings of a ''Consultation on Microbiological Criteria for Foods to be Further Processed Including by Irradiation'' held in 1989 by the International Consultative Group on Food irradiation at the Headquarters of the World Health Organization, Geneva. Difficulties in establishing reference values and defining good manufacturing practices will be pointed out. (orig.)

  12. Microbiological criteria for good manufacturing practice (GMP)

    Farkas, J. (Inst. of Preservation and Livestock Products Technology, Univ. of Horticulture and Food Industry, Budapest (Hungary)); Zukal, E. (Inst. of Preservation and Livestock Products Technology, Univ. of Horticulture and Food Industry, Budapest (Hungary))

    1992-01-01

    Good manufacturing practice (GMP) consist of an effective manufacturing operation and an effective application of food control. GMP is best supported by the Hazard Analysis Critical Control Point system (HACCP) of the preventive quality assurance, which requires that food irradiation as any food processing technology should be used only with foods of an acceptable quality and adequate handling and storage procedures should precede and follow the processing. The paper concentrates on the first element of the HACCP system for an irradiation plant: the incoming product control, i.e. whether GMP of foods to be irradiated can be assessed by establishing microbiological criteria for their previous good manufacturing practice. In this regard, it summarizes considerations and findings of a ''Consultation on Microbiological Criteria for Foods to be Further Processed Including by Irradiation'' held in 1989 by the International Consultative Group on Food irradiation at the Headquarters of the World Health Organization, Geneva. Difficulties in establishing reference values and defining good manufacturing practices will be pointed out. (orig.)

  13. Proceedings of the 17th annual day of scientific lectures and 13th annual meeting of the National Society of Black Physicists

    This report contains paper on the following topics: Evaporating Black Holes in an Inflationary Universe; Completely and Partially Polarized Signal Propagation; Is the Jet in Quasar 1038 + 064 Precessing? Physics of the Anisotropic Ferrite Wakefield Accelerator; Absolute Multilayer Characterization at High Spatial Resolution via Real Time Soft X-Ray Imaging; Update on the Status of the Superconducting Supercollider (SSC) Laboratory; The Use of Charge Coupled Devices in Remote Sensing from Space; A Scientific Visit to the USSR; Migdal-Kadanoff Study of Z5 Symmetric Systems with Generalized Action -- Part 1; Migdal-Kadanoff Study of Z5 Symmetric Systems with Generalized Action -- Part 2; and The Voyager Mission Exploration of the Solar System with Robotic Spacecraft

  14. Unusual guanylyl cyclases and cGMP signaling in Dictyostelium discoideum

    Veltman, D.M.; Bosgraaf, L.; van Haastert, P. J. M.

    2004-01-01

    cGMP is used as a second messenger in many eukaryotes. cGMP signaling requires at least three components: Guanylyl cyclases synthesize cGMP from GTP. Specific cGMP-binding proteins propagate the signal, usually by phosphorylation of their target Finally, phosphodiesterases terminate the cGMP signal

  15. [Chronic disease and health condition prevention in childhood: emphases from the 13th Symposium of Preventive Pediatrics].

    Batinica, Maja; Grgurić, Josip; Jadrijević-Cvrlje, Filip

    2013-01-01

    Chronic diseases in childhood have become an important priority, especially in developed countries, because of higher prevalence, relatively and absolutely. Besides that, inappropriate procedures a chronically ill child can result in child's growth and development disorder. According to literature data, 15-20% of children have chronic disease with the impact on their physical, mental and emotional status. Disease prevention strategies are described at the primary, secondary and tertiary level: how to avoid occurrence of disease, how to diagnose and treat existent disease in early stages, before it causes significant morbidity, and finally how to reduce negative impact of existent disease by restoring function and reducing disease-related complications - how to improve quality of life of children with chronic diseases. The new term of quaternary prevention describes methods to mitigate or avoid results of unnecessary or excessive interventions in the health system. In this paper the authors present recent attitudes about chronic diseases prevention modalities in childhood, which, at the beggining of the 21st century, have become more intriguing and represent a new challenge for pediatric health care. Thus, from preventive standpoint, the following chronic illnesses are discussed: asthma, malignant diseases, autism, epilepsy, cerebral palsy, tuberculosis, diabetes type 1, congenital heart diseases, arterial hypertension, celiac disease, and eating disorders. These emphases are from the 13th Preventive Pediatrics Symposium, which took place in Skrad, June 2nd, 2012. Further activities are planned with the aim of continuation of health care furtherance for children with other chronic illnesses. PMID:23991491

  16. Blue Fluorescent cGMP Sensor for Multiparameter Fluorescence Imaging

    Niino, Yusuke; Hotta, Kohji; Oka, Kotaro

    2010-01-01

    Cyclic GMP (cGMP) regulates many physiological processes by cooperating with the other signaling molecules such as cyclic AMP (cAMP) and Ca2+. Genetically encoded sensors for cGMP have been developed based on fluorescence resonance energy transfer (FRET) between fluorescent proteins. However, to analyze the dynamic relationship among these second messengers, combined use of existing sensors in a single cell is inadequate because of the significant spectral overlaps. A single wavelength indica...

  17. Collaborative networks in the internet of services: 13th IFIP WG 5.5 working conference on virtual enterprises, PRO-VE 2012, Bournemouth, UK, October 2012: proceedings

    L.M. Camarinha-Matos; L. Xu; H. Afsarmanesh

    2012-01-01

    This book constitutes the refereed proceedings of the 13th IFIP WG 5.5 Working Conference on Virtual Enterprises, PRO-VE 2012, held in Bournemouth, UK, in October 2012. The 61 revised papers presented were carefully selected from numerous submissions. They provide a comprehensive overview of identif

  18. Preparation of stable 125I cyclic GMP tyrosine methyl ester suitable for 3',5' cyclic GMP radioimmunoassay by HPLC

    Determination of the concentration of cyclic guanosine monophosphate (cGMP) by radioimmunoassay (RIA) depends upon the availability of suitable radiolabeled tracers and antibody to detect the product. Reverse phase chromatographic techniques can easily separate the reaction products of chloramine-T iodination of succinyl cGMP tyrosine methyl ester. The binding characteristics of the iodination reaction products to anti-cGMP antibody have been determined. Purified succinyl cyclic nucleotide 125I-tyrosine methyl ester binds to cGMP antisera identically as commercially available tracer. The tracer is stable for greater than three months

  19. Specific GMP guidelines for radiopharmaceutical products

    These guidelines are intended to complement those provided in ''Good manufacturing practices for pharmaceutical products'', as well as the GMP for sterile pharmaceutical products. The regulatory procedures necessary to control radiopharmaceutical products are in large part determined by the sources of products and methods of manufacture. Manufacturing procedures within the scope of these guidelines include: preparation of radiopharmaceuticals in hospital radiopharmacies, preparation of radiopharmaceuticals in centralized radiopharmacies, production of radiopharmaceuticals in nuclear centres, institutes or industrial manufacturers, preparation and production of radiopharmaceuticals in Positron Emission Tomography (PET) centres

  20. Sterile devices: A GMP (Good Manufacturing Practices) workshop manual

    Derision, R.; Lower, A.; Bimonte, R.

    1983-05-01

    The manual, which covers GMPs for sterilization processes, presents model procedures and forms as well as a variety of articles and reprints. It is a compilation of GMP materials that small device firms may find useful in understanding how some manufacturers have successfully compiled with the GMP requirements as they apply to the manufacture of sterile devices.

  1. Isolation and Partial Characterization of a Cyclic GMP-Dependent Cyclic GMP-Specific Phosphodiesterase from Dictyostelium discoideum

    Bulgakov, Roman; Haastert, Peter J.M. van

    1983-01-01

    The cellular slime mold, Dictyostelium discoideum, contains at least two classes of phosphodiesterase activity. One class of enzymes hydrolyses cyclic AMP (cAMP) and cyclic GMP (cGMP) with approximately equal rates. Another enzyme, which is less than 5% of the total activity, specifically hydrolyses

  2. Distinct Allostery Induced in the Cyclic GMP-binding, Cyclic GMP-specific Phosphodiesterase (PDE5) by Cyclic GMP, Sildenafil, and Metal Ions*

    Biswas, Kabir H.; Visweswariah, Sandhya S.

    2010-01-01

    The activity of many proteins orchestrating different biological processes is regulated by allostery, where ligand binding at one site alters the function of another site. Allosteric changes can be brought about by either a change in the dynamics of a protein, or alteration in its mean structure. We have investigated the mechanisms of allostery induced by chemically distinct ligands in the cGMP-binding, cGMP-specific phosphodiesterase, PDE5. PDE5 is the target for catalytic site inhibitors, s...

  3. The institutions forming the socioeconomic structure of Turkish private enterprises between the 13th and the 19th centuries

    Mehmet Özbirecikli

    2015-04-01

    principios éticos de la vida empresarial, en esencia, son los mismos. Dentro de este contexto, nos atrevemos a sugerir que las raíces del código ético de la vida empresarial turca se retrotraen en la historia a hace más de 800 años. Además, la similitud entre el funcionamiento presente y pasado indica que el origen de la formación de los aprendices para las empresas turcas tiene, igualmente, más de 800 años de historia.This study investigates three institutions forming the socioeconomic structure of Turkish private enterprises between the 13th and 19th Centuries: Akhism (13th-16th century, the Lonca System (the Guilds (16th-18th century, and the Gedik (Monopoly System (18th-20th century. The study particularly focuses on the social and economic rules, vocational training process, and organizational structure of the said institutions in order to discuss the effects of the socioeconomic structure of Turkish enterprises on economic and social development of private enterprises. The study also struggles to link between the relevant current applications and the applications in the past such as the social rules and vocational training. From economic point of view, both the statist structure of the State and the economic rules of the institutions herein caused private enterprises to remain small, and prevented them from having a competitive environment and having capital accumulation. As a result, enterprises could not benefit from new production techniques and the Turkish enterprise mentality fell behind modern developments On the other hand, although these three systems were completely abolished in the early 20th Century, it is seen that especially traces of the Akhism and Lonca systems have still been surviving. Both the most of rules of Akhism and some of the duties of the board of directors of Lonca such as keeping moral standards of production and trade remind us of professional code of ethics of today's modern business life. In other saying, there was code of

  4. Mobile as a Mainstream – Towards Future Challenges in Mobile Learning. Proceedings of the 13th World Conference on Mobile and Contextual Learning (mLearn 2014)

    Kalz, Marco; Bayyurt, Yasemin; Specht, Marcus

    2014-01-01

    This book constitutes the proceedings of the 13th World Conference on Mobile and Contextual Learning, mLearn 2014, held in Istanbul, Turkey, in November 2014. The 20 revised full papers and 17 short papers presented were carefully reviewed and selected from 65 submissions. The papers are organized in topical sections on technologies and interaction; tablets and ebook readers; learning and teaching inside and outside the classroom; learning design and design implications; evaluation and review...

  5. The cGMP system in irradiated animals

    Changes in the functioning of the cGMP system of the thymocytes and liver of mice subjected to 8 Gy roentgen irradiation were found. Within one hour after irradiation an increase in the cGMP level in thymocytes was noted; two rises in the cGMP concentration in the liver were established, at 0.5 and 24 hours after irradiation. These changes in the cGMP level were correlated to an increase in the guanylate cyclase activities in the thymocytes and liver of the mice subjected to irradiation, and to a lesser extent to changes in the activities of cGMP phosphodiesterase in these tissues. A post-irradiation increase in the rat liver guanylate cyclase activity was also observed. A decrease in cGMP phosphodiesterase activity in the liver of the irradiated mice was followed by a change in the enzymatic kinetics and an increase in cGMP phosphodiesterase thermolability. The post-irradiational rise in guanylate cyclase activity was produced by activation of the enzyme. (orig.)

  6. Multiple Degradation Pathways of Chemoattractant Mediated Cyclic GMP Accumulation in Dictyostelium

    Haastert, Peter J.M. van; Lookeren Campagne, Michiel M. van; Kesbeke, Fanja

    1983-01-01

    Chemoattractants induce a transient accumulation of cGMP levels in Dictyostelium. Intracellular cGMP levels reach a peak at 10 s and prestimulated cGMP levels are recovered at about 30 s. Intracellular and extracellular cGMP levels were detected simultaneously after stimulation of D. lacteum cells w

  7. Opposite effects of different hexachlorocyclohexane (lindane) isomers on cerebellar cyclic GMP: relation of cyclic GMP accumulation to seizure activity

    A number of different depressant and convulsant agents have been shown to alter accumulation of cerebellar cyclic GMP. Since the different hexachlorocyclohexane (HCH) isomers elicit different pharmacological responses in mammals, the authors examined their effects on the accumulation of cerebellar cyclic GMP. Mice received one of the HCH isomers and were sacrificed for determination of cyclic GMP concentrations one hour later. Gamma-HCH increased cyclic GMP while alpha and delta -HCH decreased it. In addition, alpha and delta -HCH prevented the increase in cyclic GMP due to the gamma isomer. Picrotoxin increased cyclic GMP in a manner similar to that of gamma-HCH while strychnine produced only a small increase. All three HCH isomers inhibited the binding of 3H-TBOB (a ligand for the GABA-A-receptor linked chloride channel) in mouse cerebellum. It is concluded the different HCH isomers can have different effects on cerebellar cyclic GMP accumulation and that these effects may mediated through actions at the GABA-A receptor linked chloride channel. 15 references, 1 figure, 3 tables

  8. Preface: Proceedings of the 13th Conference on Liquid and Amorphous Metals (LAM13) (Ekaterinburg, Russia, 8 14 July 2007)

    Popel, Pjotr; Gelchinskii, Boris; Sidorov, Valeriy

    2008-03-01

    The most recent developments in the field of liquid and amorphous metals and alloys are regularly updated through two complementary international conferences: the liquid and amorphous metals conference (LAM) and the rapidly quenched materials (RQ) conference. The first series of conferences started as LM1 in 1966 at Brookhaven for the basic understanding of liquid metals. The subsequent LM conferences were held in Tokyo (1972) and Bristol (1976). The conference was renewed in Grenoble (1980) as a LAM conference including amorphous metals and continued in Los Angeles (1983), Garmisch-Partenkirchen (1986), Kyoto (1989), Vienna (1992), Chicago (1995), Dortmund (1998), Yokohama (2001) and Metz (2004). The conferences are mainly devoted to liquid and amorphous metals and alloys. However, communications on some non-metallic systems such as semiconductors, quasicrystals etc, are also accepted. The conference tradition strongly encourages participation from junior researchers and graduate students. The 13th conference of the LAM series was organized in Ekaterinburg, Russia, by the Institute of Metallurgy of the Ural Branch of the Russian Academy of Sciences (IMet UB RAS) and the Ural State Pedagogical University (USPU), and held from 8-14 July 2007 under the chairmanship of Professors Pjotr Popel (USPU) and Boris Gelchinskii (IMet UB RAS). Two hundred and forty two active participants and about 60 guest participants from 20 countries attended the conference. There were no parallel sessions and all oral reports were separated into three groups: invited talks (40 min), full-scale oral reports (25 min), and brief oral reports (15 min). The program included ten sessions, ranging from purely theoretical subjects to the technological application of molten and amorphous alloys. The following sessions took place: A: Electronic structure and transport, magnetic properties; B: Phase transitions; C: Structure; D: Atomic dynamics and transport; E: Thermodynamics; F: Modelling

  9. Dexamethasone-dependent modulation of cyclic GMP synthesis in podocytes.

    Lewko, Barbara; Waszkiewicz, Anna; Maryn, Anna; Gołos, Magdalena; Latawiec, Elżbieta; Daca, Agnieszka; Witkowski, Jacek M; Angielski, Stefan; Stępiński, Jan

    2015-11-01

    Podocytes may be direct target for glucocorticoid therapy in glomerular proteinuric disease. Permeability of podocytes largely depends on their capacity to migrate which involves the contractile apparatus in their foot processes. In this study, we examined the effect of synthetic glucocorticoid dexamethasone (DEX) on the ability of podocytes to produce cyclic guanosine monophosphate (cGMP) in the presence of vasoactive factors, atrial natriuretic peptide (ANP), nitric oxide (NO), and angiotensin II (Ang II). We investigated also the effects of cGMP and DEX on podocyte motility. Primary rat podocytes and immortalized mouse podocytes were pretreated with 1 µM DEX for 4 or 24 h. Glomerular hypertension was mimicked by subjecting the cells to mechanical stress. Total and subcellular cGMP levels were determined in podocytes incubated with 0.1 µM ANP, 1 µM S-nitroso-N-acetyl penicillamine (SNAP), and 1 µM Ang II. Cell motility was estimated by a wound-healing assay. The ANP-dependent production of cGMP increased after 4 h exposition to DEX, but was attenuated after 24 h. Adversely, a 24-h pretreatment with DEX augmented the NO-dependent cGMP synthesis. Ang II suppressed the ANP-dependent cGMP production and the effect was enhanced by DEX in mechanical stress conditions. Mechanical stress reduced total cGMP production in the presence of all stimulators, whereas extracellular to total cGMP ratio increased. 8-Br cGMP enhanced podocyte migration which was accompanied by F-actin disassembly. In the presence of DEX these effects were prevented. We conclude that DEX modulates the production of cGMP in podocytes stimulated with vasoactive factors such as Ang II, ANP, and NO, and the effect is time-dependent. cGMP increases podocyte motility, which is prevented by DEX. This mechanism may account for the antiproteinuric effect of glucocorticoids. PMID:26272337

  10. Urinary cGMP concentrations in severe primary pulmonary hypertension

    M. Bogdan; Humbert, M.; Francoual, J; Claise, C.; Duroux, P.; Simonneau, G; Lindenbaum, A.

    1998-01-01

    BACKGROUND—Prognostic evaluation of patients with primary pulmonary hypertension (PPH) requires right heart catheterisation. The development of accurate non-invasive methods for monitoring these patients remains an important task. Cyclic guanosine monophosphate (cGMP) is an indicator of the action of natriuretic peptides and nitric oxide on target cells. Plasma and urinary cGMP concentrations are raised in patients with congestive heart failure in whom they correlate clos...

  11. The 13th International Conference on Micro and Nanotechnology for Power Generation and Energy Conversion Applications (PowerMEMS 2013)

    Mitcheson, Paul; Beeby, Steve

    2013-12-01

    It is a pleasure to welcome you to The Royal Society in London and the 13th International Conference on Micro- and Nano-Technology for Power Generation and Energy Conversion Applications, or PowerMEMS 2013. The objective of PowerMEMS 2013 is to catalyse innovation in miniature, micro- and nano-scale technologies for power generation and energy conversion. The conference aims to stimulate the exchange of insights and information, and the development of new ideas in the Power MEMS/NEMS field as well as at the meso-scale. It will allow the attendees to interact and network within our multidisciplinary community that includes professionals from many branches of science and engineering. The technical program is led by four invited speakers covering inductive power transfer, chip scale power sources, thermal energy harvesting and implantable biofuel cells. We received 177 abstracts and following a careful reviewing process by the Technical Program Committee a total of 137 papers were selected for presentation. These have been organised into 16 oral sessions in two parallel streams and two poster sessions that have been augmented by 10 late news papers. The oral and regular poster papers are, for the first time, being published by the Institute of Physics. We have made every effort to make PowerMEMS 2013 the busiest yet and have included for the first time the PowerMEMS School. This two-day school held at Imperial College London covered a wide range of power-MEMS topics including technologies for power generation, power transmission, energy storage, power electronics interfaces and metrology. Registrations for the School exceeded our expectations and it was full by early November. We hope this, and other activities such as the Discussion Panel and the inclusion of late news papers, will make PowerMEMS 2013 a memorable success. We have also reached out to new communities, such as those working in wireless power transfer and RF harvesting to broaden the technology remit of

  12. Role of Phosphodiesterase 5 and Cyclic GMP in Hypertension.

    Mergia, Evanthia; Stegbauer, Johannes

    2016-04-01

    Cyclic GMP (cGMP) is a ubiquitous intracellular second messenger that mediates a wide spectrum of physiologic processes in multiple cell types within the cardiovascular and nervous systems. Synthesis of cGMP occurs either by NO-sensitive guanylyl cyclases in response to nitric oxide or by membrane-bound guanylyl cyclases in response to natriuretic peptides and has been shown to regulate blood pressure homeostasis by influencing vascular tone, sympathetic nervous system, and sodium and water handling in the kidney. Several cGMPs degrading phosphodiesterases (PDEs), including PDE1 and PDE5, play an important role in the regulation of cGMP signaling. Recent findings revealed that increased activity of cGMP-hydrolyzing PDEs contribute to the development of hypertension. In this review, we will summarize recent research findings regarding the cGMP/PDE signaling in the vasculature, the central nervous system, and the kidney which are associated with the development and maintenance of hypertension. PMID:27079836

  13. Biofilms and Cyclic di-GMP (c-di-GMP) Signaling: Lessons from Pseudomonas aeruginosa and Other Bacteria.

    Valentini, Martina; Filloux, Alain

    2016-06-10

    The cyclic di-GMP (c-di-GMP) second messenger represents a signaling system that regulates many bacterial behaviors and is of key importance for driving the lifestyle switch between motile loner cells and biofilm formers. This review provides an up-to-date compendium of c-di-GMP pathways connected to biofilm formation, biofilm-associated motilities, and other functionalities in the ubiquitous and opportunistic human pathogen Pseudomonas aeruginosa This bacterium is frequently adopted as a model organism to study bacterial biofilm formation. Importantly, its versatility and adaptation capabilities are linked with a broad range of complex regulatory networks, including a large set of genes involved in c-di-GMP biosynthesis, degradation, and transmission. PMID:27129226

  14. New GMP Models for Caucasus Region

    Jorjiashvili, N.; Godoladze, T.; Tvaradze, N.; Tumanova, N.

    2014-12-01

    The Caucasus is a region of numerous natural hazards and ensuing disasters. Analysis of the losses due to past disasters indicates those most catastrophic in the region have historically been due to strong earthquakes. Estimation of expected ground motion is a fundamental earthquake hazard assessment. The most commonly used parameter for attenuation relation is peak ground acceleration because this parameter gives useful information for Seismic Hazard Assessment. Thus, many peak ground acceleration attenuation relations have been developed by different authors. However, a few attenuation relations were developed for Caucasus region: Ambraseys et al. (1996,2005) which were based on entire European region and they were not focused locally on Caucasus Region, Smit et.al.(2000) that was based on a small amount of acceleration data that really is not enough. Since 2003 construction of Georgian Digital Seismic Network has started with the help of number of International organizations, Projects and Private companies. In this study new GMP models are obtained based on new data from Georgian seismic network and also from neighboring countries. Estimation of models is obtained by classical, statistical way, regression analysis. Also site ground conditions are considered because the same earthquake recorded at the same distance may cause different damage according to ground conditions. Thus, this parameter is emphasized in the present study. Here it must be mentioned that in previous model which only one was done for Caucasus Region (Smit et. al., 2000) local conditions were not considered. Thus, it is an advantage of models from this study.

  15. The end of the nuclear power generation. On the recommendations of the ethics committee according to the 13th amendment to the Atomic Energy Act

    With the 13th Amendment to the Atomic Energy Act the nuclear consequences of the earthquake disaster in Japan will result in an end to nuclear power generation in Germany. Here, the legislature resorted to unusual methods. For the first time, the legislature received advices from the ''ethics committee reliable energy supply''. This Ethics Commission adopted its recommendations ''on behalf of the Chancellor'' in the period from 4th April to 28th May, 2011. The understanding of this development, its epochal character and speed of decision-making requires an excursion into the economic history and the establishment of nuclear power generation with their legal protection.

  16. Feeds from genetically modified plants (GMP) - Nutritional and safety assessment

    The cultivation of GMP increased worldwide from 1.7 (1996) to about 114 million ha. Currently, soybeans (60), corn (24), cotton (11) and canola (5 % of global GM area) are the most important GMP. They are modified mainly for agronomic traits. Such plants are characterized by so-called input traits (GMP of the first generation) without substantial changes in composition or nutritive value. GMP of the second generation (with output traits) should contain more valuable nutrients (e.g. amino acids, fatty acids, vitamins, enzymes etc.) or less anti-nutritive substances (e.g. mycotoxins, inhibitors, allergens etc.). Nutritional and safety assessment of feeds from GMP is a large challenge for animal nutritionists. The paper reviews feeding trials done with various animal species and categories in many countries. Up to now more than 100 feeding studies with food producing animals were published. Since 1997, 18 studies were performed at the Institute of Animal Nutrition of the FLI to determine the effect of first generation GMP feeds on the nutrition of dairy cows, growing bulls, growing and finishing pigs, laying hens, chickens for finishing, as well as with growing and laying quails. This research was recently summarized. The composition of feeds was analysed, and animal studies were used to assess nutritional qualities, including parameters such as digestibility, feed intake, health and performance of target animal species, and effects on food quality derived from the animals. Reproduction was also considered in generation studies with quails and laying hens. Both chemical analyses and the animal studies reveal no significant differences between GMP feeds and their isogenic counterparts and hence strongly support their substantial equivalence. Our results agree with more than 100 studies published in the literature and reviewed recently. Mycotoxin contamination of some GM-crops is lower than of non-GM, which may be one exception to their substantial equivalence. For

  17. JOOSE, N. Peeter (Leiden - Boston, 2004), A Syriac Encyclopaedia of Aristotelian Philosophy: Barhebraeus (13th c.), Butyrum sapientiae, Books of Ethics, Economy and Politics. A Critical Edition, with Introduction, Translation, Commentary and Glossaries

    Monferrer-Sala, J.P.

    2010-01-01

    Reseña de: A Syriac Encyclopaedia of Aristotelian Philosophy: Barhebraeus (13th c.), Butyrum sapientiae, Books of Ethics, Economy and Politics. A Critical Edition, with Introduction, Translation, Commentary and Glossaries

  18. Fluorescence-Based Reporter for Gauging Cyclic Di-GMP Levels in Pseudomonas aeruginosa

    Rybtke, Morten T.; Borlee, Bradley R.; Murakami, Keiji;

    2012-01-01

    of antipathogenic compounds. Here we describe the development of fluorescent monitors that can gauge the cellular level of cyclic di-GMP in P. aeruginosa. We have created cyclic di-GMP level reporters by transcriptionally fusing the cyclic di-GMP-responsive cdrA promoter to genes encoding green fluorescent protein....... We show that the reporter constructs give a fluorescent readout of the intracellular level of cyclic di-GMP in P. aeruginosa strains with different levels of cyclic di-GMP. Furthermore, we show that the reporters are able to detect increased turnover of cyclic di-GMP mediated by treatment of P...

  19. Pharmaceutical development and preclinical evaluation of a GMP-grade anti-inflammatory nanotherapy

    Lobatto, Mark E.; Calcagno, Claudia; Otten, Maarten J.; Millon, Antoine; Ramachandran, Sarayu; Paridaans, Maarten P M; van der Valk, Fleur M.; Storm, G; Stroes, Erik S G; Fayad, Zahi A.; Mulder, Willem J M; Metselaar, Josbert M.

    2015-01-01

    The present study describes the development of a good manufacturing practice (GMP)-grade liposomal nanotherapy containing prednisolone phosphate for the treatment of inflammatory diseases. After formulation design, GMP production was commenced which yielded consistent, stable liposomes sized 100. nm

  20. Pharmaceutical development and preclinical evaluation of a GMP-grade anti-inflammatory nanotherapy

    Lobatto, Mark E.; Calcagno, Claudia; Otten, Maarten J.; Millon, Antoine; Ramachandran, Sarayu; Paridaans, Maarten P.M.; Valk, van der Fleur M.; Storm, Gert; Stroes, Erik S.G.; Fayad, Zahi A.; Mulder, Willem J.M.; Metselaar, Josbert M.

    2015-01-01

    The present study describes the development of a good manufacturing practice (GMP)-grade liposomal nanotherapy containing prednisolone phosphate for the treatment of inflammatory diseases. After formulation design, GMP production was commenced which yielded consistent, stable liposomes sized 100 nm

  1. C-di-GMP regulates antimicrobial peptide resistance in Pseudomonas aeruginosa

    Chua, Song Lin; Tan, Sean Yang-Yi; Rybtke, Morten Theil;

    2013-01-01

    Bis-(3'-5')-cyclic dimeric GMP (c-di-GMP) is an intracellular second messenger which controls the life styles of many bacteria. A high intracellular level of c-di-GMP induces a biofilm lifestyle, whereas a low intracellular level of c-di-GMP stimulates dispersal of biofilms and promotes a plankto...... current dogma stating that dispersed cells are inevitably more susceptible to antibiotics than their sessile counterparts....

  2. Responses to Elevated c-di-GMP Levels in Mutualistic and Pathogenic Plant-Interacting Bacteria

    Pérez-Mendoza, Daniel; Aragón, Isabel M.; Prada-Ramírez, Harold A.; Romero-Jiménez, Lorena; Ramos, Cayo; Gallegos, María-Trinidad; Sanjuán, Juan

    2014-01-01

    Despite a recent burst of research, knowledge on c-di-GMP signaling pathways remains largely fragmentary and molecular mechanisms of regulation and even c-di-GMP targets are yet unknown for most bacteria. Besides genomics or bioinformatics, accompanying alternative approaches are necessary to reveal c-di-GMP regulation in bacteria with complex lifestyles. We have approached this study by artificially altering the c-di-GMP economy of diverse pathogenic and mutualistic plant-interacting bacteri...

  3. 76 FR 51395 - Draft Environmental Impact Statement for the General Management Plan (DEIS/GMP), Canaveral...

    2011-08-18

    ... National Park Service Draft Environmental Impact Statement for the General Management Plan (DEIS/GMP... Draft Environmental Impact Statement for the General Management Plan (DEIS/GMP), Canaveral National... 1969 the NPS announces the availability of a DEIS/GMP for Canaveral National Seashore, Florida....

  4. cGMP is tightly bound to bovine retinal rod phosphodiesterase

    Although the total concentration of cGMP in rod outer segments is thought to be substantially greater than the free concentration, no quantitatively relevant site for the bound cGMP has been described in mammalian photoreceptors. They have found that preparations of purified bovine rod photoreceptor cyclic nucleotide phosphodiesterase (PDE) contain 1.8 ± 0.3 mol of tightly bound cGMP per mol of PDE. When subunits of the purified PDE were separated by reverse-phase HPLC, a peak of material having spectral properties characteristic of a guanine ring was seen. This material was identified as cGMP. When incubated with 1μM [3H]cGMP, only 0.1 mol of [3H]cGMP bound per mol of purified PDE. Scatchard plots of [3H]cGMP binding have indicated that two classes of binding sites are present on the rod PDE. The observation that stoichiometric amounts of cGMP are tightly bound to PDE accounts for the inability to purify the bovine rod PDE on cGMP affinity columns or to demonstrate stoichiometric high-affinity binding sites with [3H]cGMP. More significantly, the tightly bound cGMP may resolve the apparent discrepancy between the free and total cGMP concentrations of photoreceptor outer segments

  5. Crystal Structure of PKG I:cGMP Complex Reveals a cGMP-Mediated Dimeric Interface that Facilitates cGMP-Induced Activation.

    Kim, Jeong Joo; Lorenz, Robin; Arold, Stefan T; Reger, Albert S; Sankaran, Banumathi; Casteel, Darren E; Herberg, Friedrich W; Kim, Choel

    2016-05-01

    Cyclic guanosine monophosphate (cGMP)-dependent protein kinase (PKG) is a key regulator of smooth muscle and vascular tone and represents an important drug target for treating hypertensive diseases and erectile dysfunction. Despite its importance, its activation mechanism is not fully understood. To understand the activation mechanism, we determined a 2.5 Å crystal structure of the PKG I regulatory (R) domain bound with cGMP, which represents the activated state. Although we used a monomeric domain for crystallization, the structure reveals that two R domains form a symmetric dimer where the cGMP bound at high-affinity pockets provide critical dimeric contacts. Small-angle X-ray scattering and mutagenesis support this dimer model, suggesting that the dimer interface modulates kinase activation. Finally, structural comparison with the homologous cyclic AMP-dependent protein kinase reveals that PKG is drastically different from protein kinase A in its active conformation, suggesting a novel activation mechanism for PKG. PMID:27066748

  6. Good manufacturing practices (GMP utilized on human blood irradiation process

    Cláudio Boghi

    2008-01-01

    Full Text Available Irradiation of human blood is used to avoid the TA-GVHD (transfusion-associated graft-versus-host-disease, a rare but devastating adverse effect of leukocytes present in blood components for immunocompetent transfusion recipients. Usually this irradiation practice is performed to a physical elimination of lymphocytes. The implementation of the GMP will assure that the properly dose in a range of 25Gy to 50Gy will be delivered to the blood in the bag collected in a blood tissue bank. The studies to establish the GMP were developed under the guidelines of the standard ISO 11137 - Sterilization of health care products - Requirements for validation and routine control - Radiation sterilization. In this work, two dosimetric systems were used for dose mapping during the studies of irradiator qualification, loading pattern, irradiation process validation and auditing. The CaSO4: Dy dosimeter presented difficulties concerning to uncertainty on dose measurement, stability, trace ability and calibration system. The PMMA and gafchromic dosimetric systems have shown a better performance and were adopted on establishment of GMP procedures. The irradiation tests have been done using a Gammacell 220 Irradiator. The developed GMP can be adapted for different types of gamma irradiators, allowing to set up a quality assurance program for blood irradiation.

  7. Implementation of good manufacturing practices (GMP) on human blood irradiation

    Boghi, Claudio; Napolitano, Celia M.; Ferreira, Danilo C.; Rela, Paulo Roberto [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)]. E-mails: cboghi@uol.com.br; cmnapoli@ipen.br; dancarde@ig.com.br; prela@ipen.br; Zarate, Herman S. [Comission Chilena de Energia Nuclear, Santiago (Chile)]. E-mail: hzarate@cchen.cl

    2007-07-01

    The irradiation of human blood is used to avoid the TA-GVHD (transfusion-associated graft-versus-host-disease), a rare but devastating adverse effect of leukocytes present in blood components for a immuno-competent transfusion recipients. Usually this irradiation practice is performed to a physical elimination of lymphocytes. The implementation of the GMP will assure that the properly dose in a range of 25 Gy to 50 Gy will be delivered to the blood in the bag collected in a blood tissue bank. The studies to establish the GMP were developed under the guidelines of the standard ISO 11137 - Sterilization of health care products - Requirements for validation and routine control - Radiation sterilization. In this work, two dosimetric systems were used for dose mapping during the studies of irradiator qualification, loading pattern, irradiation process validation and auditing. The CaSO{sub 4}: Dy dosimeter presented difficulties concerning to uncertainty on dose measurement, stability, trace ability and calibration system. The PMMA and gafchromic dosimetric systems have shown a better performance and were adopted on establishment of GMP procedures. The irradiation tests have been done using a Gammacell 220 Irradiator. The developed GMP can be adapted for different types of gamma irradiators, allowing to set up a quality assurance program for blood irradiation. (author)

  8. Implementation of good manufacturing practices (GMP) on human blood irradiation

    The irradiation of human blood is used to avoid the TA-GVHD (transfusion-associated graft-versus-host-disease), a rare but devastating adverse effect of leukocytes present in blood components for a immuno-competent transfusion recipients. Usually this irradiation practice is performed to a physical elimination of lymphocytes. The implementation of the GMP will assure that the properly dose in a range of 25 Gy to 50 Gy will be delivered to the blood in the bag collected in a blood tissue bank. The studies to establish the GMP were developed under the guidelines of the standard ISO 11137 - Sterilization of health care products - Requirements for validation and routine control - Radiation sterilization. In this work, two dosimetric systems were used for dose mapping during the studies of irradiator qualification, loading pattern, irradiation process validation and auditing. The CaSO4: Dy dosimeter presented difficulties concerning to uncertainty on dose measurement, stability, trace ability and calibration system. The PMMA and gafchromic dosimetric systems have shown a better performance and were adopted on establishment of GMP procedures. The irradiation tests have been done using a Gammacell 220 Irradiator. The developed GMP can be adapted for different types of gamma irradiators, allowing to set up a quality assurance program for blood irradiation. (author)

  9. Qualification of the GMP compliant FDG production and distribution unit

    Recently we installed in our laboratory a GMP-compliant compact combined FDG production and dose distribution unit. This unit, constructed by HWM, Rossendorf, consists of the mini-cell, housing the FDG synthesis module from GE, standing side-by-side with the class A shielded isolator containing remotelly controlled dose distribution module. Use of the isolator technology combined with laminar flow HEPA filtered air inside the dose distribution hot-cell allows to achieve class A environment for the aseptic filing of FDG into the opened sterile vials. Synthesis of FDG and preparation of the mother batch solution is achieved inside the hot-cells providing class C clean room environement. The whole production unit is standing in the class D laboratory. The complete set of IQ-OQ-PQ tests was performed to assure that the FDG production and distribution unit is performing accordingly to GMP requirements and is providing the product of consistent quality. The majour component in the qualification tests are the aseptic procedure validations, which consisted of the procedures recommended by GMP guidelines and adapted for the specificity of the FDG production. As a minimum for the aseptic validation we considered the Mediafil Test, Bioburden Test and test productions. All the test results assure that the FDG production unit allows to produce FDG for central distribution in accordance with the requirements of European pharmacopoeia and GMP guidelines. (author)

  10. Cyclic GMP alters Ca exchange in vascular smooth muscle

    Contraction and 42K efflux from vascular smooth muscle stimulated either by norepinephrine (NE) or by K-depolarization is dependent on an increase in cytosolic Ca concentration. The purpose of this study was to determine if cyclic GMP (cGMP) inhibited these processes and if inhibition was secondary to the action of cGMP on Ca movements. Basal cGMP content of rat aorta was 1.2 fmol/mg wet wt. Sodium nitroprusside (NP) increased cGMP ∼2-fold at 1 nM and ∼750-fold at 1 μM with no effect on cAMP levels. A 5 min pretreatment with NP (1 μM) completely prevented tension development induced by 3 μM NE. The same concentration of NP also inhibited NE-stimulated 42K and 45Ca efflux > 90 and > 80%, respectively. Removal of NP in the continued presence of NE (3 μM) caused recovery of the 42K efflux response to ∼75% of control with a half-time of ∼2.5 min. NP (1 μM) also caused a rapid relaxation of aorta contracted with 3 μM NE and a loss of the 42K efflux response with half-times of 2-3 min. In contrast, 100 μM NP produced only a 50% inhibition of contraction induced by high K (55 mM). Also, NP (1 μM) inhibited K-stimulated 42K efflux only ∼25%. These results demonstrate both a concentration- and a time-dependent relationship between increases in cGMP induced by NP and decreases in NE-stimulated contraction, 42K and 45Ca effluxes. They also indicate that the sensitivity of NE-induced contraction and 42K efflux to NP is greater than that induced by high K. These studies suggest that cGMP modulates the control sites for Ca exchange in the plasma membrane and sarcoplasmic reticulum

  11. Hydrogen peroxide is involved in cGMP modulating the lateral root development of Arabidopsis thaliana

    Li, Jisjeng; Jia, Honglei

    2013-01-01

    3′,5′-cyclic guanosine monophosphate (cGMP) and hydrogen peroxide (H2O2) function as the important signaling molecule which promote the lateral root development of Arabidopsis thaliana. In this study, interestingly, application of 8-Br-cGMP (the membrane permeable cGMP analog) promoted the endogenous H2O2 production. In addition, the decrease of endogenous H2O2 also inhibited the effect of cGMP on the lateral root development. Thus, H2O2 maybe act as a downstream signaling of cGMP molecule wh...

  12. 13th international symposium on autoradiography (13th ISA '83)

    At the conference, 39 papers were heard all of which are included in INIS. Topics covered include: trends in development of solid state track detectors, theory and methods of quantitative autoradiography, equipment for autoradiographs processing and track counting, uses of autoradiography in research of semiconductors, in metallography and in biology

  13. Signal transduction, receptors, mediators and genes: younger than ever - the 13th meeting of the Signal Transduction Society focused on aging and immunology

    Klotz Lars-Oliver

    2010-02-01

    Full Text Available Abstract The 13th meeting of the Signal Transduction Society was held in Weimar, from October 28 to 30, 2009. Special focus of the 2009 conference was "Aging and Senescence", which was co-organized by the SFB 728 "Environmentally-Induced Aging Processes" of the University of Düsseldorf and the study group 'Signal Transduction' of the German Society for Cell Biology (DGZ. In addition, several other areas of signal transduction research were covered and supported by different consortia associated with the Signal Transduction Society including the long-term associated study groups of the German Society for Immunology and the Society for Biochemistry and Molecular Biology, and for instance the SFB/Transregio 52 "Transcriptional Programming of Individual T Cell Subsets" located in Würzburg, Mainz and Berlin. The different research areas that were introduced by outstanding keynote speakers attracted more than 250 scientists, showing the timeliness and relevance of the interdisciplinary concept and exchange of knowledge during the three days of the scientific program. This report gives an overview of the presentations of the conference.

  14. Possible impacts of early-11th-, middle-12th-, and late-13th-century droughts on western Native Americans and the Mississippian Cahokians

    Benson, Larry V.; Berry, Michael S.; Jolie, Edward A.; Spangler, Jerry D.; Stahle, David W.; Hattori, Eugene M.

    2007-02-01

    One or more of three intense and persistent droughts impacted some Native American cultures in the early-11th, middle-12th and late-13th centuries, including the Anasazi, Fremont, Lovelock, and Mississippian (Cahokian) prehistorical cultures. Tree-ring-based reconstructions of precipitation and temperature indicate that warm drought periods occurred between AD 990 and 1060, AD 1135 and 1170, and AD 1276 and 1297. These droughts occurred during minima in the Pacific Decadal Oscillation and may have been associated with positive values of the Atlantic Multidecadal Oscillation. Each of the Native American cultures was supported, to a greater or lesser degree, by precipitation-dependent resources. Both the Four Corners region and Cahokia were sites of intense growth between about AD 1050 and 1130, and by AD 1150, cultures in both regions were undergoing stress. By AD 1300 the Anasazi and Fremont cultures had collapsed and their residual populations had either left their homelands or withered. In the case of Fremont populations, the AD 990-1060 drought may have had the greatest impact. This drought also may have affected the Anasazi, for it was at the end of this drought that some people from Chaco migrated to the San Juan River valley and founded the Salmon Ruin great house. Detailed data do not exist on the number of Lovelock habitation sites or populations over time; however, Lovelock populations appear to have retreated from the western Great Basin to California by AD 1300 or shortly thereafter.

  15. Results of 13th regular inspection of No.1 plant in Fukushima No.1 Nuclear Power Station, Tokyo Electric Power Co., Inc

    The 13th regular inspection of No.1 plant in Fukushima No.1 Nuclear Power Station was carried out from November 20, 1987 to April 27, 1988. The parallel operation was resumed on April 8, 1988, 141 days after the parallel-off. The facilities being the object of inspection were the reactor proper, reactor cooling system, measuring and control system, fuel facilities, radiation control facilities, waste facilities, reactor containment installation, and emergency power generation system. On these facilities being the object of inspection, the appearance, disassembling, leak, function, performance and other inspections were carried out, as the result, abnormality was not found at all. The works related to this regular inspection were accomplished within the range of allowable radiation dose based on the relevant laws. The main reconstruction works carried out during the period of this regular inspection were the charging of 52 new 8 x 8 zirconium liner fuel assemblies for improving the reliability and capacity factor, the new installation of a hollow thread film filter before the condensate desalting facility for reducing its load, the repair of a core spray piping, the installation of process radiation monitors, the replacement of a movable spatial dose rate measuring instrument, the repair of the penetration through the reactor containment vessel, the replacement of a condensate system piping and the repair of the exhaust gas condenser. (Kako, I.)

  16. A glimpse into the early origins of medieval anatomy through the oldest conserved human dissection (Western Europe, 13th c. A.D.)

    Huynh-Charlier, Isabelle; Poupon, Joël; Lancelot, Eloïse; Campos, Paula F.; Favier, Dominique; Jeannel, Gaël-François; Bonati, Maurizio Rippa; de la Grandmaison, Geoffroy Lorin; Hervé, Christian

    2013-01-01

    Introduction Medieval autopsy practice is very poorly known in Western Europe, due to a lack of both descriptive medico-surgical texts and conserved dissected human remains. This period is currently considered the dark ages according to a common belief of systematic opposition of Christian religious authorities to the opening of human cadavers. Material and methods The identification in a private collection of an autopsied human individual dated from the 13th century A.D. is an opportunity for better knowledge of such practice in this chrono-cultural context, i.e. the early origins of occidental dissections. A complete forensic anthropological procedure was carried out, completed by radiological and elemental analyses. Results The complete procedure of this body opening and internal organs exploration is explained, and compared with historical data about forensic and anatomical autopsies from this period. During the analysis, a red substance filling all arterial cavities, made of mercury sulfide (cinnabar) mixed with vegetal oil (oleic and palmitic acids) was identified; it was presumably used to highlight vascularization by coloring in red such vessels, and help in the preservation of the body. Conclusions Of particular interest for the description of early medical and anatomical knowledge, this “human preparation” is the oldest known yet, and is particularly important for the fields of history of medicine, surgery and anatomical practice. PMID:24904674

  17. Seeding the FEL of the SCSS Phase 1 Facility with the 13th Laser Harmonic of a Ti Sa Laser (61.5 nm) Produced in Xe Gas

    Lambert, G; Boutu, W; Breger, P; Couprie, M E; Garzella, D; Merdji, H; Monchicourt, P; Salieres, P

    2005-01-01

    In order to reach very short wavelengths in FEL, and to have a more compact, fully coherent and tunable source, a particular seeding configuration is foreseen to be tested as a demonstration experiment in 2006 into the SCSS phase 1 facility (Spring-8 Compact Sase Source, Japan). The external source is the 13th harmonic (61.5 nm) of a Ti: Sa laser (25 mJ, 10 Hz, 100 fs) generated in 10 Hz pulsed Xe gas cell. The harmonic generation process provides us with a intense (1 μJ) and ultra-short (50 fs) VUV beam. The design of the experiment implantation is discussed, taken into account the performances of the generation process, the focusing of the selected harmonic into the modulator, and the resistance of the optical components. Besides one should consider the vacuum needs, the geometrical problems and the mechanics for the under UHV mirrors translation. One first chamber is dedicated to the harmonic generation. A second one is used for spectral selection and adaptation of the harmonic in the modulator. F...

  18. Ca2+ current is regulated by cyclic GMP-dependent protein kinase in mammalian cardiac myocytes

    Regulation of cardiac contraction by neurotransmitters and hormones is often correlated with regulation of the L-type Ca2+-channel current (ICa) through the opposite actions for two second messengers, cyclic AMP and cyclic GMP. While cyclic AMP stimulation of ICa is mediated by the activation of cyclic AMP-dependent protein kinase, inhibition of ICa by cyclic GMP in frog heart is largely mediated by activation of cyclic AMP phosphodiesterase. The present patch-clamp study reveals that, in rat ventricular cells, cyclic GMP can also regulate ICa via activation of endogenous cyclic GMP-dependent protein kinase (cGMP-PK). Indeed, the effect of cyclic GMP on ICa was mimicked by intracellular perfusion with the proteolytic active fragment of purified cGMP-PK. Moreover, cGMP-PK immunoreactivity was detected in pure rat ventricular myocytes by using a specific polyclonal antibody. These results demonstrate a dual mechanism for the inhibitory action of cyclic GMP in heart, as well as a physiological role for cGMP-PK in the control of mammalian heart function

  19. c-di-GMP and its Effects on Biofilm Formation and Dispersion: a Pseudomonas Aeruginosa Review.

    Ha, Dae-Gon; O'Toole, George A

    2015-04-01

    Since its initial discovery as an allosteric factor regulating cellulose biosynthesis in Gluconacetobacter xylinus, the list of functional outputs regulated by c-di-GMP has grown. We have focused this article on one of these c-di-GMP-regulated processes, namely, biofilm formation in the organism Pseudomonas aeruginosa. The majority of diguanylate cyclases and phosphodiesterases encoded in the P. aeruginosa genome still remain uncharacterized; thus, there is still a great deal to be learned about the link between c-di-GMP and biofilm formation in this microbe. In particular, while a number of c-di-GMP metabolizing enzymes have been identified that participate in reversible and irreversible attachment and biofilm maturation, there is a still a significant knowledge gap regarding the c-di-GMP output systems in this organism. Even for the well-characterized Pel system, where c-di-GMP-mediated transcriptional regulation is now well documented, how binding of c-di-GMP by PelD stimulates Pel production is not understood in any detail. Similarly, c-di-GMP-mediated control of swimming, swarming and twitching also remains to be elucidated. Thus, despite terrific advances in our understanding of P. aeruginosa biofilm formation and the role of c-di-GMP in this process since the last version of this book (indeed there was no chapter on c-di-GMP!) there is still much to learn. PMID:26104694

  20. Role of nitric oxide and cyclic GMP in glutamate-induced neuronal death.

    Montoliu, C; Llansola, M; Monfort, P; Corbalan, R; Fernandez-Marticorena, I; Hernandez-Viadel, M L; Felipo, V

    2001-04-01

    Glutamate is the main excitatory neurotransmitter in mammals. However, excessive activation of glutamate receptors is neurotoxic, leading to neuronal degeneration and death. In many systems, including primary cultures of cerebellar neurons, glutamate neurotoxicity is mainly mediated by excessive activation of NMDA receptors, leading to increased intracellular calcium which binds to calmodulin and activates neuronal nitric oxide synthase (NOS), increasing nitric oxide (NO) which in turn activates guanylate cyclase and increases cGMP. Inhibition of NOS prevents glutamate neurotoxicity, indicating that NO mediates glutamate-induced neuronal death in this system. NO generating agents such as SNAP also induce neuronal death. Compounds that can act as "scavengers" of NO such as Croman 6 (CR-6) prevent glutamate neurotoxicity. The role of cGMP in the mediation of glutamate neurotoxicity remains controversial. Some reports indicate that cGMP mediates glutamate neurotoxicity while others indicate that cGMP is neuroprotective. We have studied the role of cGMP in the mediation of glutamate and NO neurotoxicity in cerebellar neurons. Inhibition of soluble guanylate cyclase prevents glutamate and NO neurotoxicity. There is a good correlation between inhibition of cGMP formation and neuroprotection. Moreover 8-Br-cGMP, a cell permeable analog of cGMP, induced neuronal death. These results indicate that increased intracellular cGMP is involved in the mechanism of neurotoxicity. Inhibitors of phosphodiesterase increased extracellular but not intracellular cGMP and prevented glutamate neurotoxicity. Addition of cGMP to the medium also prevented glutamate neurotoxicity. These results are compatible with a neurotoxic effect of increased intracellular cGMP and a neuroprotective effect of increased extracellular cGMP. PMID:14715472

  1. Design of GMP compliance radiopharmaceutical production facility in MINT

    In 1985, MINT built the only radiopharmaceutical production facility in Malaysia. The facility was designed based on IAEA (International Atomic Energy Agency) standard guidelines which provide radiation safety to the staff and the surrounding environment from radioactive contamination. Since 1999, BPFK (Biro Pengawalan Farmaseutikal Kebangsaan) has used the guidelines from Pharmaceutical Inspection Convention Scheme (PICS) to meet the requirements of the Good Manufacturing Practice (GMP) for Pharmaceutical Products. In the guidelines, the pharmaceutical production facility shall be designed based on clean room environment. In order to design a radiopharmaceutical production facility, it is important to combine the concept of radiation safety and clean room to ensure that both requirements from GMP and IAEA are met. The design requirement is necessary to set up a complete radiopharmaceutical production facility, which is safe, has high production quality and complies with the Malaysian and International standards. (Author)

  2. cGMP in ozone and NO dependent responses

    Ederli, Luisa; Meier, Stuart; Borgogni, Andrea; Reale, Lara; Ferranti, Francesco; Gehring, Chris

    2008-01-01

    We have recently reported that ozone (O3) can inhibit mitochondrial respiration and induce activation of the alternative oxidase (AOX) pathway and in particular AOX1a in tobacco. While O3 causes mitochondrial H2O2, early leaf nitric oxide (NO) as well as transient ethylene (ET) accumulation, the levels of jasmonic acid and 12-oxo-phytodienoic acid remained unchanged. It was shown that both, NO and ET dependent pathways can induce AOX1a transcription by O3. AOX plays a role in reducing reactive oxygen species (ROS) which in turn are linked to biotic and abiotic plant stresses, much like the second messengers guanosine 3′, 5′-cyclic monophosphate (cGMP). The goal is to unravel specific cGMP signatures and induction pathways downstream from O3 and NO, including transcription of AOX1a. Here we propose that some late (>3 h) responses to NO, e.g., the accumulation of phenylalanine lyase (PAL) transcripts, are critically cGMP dependent, while the early (<2 h) responses, including AOX1a induction are not. PMID:19704720

  3. Report of the Two-Day National Seminar on New Directions in Higher Education, Organized by the Kerala State Higher Education Council on 12th and 13th July 2010

    Praveen, C.

    2010-01-01

    This is a report of the Two-Day National Seminar on New Directions in Higher Education, organized by the Kerala State Higher Education Council on 12th and 13th July 2010. The objective of the seminar was to deliberate upon the reforms being undertaken by the Government of India in Higher Education. Reputed scholars from within and outside the…

  4. A study on relations between the levels of GMP-140 and microangiopathy in NIDDM

    The relations between the level of GMP-140 and microangiopathy in NIDDM for earlier diagnosis or better treatment are investigated, the level of GMP-140 in both platelet and plasma was measured. The level of GMP-140 in both platelet and plasma in 104 cases with NIDDM (55 with and 49 without microagiopathy) and 38 controls were assayed by RIA and also simultaneously with direct platelet count. The level of GMP-140 in both platelet and plasma in NIDDM was remarkably higher than that in controls (P1 = 0.69, r2 = 0.75). No differences existed in platelet count between NIDDM and controls. The level of GMP-140 and ophthalmoscopic study had no change after decreasing the concentration of blood glucose (<7.8 mmol/L) and administrating aspirin for 6 months. Microangiopathy in NIDDM had close relation with platelet function and the level of GMP-140

  5. Effect of ionizing radiation of tissue cyclic AMP and GMP in experimental rats

    Adult male Sprague Dawley rats were exposed to whole-body gamma radiation of 4 Gy and 10 Gy and cyclic AMP and GMP of several tissues were studied 4 hours, 24 hours and 72 hours after irradiation. Animals exposed to 4 Gy exhibited initial increase and subsequent decrease of plasma cyclic AMP, while opposite effects were noted in plasma cyclic GMP. Both groups of irradiated rats showed significant decrease of spleen cyclic AMP with variable responses in cyclic GMP. Initial increase with subsequent decrease of liver cyclic AMP was observed in irradiated animals with unaltered cyclic GMP values. It was interesting to note variable responses of myocardial cyclic AMP in both groups of experimental rats with a significant increase in heart cyclic GMP in rats exposed to 10 Gy only. In contrast, myocardial cyclic GMP was significantly decreased in rats exposed to 4 Gy groups. (orig.)

  6. Early Old Bulgarian Layer in the Office Menaion of Rostov Provenance of the 13th c. (RNL, F.p.I 37

    Roman N. Krivko

    2012-08-01

    Full Text Available The article is dedicated to the linguistic and textual peculiarities of the Festal Menaion F. p. I 37 preserved in Russian National Library (S.-Petersburg and dated to the beginning of the 13th c. Numerous linguistic features of the manuscript testify to Old West Bulgarian (Old Macedonian literary usage of the end of the 9th — beginning of the 10th c., on the basis of which the earliest Slavonic hymnographical translations were performed. Despite of such an archaic linguistic and textual layers observed in the manuscript, its calendar, structure and content were influenced by the monastic rite based on the Typicon of Patriarch Alexius the Studite which had been translated and introduced in Kievan Rus' at the second half of the 11th c. The linguistic features of the text version as attested by F. p. I 37 have been compared to the manuscripts testifying to the Alexius the Studite text version. It has been affirmed that adaptation of the Old Bulgarian hymnographic heritage in Kievan Rus' followed the unstable trend to neglect the most remarkable South (and South West Slavonic lingustic features and aimed at establishing “neutral” over dialectal Church Slavonic literary usage. The significant part of the article contains the edition of the earliest Slavonic, i.e., Old Bulgarian, translation of the Greek kanon Τάφῳ παρθενοδόχῳ dedicated to the Dormition of the Theotokos. The earliest Slavonic translation of this kanon has been preserved in the unique available manuscript, that is F. p. I 37. The publication of the Slavonic translation is supplied with the first critical edition of the Greek source performed on the basis of twelve manuscripts, the linguistic commentary of the Slavonic text, and remarks concerning some metric peculiarities of the original Greek version.

  7. Annual energy review 1994

    NONE

    1995-07-01

    This 13th edition presents the Energy Information Administration`s historical energy statistics. For most series, statistics are given for every year from 1949 through 1994; thus, this report is well-suited to long-term trend analyses. It covers all major energy activities, including consumption, production, trade, stocks, and prices for all major energy commodities, including fossil fuels and electricity. Statistics on renewable energy sources are also included: this year, for the first time, usage of renewables by other consumers as well as by electric utilities is included. Also new is a two-part, comprehensive presentation of data on petroleum products supplied by sector for 1949 through 1994. Data from electric utilities and nonutilities are integrated as ``electric power industry`` data; nonutility power gross generation are presented for the first time. One section presents international statistics (for more detail see EIA`s International Energy Annual).

  8. RIA of platelet GMP-140 in patients with coronary heart disease

    With monoclonal antibody SZ-51, the activated levels of the platelet granule membrane protein (GMP-140) of 94 cases with coronary heart disease (CHD) are assayed by RIA. The results show that the levels of GMP-140 with CHD are much higher than those of the controls (P<0.001), and the levels of GMP-140 with AMI 12 hours after the treatment with urokinase for thrombolysis are considerably lower than the levels one hour before the treatment. Therefore, the assay of GMP-140 with CHD is of great clinical significance in diagnosing and differentiating CHD, especially, in analysing pathogen, monitoring curative effect and predicting prognosis for AMI

  9. Study of GMP-140 RIA in the diagnosis and treatment of cerebrovascular diseases

    To investigate the relation between GMP-140 with the progress and mechanism of cerebrovascular diseases, the level of GMP-140 in both platelet and plasma in 76 cases of cerebrovascular disease and 35 normal controls were determined by radioimmunoassay (RIA). Results showed that the level of GMP-140 in both platelet and plasma of cerebrovascular diseases were remarkably higher than that in normal controls (P0.05). It was concluded that the progress, development, and prognosis of cerebrovascular diseases is strongly related to the platelet function and the level of GMP-140

  10. A study in platelet GMP-140 for coronary heart disease by RIA

    With monoclonal anti-body SZ-51, activated levels of platelet granule membrane protein (GMP-140) in coronary heart disease (CHD) were assayed by RIA. The results showed that the levels of GMP-140 with CHD were higher than that of controls (P>0.001). The levels of GMP-140 with AMI treated with urokinase for thrombolysis in 12 hours were considerably lower than that one hour before treatment. Therefore, the measurement of GMP-140 with CHD is of great clinical value in diagnosing and differentiating AMI, especially, in analysing pathogen, monitoring curative effect and predicting prognosis for AMI

  11. Intracellular photoactivation of caged cGMP induces myosin II and actin responses in motile cells.

    Pfannes, Eva K B; Anielski, Alexander; Gerhardt, Matthias; Beta, Carsten

    2013-12-01

    Cyclic GMP (cGMP) is a ubiquitous second messenger in eukaryotic cells. It is assumed to regulate the association of myosin II with the cytoskeleton of motile cells. When cells of the social amoeba Dictyostelium discoideum are exposed to chemoattractants or to increased osmotic stress, intracellular cGMP levels rise, preceding the accumulation of myosin II in the cell cortex. To directly investigate the impact of intracellular cGMP on cytoskeletal dynamics in a living cell, we released cGMP inside the cell by laser-induced photo-cleavage of a caged precursor. With this approach, we could directly show in a live cell experiment that an increase in intracellular cGMP indeed induces myosin II to accumulate in the cortex. Unexpectedly, we observed for the first time that also the amount of filamentous actin in the cell cortex increases upon a rise in the cGMP concentration, independently of cAMP receptor activation and signaling. We discuss our results in the light of recent work on the cGMP signaling pathway and suggest possible links between cGMP signaling and the actin system. PMID:24136144

  12. Atrial natriuretic peptide and oxytocin induce natriuresis by release of cGMP

    Soares, T. J.; Coimbra, T. M.; Martins, A. R.; Pereira, A. G. F.; Carnio, E. C.; Branco, L. G. S.; Albuquerque-Araujo, W. I. C.; de Nucci, G.; Favaretto, A. L. V.; Gutkowska, J.; McCann, S. M.; Antunes-Rodrigues, J.

    1999-01-01

    Our hypothesis is that oxytocin (OT) causes natriuresis by activation of renal NO synthase that releases NO followed by cGMP that mediates the natriuresis. To test this hypothesis, an inhibitor of NO synthase, l-nitroarginine methyl ester (NAME), was injected into male rats. Blockade of NO release by NAME had no effect on natriuresis induced by atrial natriuretic peptide (ANP). This natriuresis presumably is caused by cGMP because ANP also activates guanylyl cyclase, which synthesizes cGMP from GTP. The 18-fold increase in sodium (Na+) excretion induced by OT (1 μg) was accompanied by an increase in urinary cGMP and preceded by 20 min a 20-fold increase in NO3− excretion. NAME almost completely inhibited OT-induced natriuresis and increased NO3− excretion; however, when the dose of OT was increased 10-fold, a dose that markedly increases plasma ANP concentrations, NAME only partly inhibited the natriuresis. We conclude that the natriuretic action of OT is caused by a dual action: generation of NO leading to increased cGMP and at higher doses release of ANP that also releases cGMP. OT-induced natriuresis is caused mainly by decreased tubular Na+ reabsorption mediated by cGMP. In contrast to ANP that releases cGMP in the renal vessels and the tubules, OT acts on its receptors on NOergic cells demonstrated in the macula densa and proximal tubules to release cGMP that closes Na+ channels. Both ANP- and OT-induced kaliuresis also appear to be mediated by cGMP. We conclude that cGMP mediates natriuresis and kaliuresis induced by both ANP and OT. PMID:9874809

  13. Cyclic GMP protects human macrophages against peroxynitrite-induced apoptosis

    Rossi Adriano G

    2009-05-01

    Full Text Available Abstract Background Nitric oxide (NO can be both pro- and anti-apoptotic in various cell types, including macrophages. This apparent paradox may result from the actions of NO-related species generated in the microenvironment of the cell, for example the formation of peroxynitrite (ONOO-. In this study we have examined the ability of NO and ONOO- to evoke apoptosis in human monocyte-derived macrophages (MDMϕ, and investigated whether preconditioning by cyclic guanosine monophosphate (cGMP is able to limit apoptosis in this cell type. Methods Characterisation of the NO-related species generated by (Z-1- [2-(2-aminoethyl-N-(2-ammonioethylamino]diazen-1-ium-1,2-diolate (DETA/NO and 1,2,3,4-oxatriazolium, 5-amino-3-(3,4-dichlorophenyl-, chloride (GEA-3162 was performed by electrochemistry using an isolated NO electrode and electron paramagnetic resonance (EPR spectrometry. Mononuclear cells were isolated from peripheral blood of healthy volunteers and cultured to allow differentiation into MDMϕ. Resultant MDMϕ were treated for 24 h with DETA/NO (100 – 1000 μM or GEA-3162 (10 – 300 μM in the presence or absence of BAY 41–2272 (1 μM, isobutylmethylxanthine (IBMX; 1 μM, 1H- [1,2,4]oxadiazolo [4,3-a]quinoxalin-1-one (ODQ; 20 μM or 8-bromo-cGMP (1 mM. Apoptosis in MDMϕ was assessed by flow cytometric analysis of annexin V binding in combination with propidium iodide staining. Results Electrochemistry and EPR revealed that DETA/NO liberated free NO radical, whilst GEA-3162 concomitantly released NO and O2-, and is therefore a ONOO- generator. NO (DETA/NO had no effect on cell viability, but ONOO- (GEA-3162 caused a concentration-dependent induction of apoptosis in MDMϕ. Preconditioning of MDMϕ with NO in combination with the phosphodiesterase inhibitor, 3-Isobutyl-1-methylxanthine (IBMX, or the NO-independent stimulator of soluble guanylate cyclase, BAY 41–2272, significantly attenuated ONOO--induced apoptosis in a cGMP-dependent manner

  14. Methods and materials relating to IMPDH and GMP production

    Collart, Frank R.; Huberman, Eliezer

    1997-01-01

    Disclosed are purified and isolated DNA sequences encoding eukaryotic proteins possessing biological properties of inosine 5'-monophosphate dehydrogenase ("IMPDH"). Illustratively, mammalian (e.g., human) IMPDH-encoding DNA sequences are useful in transformation or transfection of host cells for the large scale recombinant production of the enzymatically active expression products and/or products (e.g., GMP) resulting from IMPDH catalyzed synthesis in cells. Vectors including IMPDH-encoding DNA sequences are useful in gene amplification procedures. Recombinant proteins and synthetic peptides provided by the invention are useful as immunological reagents and in the preparation of antibodies (including polyclonal and monoclonal antibodies) for quantitative detection of IMPDH.

  15. Captive solvent [11C]acetate synthesis in GMP conditions

    Reliable procedure for the production of 1-[11C]acetate in GMP conditions was developed based on a combination of the captive-solvent Grignard reaction conducted in the sterile catheter followed by the convenient solid-phase extraction purification on a series of ion-exchange cartridges. The described procedure proved to be reliable in more than 30 patient productions. The process provides stable radiochemical yields (65% EOB) of sodium acetate (1-[11C]) of the Ph.Eur. quality (radiochemical purity better than 95%) in a short time (5 min)

  16. A cyclic GMP signalling module that regulates gliding motility in a malaria parasite.

    Robert W Moon

    2009-09-01

    Full Text Available The ookinete is a motile stage in the malaria life cycle which forms in the mosquito blood meal from the zygote. Ookinetes use an acto-myosin motor to glide towards and penetrate the midgut wall to establish infection in the vector. The regulation of gliding motility is poorly understood. Through genetic interaction studies we here describe a signalling module that identifies guanosine 3', 5'-cyclic monophosphate (cGMP as an important second messenger regulating ookinete differentiation and motility. In ookinetes lacking the cyclic nucleotide degrading phosphodiesterase delta (PDEdelta, unregulated signalling through cGMP results in rounding up of the normally banana-shaped cells. This phenotype is suppressed in a double mutant additionally lacking guanylyl cyclase beta (GCbeta, showing that in ookinetes GCbeta is an important source for cGMP, and that PDEdelta is the relevant cGMP degrading enzyme. Inhibition of the cGMP-dependent protein kinase, PKG, blocks gliding, whereas enhanced signalling through cGMP restores normal gliding speed in a mutant lacking calcium dependent protein kinase 3, suggesting at least a partial overlap between calcium and cGMP dependent pathways. These data demonstrate an important function for signalling through cGMP, and most likely PKG, in dynamically regulating ookinete gliding during the transmission of malaria to the mosquito.

  17. Detection of plasma granule membrane protein (GMP-140) using radiolabelled monoclonal antibodies in thrombotic diseases

    Radioimmunoassay with two monoclonal antibodies to granule membrane protein (GMP-140) is used to determine whether plasma GMP-140 including its soluble and microparticle forms can be detected in patients with acute myocardial infarction (AMI) or during cardio-pulmonary bypass (CPB) and in platelet concentrates during storage. Monoclonal antibody (McAb) SZ-51 was used as a solid phase and 125I-labelled McAb S12 was used as a fluid phase. The assay showed sufficient sensitivity to detect as few as 1 ng/mL of purified GMP-140. There was only 10.0 ± 4.5 ng/mL in normal plasma (n 20) with no significant different between the male and the female controls. Ten patients undergoing CPB demonstrated a transient increase in the concentration of plasma GMP-140, especially 2 h after CPB, and its plasma level was inversely correlated with the platelet counting during bypass (r -0.81, P < 0.01). Patients with AMI (n = 16) were found to have significantly increased concentration of plasma GMP-140 after AMI, reaching the peak within 3 days and changing with the progression of AMI. The concentration of plasma GMP-140 increased progressively in platelet concentrates during storage, particularly 5 days after storage. In short, these data suggest that plasma GMP-140 can be reliably detected by radioimmunoassay with two McAbs to GMP-140 and its plasma level may serve as a useful marker of thrombosis and thrombotic diseases

  18. Sildenafil Potentiates a cGMP-Dependent Pathway to Promote Melanoma Growth

    Sandeep Dhayade

    2016-03-01

    Full Text Available Sildenafil, an inhibitor of the cGMP-degrading phosphodiesterase 5 that is used to treat erectile dysfunction, has been linked to an increased risk of melanoma. Here, we have examined the potential connection between cGMP-dependent signaling cascades and melanoma growth. Using a combination of biochemical assays and real-time monitoring of melanoma cells, we report a cGMP-dependent growth-promoting pathway in murine and human melanoma cells. We document that C-type natriuretic peptide (CNP, a ligand of the membrane-bound guanylate cyclase B, enhances the activity of cGMP-dependent protein kinase I (cGKI in melanoma cells by increasing the intracellular levels of cGMP. Activation of this cGMP pathway promotes melanoma cell growth and migration in a p44/42 MAPK-dependent manner. Sildenafil treatment further increases intracellular cGMP concentrations, potentiating activation of this pathway. Collectively, our data identify this cGMP-cGKI pathway as the link between sildenafil usage and increased melanoma risk.

  19. Structural Basis of Ligand Binding by a C-di-GMP Riboswitch

    Smith, K.; Lipchock, S; Ames, T; Wang, J; Breaker, R; Strobel, S

    2009-01-01

    The second messenger signaling molecule bis-(3{prime}-5{prime})-cyclic dimeric guanosine monophosphate (c-di-GMP) regulates many processes in bacteria, including motility, pathogenesis and biofilm formation. c-di-GMP-binding riboswitches are important downstream targets in this signaling pathway. Here we report the crystal structure, at 2.7 {angstrom} resolution, of a c-di-GMP riboswitch aptamer from Vibrio cholerae bound to c-di-GMP, showing that the ligand binds within a three-helix junction that involves base-pairing and extensive base-stacking. The symmetric c-di-GMP is recognized asymmetrically with respect to both the bases and the backbone. A mutant aptamer was engineered that preferentially binds the candidate signaling molecule c-di-AMP over c-di-GMP. Kinetic and structural data suggest that genetic regulation by the c-di-GMP riboswitch is kinetically controlled and that gene expression is modulated through the stabilization of a previously unidentified P1 helix, illustrating a direct mechanism for c-di-GMP signaling.

  20. Structural basis of ligand binding by a c-di-GMP riboswitch.

    Smith, Kathryn D; Lipchock, Sarah V; Ames, Tyler D; Wang, Jimin; Breaker, Ronald R; Strobel, Scott A

    2009-12-01

    The second messenger signaling molecule bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP) regulates many processes in bacteria, including motility, pathogenesis and biofilm formation. c-di-GMP-binding riboswitches are important downstream targets in this signaling pathway. Here we report the crystal structure, at 2.7 A resolution, of a c-di-GMP riboswitch aptamer from Vibrio cholerae bound to c-di-GMP, showing that the ligand binds within a three-helix junction that involves base-pairing and extensive base-stacking. The symmetric c-di-GMP is recognized asymmetrically with respect to both the bases and the backbone. A mutant aptamer was engineered that preferentially binds the candidate signaling molecule c-di-AMP over c-di-GMP. Kinetic and structural data suggest that genetic regulation by the c-di-GMP riboswitch is kinetically controlled and that gene expression is modulated through the stabilization of a previously unidentified P1 helix, illustrating a direct mechanism for c-di-GMP signaling. PMID:19898477

  1. The cyclic-di-GMP signaling pathway in the Lyme disease spirochete, Borrelia burgdorferi

    Elizabeth A. Novak

    2014-05-01

    Full Text Available In nature, the Lyme disease spirochete Borrelia burgdorferi cycles between the unrelated environments of the Ixodes tick vector and mammalian host. In order to survive transmission between hosts, B. burgdorferi must be able to not only detect changes in its environment, but also rapidly and appropriately respond to these changes. One manner in which this obligate parasite regulates and adapts to its changing environment is through cyclic-di-GMP (c-di-GMP signaling. c-di-GMP has been shown to be instrumental in orchestrating the adaptation of B. burgdorferi to the tick environment. B. burgdorferi possesses only one set of c-di-GMP-metabolizing genes (one diguanylate cyclase and two distinct phosphodiesterases and one c-di-GMP-binding PilZ-domain protein designated as PlzA. While studies in the realm of c-di-GMP signaling in B. burgdorferi have exploded in the last few years, there are still many more questions than answers. Elucidation of the importance of c-di-GMP signaling to B. burgdorferi may lead to the identification of mechanisms that are critical for the survival of B. burgdorferi in the tick phase of the enzootic cycle as well as potentially delineate a role (if any c-di-GMP may play in the transmission and virulence of B. burgdorferi during the enzootic cycle, thereby enabling the development of effective drugs for the prevention and/or treatment of Lyme disease.

  2. Sildenafil Potentiates a cGMP-Dependent Pathway to Promote Melanoma Growth.

    Dhayade, Sandeep; Kaesler, Susanne; Sinnberg, Tobias; Dobrowinski, Hyazinth; Peters, Stefanie; Naumann, Ulrike; Liu, He; Hunger, Robert E; Thunemann, Martin; Biedermann, Tilo; Schittek, Birgit; Simon, Hans-Uwe; Feil, Susanne; Feil, Robert

    2016-03-22

    Sildenafil, an inhibitor of the cGMP-degrading phosphodiesterase 5 that is used to treat erectile dysfunction, has been linked to an increased risk of melanoma. Here, we have examined the potential connection between cGMP-dependent signaling cascades and melanoma growth. Using a combination of biochemical assays and real-time monitoring of melanoma cells, we report a cGMP-dependent growth-promoting pathway in murine and human melanoma cells. We document that C-type natriuretic peptide (CNP), a ligand of the membrane-bound guanylate cyclase B, enhances the activity of cGMP-dependent protein kinase I (cGKI) in melanoma cells by increasing the intracellular levels of cGMP. Activation of this cGMP pathway promotes melanoma cell growth and migration in a p44/42 MAPK-dependent manner. Sildenafil treatment further increases intracellular cGMP concentrations, potentiating activation of this pathway. Collectively, our data identify this cGMP-cGKI pathway as the link between sildenafil usage and increased melanoma risk. PMID:26971999

  3. The institutions forming the socioeconomic structure of Turkish private enterprises between the 13th and the 19th centuries: Akhism, the Lonca system and the Gedik system

    Mehmet Özbirecikli

    2010-12-01

    principios éticos de la vida empresarial, en esencia, son los mismos. Dentro de este contexto, nos atrevemos a sugerir que las raíces del código ético de la vida empresarial turca se retrotraen en la historia a hace más de 800 años. Además, la similitud entre el funcionamiento presente y pasado indica que el origen de la formación de los aprendices para las empresas turcas tiene, igualmente, más de 800 años de historia.This study investigates three institutions forming the socioeconomic structure of Turkish private enterprises between the 13th and 19th Centuries: Akhism (13th-16th century, the Lonca System (the Guilds (16th-18th century, and the Gedik (Monopoly System (18th-20th century. The study particularly focuses on the social and economic rules, vocational training process, and organizational structure of the said institutions in order to discuss the effects of the socioeconomic structure of Turkish enterprises on economic and social development of private enterprises. The study also struggles to link between the relevant current applications and the applications in the past such as the social rules and vocational training. From economic point of view, both the statist structure of the State and the economic rules of the institutions herein caused private enterprises to remain small, and prevented them from having a competitive environment and having capital accumulation. As a result, enterprises could not benefit from new production techniques and the Turkish enterprise mentality fell behind modern developments. On the other hand, although these three systems were completely abolished in the early 20th Century, it is seen that especially traces of the Akhism and Lonca systems have still been surviving. Both the most of rules of Akhism and some of the duties of the board of directors of Lonca such as keeping moral standards of production and trade remind us of professional code of ethics of today’s modern business life. In other saying, there was code of

  4. Purification and characterization of cGMP binding protein-phosphodiesterase from rat lung

    The cGMP binding protein-phosphodiesterase (cG-BPP) with a phosphodiesterase specific activity of 7 μM/min/mg has been purified from rat lung by sequential chromatography on DEAE-cellulose, Blue-Sepharose, zinc chelate affinity adsorbent and HPLC-DEAE. Migration of the major band on SDS-PAGE corresponds to a MW of ∼93,000. Both cGMP phosphodiesterase activity and cGMP binding from the HPLC-DEAE profile correlate with this band. Since the authors previous work has determined the native MW to be ∼177,000, this suggests a dimeric structure comprised of two 93,000 MW subunits for the rat lung cG-BPP. At low cGMP concentrations, cGMP binding is stimulated ∼20-fold by histone and ∼5-fold by 3-isobutyl-1-methylxanthine(IBMX). The purified protein has one component of cGMP dissociation with a rate constant of 0.045/min. Photolysis of the purified protein in the presence of 32P-cGMP labels the 93,000 MW band and this labeling is increased by IBMX, indicating that the 93,000 MW band is a subunit of the cGMP-BPP. This implies that the enzyme preparation is nearly homogeneous, a conclusion also supported by a minimum [3H]-cGMP binding stoichiometry of 0.5 mol per 93,000 subunit. An additional protein band with a MW of ∼90,000 also occurs in these preparations which exhibits behavior similar to the 93,000 MW protein. N2-Hexyl-cGMP inhibits phosphodiesterase activity by competing with cGMP for hydrolysis at the catalytic site but not at the binding site. N2-Hexyl cGMP actually increases cGMP binding. This provides the first evidence that cGMP binding is increased by compounds hydrolyzed at the catalytic site. This interaction between the binding and phosphodiesterase sites could be important in the regulation of the functions of these sites in vivo

  5. The Threat of Terrorism: Putting Human Rights to the Test : Assessing the Counterterrorist Policies of France in the Aftermath of the 13th of November 2015 Terrorist Attacks 

    Poidevin, Fiore

    2016-01-01

    The terrorist attacks that occurred in Paris on the 13th of November 2015, mark a shift for counterterrorism in France. Indeed, the magnitude of the terrorist attacks persuaded the French government to call for stricter counterterrorist policies. However, the effects of these measures on human rights have been interrogated widely. Therefore, in this thesis, I wish to address the conflict between state security and individual human rights. First, I will assess if the current situation of Franc...

  6. Proceedings of the 13th International Conference on Hyperfine Interactions and 17th International Symposium on Nuclear Quadrupole Interactions (HFI/NQI 2004) Bonn, Germany, 22–27 August 2004

    Maier, K; HFI/NQI 2004

    2005-01-01

    Proceedings of the 13th International Conference on Hyperfine Interactions and 17th International Symposium on Nuclear Quadrupole Interactions, HFI/NQI 2004, held in Bonn, Germany, 22-27 August, 2004 Researchers and graduate students interested in the Mössbauer Effect and its applications will find this volume indispensable. The volume presents the most recent developments in the methodology of Mössbauer spectroscopy. Reprinted from Hyperfine Interactions (HYPE) Volume 158/159.

  7. Report on the 13th symposium on invertebrate neurobiology held 26-30 August 2015 at the Balaton Limnological Institute, MTA Centre for ecological research of the Hungarian Academy of Sciences, Tihany, Hungary.

    Crisford, Anna; Holden-Dye, Lindy; Walker, Robert J

    2016-06-01

    This report summarizes the lectures and posters presented at the International Society for Invertebrate Neurobiology's 13th symposium held 26-30 August 2015, at the Balaton Limnological Institute, MTA Centre for Ecological Research, Tihany, Hungary. The symposium provided an opportunity for scientists working on a range of topics in invertebrate neurobiology to meet and present their research and discuss ways to advance the discipline. PMID:27149972

  8. cGMP in Mouse Rods: the spatiotemporal dynamics underlying single photon responses

    Owen P. Gross

    2015-03-01

    Full Text Available Vertebrate vision begins when retinal photoreceptors transduce photons into membrane hyperpolarization, which reduces glutamate release onto second-order neurons. In rod photoreceptors, transduction of single photons is achieved by a well-understood G-protein cascade that modulates cGMP levels, and in turn, cGMP-sensitive inward current. The spatial extent and depth of the decline in cGMP during the single photon response have been major issues in phototransduction research since the discovery that single photons elicit substantial and reproducible changes in membrane current. The spatial profile of cGMP decline during the single photon response affects signal gain, and thus may contribute to reduction of trial-to-trial fluctuations in the single photon response. Here we summarize the general principles of rod phototransduction, emphasizing recent advances in resolving the spatiotemporal dynamics of cGMP during the single photon response.

  9. Fast repair activities of quercetin and rutin toward dGMP hydroxyl radical adducts

    The repair activities and mechanisms of both quercetin and rutin towards the oxidizing deoxyguanosine monophosphate (d GMP) hydroxyl radical adduct were investigated with pulse radiolytic technique. On pulse irradiation of nitrous oxide saturated 2 mm d GMP aqueous solution containing 0.1 mm quercetin, the transient absorption spectrum of the d GMP hydroxyl radical adduct decays with the formation of phenoxyl radical of quercetin within tens of microseconds. It indicates that there is a repair reaction between d GMP hydroxyl radical adduct and quercetin. The repair activity of rutin towards hydroxyl radical adducts of d GMP was also observed. The rate constants of the repair reactions were calculated to be 3.05x108 and 1.31x108 M-1 s-1for quercetin and rutin, respectively. This result together with our previous studies demonstrated that non-enzymatic, fast repair is a universal repair mechanism of phenolic antioxidants.

  10. 75 FR 17761 - Termination of an Environmental Impact Statement (EIS) for the General Management Plan (GMP) for...

    2010-04-07

    ... preparation of an EIS for the GMP for the Park, South Carolina. A Notice of Intent to prepare an EIS for the Park GMP was published in the Federal Register on October 10, 2006 (71 FR 63350), and followed by a... Plan (GMP) for Kings Mountain National Military Park (Park), South Carolina AGENCY: National...

  11. 75 FR 17756 - Availability of the Final General Management Plan and Environmental Impact Statement (GMP/EIS...

    2010-04-07

    ... (GMP/EIS) for Chattahoochee River National Recreation Area (CRNRA), GA AGENCY: National Park Service... Environmental Policy Act of 1969 the National Park Service (NPS) announces the availability of a Final GMP/EIS... CRNRA, the Final GMP/EIS describes Alternative F at the NPS preferred alternative. The...

  12. 75 FR 19422 - Availability of a Final Environmental Impact Statement for the General Management Plan (FEIS/GMP...

    2010-04-14

    ... Plan (FEIS/GMP), Tuskegee Airmen National Historic Site AGENCY: National Park Service. ACTION: Notice of availability of a Final Environmental Impact Statement for the General Management Plan (FEIS/GMP... Decision- making) the NPS announces the availability of a FEIS/GMP for the Tuskegee Airmen...

  13. Responses to elevated c-di-GMP levels in mutualistic and pathogenic plant-interacting bacteria.

    Daniel Pérez-Mendoza

    Full Text Available Despite a recent burst of research, knowledge on c-di-GMP signaling pathways remains largely fragmentary and molecular mechanisms of regulation and even c-di-GMP targets are yet unknown for most bacteria. Besides genomics or bioinformatics, accompanying alternative approaches are necessary to reveal c-di-GMP regulation in bacteria with complex lifestyles. We have approached this study by artificially altering the c-di-GMP economy of diverse pathogenic and mutualistic plant-interacting bacteria and examining the effects on the interaction with their respective host plants. Phytopathogenic Pseudomonas and symbiotic Rhizobium strains with enhanced levels of intracellular c-di-GMP displayed common free-living responses: reduction of motility, increased production of extracellular polysaccharides and enhanced biofilm formation. Regarding the interaction with the host plants, P. savastanoi pv. savastanoi cells containing high c-di-GMP levels formed larger knots on olive plants which, however, displayed reduced necrosis. In contrast, development of disease symptoms in P. syringae-tomato or P. syringae-bean interactions did not seem significantly affected by high c-di-GMP. On the other hand, increasing c-di-GMP levels in symbiotic R. etli and R. leguminosarum strains favoured the early stages of the interaction since enhanced adhesion to plant roots, but decreased symbiotic efficiency as plant growth and nitrogen contents were reduced. Our results remark the importance of c-di-GMP economy for plant-interacting bacteria and show the usefulness of our approach to reveal particular stages during plant-bacteria associations which are sensitive to changes in c-di-GMP levels.

  14. Cyclic Di-GMP modulates the disease progression of Erwinia amylovora.

    Edmunds, Adam C; Castiblanco, Luisa F; Sundin, George W; Waters, Christopher M

    2013-05-01

    The second messenger cyclic di-GMP (c-di-GMP) is a nearly ubiquitous intracellular signal molecule known to regulate various cellular processes, including biofilm formation, motility, and virulence. The intracellular concentration of c-di-GMP is inversely governed by diguanylate cyclase (DGC) enzymes and phosphodiesterase (PDE) enzymes, which synthesize and degrade c-di-GMP, respectively. The role of c-di-GMP in the plant pathogen and causal agent of fire blight disease Erwinia amylovora has not been studied previously. Here we demonstrate that three of the five predicted DGC genes in E. amylovora (edc genes, for Erwinia diguanylate cyclase), edcA, edcC, and edcE, are active diguanylate cyclases. We show that c-di-GMP positively regulates the secretion of the main exopolysaccharide in E. amylovora, amylovoran, leading to increased biofilm formation, and negatively regulates flagellar swimming motility. Although amylovoran secretion and biofilm formation are important for the colonization of plant xylem tissues and the development of systemic infections, deletion of the two biofilm-promoting DGCs increased tissue necrosis in an immature-pear infection assay and an apple shoot infection model, suggesting that c-di-GMP negatively regulates virulence. In addition, c-di-GMP inhibited the expression of hrpA, a gene encoding the major structural component of the type III secretion pilus. Our results are the first to describe a role for c-di-GMP in E. amylovora and suggest that downregulation of motility and type III secretion by c-di-GMP during infection plays a key role in the coordination of pathogenesis. PMID:23475975

  15. MRP4 Modulation of the Guanylate Cyclase-C/cGMP Pathway: Effects on Linaclotide-Induced Electrolyte Secretion and cGMP Efflux.

    Tchernychev, Boris; Ge, Pei; Kessler, Marco M; Solinga, Robert M; Wachtel, Derek; Tobin, Jenny V; Thomas, Sara R; Lunte, Craig E; Fretzen, Angelika; Hannig, Gerhard; Bryant, Alexander P; Kurtz, Caroline B; Currie, Mark G; Silos-Santiago, Inmaculada

    2015-10-01

    MRP4 mediates the efflux of cGMP and cAMP and acts as an important regulator of these secondary messengers, thereby affecting signaling events mediated by cGMP and cAMP. Immunofluorescence staining showed high MRP4 expression localized predominantly in the apical membrane of rat colonic epithelium. In vitro studies were performed using a rat colonic mucosal layer mounted in an Ussing chamber. Linaclotide activation of the guanylate cyclase-C (GC-C)/cGMP pathway induced a concentration-dependent increase in transepithelial ion current [short-circuit current (Isc)] across rat colonic mucosa (EC50: 9.2 nM). Pretreatment of colonic mucosa with the specific MRP4 inhibitor MK571 potentiated linaclotide-induced electrolyte secretion and augmented linaclotide-stimulated intracellular cGMP accumulation. Notably, pretreatment with the phosphodiesterase 5 inhibitor sildenafil increased basal Isc, but had no amplifying effect on linaclotide-induced Isc. MRP4 inhibition selectively affected the activation phase, but not the deactivation phase, of linaclotide. In contrast, incubation with a GC-C/Fc chimera binding to linaclotide abrogated linaclotide-induced Isc, returning to baseline. Furthermore, linaclotide activation of GC-C induced cGMP secretion from the apical and basolateral membranes of colonic epithelium. MRP4 inhibition blocked cGMP efflux from the apical membrane, but not the basolateral membrane. These data reveal a novel, previously unrecognized mechanism that functionally couples GC-C-induced luminal electrolyte transport and cGMP secretion to spatially restricted, compartmentalized regulation by MRP4 at the apical membrane of intestinal epithelium. These findings have important implications for gastrointestinal disorders with symptoms associated with dysregulated fluid homeostasis, such as irritable bowel syndrome with constipation, chronic idiopathic constipation, and secretory diarrhea. PMID:26216942

  16. Truncated atrial natriuretic peptide (ANP) analogs: relationship between receptor binding and cyclic GMP accumulation

    The authors have shown that cultured bovine aortic smooth muscle (BASM) cells contain high affinity receptor sites for the ANP, auriculin (ANP(4-28)). Furthermore, ANP(4-28) causes cyclic GMP (cGMP) levels to increase in BASM. In the present study, the authors synthesized a series of NH2 and/or COOH truncated ANP(4-28) analogs and examined their ability to compete for 125I-ANP(4-28) binding to BASM and increase cGMP levels. ANP-mediated cGMP responses were reduced when amino acids were deleted from the NH2 and/or COOH termini of ANP(4-28). Removal of the NH2 terminal R, R-S, or R-S-S residues resulted in a 10X decrease in potency for cGMP stimulation. Deletion of the COOH terminal R-Y and F-R-Y residues resulted in a marked decline in potency. ANP's lacking the F-R-Y tripeptide were nearly inactive in stimulating cGMP accumulation. In contrast to the cGMP effects, NH2 and/or COOH truncations of ANP(4-28) did not alter apparent receptor binding affinities (Ki(app)). All of these peptide analogs exhibited Ki(app)'s of 1-5 nM. Furthermore, peptides that bound effectively and failed to elicit cGMP responses did not antagonize ANP(4-28)-mediated cGMP increases. These binding and functional data suggest the presence of a single class of ANP receptors on BASM is insufficient to explain the actions of ANP's in these cells

  17. Modulation by cyclic GMP of the odour sensitivity of vertebrate olfactory receptor cells

    Leinders-Zufall, T.; Shepherd, G. M.; Zufall, F.

    1996-01-01

    Recent evidence has indicated a significant role for the cGMP second messenger system in vertebrate olfactory transduction but no clear functions have been identified for cGMP so far. Here, we have examined the effects of 8-Br-cGMP and carbon monoxide (CO) on odour responses of salamander olfactory receptor neurons using perforated patch recordings. We report that 8-Br-cGMP strongly down-regulates the odour sensitivity of the cells, with a K1/2 of 460 nM. This adaptation-like effect can be mimicked by CO, an activator of soluble guanylyl cyclase, with a K1/2 of 1 microM. Sensitivity modulation is achieved through a regulatory chain of events in which cGMP stimulates a persistent background current due to the activation of cyclic nucleotide-gated channels. This in turn leads to sustained Ca2+ entry providing a negative feedback signal. One consequence of the Ca2+ entry is a shift to the right of the stimulus-response curve and a reduction in saturating odour currents. Together, these two effects can reduce the sensory generator current by up to twenty-fold. Thus, cGMP functions to control the gain of the G-protein coupled cAMP pathway. Another consequence of the action of cGMP is a marked prolongation of the odour response kinetics. The effects of CO/cGMP are long-lasting and can continue for minutes. Hence, we propose that cGMP helps to prevent saturation of the cell's response by adjusting the operational range of the cAMP cascade and contributes to olfactory adaptation by decreasing the sensitivity of olfactory receptor cells to repeated odour stimuli.

  18. A cGMP-signaling pathway in a subset of olfactory sensory neurons

    Meyer, M.R.; Angele, A.; Kremmer, E; Kaupp, U B; F. Müller

    2000-01-01

    It is well established that signal transduction in sensory neurons of the rat olfactory epithelium involves a cAMP-signaling pathway. However, a small number of olfactory neurons specifically express cGMP-signaling components, namely a guanylyl cyclase (GC-D) and a cGMP-stimulated phosphodiesterase (PDE2). Here, we show that this subset of olfactory neurons expressing GC-D and PDE2 does also express the subunit of a cGMP-selective cyclic nucleotide-gated (CNG) channel that has been previously...

  19. Radioimmunoassay for cyclic GMP with femtomole sensitivity using tritiated label and acetylated ligands

    An economical assay using [3H]cyclic GMP is described which will be of use to those laboratories either restricted from using γ isotopes or who do not possess gamma-counting facilities. In this assay, label, standards, and samples are acetylated, and interacted with an antibody raised against the succinyl derivative of cyclic GMP prepared in the usual way. Using this assay procedure 20 fmol of cyclic GMP are detectable. The optimal assay conditions were calculated on theoretical grounds. The use of acetylated ligands might also be applied to the assay of cyclic AMP

  20. GEp/GMp ratio by polarization transfer in e --> p --> ep -->

    The ratio of the proton's elastic electromagnetic form factors, GEp/GMp was obtained by measuring Pi and Pl, the transverse and the longitudinal recoil proton polarization, respectively. For elastic ep to ep, GEp/GMp is proportional to Pt/Pl. Simultaneous measurement of Pt and Pl in a polarimeter provides good control of the systematic uncertainty. The results for the ratio GEp/GMp show a systematic decrease as Q2 increases from 0.5 to 3.5 GeV2, indicating for the first time a definite difference in the spatial distribution of charge and magnetization currents in the proton

  1. GEp/GMp ratio by polarization transfer in ep → ep

    The ratio of the proton's elastic electromagnetic form factors, GEp/GMp was obtained by measuring Pt and P(ell), the transverse and the longitudinal recoil proton polarization, respectively. For elastic ep → ep, GEp/GMp is proportional to Pt/P(ell). Simultaneous measurement of Pt and P(ell) in a polarimeter provides good control of the systematic uncertainty. The results for the ratio GEp/GMp show a systematic decrease as Q2 increases from 0.5 to 3.5 GeV2, indicating for the first time a definite difference in the spatial distribution of charge and magnetization currents in the proton

  2. Mechanism of activation of bacterial cellulose synthase by cyclic-di-GMP

    Morgan, Jacob L.W.; McNamara, Joshua T.; Zimmer, Jochen

    2014-01-01

    The bacterial signaling molecule cyclic-di-GMP stimulates the synthesis of bacterial cellulose, frequently found in biofilms. Bacterial cellulose is synthesized and translocated across the inner membrane by a complex of the cellulose synthase BcsA and BcsB subunits. Here we present crystal structures of the cyclic-di-GMP-activated BcsA–B complex. The structures reveal that cyclic-di-GMP releases an auto-inhibited state of the enzyme by breaking a salt bridge which otherwise tethers a conserve...

  3. Pathophysiology of Hypertension in the Absence of Nitric Oxide/Cyclic GMP Signaling

    Thoonen, Robrecht; Sips, Patrick; Bloch, Kenneth Daniel; Buys, Emmanuel

    2012-01-01

    The nitric oxide (NO)-cyclic guanosine monophosphate (cGMP) signaling system is a well-characterized modulator of cardiovascular function, in general, and blood pressure, in particular. The availability of mice mutant for key enzymes in the NO-cGMP signaling system facilitated the identification of interactions with other blood pressure modifying pathways (e.g. the renin-angiotensin-aldosterone system) and of gender-specific effects of impaired NO-cGMP signaling. In addition, recent genome-wi...

  4. Addendum to papers from the GANIL (Grand Accelerateur National d'Ions Lourds) GANISOL Team, published in Proceedings of the 13th International Conference on Ion Sources, Gatlinburg, Tennessee, September 2009

    This addendum applies to the paper authored by contributors from the Grand Accelerateur National d'Ions Lourds (GANIL) published in the February 2010 issue of Review of Scientific Instruments, within the Proceedings of the 13th International Conference on Ion Sources, Gatlinburg, Tennessee, September 2009 (Key Scientific coordinator: P. Delahaye; Technical coordinator: P. Jardin; SPIRAL 2 coordinator: R. Leroy and H. Fraanberg-Delahaye; GPI leader: P. Leherissier; and Direction correspondant: P. Roussel-Chomaz). This addendum provides the full list of GANISOL contributors and their affiliations.

  5. A calcium-permeable cGMP-activated cation conductance in hippocampal neurons

    Leinders-Zufall, T.; Rosenboom, H.; Barnstable, C. J.; Shepherd, G. M.; Zufall, F.

    1995-01-01

    Whole-cell patch clamp recordings detected a previously unidentified cGMP-activated membrane conductance in cultured rat hippocampal neurons. This conductance is nonselectively permeable for cations and is completely but reversibly blocked by external Cd2+. The Ca2+ permeability of the hippocampal cGMP-activated conductance was examined in detail, indicating that the underlying ion channels display a high relative permeability for Ca2+. The results indicate that hippocampal neurons contain a cGMP-activated membrane conductance that has some properties similar to the cyclic nucleotide-gated channels previously shown in sensory receptor cells and retinal neurons. In hippocampal neurons this conductance similarly could mediate membrane depolarization and Ca2+ fluxes in response to intracellular cGMP elevation.

  6. Determination of reactivity of M-cholinergic receptor/cGMP system of peripheral lymphocytes

    Lymphocytes were separated from peripheral blood and incubated in Hanks' solution with calcium or buffer solution containing phosphodiesterase inhibitor IBMX. When carbachol at 1-10 mmol/L was added to the incubation medium, the intracellular cGMP content as determined by high-sensitivity RIA was elevated by 66%-80%. Such an elevation could be blocked by atropine and hence is a specific reaction mediated through M-receptors. A method based on this reaction was thus established to reflect the reactivity of M-receptor/cGMP system. Preliminary experiment revealed that the base level of lymphocyte cGMP as well as the elevation of cGMP induced by carbachol in aged (24-26 months old) rats were significantly lower than young rats. The methodology, especially the influence of incubation conditions was discussed

  7. The role of cGMP hydrolysing phosphodiesterases 1 and 5 in cerebral artery dilatation

    Kruuse, Christina; Rybalkin, S D; Khurana, T S; Jansen-Olesen, I; Olesen, J.; Edvinsson, L

    2001-01-01

    The aim was to investigate the presence and activity of cGMP hydrolysing phosphodiesterases in guinea pig basilar arteries and the effect of selective and non-selective phosphodiesterase inhibitors on cerebral artery dilatation involving the nitric oxide (NO)-guanosine cyclic 3'5-monophosphate (cGMP......M) pretreatment, indicating a close relation to the nitric oxide-cGMP pathway. The responses to zaprinast and dipyridamole, however, were not only moderately affected, but also restored by sodium nitroprusside (0.1 microM) pretreatment. At high concentrations, the dilatory effects of zaprinast and dipyridamole...... were partly caused by cGMP-independent mechanisms. Targeting the phosphodiesterases present in cerebral arteries, with selective inhibitors or activators of phosphodiesterase, may be a possible new way of treating cerebrovascular disease....

  8. Compilation of key GMP requirements in us and Japan for Tablet Manufacturing

    Sujith Kumar . M

    2011-12-01

    Full Text Available The tablet is most popular dosage form in the world. The objective of this work helps in bringing the awareness about the manufacturing requirements as per USFDA and Japan. GMP covers all aspects of production, from the starting materials, premises, equipment and training and personal hygiene to staff. GMP of tablet manufacturing was designed to ensure that all is well. Implementation of GMP is an investment in good quality medicines. It demonstrates industry and regulatory authority’s support for an effective pharmaceutical quality system to enhance the quality and availability of medicines around the world in the interest of public health. Implementation of GMP throughout the product lifecycle should facilitate innovation and continual improvement and strengthen the link between pharmaceutical development and manufacturing activities. It also helps in increasing the process efficiency and product quality by adopting the current, risk-based manufacturing approach and in turn optimizes manufacturing process and improves quality of the end-product.

  9. Preparation of 2'-O-succinyl[8-3H]cGMP and of 2'-O-([2,3-3H]succinyl)cGMP

    The methods are described of the preparation of 2'-O-succinyl[8-3H]cGMP of a molar activity of 416 GBq mmol-1 and 2'-O-([2,3-3H]succinyl)cGMP of a molar activity of 1.722 TBq mmol-1. In tracer tests the effect was studied of molar concentration of succinic anhydride on reaction yield. Also identified were reaction conditions under which the isotopic exchange of tritium in [8-3H]cGMP is limited [2,3-3H]succinic anhydride was prepared by tritiation of maleinic anhydride in anhydrous dioxane catalyzed by 10% Pd/C. (author). 1 fig., 7 tabs., 6 refs

  10. Expression and Genetic Activation of Cyclic Di-GMP-Specific Phosphodiesterases in Escherichia coli

    Reinders, Alberto; Hee, Chee-Seng; Ozaki, Shogo; Mazur, Adam; Boehm, Alex; Schirmer, Tilman

    2015-01-01

    ABSTRACT Intracellular levels of the bacterial second messenger cyclic di-GMP (c-di-GMP) are controlled by antagonistic activities of diguanylate cyclases and phosphodiesterases. The phosphodiesterase PdeH was identified as a key regulator of motility in Escherichia coli, while deletions of any of the other 12 genes encoding potential phosphodiesterases did not interfere with motility. To analyze the roles of E. coli phosphodiesterases, we demonstrated that most of these proteins are expressed under laboratory conditions. We next isolated suppressor mutations in six phosphodiesterase genes, which reinstate motility in the absence of PdeH by reducing cellular levels of c-di-GMP. Expression of all mutant alleles also led to a reduction of biofilm formation. Thus, all of these proteins are bona fide phosphodiesterases that are capable of interfering with different c-di-GMP-responsive output systems by affecting the global c-di-GMP pool. This argues that E. coli possesses several phosphodiesterases that are inactive under laboratory conditions because they lack appropriate input signals. Finally, one of these phosphodiesterases, PdeL, was studied in more detail. We demonstrated that this protein acts as a transcription factor to control its own expression. Motile suppressor alleles led to a strong increase of PdeL activity and elevated pdeL transcription, suggesting that enzymatic activity and transcriptional control are coupled. In agreement with this, we showed that overall cellular levels of c-di-GMP control pdeL transcription and that this control depends on PdeL itself. We thus propose that PdeL acts both as an enzyme and as a c-di-GMP sensor to couple transcriptional activity to the c-di-GMP status of the cell. IMPORTANCE Most bacteria possess multiple diguanylate cyclases and phosphodiesterases. Genetic studies have proposed that these enzymes show signaling specificity by contributing to distinct cellular processes without much cross talk. Thus, spatial

  11. Discrete cyclic di-GMP-dependent control of bacterial predation versus axenic growth in Bdellovibrio bacteriovorus

    Laura Hobley; Fung, Rowena K. Y.; Carey Lambert; Maximilian A T S Harris; Dabhi, Jayesh M.; King, Simon S.; Basford, Sarah M; Kaoru Uchida; Robert Till; Rashidah Ahmad; Shin-Ichi Aizawa; Mark Gomelsky; R Elizabeth Sockett

    2012-01-01

    Bdellovibrio bacteriovorus is a Delta-proteobacterium that oscillates between free-living growth and predation on Gram-negative bacteria including important pathogens of man, animals and plants. After entering the prey periplasm, killing the prey and replicating inside the prey bdelloplast, several motile B. bacteriovorus progeny cells emerge. The B. bacteriovorus HD100 genome encodes numerous proteins predicted to be involved in signalling via the secondary messenger cyclic di-GMP (c-di-GMP)...

  12. Clinical relevance of cyclic GMP modulators: A translational success story of network pharmacology.

    Oettrich, J M; Dao, V T; Frijhoff, J; Kleikers, Pwm; Casas, A I; Hobbs, A J; Schmidt, Hhhw

    2016-04-01

    Therapies that modulate cyclic guanosine-3'-5'-monophosphate (cGMP) have emerged as one of the most successful areas in recent drug discovery and clinical pharmacology. Historically, their focus has been on cardiovascular disease phenotypes; however, cGMP's relevance is likely to go beyond this rather limited organ-based set of indications. Moreover, the multitude of targets and their apparent interchangeability is a proof-of-concept of network pharmacology. PMID:26765222

  13. Design of GMP compliance radiopharmaceutical production facility in MINT

    Full text: In 1985, MINT built the only radiopharmaceutical production facility in Malaysia. The facility was designed based on IAEA (International Atomic Energy Agency) standard guidelines, which provides radiation safety to the operator and the surrounding environment from radioactive contamination. In 1999, BPFK (Malaysia National Pharmaceutical Control Bureau) enforced the guidelines from Pharmaceutical Inspection Convention Scheme (PICS) to meet the requirements of the Good Manufacturing Practice (GMP) for Pharmaceutical Products. Among others, the guidelines require the pharmaceutical production facility to be designed based on cleanroom environment. In order to meet this requirement, the design of a radiopharmaceutical production facility shall combine the concept of radiation safety and cleanroom to ensure that both requirements from GMP and IAEA are met. This design requirement is necessary to ensure a radiopharmaceutical production facility which is safe, has high production quality and in compliance with the Malaysian and International standards. In order to obtain the license to continue producing radiopharmaceutical products, MINT shall closely follow the guidelines from PICS: GMP and nuclear facility requirements. To meet those requirements, the layout of MINT facility was redesigned to ensure the contaminants or particulates from outside area will not enter the production area. The flow path of personnel to the production room is equiped with air lock and change room in order to avoid contamination from outside environment. The airlock and changing room provide the physical separation of different stages of cleanroom class and between clean area and non-clean area. The cleanliness level or class for each room is determined by the criticality of exposure of the process and products to the environment. The clean class which is grade A is used for critical process and the other grade is for non-critical process or as a background. The concepts of

  14. The Nitric Oxide/Cyclic GMP Pathway in Organ Transplantation: Critical Role in Successful Lung Preservation

    Pinsky, David J.; Naka, Yoshifumi; Chowdhury, Nepal C.; Liao, Hui; Oz, Mehmet C.; Michler, Robert E.; Kubaszewski, Eugeniusz; Malinski, Tadeusz; Stern, David M.

    1994-12-01

    Reestablishment of vascular homeostasis following ex vivo preservation is a critical determinant of successful organ transplantation. Because the nitric oxide (NO) pathway modulates pulmonary vascular tone and leukocyte/endothelial interactions, we hypothesized that reactive oxygen intermediates would lead to decreased NO (and hence cGMP) levels following pulmonary reperfusion, leading to increased pulmonary vascular resistance and leukostasis. Using an orthotopic rat model of lung transplantation, a porphyrinic microsensor was used to make direct in vivo measurements of pulmonary NO. NO levels measured at the surface of the transplanted lung plummeted immediately upon reperfusion, with levels moderately increased by topical application of superoxide dismutase. Because cGMP levels declined in preserved lungs after reperfusion, this led us to buttress the NO pathway by adding a membrane-permeant cGMP analog to the preservation solution. Compared with grafts stored in its absence, grafts stored with supplemental 8-Br-cGMP and evaluated 30 min after reperfusion demonstrated lower pulmonary vascular resistances with increased graft blood flow, improved arterial oxygenation, decreased neutrophil infiltration, and improved recipient survival. These beneficial effects were dose dependent, mimicked by the type V phosphodiesterase inhibitor 2-o-propoxyphenyl-8-azapurin-6-one, and inhibited by a cGMP-dependent protein kinase antagonist, the R isomer of 8-(4-chlorophenylthio)guanosine 3',5'-cyclic monophosphorothioate. Augmenting the NO pathway at the level of cGMP improves graft function and recipient survival following lung transplantation.

  15. CPT-cGMP Is A New Ligand of Epithelial Sodium Channels.

    Ji, Hong-Long; Nie, Hong-Guang; Chang, Yongchang; Lian, Qizhou; Liu, Shan-Lu

    2016-01-01

    Epithelial sodium channels (ENaC) are localized at the apical membrane of the epithelium, and are responsible for salt and fluid reabsorption. Renal ENaC takes up salt, thereby controlling salt content in serum. Loss-of-function ENaC mutations lead to low blood pressure due to salt-wasting, while gain-of-function mutations cause impaired sodium excretion and subsequent hypertension as well as hypokalemia. ENaC activity is regulated by intracellular and extracellular signals, including hormones, neurotransmitters, protein kinases, and small compounds. Cyclic nucleotides are broadly involved in stimulating protein kinase A and protein kinase G signaling pathways, and, surprisingly, also appear to have a role in regulating ENaC. Increasing evidence suggests that the cGMP analog, CPT-cGMP, activates αβγ-ENaC activity reversibly through an extracellular pathway in a dose-dependent manner. Furthermore, the parachlorophenylthio moiety and ribose 2'-hydroxy group of CPT-cGMP are essential for facilitating the opening of ENaC channels by this compound. Serving as an extracellular ligand, CPT-cGMP eliminates sodium self-inhibition, which is a novel mechanism for stimulating salt reabsorption in parallel to the traditional NO/cGMP/PKG signal pathway. In conclusion, ENaC may be a druggable target for CPT-cGMP, leading to treatments for kidney malfunctions in salt reabsorption. PMID:27019621

  16. Bacterial Signal Transduction by Cyclic Di-GMP and Other Nucleotide Second Messengers.

    Hengge, Regine; Gründling, Angelika; Jenal, Urs; Ryan, Robert; Yildiz, Fitnat

    2016-01-01

    The first International Symposium on c-Di-GMP Signaling in Bacteria (22 to 25 March 2015, Harnack-Haus, Berlin, Germany)brought together 131 molecular microbiologists from 17 countries to discuss recent progress in our knowledge of bacterial nucleotide second messenger signaling. While the focus was on signal input, synthesis, degradation, and the striking diversity of the modes of action of the current second messenger paradigm, i.e., cyclic di-GMP (c-di-GMP), “classics” like cAMP and (p)ppGpp were also presented, in novel facets, and more recent “newcomers,” such as c-di-AMP and c-AMP-GMP, made an impressive appearance. A number of clear trends emerged during the 30 talks, on the 71 posters, and in the lively discussions, including (i)c-di-GMP control of the activities of various ATPases and phosphorylation cascades, (ii) extensive cross talk between c-di-GMP and other nucleotide second messenger signaling pathways, and (iii) a stunning number of novel effectors for nucleotide second messengers that surprisingly include some long-known master regulators of developmental pathways. Overall, the conference made it amply clear that second messenger signaling is currently one of the most dynamic fields within molecular microbiology,with major impacts in research fields ranging from human health to microbial ecology. PMID:26055111

  17. NO, nitrotyrosine, and cyclic GMP in signal transduction

    Hanafy, K. A.; Krumenacker, J. S.; Murad, F.

    2001-01-01

    Over the past 25 years, the role of nitric oxide (NO) in biology has evolved from being recognized as an environmental pollutant to an endogenously produced substance involved in cell communication and signal transduction. NO is produced by a family of enzymes called nitric oxide synthases (NOSs), which can be stimulated by a variety of factors that mediate responses to various stimuli. NO can initiate its biological effects through activation of the heterodimeric enzyme, soluble guanylyl cyclase (sGC), or through several other chemical reactions. Activation of sGC results in the production of 3',5'-cyclic guanosine monophosphate (cGMP), an intracellular second messenger signaling molecule, which can subsequently mediate such diverse physiological events such as vasodilatation and immunomodulation. Chemically reactive NO can affect physiological changes through modifications to cellular proteins, one of which is tyrosine nitration. The demonstration that NO is involved in so many biological pathways indicates the importance of this endogenously produced substance, and suggests that there is much more to be discovered about its role in biology in years to come.

  18. In Vivo Reconstitution of the Negative Feedback in Nitric Oxide/cGMP Signaling: Role of Phosphodiesterase Type 5 Phosphorylation

    Mullershausen, Florian; Russwurm, Michael; Koesling, Doris; Friebe, Andreas

    2004-01-01

    Most effects of the messenger molecule nitric oxide (NO) are mediated by cGMP, which is formed by NO-sensitive guanylyl cyclase (GC) and degraded by phosphodiesterases (PDEs). In platelets, NO elicits a spike-like cGMP response and causes a sustained desensitization. Both characteristics have been attributed to PDE5 activation caused by cGMP binding to its regulatory GAF domain. Activation is paralleled by phosphorylation whose precise function remains unknown. Here, we report reconstitution ...

  19. A Cyclic GMP-Dependent K+ Channel in the Blastocladiomycete Fungus Blastocladiella emersonii.

    Avelar, Gabriela Mól; Glaser, Talita; Leonard, Guy; Richards, Thomas A; Ulrich, Henning; Gomes, Suely L

    2015-09-01

    Phototaxis in flagellated zoospores of the aquatic fungus Blastocladiella emersonii depends on a novel photosensor, Blastocladiella emersonii GC1 (BeGC1), comprising a type I (microbial) rhodopsin fused to a guanylyl cyclase catalytic domain, that produces the conserved second messenger cyclic GMP (cGMP). The rapid and transient increase in cGMP levels during the exposure of zoospores to green light was shown to be necessary for phototaxis and dependent on both rhodopsin function and guanylyl cyclase activity. It is noteworthy that BeGC1 was localized to the zoospore eyespot apparatus, in agreement with its role in the phototactic response. A putative cyclic nucleotide-gated channel (BeCNG1) was also identified in the genome of the fungus and was implicated in flagellar beating via the action of a specific inhibitor (l-cis-diltiazem) that compromised zoospore motility. Here we show that B. emersonii expresses a K(+) channel that is activated by cGMP. The use of specific channel inhibitors confirmed the activation of the channel by cGMP and its K(+) selectivity. These characteristics are consistent with the function of an ion channel encoded by the BeCNG1 gene. Other blastocladiomycete fungi, such as Allomyces macrogynus and Catenaria anguillulae, possess genes encoding a similar K(+) channel and the rhodopsin-guanylyl cyclase fusion protein, while the genes encoding both these proteins are absent in nonflagellated fungi. The presence of these genes as a pair seems to be an exclusive feature of blastocladiomycete fungi. Taken together, these data demonstrate that the B. emersonii cGMP-activated K(+) channel is involved in the control of zoospore motility, most probably participating in the cGMP-signaling pathway for the phototactic response of the fungus. PMID:26150416

  20. Cyclic di-GMP stimulates biofilm formation and inhibits virulence of Francisella novicida.

    Zogaj, Xhavit; Wyatt, Geoff C; Klose, Karl E

    2012-12-01

    Francisella tularensis is a gram-negative bacterium that is highly virulent in humans, causing the disease tularemia. F. novicida is closely related to F. tularensis and exhibits high virulence in mice, but it is avirulent in healthy humans. An F. novicida-specific gene cluster (FTN0451 to FTN0456) encodes two proteins with diguanylate cyclase (DGC) and phosphodiesterase (PDE) domains that modulate the synthesis and degradation of cyclic di-GMP (cdGMP). No DGC- or PDE-encoding protein genes are present in the F. tularensis genome. F. novicida strains lacking either the two DGC/PDE genes (cdgA and cdgB) or the entire gene cluster (strain KKF457) are defective for biofilm formation. In addition, expression of CdgB or a heterologous DGC in strain KKF457 stimulated F. novicida biofilms, even in a strain lacking the biofilm regulator QseB. Genetic evidence suggests that CdgA is predominantly a PDE, while CdgB is predominantly a DGC. The F. novicida qseB strain showed reduced cdgA and cdgB transcript levels, demonstrating an F. novicida biofilm signaling cascade that controls cdGMP levels. Interestingly, KKF457 with elevated cdGMP levels exhibited a decrease in intramacrophage replication and virulence in mice, as well as increased growth yields and biofilm formation in vitro. Microarray analyses revealed that cdGMP stimulated the transcription of a chitinase (ChiB) known to contribute to biofilm formation. Our results indicate that elevated cdGMP in F. novicida stimulates biofilm formation and inhibits virulence. We suggest that differences in human virulence between F. novicida and F. tularensis may be due in part to the absence of cdGMP signaling in F. tularensis. PMID:22988021

  1. Structural and Biochemical Determinants of Ligand Binding by the c-di-GMP Riboswitch

    The bacterial second messenger c-di-GMP is used in many species to control essential processes that allow the organism to adapt to its environment. The c-di-GMP riboswitch (GEMM) is an important downstream target in this signaling pathway and alters gene expression in response to changing concentrations of c-di-GMP. The riboswitch selectively recognizes its second messenger ligand primarily through contacts with two critical nucleotides. However, these two nucleotides are not the most highly conserved residues within the riboswitch sequence. Instead, nucleotides that stack with c-di-GMP and that form tertiary RNA contacts are the most invariant. Biochemical and structural evidence reveals that the most common natural variants are able to make alternative pairing interactions with both guanine bases of the ligand. Additionally, a high-resolution (2.3 (angstrom)) crystal structure of the native complex reveals that a single metal coordinates the c-di-GMP backbone. Evidence is also provided that after transcription of the first nucleotide on the 3(prime)-side of the P1 helix, which is predicted to be the molecular switch, the aptamer is functional for ligand binding. Although large energetic effects occur when several residues in the RNA are altered, mutations at the most conserved positions, rather than at positions that base pair with c-di-GMP, have the most detrimental effects on binding. Many mutants retain sufficient c-di-GMP affinity for the RNA to remain biologically relevant, which suggests that this motif is quite resilient to mutation.

  2. Nucleotide binding by the widespread high-affinity cyclic di-GMP receptor MshEN domain.

    Wang, Yu-Chuan; Chin, Ko-Hsin; Tu, Zhi-Le; He, Jin; Jones, Christopher J; Sanchez, David Zamorano; Yildiz, Fitnat H; Galperin, Michael Y; Chou, Shan-Ho

    2016-01-01

    C-di-GMP is a bacterial second messenger regulating various cellular functions. Many bacteria contain c-di-GMP-metabolizing enzymes but lack known c-di-GMP receptors. Recently, two MshE-type ATPases associated with bacterial type II secretion system and type IV pilus formation were shown to specifically bind c-di-GMP. Here we report crystal structure of the MshE N-terminal domain (MshEN1-145) from Vibrio cholerae in complex with c-di-GMP at a 1.37 Å resolution. This structure reveals a unique c-di-GMP-binding mode, featuring a tandem array of two highly conserved binding motifs, each comprising a 24-residue sequence RLGxx(L/V/I)(L/V/I)xxG(L/V/I)(L/V/I)xxxxLxxxLxxQ that binds half of the c-di-GMP molecule, primarily through hydrophobic interactions. Mutating these highly conserved residues markedly reduces c-di-GMP binding and biofilm formation by V. cholerae. This c-di-GMP-binding motif is present in diverse bacterial proteins exhibiting binding affinities ranging from 0.5 μM to as low as 14 nM. The MshEN domain contains the longest nucleotide-binding motif reported to date. PMID:27578558

  3. Inhibition of cGMP-dependent protein kinase by (Rp)-guanosine 3',5'-monophosphorothioates.

    Butt, E; van Bemmelen, M; Fischer, L; Walter, U; Jastorff, B

    1990-04-01

    The activation of the cGMP-dependent protein kinase and cAMP-dependent protein kinase by the diastereomers of guanosine 3',5'-monophosphorothioate, (Sp)-cGMPS and (Rp)-cGMPS, and 8-chloroguanosine 3',5'-monophosphorothioate, (Sp)-8-Cl-cGMPS and (Rp)-8-Cl-cGMPS, was investigated using the peptide Kemptide as substrate. The (Sp)-diastereomers, which have an axial exocyclic sulfur atom, bound to the cGMP-dependent protein kinase and stimulated its phosphotransferase activity. In contrast, the (Rp)-isomers, which have an equatorial exocyclic sulfur atom, bound to the enzyme without stimulation of its activity. (Rp)-cGMPS and (Rp)-8-Cl-cGMPS antagonized the activation of the cGMP-dependent protein kinase with a Ki of 20 microM and 1.5 microM, respectively. (Rp)-cGMPS also antagonized the activation of cAMP-dependent protein kinase with a Ki of 20 microM. In contrast, (Rp)-8-cGMPS ws a weak inhibitor of the cAMP-dependent protein kinase with a Ki of 100 microM. (Rp)-8-Cl-cGMPS appears to be a rather selective inhibitor of the cGMP-dependent protein kinase and may be a useful tool for studying the role of cGMP in broken and intact cell systems. PMID:2158906

  4. Acute stress-induced antinociception is cGMP-dependent but heme oxygenase-independent

    Carvalho-Costa, P.G. [Programa de Graduação em Psicobiologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil); Branco, L.G.S. [Departamento de Morfologia, Fisiologia e Patologia Básica, Faculdade de Odontologia de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil); Leite-Panissi, C.R.A. [Programa de Graduação em Psicobiologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil); Departamento de Morfologia, Fisiologia e Patologia Básica, Faculdade de Odontologia de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil)

    2014-09-19

    Endogenous carbon monoxide (CO), which is produced by the enzyme heme oxygenase (HO), participates as a neuromodulator in physiological processes such as thermoregulation and nociception by stimulating the formation of 3′,5′-cyclic guanosine monophosphate (cGMP). In particular, the acute physical restraint-induced fever of rats can be blocked by inhibiting the enzyme HO. A previous study reported that the HO-CO-cGMP pathway plays a key phasic antinociceptive role in modulating noninflammatory acute pain. Thus, this study evaluated the involvement of the HO-CO-cGMP pathway in antinociception induced by acute stress in male Wistar rats (250-300 g; n=8/group) using the analgesia index (AI) in the tail flick test. The results showed that antinociception induced by acute stress was not dependent on the HO-CO-cGMP pathway, as neither treatment with the HO inhibitor ZnDBPG nor heme-lysinate altered the AI. However, antinociception was dependent on cGMP activity because pretreatment with the guanylate cyclase inhibitor 1H-[1,2,4] oxadiazolo [4,3-a] quinoxaline-1-one (ODQ) blocked the increase in the AI induced by acute stress.

  5. Effects of radioprotectors on the cAMP and cGMP systems

    The sulphur-containing radioprotectors mercaptoethylamine (MEA), aminoethylisothiourea (AET), 2-aminothiazoline, 4-oxo-2-amino-thiazoline, and S-S-oxapentane-1,5-diisorthiourea, and the radioprotective biogenic amines serotonin, histamine, and dopamine, caused the elevation of cAMP content and intensified the rate of cAMP-dependent protein phosphorylation in tissues of animals following intraperitoneal injection at radioprotective doses. Biogenic amines stimulated the adenylate cyclase activity in membrane preparations from liver, spleen, and small-intestine mucosa; sulphur-containing radioprotectors caused no such effects. None of the radioprotectors affected CAMP and cGMP phosphodiesterases in vitro. AET and MEA inhibited guanylate cyclase in vitro, whereas serotonin and dopamine stimulated the enzyme. A biphasic change in the level of cGMP was observed in tissues after the administration of MEA and AET (more than 2-fold fally by 1-3 min after the administration of drug and 1.4-fold rise after 15-20 min); serotonin and dopamine caused a slow rise in the cGMP level, the cAMP/cGMP ratio in liver showed biphasic changes in level during the 20 min following injection of serotonin. The data obtained support the conclusion that the action of radioprotectors on cellular metabolism in animals may be mediated by the cAMP system.The reciprocal regulation of radioresistance by cAMP and cGMP is unlikely to exist. (orig.)

  6. Acute stress-induced antinociception is cGMP-dependent but heme oxygenase-independent

    Endogenous carbon monoxide (CO), which is produced by the enzyme heme oxygenase (HO), participates as a neuromodulator in physiological processes such as thermoregulation and nociception by stimulating the formation of 3′,5′-cyclic guanosine monophosphate (cGMP). In particular, the acute physical restraint-induced fever of rats can be blocked by inhibiting the enzyme HO. A previous study reported that the HO-CO-cGMP pathway plays a key phasic antinociceptive role in modulating noninflammatory acute pain. Thus, this study evaluated the involvement of the HO-CO-cGMP pathway in antinociception induced by acute stress in male Wistar rats (250-300 g; n=8/group) using the analgesia index (AI) in the tail flick test. The results showed that antinociception induced by acute stress was not dependent on the HO-CO-cGMP pathway, as neither treatment with the HO inhibitor ZnDBPG nor heme-lysinate altered the AI. However, antinociception was dependent on cGMP activity because pretreatment with the guanylate cyclase inhibitor 1H-[1,2,4] oxadiazolo [4,3-a] quinoxaline-1-one (ODQ) blocked the increase in the AI induced by acute stress

  7. Auxin-induced nitric oxide, cGMP and gibberellins were involved in the gravitropism

    Cai, Weiming; Hu, Liwei; Hu, Xiangyang; Cui, Dayong; Cai, Weiming

    Gravitropism is the asymmetric growth or curvature of plant organs in response to gravistimulation. There is a complex signal transduction cascade which involved in the differential growth of plants in response to changes in the gravity vector. The role of auxin in gravitropism has been demonstrated by many experiments, but little is known regarding the molecular details of such effects. In our studies before, mediation of the gravitropic bending of soybean roots and rice leaf sheath bases by nitric oxide, cGMP and gibberellins, are induced by auxin. The asymmetrical distribution of nitric oxide, cGMP and gibberellins resulted from the asymmetrical synthesis of them in bending sites. In soybean roots, inhibitions of NO and cGMP synthesis reduced differential NO and cGMP accumulation respectively, which both of these effects can lead to the reduction of gravitropic bending. Gibberellin-induced OsXET, OsEXPA4 and OsRWC3 were also found involved in the gravitropic bending. These data indicated that auxin-induced nitric oxide, cGMP and gibberellins were involved in the gravitropism. More experiments need to prove the more detailed mechanism of them.

  8. Bacterial c-di-GMP Affects Hematopoietic Stem/Progenitors and Their Niches through STING

    Hiroshi Kobayashi

    2015-04-01

    Full Text Available Upon systemic bacterial infection, hematopoietic stem and progenitor cells (HSPCs migrate to the periphery in order to supply a sufficient number of immune cells. Although pathogen-associated molecular patterns reportedly mediate HSPC activation, how HSPCs detect pathogen invasion in vivo remains elusive. Bacteria use the second messenger bis-(3′-5′-cyclic dimeric guanosine monophosphate (c-di-GMP for a variety of activities. Here, we report that c-di-GMP comprehensively regulated both HSPCs and their niche cells through an innate immune sensor, STING, thereby inducing entry into the cell cycle and mobilization of HSPCs while decreasing the number and repopulation capacity of long-term hematopoietic stem cells. Furthermore, we show that type I interferon acted as a downstream target of c-di-GMP to inhibit HSPC expansion in the spleen, while transforming growth factor-β was required for c-di-GMP-dependent splenic HSPC expansion. Our results define machinery underlying the dynamic regulation of HSPCs and their niches during bacterial infection through c-di-GMP/STING signaling.

  9. Nutrition at school: preparing for the future. The Thirteenth Annual Nutrition Symposium.

    1994-01-01

    Two of the best ways to improve the quality of childhood nutrition are (a) more collaboration at the national, State, and local levels and (b) adoption of innovative and multimedia learning methods, according to the leaders of nutrition education. These themes were discussed at the 13th Annual Nutrition Symposium, which was held March 9, 1994, in Washington, DC. In recognition of National Nutrition Month, a collaborative effort of the Public Health Service's Office of Disease Prevention and H...

  10. Differential modulation of the glutamate-nitric oxide-cyclic GMP pathway by distinct neurosteroids in cerebellum in vivo.

    Cauli, O; González-Usano, A; Agustí, A; Felipo, V

    2011-09-01

    The glutamate-nitric oxide (NO)-cGMP pathway mediates many responses to activation of N-methyl-d-aspartate (NMDA) receptors, including modulation of some types of learning and memory. The glutamate-NO-cGMP pathway is modulated by GABAergic neurotransmission. Activation of GABA(A) receptors reduces the function of the pathway. Several neurosteroids modulate the activity of GABA(A) and/or NMDA receptors, suggesting that they could modulate the function of the glutamate-NO-cGMP pathway. The aim of this work was to assess, by in vivo microdialysis, the effects of several neurosteroids with different effects on GABA(A) and NMDA receptors on the function of the glutamate-NO-cGMP pathway in cerebellum in vivo. To assess the effects of the neurosteroids on the glutamate-NO-cGMP pathway, they were administered through the microdialysis probe before administration of NMDA and the effects on NMDA-induced increase in extracellular cGMP were analyzed. We also assessed the effects of the neurosteroids on basal levels of extracellular cGMP. To assess the effects of neurosteroids on nitric oxide synthase (NOS) activity and on NMDA-induced activation of NOS, we also measured the effects of the neurosteroids on extracellular citrulline. Pregnanolone and tetrahydrodeoxy-corticosterone (THDOC) behave as agonists of GABA(A) receptors and completely block NMDA-induced increase in cGMP. Pregnanolone but not THDOC also reduced basal levels of extracellular cGMP. Pregnenolone did not affect extracellular cGMP or its increase by NMDA administration. Pregnenolone sulfate increased basal extracellular cGMP and potentiated NMDA-induced increase in cGMP, behaving as an enhancer of NMDA receptors activation. Allopregnanolone and dehydroepiandrosterone sulphate behave as antagonists of NMDA receptors, increasing basal cGMP and blocking completely NMDA-induced increase in cGMP. Dehydroepiandrosterone sulphate seems to do this by activating sigma receptors. These data support the concept that, at

  11. Regulation of biofilm formation and cellular buoyancy through modulating intracellular cyclic di-GMP levels in engineered cyanobacteria.

    Agostoni, Marco; Waters, Christopher M; Montgomery, Beronda L

    2016-02-01

    The second messenger cyclic dimeric (3'→5') GMP (cyclic di-GMP or c-di-GMP) has been implicated in the transition between motile and sessile lifestyles in bacteria. In this study, we demonstrate that biofilm formation, cellular aggregation or flocculation, and cellular buoyancy are under the control of c-di-GMP in Synechocystis sp. PCC 6803 (Synechocystis) and Fremyella diplosiphon. Synechocystis is a unicellular cyanobacterium and displays lower levels of c-di-GMP; F. diplosiphon is filamentous and displays higher intracellular c-di-GMP levels. We transformed Synechocystis and F. diplosiphon with a plasmid for constitutive expression of genes encoding diguanylate cylase (DGC) and phosphodiesterase (PDE) proteins from Vibrio cholerae or Escherichia coli, respectively. These engineered strains allowed us to modulate intracellular c-di-GMP levels. Biofilm formation and cellular deposition were induced in the DGC-expressing Synechocystis strain which exhibited high intracellular levels of c-di-GMP; whereas strains expressing PDE in Synechocystis and F. diplosiphon to drive low intracellular levels of c-di-GMP exhibited enhanced cellular buoyancy. In addition, the PDE-expressing F. diplosiphon strain showed elevated chlorophyll levels. These results imply roles for coordinating c-di-GMP homeostasis in regulating native cyanobacterial phenotypes. Engineering exogenous DGC or PDE proteins to regulate intracellular c-di-GMP levels represents an effective tool for uncovering cryptic phenotypes or modulating phenotypes in cyanobacteria for practical applications in biotechnology applicable in photobioreactors and in green biotechnologies, such as energy-efficient harvesting of cellular biomass or the treatment of metal-containing wastewaters. PMID:26192200

  12. 国际钨协第13届年会市场报告%A Market Report for the 13th Annual General Meeting of International Tungsten Industry Association

    Maby,M; 邱银兰

    2000-01-01

    @@ 0 前言 本报告是为在美国华盛顿召开的国际钨协第13届年度大会准备的,内备有最新统计数据(截止2000年6月)对原钨的供应和需求进行了评估.所有的数据均以钨含量为单位.感谢所有提供有关统计数据和资料的会员们的合作,特别是感谢欧洲金属协会和美国地质调查局给予的支持和帮助.

  13. Perspectives on the Economy and Monetary Policy The 13th Annual Economic Outlook The School of Business at La Salle University, Union League of Philadelphia Business Network, January 14, 2014, Philadelphia, PA

    Charles I. Plosser

    2014-01-01

    Presented by Charles I. Plosser, President and Chief Executive Officer, Federal Reserve Bank of Philadelphia The School of Business at La Salle University, Union League of Philadelphia Business Network, January 14, 2014, Philadelphia, PA President Plosser discusses his forecast for growth, prospects for the labor market, and inflation expectations, as well as the extraordinary policy actions the Federal Reserve has taken to support economic recovery. He will also speak about the history of th...

  14. Larval Fish Recruitment and Research in the Americas: Proceedings of the Thirteenth Annual Larval Fish Conference Merida, Mexico, 21-26 May 1989

    1991-01-01

    The 13th Annual Larval Fish Conference and Annual Meeting of the American Fisheries Society Early Life History Section cohosted by Mote Marine Laboratory, United States, and the Instituto Nacional de la Pesca, Mexico, were held 21-26 May 1989, in Merida, Yucatan, Mexico. The purpose of holding the meeting in Mexico was to encourage the participation of our Latin American and Caribbean colleagues and to provide a forum for the exchange of ideas and information among researchers working in the ...

  15. Role of nitric oxide and cyclic GMP signaling in melanocyte response to hypergravity

    Ivanova, Krassimira; Lambers, Britta; Tsiockas, Wasiliki; Block, Ingrid; Gerzer, Rupert

    Nitric oxide (NO) has a prominent role in many (patho)physiological processes in the skin including erythema, inflammation, and cancerogenesis. The soluble guanylyl cyclase (sGC), a key transducer in NO signaling, catalyzes the formation of the second messenger guanosine 3´,5´-cyclic monophosphate (cyclic cGMP or cGMP). For human melanocytes, which are responsible for skin pigmentation by synthesizing the pigment melanin, it has been reported that the NO/sGC/cGMP pathway is involved in UVB-induced melanogenesis. Melanin acts as a scavenger for free radicals that may arise during metabolic stress. It may also act as a photosensitizer that generates active oxygen species upon UV irradiation, which may initiate hypopigmentary disorders (e.g., vitiligo) as well as UV-induced oncogene cell transformation. In addition, melanoma, a deadly skin cancer, which arises from transformed melanocytes, is characterized by a resistance to chemotherapy. In our studies we have shown that NO can induce perturbation of melanocyte-extracellular matrix component interactions, which may contribute to loss of melanocytes or melanoma metastasis. Such NO effects appear to be modulated partly via cGMP. Moreover, we found that different guanylyl cyclase isoforms are responsible for cGMP synthesis in melanocytic cells. Normal human melanocytes and nonmetastatic melanoma cells predominantly express sGC, which appears to be associated with melanogenesis, whereas absence of NO-sensitive GC, but up-regulated activities of the natriuretic peptide-sensitive membrane guanylyl cyclase isoforms were found in highly metastatic phenotypes. Due to the growing interest in the regulation of signaling activities in normal and transformed cells under altered gravity conditions, we have further investigated whether the NO/cGMP signaling is involved in melanocyte response to gravitational stress. We found that normal human melanocytes and non-metastatic melanoma cell lines, but not highly metastatic cells

  16. Effect of luteinizing hormone on the concentration of cyclic GMP in rabbit ovaries

    It has been shown that luteinizing hormone (LH) causes a rapid increase in cyclic AMP in the ovarian tissue of rabbits which precedes the LH-induced increase in steroidogenesis. Investigations have been made to ascertain the effect of LH on cyclic GMP, using radioimmunoassay techniques. It was found that the concentration of cyclic GMP remained unaffected up to four hours after LH stimulation, but between 4 and 8 hours it was significantly reduced and then returned to the control level. It is suggested that cyclic GMP may have no role in ovarian events in the first few hours after LH treatment. The diminished levels in the later periods may conceivably be related to later effects of LH, such as stimulation of prostaglandin biosynthesis. (U.K.)

  17. Effects of corydalis amabilis Migo total alkaloids on platelet cyclic AMP and cyclic GMP

    Corydalis amabilis Migo total alkaloids (COAMTA), isolated from the root of Corydalis decumbens (Thunb) Pers, possessed inhibitory action on ADP-induced blood platelet aggregation and the formation of thrombus in animal experiments. Its effects on the platelet cyclic AMP and cyclic GMP in incubated rabbit platelet were measured by radioimmunoassay. cAMP was significantly elevated when COAMTA was given 0.08-0.16 mg/ml PRP. In concentration up to 0.16 mg/ml PRP, COAMTA elevated cGMP, but the ratio value of cAMP/cGMP was much higher than that of control group. These results suggest that the anti-platelet activity of COAMTA were mediated through elevation of cAMP levels

  18. Special section containing papers presented at the 13th IAEA Technical Meeting on Energetic Particles in Magnetic Confinement Systems (Beijing, China, 17-20 September 2013) Special section containing papers presented at the 13th IAEA Technical Meeting on Energetic Particles in Magnetic Confinement Systems (Beijing, China, 17-20 September 2013)

    Lin, Z.

    2014-10-01

    In magnetic fusion plasmas, a significant fraction of the kinetic pressure is contributed by superthermal charged particles produced by auxiliary heating (fast ions and electrons) and fusion reactions (a-particles). Since these energetic particles are often far away from thermal equilibrium due to their non-Maxwellian distribution and steep pressure gradients, the free energy can excite electromagnetic instabilities to intensity levels well above the thermal fluctuations. The resultant electromagnetic turbulence could induce large transport of energetic particles, which could reduce heating efficiency, degrade overall plasma confinement, and damage fusion devices. Therefore, understanding and predicting energetic particle confinement properties are critical to the success of burning plasma experiments such as ITER since the ignition relies on plasma self-heating by a-particles. To promote international exchanges and collaborations on energetic particle physics, the biannual conference series under the auspices of the International Atomic Energy Agency (IAEA) were help in Kyiv (1989), Aspenas (1991), Trieste (1993), Princeton (1995), JET/Abingdon (1997), Naka (1999), Gothenburg (2001), San Diego (2003), Takayama (2005), Kloster Seeon (2007), Kyiv (2009), and Austin (2011). The papers in this special section were presented at the most recent meeting, the 13th IAEA Technical Meeting on Energetic Particles in Magnetic Confinement Systems, which was hosted by the Fusion Simulation Center, Peking University, Beijing, China (17-20 September 2013). The program of the meeting consisted of 71 presentations, including 13 invited talks, 26 oral contributed talks, 30 posters, and 2 summary talks, which were selected by the International Advisory Committee (IAC). The IAC members include H. Berk, L.G. Eriksson, A. Fasoli, W. Heidbrink, Ya. Kolesnichenko, Ph. Lauber, Z. Lin, R. Nazikian, S. Pinches, S. Sharapov, K. Shinohara, K. Toi, G. Vlad, and X.T. Ding. The conference program

  19. EDITORIAL: Invited papers from the 15th International Congress on Plasma Physics combined with the 13th Latin American Workshop on Plasma Physics Invited papers from the 15th International Congress on Plasma Physics combined with the 13th Latin American Workshop on Plasma Physics

    Soto, Leopoldo

    2011-07-01

    The International Advisory Committee of the 15th International Congress on Plasma Physics (ICPP 2010) and the International Advisory Committee of the 13th Latin American Workshop on Plasma Physics (LAWPP 2010) both agreed to hold this combined meeting ICPP-LAWPP-2010 in Santiago de Chile, 8-13 August 2010, considering the celebration of the Bicentennial of Chilean Independence. ICPP-LAWPP-2010 was organized by the Thermonuclear Plasma Department of the Chilean Nuclear Energy Commission (CCHEN) as part of its official program, within the framework of the Chilean Bicentennial activities. This event was also a scientific and academic activity of the project `Center for Research and Applications in Plasma Physics and Pulsed Power, P4', supported by the National Scientific and Technological Commission, CONICYT-Chile, under grant ACT-26. The International Congress on Plasma Physics was first held in Nagoya in 1980, and was followed by: Gothenburg (1982), Lausanne (1984), Kiev (1987), New Delhi (1989), Innsbruck (1992), Foz do Iguacu (1994), Nagoya (1996), Prague (1998), Quebec City (2000), Sydney (2002), Nice (2004), Kiev (2006) and Fukuoka (2008). The purpose of the Congress is to discuss recent progress and outlooks in plasma science, covering fundamental plasma physics, fusion plasmas, astrophysical plasmas, plasma applications, etc. The Latin American Workshop on Plasma Physics was first held in 1982 in Cambuquira, Brazil, followed by: Medellín (1985), Santiago (1988), Buenos Aires (1990), Mexico City (1992), Foz do Iguacu (1994, also combined with ICPP), Caracas (1997), Tandil (1998), La Serena (2000), Sao Pedro (2003), Mexico City (2005) and Caracas (2007). The purpose of the Latin American Workshop on Plasma Physics is to provide a forum in which the achievements of the Latin American plasma physics communities can be displayed, as well as to foster collaboration between plasma scientists within the region and elsewhere. The Program of ICPP-LAWPP-2010 included

  20. Receptors and cGMP signalling mechanism for E. coli enterotoxin in opossum kidney

    Receptors for the heat-stable enterotoxin produced by Escherichia coli were found in the kidney and intestine of the North American opossum and in cultured renal cell lines. The enterotoxin markedly increased guanosine 3',5'-cyclic monophosphate (cGMP) production in slices of kidney cortex and medulla, in suspensions of intestinal mucosa, and in the opossum kidney (OK) and rat kangaroo kidney (PtK-2) cell lines. In contrast, atrial natriuretic factor elicited much smaller increases in cGMP levels of kidney, intestine, or cultured kidney cell lines. The enterotoxin receptors in OK cells had a molecular mass of approximately 120 kDa when measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of receptors crosslinked with 125I-enterotoxin. The occurrence of receptors for the E. coli peptide in OK implies that these receptors may be involved in the regulation of renal tubular function in the opossum. E. coli enterotoxin caused a much larger increase in urine cGMP excretion than did atrial natriuretic factor when these peptides were injected intravenously into opossums. However, atrial natriuretic factor elicited a marked diuresis, natriuresis, and increased urinary excretion of calcium, phosphate, potassium, and magnesium. In contrast, the enterotoxin did not acutely influence OK fluid and electrolyte excretion. Thus the substantial increase in cGMP synthesis produced by the bacterial peptide in OK cortex and medulla in vitro and the increased renal excretion of cGMP in vivo were not associated with changes in electrolyte or water excretion. Whether cGMP represents a second messenger molecule in the kidney is an interesting question that was raised but not answered in this series of experiments

  1. Discrete cyclic di-GMP-dependent control of bacterial predation versus axenic growth in Bdellovibrio bacteriovorus.

    Laura Hobley

    2012-02-01

    Full Text Available Bdellovibrio bacteriovorus is a Delta-proteobacterium that oscillates between free-living growth and predation on Gram-negative bacteria including important pathogens of man, animals and plants. After entering the prey periplasm, killing the prey and replicating inside the prey bdelloplast, several motile B. bacteriovorus progeny cells emerge. The B. bacteriovorus HD100 genome encodes numerous proteins predicted to be involved in signalling via the secondary messenger cyclic di-GMP (c-di-GMP, which is known to affect bacterial lifestyle choices. We investigated the role of c-di-GMP signalling in B. bacteriovorus, focussing on the five GGDEF domain proteins that are predicted to function as diguanylyl cyclases initiating c-di-GMP signalling cascades. Inactivation of individual GGDEF domain genes resulted in remarkably distinct phenotypes. Deletion of dgcB (Bd0742 resulted in a predation impaired, obligately axenic mutant, while deletion of dgcC (Bd1434 resulted in the opposite, obligately predatory mutant. Deletion of dgcA (Bd0367 abolished gliding motility, producing bacteria capable of predatory invasion but unable to leave the exhausted prey. Complementation was achieved with wild type dgc genes, but not with GGAAF versions. Deletion of cdgA (Bd3125 substantially slowed predation; this was restored by wild type complementation. Deletion of dgcD (Bd3766 had no observable phenotype. In vitro assays showed that DgcA, DgcB, and DgcC were diguanylyl cyclases. CdgA lacks enzymatic activity but functions as a c-di-GMP receptor apparently in the DgcB pathway. Activity of DgcD was not detected. Deletion of DgcA strongly decreased the extractable c-di-GMP content of axenic Bdellovibrio cells. We show that c-di-GMP signalling pathways are essential for both the free-living and predatory lifestyles of B. bacteriovorus and that obligately predatory dgcC- can be made lacking a propensity to survive without predation of bacterial pathogens and thus possibly

  2. Cyclic Di-GMP Stimulates Biofilm Formation and Inhibits Virulence of Francisella novicida

    Zogaj, Xhavit; Wyatt, Geoff C.; Klose, Karl E.

    2012-01-01

    Francisella tularensis is a Gram-negative bacterium that is highly virulent in humans, causing the disease tularemia. F. novicida is closely related to F. tularensis and exhibits high virulence in mice, but it is avirulent in healthy humans. An F. novicida-specific gene cluster (FTN0451 to FTN0456) encodes two proteins with diguanylate cyclase (DGC) and phosphodiesterase (PDE) domains that modulate the synthesis and degradation of cyclic di-GMP (cdGMP). No DGC- or PDE-encoding protein genes a...

  3. Study on design of GMP facility for radio-pharmaceuticals production

    For production and research for practical use of radio-pharmaceuticals using for medical treatment and diagnosis, the complex facility offer shield and clean environment is basically required for protecting personnel from radiation, and protecting the product from contamination. This facility should be designed to comply with GMP(Good Manufacturing Practice). In this study, technical requirements and guide for hot cell and clean room were investigated, and design concept and requirements of facility for radio-pharmaceuticals handling are suggested. And also, GMP facility for production of Tc-99m generator and other radio-pharmaceuticals(Ho-166 etc.) was designed to comply with this design concept and requirements

  4. Biophysical Techniques for Detection of cAMP and cGMP in Living Cells

    Viacheslav O. Nikolaev

    2013-04-01

    Full Text Available Cyclic nucleotides cAMP and cGMP are ubiquitous second messengers which regulate myriads of functions in virtually all eukaryotic cells. Their intracellular effects are often mediated via discrete subcellular signaling microdomains. In this review, we will discuss state-of-the-art techniques to measure cAMP and cGMP in biological samples with a particular focus on live cell imaging approaches, which allow their detection with high temporal and spatial resolution in living cells and tissues. Finally, we will describe how these techniques can be applied to the analysis of second messenger dynamics in subcellular signaling microdomains.

  5. Polymerization of wheat gluten and the changes of glutenin macropolymer (GMP) during the production of Chinese steamed bread.

    Wang, Xiang-Yu; Guo, Xiao-Na; Zhu, Ke-Xue

    2016-06-15

    Polymerization of gluten and the changes of glutenin macropolymer (GMP) during the production of Chinese steamed bread (CSB) were investigated, providing a theoretical basis to improve and regulate the quality of CSB. Protein extractability and free sulfhydryl (SH) contents increased to some degree during the dough preparation stage, but significantly (PSDS-PAGE) patterns. The microstructure study of the gas cell and the protein network by confocal laser scanning microscopy (CLSM) further revealed the formation of a continuous and three-dimensional gluten network. The loss and recovery of GMP wet weight during dough processing were significant (P<0.05). Glutenin depolymerization negatively correlated with GMP wet weight and the contents of high molecular weight glutenin subunits (HMW-GS) and low molecular weight glutenin subunits (LMW-GS). Gluten polymerization led to a decrease in G' and G″ of GMP while gluten depolymerization induced a slight recovery in G' and G″ of GMP. PMID:26868577

  6. Studies on the interactions between the cyclic nucleotide-binding sites of cGMP-dependent protein kinase

    The rate and equilibrium kinetics of [3H]cGMP binding to the two rapidly exchanging and two slowly exchanging sites of dimeric cGMP-dependent protein kinase from bovine lung were studied. As observed by McCune and Gill, unlabeled cGMP retarded the dissociation of [3H]cGMP bound to the slow site. This effect was due to interaction of unlabeled cGMP with the rapid rather than the slow site. First, the potencies of unlabeled cGMP and a number of cGMP analogs correlated nearly perfectly with their affinities for the rapid site. Second, the rate of dissociation in the absence of unlabeled ligand was independent of the degree of saturation of the slow sites. Third, unlabeled ligand inhibited the rate of dissociation more (about 10-fold) than theoretically predicted (maximum 2-fold) from interaction between two similar sites in one macromolecule. A favorable free energy coupling appeared to exist between the rapid and slow sites but not between the slow sites. cGMP associated faster to the slow site than the rapid site. Mg/ATP decreased the rate of association to either site by 50% and increased about ten-fold the rate of dissociation from the slow site. The dissociation of cGMP from the slow site could be described by a single activation energy (Ea = 71 kJ X mol-1) for the whole temperature range (0-37 degrees C) tested. These data indicated that the cyclic nucleotide-binding sites of the cGMP-kinase are kinetically more homologous to those in the cAMP-dependent protein kinases than previously recognized

  7. 48th Annual Convention of Computer Society of India

    Avadhani, P; Udgata, Siba; Lakshminarayana, Sadasivuni; ICT and Critical Infrastructure

    2014-01-01

      This volume contains 85 papers presented at CSI 2013: 48th Annual Convention of Computer Society of India with the theme “ICT and Critical Infrastructure”. The convention was held during 13th –15th December 2013 at Hotel Novotel Varun Beach, Visakhapatnam and hosted by Computer Society of India, Vishakhapatnam Chapter in association with Vishakhapatnam Steel Plant, the flagship company of RINL, India. This volume contains papers mainly focused on Data Mining, Data Engineering and Image Processing, Software Engineering and Bio-Informatics, Network Security, Digital Forensics and Cyber Crime, Internet and Multimedia Applications and E-Governance Applications.

  8. Activated G-protein releases cGMP from high affinity binding sites on PDE from toad rod outer segments (ROS)

    cGMP binding proteins in toad ROS were identified by direct photoaffinity labeling (PAL) with 32P-cGMP and quantified by retention of complexes on nitrocellulose filters. By PAL, high affinity sites were present on the α and β subunits of the cGMP-specific phosphodiesterase (PDE) which have MW/sub app/ of 94 and 90 kDa. A doublet was deduced from its photolabeling properties to represent PDE/sub γ/ photocrosslinked with PDE/sub α/ or PDE/sub β/, respectively. cGMP prebound to these high affinity sites was released by light-activated G-protein or its α subunit complexed with GTPγS; this inhibition of cGMP binding to PDE did not result from decreased cGMP availability due to enhanced hydrolysis. A low affinity cGMP binding component identified by PAL is tightly associated with ROS membranes. Apparent ATP/light-dependent stimulation of cGMP binding was shown to result from light activated cGMP hydrolysis in conjunction with ATP-promoted conversion of GMP to GDP/GTP and increased GDP/GTP binding. These findings coincide with a model for light-related regulation of cGMP binding and metabolism predicted from intact and cellfree kinetic measurements: in the dark state the cGMP hydrolic rate is constrained by the availability of cGMP because of its binding to high affinity sites on PDE. Light activated G-protein releases cGMP from these sites and allows for its redistribution to lower affinity sites represented by PDE catalytic site(s) and possible cGMP-dependent membrane cation channels

  9. FOREWORD: 13th International Workshop on Plasma-Facing Materials and Components for Fusion Applications/1st International Conference on Fusion Energy Materials Science 13th International Workshop on Plasma-Facing Materials and Components for Fusion Applications/1st International Conference on Fusion Energy Materials Science

    Jacob, Wolfgang; Linsmeier, Christian; Rubel, Marek

    2011-12-01

    The 13th International Workshop on Plasma-Facing Materials and Components (PFMC-13) jointly organized with the 1st International Conference on Fusion Energy Materials Science (FEMaS-1) was held in Rosenheim (Germany) on 9-13 May 2011. PFMC-13 is a successor of the International Workshop on Carbon Materials for Fusion Applications series. Between 1985 and 2003 ten 'Carbon Workshops' were organized in Jülich, Stockholm and Hohenkammer. Then it was time for a change and redefinition of the scope of the symposium to reflect the new requirements of ITER and the ongoing evolution in the field. Under the new name (PFMC-11), the workshop was first organized in 2006 in Greifswald, Germany and PFMC-12 took place in Jülich in 2009. Initially starting in 1985 with about 40 participants as a 1.5 day workshop, the event has continuously grown to about 220 participants at PFMC-12. Due to the joint organization with FEMaS-1, PFMC-13 set a new record with more than 280 participants. The European project Fusion Energy Materials Science, FEMaS, coordinated by the Max-Planck-Institut für Plasmaphysik (IPP), organizes and stimulates cooperative research activities which involve large-scale research facilities as well as other top-level materials characterization laboratories. Five different fields are addressed: benchmarking experiments for radiation damage modelling, the application of micro-mechanical characterization methods, synchrotron and neutron radiation-based techniques and advanced nanoscopic analysis based on transmission electron microscopy. All these fields need to be exploited further by the fusion materials community for timely materials solutions for a DEMO reactor. In order to integrate these materials research fields, FEMaS acted as a co-organizer for the 2011 workshop and successfully introduced a number of participants from research labs and universities into the PFMC community. Plasma-facing materials experience particularly hostile conditions as they are

  10. From bedside to bench--meeting report of the 7th International Conference on cGMP "cGMP: generators, effectors and therapeutic implications" in Trier, Germany, from June 19th to 21st 2015.

    Friebe, Andreas; Sandner, Peter; Seifert, Roland

    2015-12-01

    During the past decade, our knowledge on the physiology, pathophysiology, basic pharmacology, and clinical pharmacology of the second messenger (cGMP) has increased tremendously. It is now well-established that cGMP, generated by soluble and particulate guanylate cyclases, is highly compartmentalized in cells and regulates numerous body functions. New cGMP-regulated physiological functions include meiosis and temperature perception. cGMP is involved in the genesis of numerous pathologies including cardiovascular, pulmonary, endocrine, metabolic, neuropsychiatric, eye, and tumor diseases. Several new clinical uses of stimulators and activators of soluble guanylate cyclase and of phosphodiesterase inhibitors such as heart failure, kidney failure, cognitive disorders, obesity bronchial asthma, and osteoporosis are emerging. The combination of neprilysin inhibitors-enhancing stimulation of the particulate guanylate cyclase pathway by preventing natriuretic peptide degradation-with angiotensin AT1 receptor antagonists constitutes a novel promising strategy for heart failure treatment. The role of oxidative stress in cGMP signaling, application of cGMP sensors, and gene therapy for degenerative eye diseases are emerging topics. It is anticipated that cGMP research will further prosper over the next years and reach out into more and more basic and clinical disciplines. PMID:26486926

  11. MECHANISM FOR INTRODUCTION OF GMP INTERNATIONAL STANDARDS INTO THE PHARMACEUTICAL SECTOR OF UKRAINIAN ECONOMY

    Ovcharenko, T.; Yermolayeva, V.

    2009-01-01

    In the article the modern consisting and problems of the introduce international standards of GMP in pharmaceutical sector of the national economy is analyzed. The article suggests the ways for improving the international standards system that positively influences on the medicine quality of provision in Ukraine.

  12. Opposing actions of dibutyryl cyclic AMP and GMP on temperature in conscious guinea-pigs

    Kandasamy, S. B.; Williaes, B. A.

    1983-01-01

    It is shown that the intracerebroventricular administration of dibutyryl cyclic AMP (Db-cAMP) induced hyperthermia in guinea pigs which was not mediated through prostaglandins or norepinephrine since a prostaglandin synthesis inhibitor and an alpha-adrenergic receptor blocking agent did not antagonize the hyperthermia. However, the hyperthermic response to Db-cAMP was attenuated by the central administration of a beta-adrenergic receptor antagonist, which indicates that cAMP may be involved, through beta-adrenergic receptors, in the central regulation of heat production and conservation. The central administration of Db-cGMP produced hypothermia which was not mediated via histamine H1 or H2 receptors and serotonin. The antagonism of hypothermia induced by Db-cGMP and acetylcholine + physostigmine by central administration of a cholinergic muscarine receptor antagonist and not by a cholinergic nicotinic receptor antagonist suggests that cholinoceptive neurons and endogenous cGMP may regulate heat loss through cholinergic muscarine receptors. It is concluded that these results indicate a regulatory role in thermoregulation provided by a balance between opposing actions of cAMP and cGMP in guinea pigs.

  13. Atrial natriuretic peptide receptor heterogeneity and effects on cyclic GMP accumulation

    The effects of atrial natriuretic peptide (ANP), oxytocin (OT) and vasopressin (AVP) on guanylate cyclase activity and cyclic GMP accumulation were examined, since these hormones appear to be intimately associated with blood pressure and intravascular volume homeostasis. ANP was found to increase cyclic GMP accumulation in ten cell culture systems, which were derived from blood vessels, adrenal cortex, kidney, lung, testes and mammary gland. ANP receptors were characterized in intact cultured cells using 125I-ANP8-33. Specific 125I-ANP binding was saturable and of high affinity. Scratchard analysis of the binding data for all cell types exhibited a straight line, indicating that these cells possessed a single class of binding sites. Despite the presence of linear Scatchard plots, these studies demonstrated that cultured cells possess two functionally and physically distinct ANP-binding sites. Most of the ANP-binding sites in cultured cells have a molecular size of 66,000 daltons under reducing conditions. The identification of cultured cell types in which hormones (ANP and oxytocin) regulate guanylate cyclase activity and increase cyclic GMP synthesis will provide valuable systems to determine the mechanisms of hormone-receptor coupling to guanylate cyclase and the cellular processes regulated by cyclic GMP

  14. Connection between the elastic GEp/GMp and P to Delta form factors

    Stoler, P

    2003-01-01

    It is suggested that the falloff in Qsq of the P to Delta magnetic form factor GM* is related to the recently observed falloff of the elastic electric form factor GEp/GMp. Calculation is carried out in the framework of a two-body GPD mechanism.

  15. Plasma levels of cAMP, cGMP and CGRP in sildenafil-induced headache

    Kruuse, Christina Rostrup; Frandsen, E; Schifter, S; Thomsen, L L; Birk, S; Olesen, J.

    2004-01-01

    Sildenafil, a selective inhibitor of the cyclic guanosine monophosphate (cGMP) degrading phosphodiestrase 5 (PDE5), induced migraine without aura in 10 of 12 migraine patients and in healthy subjects it induced significantly more headache than placebo. The aim of the present study was to determin...

  16. Atrial natriuretic peptide receptor heterogeneity and effects on cyclic GMP accumulation

    Leitman, D.C.

    1988-01-01

    The effects of atrial natriuretic peptide (ANP), oxytocin (OT) and vasopressin (AVP) on guanylate cyclase activity and cyclic GMP accumulation were examined, since these hormones appear to be intimately associated with blood pressure and intravascular volume homeostasis. ANP was found to increase cyclic GMP accumulation in ten cell culture systems, which were derived from blood vessels, adrenal cortex, kidney, lung, testes and mammary gland. ANP receptors were characterized in intact cultured cells using {sup 125}I-ANP{sub 8-33}. Specific {sup 125}I-ANP binding was saturable and of high affinity. Scratchard analysis of the binding data for all cell types exhibited a straight line, indicating that these cells possessed a single class of binding sites. Despite the presence of linear Scatchard plots, these studies demonstrated that cultured cells possess two functionally and physically distinct ANP-binding sites. Most of the ANP-binding sites in cultured cells have a molecular size of 66,000 daltons under reducing conditions. The identification of cultured cell types in which hormones (ANP and oxytocin) regulate guanylate cyclase activity and increase cyclic GMP synthesis will provide valuable systems to determine the mechanisms of hormone-receptor coupling to guanylate cyclase and the cellular processes regulated by cyclic GMP.

  17. A novel crosstalk between Alk7 and cGMP signaling differentially regulates brown adipocyte function

    Aileen Balkow

    2015-08-01

    Conclusions: We found a so far unknown crosstalk between cGMP and Alk7 signaling pathways. Tight regulation of Alk7 is required for efficient differentiation of brown adipocytes. Alk7 has differential effects on adipogenic differentiation and the development of the thermogenic program in brown adipocytes.

  18. PREFACE: 13th International Conference on Electrostatics

    Taylor, D. Martin

    2011-06-01

    Electrostatics 2011 was held in the city of Bangor which is located in North West Wales in an area of outstanding natural beauty close to the Snowdonia mountain range and bordering the Irish Sea. The history of the area goes back into the mists of times, but a continuous technological thread can be traced from the stone- and bronze-age craftsmen, who inhabited the area several thousand years ago, via the civil engineering and fortifications of the Romans and Edward I of England, through Marconi's long-wave trans-Atlantic transmitter near Caernarfon to the conference host. The School of Electronic Engineering at Bangor University has contributed much to the discipline of Electrostatics not only in teaching and research but also in supporting industry. It was a great pleasure for me, therefore, to have the pleasure of welcoming the world's experts in Electrostatics to Bangor in April 2011. In my preface to the Proceedings of Electrostatics 1999, I reported that almost 90 papers were presented. Interestingly, a similar number were presented in 2011 testifying to the importance and endurance of the subject. The all-embracing nature of electrostatics is captured in the pictorial depiction used for the conference logo: a hand-held plasma ball with its close link to gaseous discharges and the superimposed Antarctic aurora highlighting the featured conference themes of atmospheric, planetary and environmental electrostatics. Leading these themes were three invited contributions, the first by Giles Harrison who delivered the Bill Bright Memorial Lecture 'Fair weather atmospheric electricity', Carlos Calle on 'The electrostatic environments of Mars and the Moon' and Istvan Berta on 'Lightning protection - challenges, solutions and questionable steps in the 21st century'. Leading other key sessions were invited papers by Atsushi Ohsawa on 'Statistical analysis of fires and explosions attributed to static electricity over the last 50 years in Japanese industry' and Antonio Ramos on 'Electrohydrodynamic pumping in microsystems'. Of the papers submitted for publication 69 passed through the thorough review process and I take this opportunity to warmly thank the reviewers for their constructive criticism and rapid turnaround which has allowed the Proceedings to be delivered to the publisher on time. It is a pleasure also to thank members of the International Advisory Panel, and the Organizing and Programme Committees for their guidance and suggestions and especially Claire Garland and her team at the Institute of Physics for their support, all of which ensured a successful and enjoyable conference. Special thanks are due to Jeremy Smallwood for organising the pre-conference workshop, to Tom Jones, Martin Glor and Dave Swenson for their highly informative and educational contributions at the workshop, to CST for organising the simulation workshop, and to CST and JCI Chilworth for their much appreciated sponsorship of the conference. I am sure you will enjoy reading this record of Electrostatics 2011, covering as it does the wide range of subjects upon which static electricity impinges. Especially important is the development of electrostatic-based methods for reducing atmospheric pollution. In this context it is interesting to see how Masuda's work on the surface-discharge-based Boxer charger, first reported over 30 years ago, has now developed into dielectric barrier discharge (DBD) systems for the removal of noxious molecules from industrial and vehicle exhaust gases. Thanks to our hard working conference chairman, Paul Holdstock, the conference retained its now well-established reputation for providing a friendly, sociable atmosphere for discussing the newest developments in this important scientific area. Finally, my sincere thanks go to all the presenters and to all those who attended and contributed to another successful conference. Professor D. Martin TaylorProceedings EditorBangor, May 2011

  19. 13th Techtextil Closes with Record Figures

    2009-01-01

    @@ The technical textile and nonwovens sector came in droves to its leading international trade fair, with numbers never before seen. "Precisely because these are times of economic uncertainty, companies have been setting themselves new targets with innovative products and areas of application -and have in part already managed to achieve them at Techtextil. Some eight percent more exhibitors and two percent more trade visitors represent a double plus for Techtextil - a fantastic outcome," summarizes Detlef Braun, Member of the Board of Management of Messe Frankfurt, looking back over the days of the trade fair.

  20. 13th Radiochemical Conference. Booklet of Abstracts

    The Conference included the following sessions: (i) Opening plenary presentations (6 contributions); (ii) Chemistry of natural radionuclides, discovery of radium and polonium (6 verbal presentations + 5 poster presentations); (iii) Radionuclides in the environment, radioecology (29 + 48); (iv) Activation analysis and other radioanalytical methods (36 + 49); (v) Ionizing radiation in science and technology (12 + 12); (vi) Chemistry of actinide and trans-actinide elements (11 + 14); (vii) Separation methods, speciation (18 + 41); (viii) Production and application of radionuclides (14 + 29); and (ix) Radiochemical problems in nuclear waste management (12 + 22). The majority of verbal presentations has been input to INIS, mostly in the form of the full authors' abstracts. (P.A.)

  1. Construction Conceptions and Key Technologies about Water Resources Informatization during 13th Five-Year%全国水利信息化“十三五”建设构想与关键技术

    李臣明; 曾焱; 王慧斌; 张丽丽

    2015-01-01

    13th Five-Year is one important stage to push forward water resources modernization process and promote water safety guarantee capability. Based on major changes of informatization strategy and water control policy of China, and requirement about water resources informatization which is put forward by water deepening reform, it is necessary to anew comb and plan technology system and strategy of informatization construction combining with developmental feature of latest information technology. This paper analyzes the relationship between information developing process and capability of information ability raise, and obtains the meaning of water resources informatization construction in the current emergency of the improvement of the information capability. It also states 13th Five-Year water resources informatization construction and technical ideas whose central content is informatization resource management and service, and gives some suggestions on how to construct and implement about the important parts in detail.%“十三五”是推进水利现代化进程、提升水安全保障能力的重要时期,根据国家信息化战略和治水方略的重大调整,以及水利深化改革对水利信息化提出的要求,有必要结合最新信息技术发展特点,重新梳理和规划信息化建设中的技术体系和策略。分析信息化发展过程与信息能力提升之间的关系,明确当前水利信息化正处于以提升信息能力为核心的水利信息化建设内涵,基于信息能力视角,论述以信息化资源管理与服务为中心内容的“十三五”水利信息化建设体系和技术构想,并对建设重点和实施的关键技术进行阐述说明。

  2. "十三五"时期创新开放发展模式的思考与建议%Thoughts and Suggestions of Innovative Opening Development Mode During the 13th Five-year Plan

    周军伟

    2015-01-01

    本研究以郑州市为例,从分析郑州市对外开放现状、存在问题和面临形势着手,重点研究"一带一路"对未来对外开放新格局的影响,推动更高水平、更宽领域、更大规模对外开放,深度融入国家"一带一路"战略;如何以开放促改革、促发展,加快政府管理和监管体制创新,营造国际化、法制化的营商环境,更好地适应构建开放型经济新体制新要求,使投资更加便利、贸易更加自由、经济更加开放.因此,从创新定位理念、创新发展路径、创新体制机制等方面,提出郑州市"十三五"期间创新开放发展模式、加快对外开放工作的政策措施建议,为当前正在编制"十三五"规划的相关同类地市提供决策参考.%The research takes Zhengzhou as an example, analyzes its opening status, the existing problems and the present situation. It focuses on the impacts of"One belt, one road"on the new opening pattern in the fu?ture, which may promote the reform and opening at higher levels, wider areas, and larger scales, deeply integrate into the country's"One belt, one road"strategy. How to promote reform and development through opening, ac?celerate innovation of the management and the supervision system of the local government, and create an interna?tional and legal business environment, better adapt to the new requirements of build a new opening economic system to make the investments more convenient, the trades much free, and the economy more open. Therefore, from the aspects of the innovation-oriented concepts, paths and mechanisms, the essay proposes the mode of in?novative and opening development during the 13th five-year plan and puts forward the suggestions of speeding upopening policy measures, providing a reference to other same-level cities when they make policies of the 13th five-year plan.

  3. Pro-survival effects of 17β-estradiol on osteocytes are mediated by nitric oxide/cGMP via differential actions of cGMP-dependent protein kinases I and II.

    Marathe, Nisha; Rangaswami, Hema; Zhuang, Shunhui; Boss, Gerry R; Pilz, Renate B

    2012-01-01

    Estrogens promote bone health in part by increasing osteocyte survival, an effect that requires activation of the protein kinases Akt and ERK1/2, but the molecular mechanisms involved are only partly understood. Because estrogens increase nitric oxide (NO) synthesis and NO can have anti-apoptotic effects, we examined the role of NO/cGMP signaling in estrogen regulation of osteocyte survival. Etoposide-induced death of MLO-Y4 osteocyte-like cells, assessed by trypan blue staining, caspase-3 cleavage, and TUNEL assays, was completely prevented when cells were pre-treated with 17β-estradiol. This protective effect was mimicked when cells were pre-treated with a membrane-permeable cGMP analog and blocked by pharmacological inhibitors of NO synthase, soluble guanylate cyclase, or cGMP-dependent protein kinases (PKGs), supporting a requirement for NO/cGMP/PKG signaling downstream of 17β-estradiol. siRNA-mediated knockdown and viral reconstitution of individual PKG isoforms demonstrated that the anti-apoptotic effects of estradiol and cGMP were mediated by PKG Iα and PKG II. Akt and ERK1/2 activation by 17β-estradiol required PKG II, and cGMP mimicked the effects of estradiol on Akt and ERK, including induction of ERK nuclear translocation. cGMP induced BAD phosphorylation on several sites, and experiments with phosphorylation-deficient BAD mutants demonstrated that the anti-apoptotic effects of cGMP and 17β-estradiol required BAD phosphorylation on Ser(136) and Ser(155); these sites were targeted by Akt and PKG I, respectively, and regulate BAD interaction with Bcl-2. In conclusion, 17β-estradiol protects osteocytes against apoptosis by activating the NO/cGMP/PKG cascade; PKG II is required for estradiol-induced activation of ERK and Akt, and PKG Iα contributes to pro-survival signaling by directly phosphorylating BAD. PMID:22117068

  4. Cyclic di-GMP is essential for the survival of the lyme disease spirochete in ticks.

    Ming He

    2011-06-01

    Full Text Available Cyclic dimeric GMP (c-di-GMP is a bacterial second messenger that modulates many biological processes. Although its role in bacterial pathogenesis during mammalian infection has been documented, the role of c-di-GMP in a pathogen's life cycle within a vector host is less understood. The enzootic cycle of the Lyme disease pathogen Borrelia burgdorferi involves both a mammalian host and an Ixodes tick vector. The B. burgdorferi genome encodes a single copy of the diguanylate cyclase gene (rrp1, which is responsible for c-di-GMP synthesis. To determine the role of c-di-GMP in the life cycle of B. burgdorferi, an Rrp1-deficient B. burgdorferi strain was generated. The rrp1 mutant remains infectious in the mammalian host but cannot survive in the tick vector. Microarray analyses revealed that expression of a four-gene operon involved in glycerol transport and metabolism, bb0240-bb0243, was significantly downregulated by abrogation of Rrp1. In vitro, the rrp1 mutant is impaired in growth in the media containing glycerol as the carbon source (BSK-glycerol. To determine the contribution of the glycerol metabolic pathway to the rrp1 mutant phenotype, a glp mutant, in which the entire bb0240-bb0243 operon is not expressed, was generated. Similar to the rrp1 mutant, the glp mutant has a growth defect in BSK-glycerol medium. In vivo, the glp mutant is also infectious in mice but has reduced survival in ticks. Constitutive expression of the bb0240-bb0243 operon in the rrp1 mutant fully rescues the growth defect in BSK-glycerol medium and partially restores survival of the rrp1 mutant in ticks. Thus, c-di-GMP appears to govern a catabolic switch in B. burgdorferi and plays a vital role in the tick part of the spirochetal enzootic cycle. This work provides the first evidence that c-di-GMP is essential for a pathogen's survival in its vector host.

  5. cGMP modulates stem cells differentiation to neurons in brain in vivo.

    Gómez-Pinedo, U; Rodrigo, R; Cauli, O; Herraiz, S; Garcia-Verdugo, J-M; Pellicer, B; Pellicer, A; Felipo, V

    2010-02-17

    During brain development neural stem cells may differentiate to neurons or to other cell types. The aim of this work was to assess the role of cGMP (cyclic GMP) in the modulation of differentiation of neural stem cells to neurons or non-neuronal cells. cGMP in brain of fetuses was reduced to 46% of controls by treating pregnant rats with nitroarginine-methylester (L-NAME) and was restored by co-treatment with sildenafil.Reducing cGMP during brain development leads to reduced differentiation of stem cells to neurons and increased differentiation to non-neuronal cells. The number of neurons in the prefrontal cortex originated from stem cells proliferating on gestational day 14 was 715+/-14/mm(2) in control rats and was reduced to 440+/-29/mm(2) (61% of control) in rats treated with L-NAME. In rats exposed to L-NAME plus sildenafil, differentiation to neurons was completely normalized, reaching 683+/-11 neurons/mm(2). In rats exposed to sildenafil alone the number of cells labelled with bromodeoxyuridine (BrdU) and NeuN was 841+/-16/mm(2). In prefrontal cortex of control rats 48% of the neural stem cells proliferating in gestational day 14 differentiate to neurons, but only 24% in rats exposed to L-NAME. This was corrected by sildenafil, 40% of cells differentiate to neurons. Similar results were obtained for neurons proliferating during all developmental period. Treatment with L-NAME did not reduce the total number of cells labelled with BrdU, further supporting that L-NAME reduces selectively the differentiation of stem cells to neurons. Similar results were obtained in hippocampus. Treatment with L-NAME reduced the differentiation of neural stem cells to neurons, although the effect was milder than in prefrontal cortex. These results support that cGMP modulates the fate of neural stem cells in brain in vivo and suggest that high cGMP levels promote its differentiation to neurons while reduced cGMP levels promote differentiation to non-neuronal cells. PMID:19958812

  6. Involvement of cyclic GMP in phytochrome-controlled flowering of Pharbitis nil.

    Szmidt-Jaworska, Adriana; Jaworski, Krzysztof; Kopcewicz, Jan

    2008-05-26

    Light is one of the most important environmental factors influencing the induction of flowering in plants. Light is absorbed by specific photoreceptors--the phytochromes and cryptochromes system--which fulfil a sensory and a regulatory function in the process. The absorption of light by phytochromes initiates a cascade of related biochemical events in responsive cells, and subsequently changes plant growth and development. Induction of flowering is controlled by several paths. One is triggered by the guanosine-3':5'-cyclic monophosphate (cGMP) level. Thus, the aim of our study was to investigate the role of cGMP in phytochrome-controlled flowering. It is best to conduct such research on short-day plants because the photoperiodic reactions of only these plants are totally unequivocal. The most commonly used plant is the model short-day plant Pharbitis nil. The seedlings of P. nil were cultivated under special photoperiodic conditions: 72-h-long darkness, 24-h-long white light with low intensity and 24-h-long inductive night. Such light conditions cause a degradation of the light-labile phytochrome. Far red (FR) treatment before night causes inactivation of the remaining light-stable phytochrome. During the 24-h-long inductive darkness period, the total amount of cGMP in cotyledons underwent fluctuations, with maxima at the 4th, 8th and 14th hours. When plants were treated with FR before the long night, fluctuations were not observed. A red light pulse given after FR treatment could reverse the effect induced by FR, and the oscillation in the cGMP level was observed again. Because the intracellular level of cGMP is controlled by the opposite action of guanylyl cyclases (GCs) and phosphodiesterases (PDEs), we first tested whether accumulation of the nucleotide in P. nil tissue may be changed after treatment with a GC stimulator or PDE inhibitor. Accumulation of the nucleotide in P. nil cotyledons treated with a stimulator of cGMP synthesis (sodium nitroprusside) was

  7. Changes of platelet GMP-140 in diabetic nephropathy and its multi-factor regression analysis

    The relation of platelet GMP-140 and its related factors with diabetic nephropathy was studied. 144 patients of diabetic mellitus without nephropathy (group without DN, mean suffering duration of 25.5 +- 18.6 months); 80 with diabetic nephropathy (group DN, mean suffering duration of 58.7 +- 31.6 months) and 50 normal controls were chosen in the research. Platelet GMP-140, plasma α1-MG, β2-MG, and 24 hour urine albumin (ALB), IgG, α1-MG, β2-MG were detected by RIA, while HBA1C via chromatographic separation and FBG, PBG, Ch, TG, HDL, FG via biochemical methods. All the data had been processed with software on computer with t-test and linear regression, and multi-factor analysis were done also. The levels of platelet GMP-140, FG, DBP, TG, HBA1C and PBG in group DN were significantly higher than those of group without DN and normal control (P 0.05), while they were higher than those of normal controls. Multi-factor analysis of platelet GMP-140 with TG, DBP and HBA1C were performed in 80 patients with DN (P 1C are the independent factors enhancing the activation of platelets. The disturbance of lipid metabolism in type II diabetic mellitus may also enhance the activation of platelets. Elevation of blood pressure may accelerate the initiation and deterioration of DN in which change of platelet GMP-140 is an independent factor. Elevation of HBA1C and blood glucose are related closely to the diabetic nephropathy

  8. Intestinal GPS: bile and bicarbonate control cyclic di-GMP to provide Vibrio cholerae spatial cues within the small intestine.

    Koestler, Benjamin J; Waters, Christopher M

    2014-01-01

    The second messenger cyclic di-GMP (c-di-GMP) regulates numerous phenotypes in response to environmental stimuli to enable bacteria to transition between different lifestyles. Here we discuss our recent findings that the human pathogen Vibrio cholerae recognizes 2 host-specific signals, bile and bicarbonate, to regulate intracellular c-di-GMP. We have demonstrated that bile acids increase intracellular c-di-GMP to promote biofilm formation. We have also shown that this bile-mediated increase of intracellular c-di-GMP is negated by bicarbonate, and that this interaction is dependent on pH, suggesting that V. cholerae uses these 2 environmental cues to sense and adapt to its relative location in the small intestine. Increased intracellular c-di-GMP by bile is attributed to increased c-di-GMP synthesis by 3 diguanylate cyclases (DGCs) and decreased expression of one phosphodiesterase (PDE) in the presence of bile. The molecular mechanisms by which bile controls the activity of the 3 DGCs and the regulators of bile-mediated transcriptional repression of the PDE are not yet known. Moreover, the impact of varying concentrations of bile and bicarbonate at different locations within the small intestine and the response of V. cholerae to these cues remains unclear. The native microbiome and pharmaceuticals, such as omeprazole, can impact bile and pH within the small intestine, suggesting these are potential unappreciated factors that may alter V. cholerae pathogenesis. PMID:25621620

  9. Structural Analysis of the GGDEF-EAL Domain-Containing c-di-GMP Receptor FimX

    Navarro, M.; De, N; Bae, N; Wang, Q; Sondermann, H

    2009-01-01

    Bacterial pathogenesis involves social behavior including biofilm formation and swarming, processes that are regulated by the bacterially unique second messenger cyclic di-GMP (c-di-GMP). Diguanylate cyclases containing GGDEF and phosphodiesterases containing EAL domains have been identified as the enzymes controlling cellular c-di-GMP levels, yet less is known regarding signal transmission and the targets of c-di-GMP. FimX, a protein from Pseudomonas aeruginosa that governs twitching motility, belongs to a large subfamily containing both GGDEF and EAL domains. Biochemical and structural analyses reveals its function as a high-affinity receptor for c-di-GMP. A model for full-length FimX was generated combining solution scattering data and crystal structures of the degenerate GGDEF and EAL domains. Although FimX forms a dimer in solution via the N-terminal domains, a crystallographic EAL domain dimer suggests modes for the regulation of FimX by c-di-GMP binding. The results provide the structural basis for c-di-GMP sensing via degenerate phosphodiesterases.

  10. Inhibitory effect of dibutyryl cyclic GMP on potassium-stimulated 45Ca uptake by synaptosomes from rat brain

    The effects of dibutyryl cyclic GMP (db-cGMP) and dibutyryl cyclic AMP (db-cAMP) on potassium-stimulated 45Ca uptake by the P2 fraction of Gray and Whittaker were investigated with the following results. (1)db-cGMP inhibited initial rate of potassium-stimulated 45Ca-uptake in a dose-dependent manner in 0.1 mM Ca0 medium, but had no effect on the uptake in low K+ medium. In 0.1 mM Ca0 medium, the concentration of db-cGMP causing 50% inhibition was about 3 mM. db-cAMP (5mM) had no effect on the uptake. (2) db-cGMP inhibited potassium-stimulated 45Ca uptake in 1 mM Ca0 medium, though less than in 0.1 mM Ca0 medium. (3) db-cGMP inhibited potassium-stimulated 45Ca uptake by synaptosomes (pinched-off nerve terminals) more than the uptakes by other subfractions of P2 fraction. It is suggested from the results that cGMP inhibits the Ca influx resulting from depolarization of the nerve endings in situ. (Auth.)

  11. The participation of elevated levels of cyclic GMP in the recovery from radiation-induced mitotic delay

    The levels of cyclic AMP and cyclic GMP have been measured in Physarum plasmodia before and after treatment with gamma-radiation, 2 mM caffeine, or combinations of the two agents compared to the length of the radiation-induced mitotic delay. Caffeine alone produces a rapid transient elevation of cyclic AMP and a slower delayed elevation of cyclic GMP. Irradiation elicits an immediate transient increase in cyclic AMP and a later cyclic GMP increase which accompanies or precedes the delayed mitosis. A composite pattern is produced by combinations of radiation and caffeine, a distinctive feature of which is an elevated level of cyclic GMP near the time of the radiation-delayed and caffeine-promoted mitosis. With pretreatment by caffeine, the least radiation-induced mitotic delay occurs when plasmodia are irradiated during the caffeine-elicited increase in cyclic GMP. The plasmodium becomes refractory to the reduction of mitotic delay by caffeine at approximately the time it becomes refractory to the further elevation of cyclic GMP by caffeine. The data support a role for cyclic AMP in the onset of and for cyclic GMP in the recovery from mitotic delay induced by ionizing radiation. (author)

  12. A Minimal Threshold of c-di-GMP Is Essential for Fruiting Body Formation and Sporulation in Myxococcus xanthus.

    Skotnicka, Dorota; Smaldone, Gregory T; Petters, Tobias; Trampari, Eleftheria; Liang, Jennifer; Kaever, Volkhard; Malone, Jacob G; Singer, Mitchell; Søgaard-Andersen, Lotte

    2016-05-01

    Generally, the second messenger bis-(3'-5')-cyclic dimeric GMP (c-di-GMP) regulates the switch between motile and sessile lifestyles in bacteria. Here, we show that c-di-GMP is an essential regulator of multicellular development in the social bacterium Myxococcus xanthus. In response to starvation, M. xanthus initiates a developmental program that culminates in formation of spore-filled fruiting bodies. We show that c-di-GMP accumulates at elevated levels during development and that this increase is essential for completion of development whereas excess c-di-GMP does not interfere with development. MXAN3735 (renamed DmxB) is identified as a diguanylate cyclase that only functions during development and is responsible for this increased c-di-GMP accumulation. DmxB synthesis is induced in response to starvation, thereby restricting DmxB activity to development. DmxB is essential for development and functions downstream of the Dif chemosensory system to stimulate exopolysaccharide accumulation by inducing transcription of a subset of the genes encoding proteins involved in exopolysaccharide synthesis. The developmental defects in the dmxB mutant are non-cell autonomous and rescued by co-development with a strain proficient in exopolysaccharide synthesis, suggesting reduced exopolysaccharide accumulation as the causative defect in this mutant. The NtrC-like transcriptional regulator EpsI/Nla24, which is required for exopolysaccharide accumulation, is identified as a c-di-GMP receptor, and thus a putative target for DmxB generated c-di-GMP. Because DmxB can be-at least partially-functionally replaced by a heterologous diguanylate cyclase, these results altogether suggest a model in which a minimum threshold level of c-di-GMP is essential for the successful completion of multicellular development in M. xanthus. PMID:27214040

  13. Characterization of particulate cyclic nucleotide phosphodiesterases from bovine brain: Purification of a distinct cGMP-stimulated isoenzyme

    In the absence of detergent, ∼80-85% of the total cGMP-stimulated phosphodiesterase (PDE) activity in bovine brain was associated with washed particulate fractions; ∼85-90% of the calmodulin-sensitive PDE was soluble. Particulate cGMP-stimulated PDE was higher in cerebral cortical gray matter than in other regions. Homogenization of the brain particulate fraction in 1% Lubrol increased cGMP-stimulated activity ∼100% and calmodulin-stimulated ∼400-500%. Although 1% Lubrol readily solubilized these PDE activities, ∼75% of the cAMP PDE activity (0.5 μM [3H]cAMP) that was not affected by cGMP was not solubilized. This cAMP PDE activity was very sensitive to inhibition by Rolipram but not cilostamide. Thus, three different PDE types, i.e., cGMP stimulated, calmodulin sensitive, and Rolipram inhibited, are associated in different ways with crude bovine brain particulate fractions. The brain enzyme exhibited a slightly greater subunit Mr than did soluble forms from calf liver or bovine brain, as evidenced by protein staining or immunoblotting after polyacrylamide gel electrophoresis under denaturing conditions. Incubation of brain particulate and liver soluble cGMP-stimulated PDEs with V8 protease produced several peptides of similar size, as well as at least two distinct fragments of ∼27 kDa from the brain and ∼23 kDa from the liver enzyme. After photolabeling in the presence of [32P]cGMP and digestion with V8 protease, [32P]cGMP in each PDE was predominantly recovered with a peptide of ∼14 kDa. All of these observations are consistent with the existence of at least two discrete forms (isoenzymes) of cGMP-stimulated PDE

  14. PDE5 is converted to an activated state upon cGMP binding to the GAF A domain

    Rybalkin, Sergei D.; Rybalkina, Irina G.; Shimizu-Albergine, Masami; Tang, Xiao-Bo; Beavo, Joseph A.

    2003-01-01

    cGMP-specific, cGMP-binding phosphodiesterase (PDE5) regulates such physiological processes as smooth muscle relaxation and neuronal survival. PDE5 contains two N-terminal domains (GAF A and GAF B), but the functional roles of these domains have not been determined. Here we show that recombinant PDE5 is activated directly upon cGMP binding to the GAF A domain, and this effect does not require PDE5 phosphorylation. PDE5 exhibited time- and concentration-dependent reversible activation in respo...

  15. 13th February 2012 - German CEO Barmenia Insurance Group and Chair of the Hochschulrat Board of Governors of the Bergische Universitaet Wuppertal J. Beutelmann visiting ATLAS experimental area and signing the guest book with CERN Director-General R. Heuer and Advise R. Voss.

    Maximilien Brice

    2012-01-01

    13th February 2012 - German CEO Barmenia Insurance Group and Chair of the Hochschulrat Board of Governors of the Bergische Universitaet Wuppertal J. Beutelmann visiting ATLAS experimental area and signing the guest book with CERN Director-General R. Heuer and Advise R. Voss.

  16. GMP compliant radiosynthesis of11C and18F-labeled PET radiopharmaceuticals with a modular disposable cassette system

    Hessels-Scheper, J.G.; Maarsingh, P.; Kwizera, C.; Zijlma, R.; Maas, B.; De Vries, A.M.T.; Antunes, I.F.; Lub-de Hooge, M.N.; Boersma, H.H.; Dierckx, R.A.J.O.; De Vries, E.F.J.; Luurtsema, G.; Elsinga, P.H.

    2014-01-01

    Background Many nuclear medicine departments have an extensive radiopharmaceutical portfolio. Consequently, these multiple PET radiopharmaceuticals have to be produced with the same synthesis module. An important consideration in GMP-compliant PET production is to avoid potential cross-contamination

  17. Genetic locus (stmF) associated with cyclic GMP phosphodiesterase activity in Dictyostelium discoideum maps in linkage group II.

    Coukell, M. B.; Cameron, A M

    1985-01-01

    Previous attempts to map the stmF locus in Dictyostelium discoideum, by using only clone morphology as a marker, have led to equivocal results. Since strains carrying mutations at the stmF locus possess very low cyclic GMP phosphodiesterase activity, we have remapped this locus using both morphological and biochemical markers. Our results indicate that mutations producing a stable "streamer" phenotype and reduced cyclic GMP phosphodiesterase activity are located in linkage group II, probably ...

  18. Stress-dependent dilated cardiomyopathy in mice with cardiomyocyte-restricted inactivation of cyclic GMP-dependent protein kinase I

    Frantz, Stefan; Klaiber, Michael; Baba, Hideo A.; Oberwinkler, Heinz; Völker, Katharina; Gaßner, Birgit; Bayer, Barbara; Abeßer, Marco; Schuh, Kai; Feil, Robert; Hofmann, Franz; Kuhn, Michaela

    2016-01-01

    Aims: Cardiac hypertrophy is a common and often lethal complication of arterial hypertension. Elevation of myocyte cyclic GMP levels by local actions of endogenous atrial natriuretic peptide (ANP) and C-type natriuretic peptide (CNP) or by pharmacological inhibition of phosphodiesterase-5 was shown to counter-regulate pathological hypertrophy. It was suggested that cGMP-dependent protein kinase I (cGKI) mediates this protective effect, although the role in vivo is under debate. Here, we inves...

  19. Regulation of hippocampal cGMP levels as a candidate to treat cognitive deficits in Huntington's disease.

    Ana Saavedra

    Full Text Available Huntington's disease (HD patients and mouse models show learning and memory impairment associated with hippocampal dysfunction. The neuronal nitric oxide synthase/3',5'-cyclic guanosine monophosphate (nNOS/cGMP pathway is implicated in synaptic plasticity, and in learning and memory processes. Here, we examined the nNOS/cGMP pathway in the hippocampus of HD mice to determine whether it can be a good therapeutic target for cognitive improvement in HD. We analyzed hippocampal nNOS and phosphodiesterase (PDE 5 and 9 levels in R6/1 mice, and cGMP levels in the hippocampus of R6/1, R6/2 and Hdh(Q7/Q111 mice, and of HD patients. We also investigated whether sildenafil, a PDE5 inhibitor, could improve cognitive deficits in R6/1 mice. We found that hippocampal cGMP levels were 3-fold lower in 12-week-old R6/1 mice, when they show deficits in object recognition memory and in passive avoidance learning. Consistent with hippocampal cGMP levels, nNOS levels were down-regulated, while there were no changes in the levels of PDE5 and PDE9 in R6/1 mice. A single intraperitoneal injection of sildenafil (3 mg/Kg immediately after training increased cGMP levels, and improved memory in R6/1 mice, as assessed by using the novel object recognition and the passive avoidance test. Importantly, cGMP levels were also reduced in R6/2 mouse and human HD hippocampus. Therefore, the regulation of hippocampal cGMP levels can be a suitable treatment for cognitive impairment in HD.

  20. Identification of a cyclic-di-GMP-modulating response regulator that impacts biofilm formation in a model sulfate reducing bacterium

    AindrilaMukhopadhyay

    2014-07-01

    Full Text Available We surveyed the eight putative cyclic-di-GMP-modulating response regulators (RRs in Desulfovibrio vulgaris Hildenborough that are predicted to function via two-component signaling. Using purified proteins, we examined cyclic-di-GMP production or turnover in vitro of all eight proteins. The two RRs containing only GGDEF domains (DVU2067, DVU0636 demonstrated cyclic-di-GMP production activity in vitro. Of the remaining proteins, three RRs with HD-GYP domains (DVU0722, DVUA0086 and DVU2933 were confirmed to be Mn2+ dependent phosphodiesterases in vitro and converted cyclic-di-GMP to its linear form, pGpG. DVU0408, containing both cyclic-di-GMP production (GGDEF and degradation domains (EAL, showed cyclic-di-GMP turnover activity in vitro also with production of pGpG. No cyclic-di-GMP related activity could be assigned to the RR DVU0330, containing a metal-dependent phosphohydrolase HD-OD domain, or to the HD-GYP domain RR, DVU1181. Studies included examining the impact of overexpressed cyclic-di-GMP-modulating RRs in the heterologous host E. coli and led to the identification of one RR, DVU0636, with increased cellulose production. Evaluation of a transposon mutant in DVU0636 indicated that the strain was impaired in biofilm formation and demonstrated an altered carbohydrate:protein ratio relative to the D. vulgaris wild type biofilms. However, grown in liquid lactate/sulfate medium, the DVU0636 transposon mutant showed no growth impairment relative to the wild-type strain. Among the eight candidates, only the transposon disruption mutant in the DVU2067 RR presented a growth defect in liquid culture. Our results indicate that, of the two diguanylate cyclases that function as part of two-component signaling, DVU0636 plays an important role in biofilm formation while the function of DVU2067 has pertinence in planktonic growth.

  1. Endothelin and a Ca2+ ionophore raise cyclic GMP levels in a neuronal cell line via formation of nitric oxide.

    Reiser, G.

    1990-01-01

    1. The vasoconstrictor peptide endothelin-1 caused a fast, transient rise in guanosine 3':5'-cyclic monophosphate (cyclic GMP) levels in a neuronal cell line (mouse neuroblastoma x rat glioma hybrid cells 108CC15). The mechanism of activation of guanylate cyclase by endothelin-1 was investigated. The endothelin-1-induced rise depended on the release of internal Ca2+. 2. The stimulation of cyclic GMP synthesis induced by endothelin-1 was suppressed after preincubating the cells in medium conta...

  2. Genetic reductionist approach for dissecting individual roles of GGDEF proteins within the c-di-GMP signaling network in Salmonella

    Solano, Cristina; García, Begoña; Latasa, Cristina; Toledo-Arana, Alejandro; Zorraquino, Violeta; Valle, Jaione; Casals, Joan; Pedroso, Enrique; Lasa, Iñigo

    2009-01-01

    Bacteria have developed an exclusive signal transduction system involving multiple diguanylate cyclase and phosphodiesterase domain-containing proteins (GGDEF and EAL/HD-GYP, respectively) that modulate the levels of the same diffusible molecule, 3′-5′-cyclic diguanylic acid (c-di-GMP), to transmit signals and obtain specific cellular responses. Current knowledge about c-di-GMP signaling has been inferred mainly from the analysis of recombinant bacteria that either lack or overproduce individ...

  3. Retinal degeneration is rescued in transgenic rd mice by expression of the cGMP phosphodiesterase beta subunit.

    Lem, J.; Flannery, J. G.; Li, T; Applebury, M L; Farber, D B; Simon, M. I.

    1992-01-01

    The beta subunit of the cGMP phosphodiesterase (PDE) gene has been identified as the candidate gene for retinal degeneration in the rd mouse. To study the molecular mechanisms underlying degeneration and the potential for gene repair, we have expressed a functional bovine cGMP PDE beta subunit in transgenic rd mice. One transgenic mouse line showed complete photoreceptor rescue across the entire span of the retina. A second independently derived line showed partial rescue in which photorecept...

  4. Retinal degeneration is rescued in transgenic rd mice by expression of the cGMP phosphodiesterase ß subunit

    Lem, Janis; Flannery, John G.; Li, Tiansen; Applebury, Meredithe L.; Farber, Debora B.; Simon, Melvin I.

    1992-01-01

    The ß subunit of the cGMP phosphodiesterase (PDE) gene has been identified as the candidate gene for retinal degeneration in the rd mouse. To study the molecular mechanisms underlying degeneration and the potential for gene repair, we have expressed a functional bovine cGMP PDE ß subunit in transgenic rd mice. One transgenic mouse line showed complete photoreceptor rescue across the entire span of the retina. A second independently derived line showed partial rescue in which photoreceptors in...

  5. Clinical Grade Treg: GMP Isolation, Improvement of Purity by CD127pos Depletion, Treg Expansion, and Treg Cryopreservation

    Peters, Jorieke H; Preijers, Frank W.; Rob Woestenenk; Hilbrands, Luuk B; Koenen, Hans J.P.M.; Irma Joosten

    2008-01-01

    BACKGROUND: Treg based immunotherapy is of great interest to facilitate tolerance in autoimmunity and transplantation. For clinical trials, it is essential to have a clinical grade Treg isolation protocol in accordance with Good Manufacturing Practice (GMP) guidelines. To obtain sufficient Treg for immunotherapy, subsequent ex vivo expansion might be needed. METHODOLOGY/PRINCIPAL FINDINGS: Treg were isolated from leukapheresis products by CliniMACS based GMP isolation strategies, using anti-C...

  6. Tetrameric assembly of hGBP1 is crucial for both stimulated GMP formation and antiviral activity.

    Pandita, Esha; Rajan, Sudeepa; Rahman, Safikur; Mullick, Ranajoy; Das, Saumitra; Sau, Apurba Kumar

    2016-06-15

    Interferon-γ inducible human guanylate binding protein-1 (hGBP1) shows a unique characteristic that hydrolyses GTP to a mixture of GDP and GMP through successive cleavages, with GMP being the major product. Like other large GTPases, hGBP1 undergoes oligomerization upon substrate hydrolysis, which is essential for the stimulation of activity. It also exhibits antiviral activity against many viruses including hepatitis C. However, which oligomeric form is responsible for the stimulated activity leading to enhanced GMP formation and its influence on antiviral activity, are not properly understood. Using mutant and truncated proteins, our data indicate that transition-state-induced tetramerization is associated with higher rate of GMP formation. This is supported by chimaeras that are defective in both tetramerization and enhanced GMP formation. Unlike wild-type protein, chimaeras did not show allosteric interactions, indicating that tetramerization and enhanced GMP formation are allosterically coupled. Hence, we propose that after the cleavage of the first phosphoanhydride bond GDP·Pi-bound protein dimers transiently associate to form a tetramer that acts as an allosteric switch for higher rate of GMP formation. Biochemical and biophysical studies reveal that sequential conformational changes and interdomain communications regulate tetramer formation via dimer. Our studies also show that overexpression of the mutants, defective in tetramer formation in Rep2a cells do not inhibit proliferation of hepatitis C virus, indicating critical role of a tetramer in the antiviral activity. Thus, the present study not only highlights the importance of hGBP1 tetramer in stimulated GMP formation, but also demonstrates its role in the antiviral activity against hepatitis C virus. PMID:27071416

  7. Lanthanum-containing ternary solid solutions with the NaZn13-, ThMn12-and Th2Zn17-type crystal structures. Pt. I. Lattice dimensions and magnetic property analysis

    Phase equilibria and extensions of solid solutions based on the binary La-phases of the NaZn13, ThMn12, and Th2Zn17 structure types in several ternary systems are described. In the La-Fe-Al system, besides the ternary phases with the above given structure types, three other ternary phases of unresolved structures are obtained at compositions La5Fe6Al4, LaFe2Al10 and La2Fe2Al15. The phase LaCo13 (NaZn13) dissolves up to 5.5 atoms of Fe per formula unit in place of Co leading to a steady increase of lattice parameter. Ternary phases with unresolved crystal structures are encountered in the La-Co-Fe alloys, one near LaCo5 and another near the 2:17 stoichiometry. Likewise, LaCo13 dissolves up to 3 atoms of Al per formula unit in place of Co. However, it dissolves very little, 2Zn17-type phases, one on the Co-rich side and the other on the Mn-rich side, occur in the La-Co-Mn alloys. Addition of Fe to LaCu13 (NaZn13) possibly leads to structural distortions. LaFe9Al4 (NaZn13) is softly ferromagnetic at 77 K; its Curie temperature is around 170 K. The La(Co, X)13 solid solution phases with Fe and Al have much stronger ferro-magnetic moments even at room temperature. The ThMn12-type phase in the La-Fe-Al system and the Mn-rich Th2Zn17-type phase in the La-Co-Mn system are weakly paramagnetic at room temperature. (orig.)

  8. Improvements in the automated radioimmunoassay for cAMP or cGMP

    The work others in developing antibodies and the original radioimmunoassay for cyclic nucleotides provides the basis for these sensitive assays. The acetylation radioimmunoassay for cyclic nucleotides has enabled the measurement of cyclic AMP and cyclic GMP in very small biological samples. This is because accurate determinations can be made in samples containing less than 1 fmol of cyclic AMP or cyclic GMP. The Gamma-Flo automated radioimmunoassay system has been adapted to these assays such that cyclic nucleotides can be automatically measured at a rate of about 60 samples/hr. The Gamma-Flo instrument provides high-precision assays and eliminates human intervention in all steps of the radioimmunoassay. The automated assay has been in continuous operation in our laboratory over the last 10 years and this chapter summarizes the methodology and delineates improvements which have occurred over that time frame. Details for the preparation of the radioligands apply also to the manual acetylated radioimmunoassay for cyclic nucleotides

  9. Electrocatalytic activity of oxidation products of guanine and 5'-GMP towards the oxidation of NADH

    We have studied the potential electrocatalytic activity towards the oxidation of NADH of several oxidation products of guanine and its derivative guanosine-5'-monophosphate (5'-GMP) on pyrolytic graphite electrodes (PGE). The distribution of products generated strongly depends on the experimental conditions. Our investigations focused on the oxidation products that are adsorbed on the electrode surface, are redox active and, exhibited electrocatalytic activity toward the oxidation of NADH. These compounds were electrochemically and kinetically characterized in terms of dependence of the formal potential on pH and electron transfer rate constant (ks). The voltammetric and catalytic behavior of both guanine and 5'-GMP oxidation products was compared with that of other guanine derivatives we have previously studied. Some mechanistic aspects concerning the generation of the catalysts are also discussed

  10. Direct ionization effects of purine derivatives: ESR and ENDOR studies of 5'GMP

    In this study the free acid of 5' GMP was investigated (in the form of 5'GMP.3H/sub 2/O crystals). Irradiation and study of the molecular system below 10K gave evidence for at least three products in the ESR. Product I was also studied by ENDOR, and has been tentatively identified as the result of the anion protonated at 06. From the experimental data the spin distribution in product I is: 30% at C8; 12% at N1; 10% at N7. Product II was identified from its characteristic ESR as the result of hydrogen addition at C8. Product III was evident in the ESR but has not yet been identified

  11. Design and package of a 14CO2 field analyzer: the Global Monitor Platform (GMP)

    Marino, Bruno D. V.; Bright, Michelle; Gronniger, Glen

    2011-09-01

    Carbon Capture and Sequestration (CCS) is widely accepted as a means to reduce and eliminate the fossil fuel CO2 (ff- CO2) emissions from coal fired power plants. Success of CCS depends on near zero leakage rates over decadal time scales. Currently no commercial methods to determine leakage of ff-CO2 are available. The Global Monitor Platform (GMP) field analyzer provides high precision analysis of CO2 isotopes [12C (99%), 13C (Kyoto Protocol) and to characterize the nuclear fuel cycle. Planetary Emissions Management (PEM), working with the National Secure Manufacturing Center (NSMC), has the goal of designing, ruggedizing and packaging the GMP for field deployment. The system will conduct atmosphere monitoring then adapt to water and soil evaluations. Measuring 14CO2 in real time will provide quantitative concentration data for ff-CO2 in the atmosphere and CCS leakage detection. Initial results will be discussed along with design changes for improved detection sensitivity and manufacturability.

  12. Clinical significance of changes of plasma ADM, ET-1 and GMP-140 levels in patients with acute coronary syndrome (ACS)

    Objective: To investigate the clinical significance of changes of plasma levels of adrenomedullin (ADM), ET-1 and α-granule membrane protein (GMP-140) levels in the pathogenesis of acute coronary syndrome (ACS). Methods: Plasma ADM, ET-1 and GMP-140 levels were measured with RIA in (1) 31 cases of ACS with unstable angina pectoris (UAP) or non-ST segment elevation myocardial infarction (NSTEMI), (2) 32 cases of ACS with ST segment elevation myocardial infarction (STEMI), (3) 31 patients with stable angina pectoris (SAP) and, (4) 33 controls. Results: The plasma ADM, ET-1 and GMP-140 levels in both patient groups with UAP/NSTEMI and STEMI were significantly higher than those in controls (P0.05). However, the plasma levels of ADM, ET-1 and GMP-140 in the three groups of patients (2 groups of ACS and 1 group of SAP) were significantly different from each other (P< 0.01). Plasma levels of ADM, ET-1 and GMP-140 were mutually positively correlated. Conclusion: Changes of plasma levels of ADM, ET-1 and GMP-140 might reflect the severity of acute coronary syndrome and might serve as a marker for instability of the atheromatous plaques. (authors)

  13. Detection of plasma α-granule membrane protein (GMP-140) using 125I labeled monoclonal antibodies and its clinic applications

    Radioimmunoassay with two monoclonal antibodies to different epitopes of granule membrane protein (GMP-140) was used to measure the plasma GMP-140 level in patients suffering from acute myocardial infarction (AMI) or cerebral thrombosis and in patients during cardiopulmonary bypass (CPB). Monoclonal antibody (McAb) SZ-51 was used as a solid phase and 125I-labeled McAb S12 as a fluid phase. The assay was so sensitive that it could detect as little as 1 μg/L of purified GMP-140. The intra- and inter-assay coefficients of variation were 4.2% and 7.1% respectively. The concentration of plasma GMP-140 was found to be 10 +- 4.5 μg/L in normal subjects (n = 20). Patients undergoing CPB (n = 10) and patients with AMI (n = 16) or with acute cerebral thrombosis (n = 22) were also studied. The results suggest that plasma GMP-140 can be reliable detected by radioimmunoassay with two McAbs and plasma GMP-140 may provide a useful marker for thrombotic diseases

  14. A Cyclic di-GMP-binding Adaptor Protein Interacts with Histidine Kinase to Regulate Two-component Signaling.

    Xu, Linghui; Venkataramani, Prabhadevi; Ding, Yichen; Liu, Yang; Deng, Yinyue; Yong, Grace Lisi; Xin, Lingyi; Ye, Ruijuan; Zhang, Lianhui; Yang, Liang; Liang, Zhao-Xun

    2016-07-29

    The bacterial messenger cyclic di-GMP (c-di-GMP) binds to a diverse range of effectors to exert its biological effect. Despite the fact that free-standing PilZ proteins are by far the most prevalent c-di-GMP effectors known to date, their physiological function and mechanism of action remain largely unknown. Here we report that the free-standing PilZ protein PA2799 from the opportunistic pathogen Pseudomonas aeruginosa interacts directly with the hybrid histidine kinase SagS. We show that PA2799 (named as HapZ: histidine kinase associated PilZ) binds directly to the phosphoreceiver (REC) domain of SagS, and that the SagS-HapZ interaction is further enhanced at elevated c-di-GMP concentration. We demonstrate that binding of HapZ to SagS inhibits the phosphotransfer between SagS and the downstream protein HptB in a c-di-GMP-dependent manner. In accordance with the role of SagS as a motile-sessile switch and biofilm growth factor, we show that HapZ impacts surface attachment and biofilm formation most likely by regulating the expression of a large number of genes. The observations suggest a previously unknown mechanism whereby c-di-GMP mediates two-component signaling through a PilZ adaptor protein. PMID:27231351

  15. Pharmacological manipulation of cyclic GMP levels in brain restores learning ability in animal models of hepatic encephalopathy: therapeutic implications

    Rodrigo, Regina; Monfort, Pilar; Cauli, Omar; Erceg, Slaven; Felipo, Vicente

    2006-01-01

    Hepatic encephalopathy (HE) is a complex neuropsychiatric syndrome present in patients with liver disease that includes impaired intellectual function. To develop therapeutic treatments to restore cognitive function, it is important to understand the molecular mechanisms that impair cognitive function in HE. This review summarizes data showing that: (a) cognitive function and learning are impaired in patients with liver disease and in animal models of chronic liver failure or hyperammonemia; (b) the glutamate–NO–cGMP pathway modulates some forms of learning; and (c) the function of this pathway is impaired in brain in vivo in rats with chronic hyperammonemia or liver failure and from patients who died from HE. Learning ability of hyperammonemic rats was restored by increasing cGMP by: (1) continuous intracerebral administration of zaprinast, an inhibitor of the cGMP-degrading phosphodiesterase; (2) chronic oral administration of sildenafil, an inhibitor of the phosphodiesterase that crosses the blood–brain barrier; and (3) continuous intracerebral administration of cGMP. The data summarized indicate that impairment of learning ability in rats with chronic liver failure or hyperammonemia is due to impairment of the glutamate–NO–cGMP pathway. Moreover, increasing extracellular cGMP by pharmacological means may be a new therapeutic approach to improve cognitive function in patients with HE. PMID:19412446

  16. Purine salvage pathways of Bacillus subtilis and effect of guanine on growth of GMP reductase mutants.

    Endo, T.; Uratani, B; Freese, E.

    1983-01-01

    We have isolated numerous mutants containing mutations in the salvage pathways of purine synthesis. The mutations cause deficiencies in adenine phosphoribosyltransferase (adeF), in hypoxanthine-guanine phosphoribosyltransferase (guaF), in adenine deaminase (adeC), in inosine-guanosine phosphorylase, (guaP), and in GMP reductase (guaC). The physiological properties of mutants containing one or more of these mutations and corresponding enzyme measurements have been used to derive a metabolic ch...

  17. Importance and globalization status of good manufacturing practice (GMP) requirements for pharmaceutical excipients

    Abdellah, Abubaker; Noordin, Mohamed Ibrahim; Wan Ismail, Wan Azman

    2013-01-01

    Pharmaceutical excipients are no longer inert materials but it is effective and able to improve the characteristics of the products’ quality, stability, functionality, safety, solubility and acceptance of patients. It can interact with the active ingredients and alter the medicament characteristics. The globalization of medicines’ supply enhances the importance of globalized good manufacturing practice (GMP) requirements for pharmaceutical excipients. This review was intended to assess the gl...

  18. The specific cGMP-dependent protein kinase substrate stop protein, is phosphorylated in nerve terminals

    Full text: cGMP-dependent protein kinase (PKG) plays a key role to transducing cGMP action such as neurotransmission, learning and memory and neuronal differentiation. PKG achieves its function by phosphorylating its substrates. Despite its apparent importance, however, remarkably few specific PKG substrates in nerve terminals have been identified. We have demonstrated that there are more than 10 specific PKG substrates in nerve terminals (synaptosomes) of rat brain. Among them, we purified a 130 kDa (P130) protein and identified it as STOP (stable tubule-only polypeptide) protein by protein micro-sequencing. STOP binds to cold stable microtubules and prevents them from disassembly. STOP was phosphorylated by PKG on a serine residue in vitro but not by cAMP-dependent protein kinase or protein kinase C. The phosphorylation of STOP by PKG only occurred at lower Mg2+ concentration (1 mM) and not at 3 or 10 mM. STOP was also phosphorylated in living nerve terminals, as demonstrated by immunoprecipitation, from 32Pi-labeled synaptosomes using a synthetic peptide antibody we raised against rat brain STOP. The 32Pi-labeled STOP from synaptosomes comigrated on SDS gels with purified STOP phosphorylated by PKG in vitro. Studies of the possible effect of cGMP analogues or nitric oxide donor on STOP phosphorylation and neurotransmission are underway. STOP represents the third identified nerve terminal PKG substrates (after G-substrate and DARPP-32) and hence may impact on our understanding of the role of cGMP signalling in neuronal function. Copyright (1998) Australian Neuroscience Society

  19. The plant natriuretic peptide receptor is a guanylyl cyclase and enables cGMP-dependent signaling.

    Turek, Ilona; Gehring, Chris

    2016-06-01

    The functional homologues of vertebrate natriuretic peptides (NPs), the plant natriuretic peptides (PNPs), are a novel class of peptidic hormones that signal via guanosine 3',5'-cyclic monophosphate (cGMP) and systemically affect plant salt and water balance and responses to biotrophic plant pathogens. Although there is increasing understanding of the complex roles of PNPs in plant responses at the systems level, little is known about the underlying signaling mechanisms. Here we report isolation and identification of a novel Leucine-Rich Repeat (LRR) protein that directly interacts with A. thaliana PNP, AtPNP-A. In vitro binding studies revealed that the Arabidopsis AtPNP-A binds specifically to the LRR protein, termed AtPNP-R1, and the active region of AtPNP-A is sufficient for the interaction to occur. Importantly, the cytosolic part of the AtPNP-R1, much like in some vertebrate NP receptors, harbors a catalytic center diagnostic for guanylyl cyclases and the recombinant AtPNP-R1 is capable of catalyzing the conversion of guanosine triphosphate to cGMP. In addition, we show that AtPNP-A causes rapid increases of cGMP levels in wild type (WT) leaf tissue while this response is significantly reduced in the atpnp-r1 mutants. AtPNP-A also causes cGMP-dependent net water uptake into WT protoplasts, and hence volume increases, whereas responses of the protoplasts from the receptor mutant are impaired. Taken together, our results suggest that the identified LRR protein is an AtPNP-A receptor essential for the PNP-dependent regulation of ion and water homeostasis in plants and that PNP- and vertebrate NP-receptors and their signaling mechanisms share surprising similarities. PMID:26945740

  20. The proconvulsant effect of sildenafil in mice: role of nitric oxide–cGMP pathway

    Riazi, Kiarash; Roshanpour, Maryam; Rafiei-Tabatabaei, Neda; Homayoun, Houman; Ebrahimi, Farzad; Dehpour, Ahmad Reza

    2006-01-01

    Recent evidence indicates that sildenafil may exert some central effects through enhancement of nitric oxide (NO)-mediated effects. NO is known to have modulatory effects on seizure threshold, raising the possibility that sildenafil may alter seizure susceptibility through NO-mediated mechanisms. This study was performed to examine whether sildenafil influences the threshold of clonic and/or generalized tonic seizures through modulation of nitric oxide (NO)–cGMP pathway.The effect of sildenaf...

  1. The Optimization of GMP/GLP Laboratories: : The laboratory utilization and work safety improvement project

    Suuronen, Heli

    2011-01-01

    The pharmaceutical industry is regulated by the Good Manufacturing Practices (GMP) and Good Laboratory Practices (GLP) in order to minimize the risks that might have an impact on the safety of the patients. The purpose of the regulations is to assure, that the pharmaceutical products meet the safety requirements and have the intended product identity, quality and purity characteristics. Regulations are regulating and covering the whole manufacturing process from the used premises and starting...

  2. The implementation of tissue banking experiences for setting up a cGMP cell manufacturing facility.

    Arjmand, Babak; Emami-Razavi, Seyed Hassan; Larijani, Bagher; Norouzi-Javidan, Abbas; Aghayan, Hamid Reza

    2012-12-01

    Cell manufacturing for clinical applications is a unique form of biologics manufacturing that relies on maintenance of stringent work practices designed to ensure product consistency and prevent contamination by microorganisms or by another patient's cells. More extensive, prolonged laboratory processes involve greater risk of complications and possibly adverse events for the recipient, and so the need for control is correspondingly greater. To minimize the associate risks of cell manufacturing adhering to international quality standards is critical. Current good tissue practice (cGTP) and current good manufacturing practice (cGMP) are examples of general standards that draw a baseline for cell manufacturing facilities. In recent years, stem cell researches have found great public interest in Iran and different cell therapy projects have been started in country. In this review we described the role of our tissue banking experiences in establishing a new cGMP cell manufacturing facility. The authors concluded that, tissue banks and tissue banking experts can broaden their roles from preparing tissue grafts to manufacturing cell and tissue engineered products for translational researches and phase I clinical trials. Also they can collaborate with cell processing laboratories to develop SOPs, implement quality management system, and design cGMP facilities. PMID:21870140

  3. cGMP-Compliant Expansion of Human iPSC Cultures as Adherent Monolayers.

    Parr, Ann M; Walsh, Patrick J; Truong, Vincent; Dutton, James R

    2016-01-01

    Therapeutic uses of cells differentiated from human pluripotent stem cells (hPSCs), either embryonic stem (ES) cells or induced pluripotent stem cells (iPSCs), are now being tested in clinical trials, and it is likely that this will lead to increased commercial interest in the clinical translation of promising hPSC research. Recent technical advances in the use of defined media and culture substrates have significantly improved both the simplicity and predictability of growing hPSCs, allowing a much more straightforward application of current good manufacturing practices (cGMP) to the culture of these cells. In addition, the adoption of cGMP-compliant techniques in research environments will both improve the replication of results and make the transition of promising investigations to the commercial sector significantly less cumbersome. However, passaging methods for hPSCs are inherently unpredictable and rely on operator experience and expertise. This is problematic for the cell manufacturing process where operator time and process predictability are often determining cost drivers. We have adopted a human iPSC system using defined media and a recombinant substrate that employs cell dissociation with a hypertonic citrate solution which eliminates variability during hPSC cell expansion and provides a simple cGMP-compliant technique for hiPSC cultivation that is appropriate in both research and commercial applications. PMID:25863788

  4. A proposed impact assessment method for genetically modified plants (AS-GMP Method)

    An essential step in the development of products based on biotechnology is an assessment of their potential economic impacts and safety, including an evaluation of the potential impact of transgenic crops and practices related to their cultivation on the environment and human or animal health. The purpose of this paper is to provide an assessment method to evaluate the impact of biotechnologies that uses quantifiable parameters and allows a comparative analysis between conventional technology and technologies using GMOs. This paper introduces a method to perform an impact analysis associated with the commercial release and use of genetically modified plants, the Assessment System GMP Method. The assessment is performed through indicators that are arranged according to their dimension criterion likewise: environmental, economic, social, capability and institutional approach. To perform an accurate evaluation of the GMP specific indicators related to genetic modification are grouped in common fields: genetic insert features, GM plant features, gene flow, food/feed field, introduction of the GMP, unexpected occurrences and specific indicators. The novelty is the possibility to include specific parameters to the biotechnology under assessment. In this case by case analysis the factors of moderation and the indexes are parameterized to perform an available assessment.

  5. Simulation of an Aspheric Glass Lens Forming Behavior in Progressive GMP Process

    Recently, GMP(Glass Molding Press) process is mainly used to produce aspheric glass lenses. Because glass lens is heated at high temperature above Tg (Transformation Temperature) for forming the glass, the quality of aspheric glass lens is deteriorated by residual stresses which are generated in a aspheric glass lens after forming. In this study, as a fundamental study to develop the mold for progressive GMP process, we conducted a aspheric glass lens forming simulation. Prior to a aspheric glass lens forming simulation, compression and thermal conductivity tests were carried out to obtain mechanical and thermal properties of K-PBK40 which is newly developed material for precision molding, and flow characteristics of K-PBK40 were obtained at high temperature. Then, using the flow characteristics obtained, compression simulation was carried out and compared with the experimental result for the purpose of verifying the obtained flow characteristics. Finally, a glass lens press simulation in progressive GMP process was carried out and we could forecast the shape of deformed glass lenses and residual stresses contribution in the structure of deformed glass lenses after forming

  6. The Cross Talk between cGMP Signal Pathway and PKC in Pulmonary Endothelial Cell Angiogenesis

    Zhen Zeng

    2014-06-01

    Full Text Available Angiogenic proliferation of vascular endothelial cells is believed to play an important role in pulmonary vascular remodeling in pulmonary arterial hypertension. In the present study, we found that c-GMP (cyclic guanosine monophosphate inhibited the proliferation and tube formation of pulmonary vascular endothelial cells induced by TGF-β1, and that this process was reversed by PKG (protein kinase G inhibitor and PKC (protein kinase C inhibitor. In addition, small interfering RNA (siRNA targeting ERK also reduced cellular proliferation. Furthermore, western blotting showed that cGMP down-regulated the phosphorylation level of ERK1/2, which was reversed not only by PKG inhibitor but also by PKC inhibitor. Silencing different PKC isoforms showed that PKCΔ, PKCγ and PKCα were involved in ERK phosphorylation, suggesting that PKC kinases have a permissive action. Three subtypes, PKCΔ, PKCγ and PKCα are likely to be involved the phosphorylation suppression of ERK included cGMP. Taken together, these data suggest that ERK phosphorylation mediates the proliferation of pulmonary vascular endothelial cells, and PKC kinases have a permissive action in this process.

  7. Chloroquine-induced scratching is mediated by NO/cGMP pathway in mice.

    Foroutan, Arash; Haddadi, Nazgol Sadat; Ostadhadi, Sattar; Sistany, Narges; Dehpour, Ahmad Reza

    2015-07-01

    Chloroquine (CQ), a 4-aminoquinoline drug, has long been used in the treatment and prevention of malaria. However its side effect generalized pruritus contributes to treatment failures, and consequently results in the development of chloroquine resistant strains of Plasmodium falciparum. It was proposed that the administration of CQ correlated with increase in nitric oxide (NO) production. Nitric oxide is involved in some pruritic disorders such as atopic dermatitis, psoriasis and scratching behavior evoked by pruritogens like substance P. Therefore, the aim of this study was to investigate the involvement of NO/cGMP pathway in CQ-induced scratching in mice. Scratching behaviors were recorded by a camera after intradermal (ID) injection of CQ in the shaved rostral back of the mice. The results obtained show that CQ elicited scratching in a dose-dependent manner with a peak effective dose of 400μg/site. Injection of non-specific NOS inhibitor, N-nitro-l-arginine methyl ester or neuronal NOS selective inhibitor and 7-nitroindazole, reduced CQ-induced scratching significantly. On the other hand, administration of aminoguanidine as inducible NOS inhibitor has no inhibitory effect on this behavior. Also, injection of l-arginine as a precursor of NO significantly increased this response. Conversely, accumulation of cGMP by sildenafil as a selective phosphodiesterase type 5 inhibitor, potentiated the scratching behavior by CQ. This study therefore shows that CQ-induced scratching behavior is mediated by the NO/cGMP pathway. PMID:25957523

  8. Gibberellic acid and cGMP-dependent transcriptional regulation in arabidopsis thaliana

    Bastian, René

    2010-03-01

    An ever increasing amount of transcriptomic data and analysis tools provide novel insight into complex responses of biological systems. Given these resources we have undertaken to review aspects of transcriptional regulation in response to the plant hormone gibberellic acid (GA) and its second messenger guanosine 3\\',5\\'-cyclic monophosphate (cGMP) in Arabidopsis thaliana, both wild type and selected mutants. Evidence suggests enrichment of GA-responsive (GARE) elements in promoters of genes that are transcriptionally upregulated in response to cGMP but downregulated in a GA insensitive mutant (ga1-3). In contrast, in the genes upregulated in the mutant, no enrichment in the GARE is observed suggesting that GARE motifs are diagnostic for GA-induced and cGMP-dependent transcriptional upregulation. Further, we review how expression studies of GA-dependent transcription factors and transcriptional networks based on common promoter signatures derived from ab initio analyses can contribute to our understanding of plant responses at the systems level. © 2010 Landes Bioscience.

  9. Identification, activity and disulfide connectivity of C-di-GMP regulating proteins in Mycobacterium tuberculosis.

    Kajal Gupta

    Full Text Available C-di-GMP, a bacterial second messenger plays a key role in survival and adaptation of bacteria under different environmental conditions. The level of c-di-GMP is regulated by two opposing activities, namely diguanylate cyclase (DGC and phosphodiesterase (PDE-A exhibited by GGDEF and EAL domain, respectively in the same protein. Previously, we reported a bifunctional GGDEF-EAL domain protein, MSDGC-1 from Mycobacterium smegmatis showing both these activities (Kumar and Chatterji, 2008. In this current report, we have identified and characterized the homologous protein from Mycobacterium tuberculosis (Rv 1354c named as MtbDGC. MtbDGC is also a bifunctional protein, which can synthesize and degrade c-di-GMP in vitro. Further we expressed Mtbdgc in M. smegmatis and it was able to complement the MSDGC-1 knock out strain by restoring the long term survival of M. smegmatis. Another protein Rv 1357c, named as MtbPDE, is an EAL domain protein and degrades c-di-GMP to pGpG in vitro. Rv1354c and 1357c have seven cysteine amino acids in their sequence, distributed along the full length of the protein. Disulfide bonds play an important role in stabilizing protein structure and regulating protein function. By proteolytic digestion and mass spectrometric analysis of MtbDGC, connectivity between cysteine pairs Cys94-Cys584, Cys2-Cys479 and Cys429-Cys614 was determined, whereas the third cysteine (Cys406 from N terminal was found to be free in MtbDGC protein, which was further confirmed by alkylation with iodoacetamide labeling. Bioinformatics modeling investigations also supported the pattern of disulfide connectivity obtained by Mass spectrometric analysis. Cys406 was mutated to serine by site directed mutagenesis and the mutant MtbC406S was not found to be active and was not able to synthesize or degrade c-di-GMP. The disulfide connectivity established here would help further in understanding the structure - function relationship in MtbDGC.

  10. c-di-GMP-abhängige Signal-transduktion bei der Kontrolle der Cellulose-Synthese in Escherichia coli Biofilmen

    Richter, Anja

    2016-01-01

    c-di-GMP stellt einen wichtigen Regulator bei der Kontrolle von Motilität, Virulenz und der Biofilmbildung in Bakterien dar. Aufgrund der Vielfalt c-di-GMP-synthetisierender (Diguanylatzyklasen, DGC) bzw. abbauender (Phosphodiesterasen, PDE) Enzyme, c-di-GMP-bindender Effektoren und zellulärer Antworten etablierte sich die Theorie paralleler Regulationsmodule, die sich aus DGC, PDE, Effektor und Zielmolekül zusammensetzen und spezifische Prozesse wie die Cellulose-Synthese kontrollieren. Währ...

  11. Compartmentation and compartment-specific regulation of PDE5 by protein kinase G allows selective cGMP-mediated regulation of platelet functions

    Wilson, Lindsay S.; Elbatarny, Hisham S.; Crawley, Scott W.; Bennett, Brian M.; Maurice, Donald H.

    2008-01-01

    It is generally accepted that nitric oxide (NO) donors, such as sodium nitroprusside (SNP), or phosphodiesterase 5 (PDE5) inhibitors, including sildenafil, each impact human platelet function. Although a strong correlation exists between the actions of NO donors in platelets and their impact on cGMP, agents such as sildenafil act without increasing global intra-platelet cGMP levels. This study was undertaken to identify how PDE5 inhibitors might act without increasing cGMP. Our data identify ...

  12. The vasodilatory effect of sulfur dioxide via SGC/cGMP/PKG pathway in association with sulfhydryl-dependent dimerization.

    Yao, Qiuyu; Huang, Yaqian; Liu, Angie Dong; Zhu, Mingzhu; Liu, Jia; Yan, Hui; Zhang, Qingyou; Geng, Bin; Gao, Yuansheng; Du, Shuxu; Huang, Pan; Tang, Chaoshu; Du, Junbao; Jin, Hongfang

    2016-06-01

    The present study was designed to explore the role of soluble guanylate cyclase (sGC)/cyclic guanosine monophosphate (cGMP)/PKG pathway in sulfur dioxide (SO2)-induced vasodilation. We showed that SO2 induced a concentration-dependent relaxation of phenylephrine (PE)-precontracted rat aortic rings in association with an increase in cGMP concentration, whereas l-aspartic acid β-hydroxamate (HDX), an inhibitor of SO2 synthase, contracted rings in a dose-dependent manner. Pretreatment of aortic rings with the sGC inhibitor ODQ (30 μM) attenuated the vasodilatory effects of SO2, suggesting the involvement of cGMP pathway in SO2-induced vasodilation. Mechanistically, SO2 upregulated the protein levels of sGC and PKG dimers, while HDX inhibited it, indicating SO2 could promote cGMP synthesis through sGC activation. Furthermore, the dimerization of sGC and PKG and vasodilation induced by SO2 in precontracted rings were significantly prevented by thiol reductants dithiothreitol (DTT). In addition, SO2 reduced the activity of phosphodiesterase type 5 (PDE5), a cGMP-specific hydrolytic enzyme, implying that SO2 elevated cGMP concentration by inhibiting its hydrolysis. Hence, SO2 exerted its vasodilatory effects at least partly by promoting disulfide-dependent dimerization of sGC and PKG, resulting in an activated sGC/cGMP/PKG pathway in blood vessels. These findings revealed a new mode of action and mechanisms by which SO2 regulated the vascular tone. PMID:27009048

  13. GMP-140 binds to a glycoprotein receptor on human neutrophils: Evidence for a lectin-like interaction

    GMP-140 is a rapidly inducible receptor for neutrophils and monocytes expressed on activated platelets and endothelial cells. It is a member of the selectin family of lectin-like cell surface molecules that mediate leukocyte adhesion. We used a radioligand binding assay to characterize the interaction of purified GMP-140 with human neutrophils. Unstimulated neutrophils rapidly bound [125I]GMP-140 at 4 degrees C, reaching equilibrium in 10-15 min. Binding was Ca2+ dependent, reversible, and saturable at 3-6 nM free GMP-140 with half-maximal binding at approximately 1.5 nM. Receptor density and apparent affinity were not altered when neutrophils were stimulated with 4 beta-phorbol 12-myristate 13-acetate. Treatment of neutrophils with proteases abolished specific binding of [125I]GMP-140. Binding was also diminished when neutrophils were treated with neuraminidase from Vibrio cholerae, which cleaves alpha 2-3-, alpha 2-6-, and alpha 2-8-linked sialic acids, or from Newcastle disease virus, which cleaves only alpha 2-3- and alpha 2-8-linked sialic acids. Binding was not inhibited by an mAb to the abundant myeloid oligosaccharide, Lex (CD15), or by the neoglycoproteins Lex-BSA and sialyl-Lex-BSA. We conclude that neutrophils constitutively express a glycoprotein receptor for GMP-140, which contains sialic acid residues that are essential for function. These findings support the concept that GMP-140 interacts with leukocytes by a lectin-like mechanism

  14. Regulation of cyclic GMP metabolism in toad photoreceptors. Definition of the metabolic events subserving photoexcited and attenuated states

    Photoreceptor metabolism of cGMP and its regulation were characterized in isolated toad retinas by determining the intensity and time dependence of light-induced changes in the following metabolic parameters: cGMP hydrolytic flux determined by the rate of 18O incorporation from 18O-water into retinal guanine nucleotide alpha-phosphoryls; changes in the total concentrations of the guanine nucleotide metabolic intermediates; and changes in the concentration of metabolic GDP calculated from the fraction of the alpha-GDP that undergoes labeling with 18O. With narrow band 500 nm light that preferentially stimulates red rod photoreceptors, a range of intensities covering approximately 5 log units produced increases of over 10-fold in cGMP metabolic flux. However, the characteristics of the cGMP metabolic response over the first 2.5 log units of intensity are readily distinguishable from those at higher intensities which exhibit progressive attenuation by an intensity- and time-dependent process. Over the range of low intensities the metabolic response is characterized by 1) increases in cGMP hydrolytic flux of up to 8-fold as a logarithmic function of intensity of photic stimulation that are sustained for at least 200 s; 2) small increases or no change in the concentration of total cGMP; 3) large increases of up to 10-fold in the concentration of metabolically active GDP as a linear function of intensity with no significant change in the tissue concentrations of total GDP or GTP; and 4) amplification of the photosignal by the metabolism of approximately 10,000 molecules of cGMP per photoisomerization with the major site of amplification at the level of the interaction of bleached rhodopsin with G-protein

  15. Magnesium Lithospermate B, an Active Extract of Salvia miltiorrhiza, Mediates sGC/cGMP/PKG Translocation in Experimental Vasospasm

    Chih-Zen Chang

    2014-01-01

    Full Text Available Background. Soluble guanylyl cyclases (sGCs and Ras homolog gene family, member A (rhoA/Ras homolog gene family kinase(rho-kinase plays a role in vascular smooth muscle relaxation in subarachnoid hemorrhage (SAH. It is of interest to examine the effect of MLB on rhoA/ROCK and sGC/cGMP/PKG expression. Methods. A rodent SAH model was employed. Tissue samples were for sGCα1, sGCβ1, PKG, rhoA, ROCK (Western blot, and cGMP (ELISA measurement. Results. MLB morphologically improved convolution of the internal elastic lamina, distortion of endothelial wall, and necrosis of the smooth muscle in the SAH rats. Expressed cGMP, sGCα1, sGCβ1, and PKG in the SAH groups were reduced (P<0.01, and MLB precondition significantly induced cGMP, sGCα1, sGCβ1, and PKG. L-NAME reversed the vasodilation effect of MLB, reduced the bioexpression of PKG and cGMP (P<0.01, and tends to reduce sGCα1 level and induce rhoA, ROCK level in MLB precondition + SAH groups. Conclusion. These results demonstrate that sGC/cGMP/PKG and NO/ET pathways play pivotal roles in SAH-induced vasospasm. Through activating sGC/cGMP/PKG pathway and partially by inactivating rho-kinase in a NO-dependent mechanism, MLB shows promise to be an effective strategy for the treatment of this disease entity.

  16. 11-13世纪北欧海上贸易网络的复兴%Revival of Nordic Marine Trade Network in the 11th-13th Centuries

    刘程

    2013-01-01

    The Baltic region had created a chance for Nordic businessmen to trade because of its geographical location and natural conditions, before the German Hanseatic League formed a monopoly in the North Sea-Baltic trade in the 13th century.The Scandinavians and Frisians had opened up some trade routes to eastward and westward:the Swedes went to the Rus region and cre-ated the famous Varangian trade route, establishing the trade relationship with Byzantium and the Muslim world; Danes and Fri-sians established the trade contact with England and Netherland .After Large-scale Entrepot trade had been finished in North Eu-rope, German merchants replaced the Scandinavians and Frisians and established a frequent and stable link between the land and the sea.A stable North Sea-Baltic Trade Network was initially established .After the formation of the Hanseatic League , the trade network became one of the central areas of the European trade.%  在13世纪汉萨同盟垄断北海-波罗的海贸易之前,波罗的海地区因其便于贸易的地理位置和自然条件,为北欧商人进行早期的贸易开拓提供了可能。斯堪的纳维亚人和弗里斯兰人此时正分别向东和向西寻求海上贸易:瑞典人进入罗斯地区,开创了著名的瓦兰琴商路,建立起与拜占庭和穆斯林世界的贸易联系;丹麦人和弗里斯兰人则向西开拓至英格兰和尼德兰。大宗转运贸易方式在北欧确立后,德意志人替代了斯堪的纳维亚人和弗里斯兰人的贸易地位,建立起频繁稳定的海陆联系,并初步确立起一个稳固的北海-波罗的海贸易区。在汉萨同盟形成后,这一贸易网络成为欧洲贸易的中心区域之一。

  17. Chemical and functional properties of glycomacropeptide (GMP) and its role in the detection of cheese whey adulteration in milk: a review.

    Neelima; Sharma, Rajan; Rajput, Yudhishthir Singh; Mann, Bimlesh

    2013-01-01

    Glycomacropeptide (GMP) is a C-terminal part (f 106-169) of kappa-casein which is released in whey during cheese making by the action of chymosin. GMP being a biologically active component has gained much attention in the past decade. It also has unique chemical and functional properties. Many of the biological properties have been ascribed to the carbohydrate moieties attached to the peptide. The unique set of amino acids in GMP makes it a sought-after ingredient with nutraceutical properties. Besides its biological activity, GMP has several interesting techno-functional properties such as wide pH range solubility, emulsifying properties as well as foaming abilities which are shown to be promising for applications in food and nutrition industry. These properties of GMP have given new dimension for the profitable utilization of cheese whey to the dairy industry. A number of protocols for isolation of GMP from cheese whey have been reported. Moreover, its role in detection of sweet/rennet whey adulteration in milk and milk products has also attracted attention of various researchers, and many GMP-specific analytical methods have been proposed. This review discusses the chemico-functional properties of GMP and its role in the detection methods for checking cheese or sweet whey adulteration in milk. Recent concepts used in the isolation of GMP from cheese whey have also been discussed. PMID:23396893

  18. E88, a new cyclic-di-GMP class I riboswitch aptamer from Clostridium tetani, has a similar fold to the prototypical class I riboswitch, Vc2, but differentially binds to c-di-GMP analogs.

    Luo, Yiling; Chen, Bin; Zhou, Jie; Sintim, Herman O; Dayie, T Kwaku

    2014-03-01

    C-di-GMP has emerged as a ubiquitous second messenger, which regulates the transition between sessile and motile lifestyles and virulence factor expression in many pathogenic bacteria using both RNA riboswitches and protein effectors. We recently showed that two additional class I c-di-GMP riboswitch aptamers (Ct-E88 and Cb-17B) bind c-di-GMP with nanomolar affinity, and that Ct-E88 RNA binds 2'-F-c-di-GMP 422 times less tightly than class I Vc2 RNA. Based on sequence comparison, it was concluded that the global folds of Ct-E88 and Vc2 RNAs were similar and that differences in ligand binding were probably due to differences in binding site architectures. Herein, we utilized EMSA, aptamer sensing spinach modules, SAXS and 1D NMR titration to study the conformational transitions of Ct-E88. We conclude that whereas the global folds of the bound states of Vc2 and Ct-E88 RNAs are similar, the unbound states are different and this could explain differences in ligand affinities between these class I c-di-GMP riboswitches. PMID:24430255

  19. Ristripatsid Eesti 12.–13. sajandi laibakalmistutes: kas ehted või usu tunnused? / The cross-shaped pendants of inhumation burials in 12th-13th century Estonia: adornments or signs of belief?

    Tuuli Kurisoo

    2012-01-01

    Full Text Available Cross-shaped pendants are one of the most discussed symbolically laden artefacts among Estonian archaeological material. The beginning of the distribution of cross-shaped pendants dates back to 11th century when the artefacts, though scarce, appear. The number of such pendants increases noticeably at the very end of the Iron Age. Due to the ambiguous entity of cross and time context, cross-pendants have been interpreted either as symbols of Christian faith, as pieces of adornment not affiliated with Christianity, or as objects with magical qualities. In this paper I study pendants found in the inhumation cemeteries of Estonia. Only those provide us with an archaeological context which shows precisely who could have worn them and how. The analyzed cross-pendants were found at Pada, Kaberla, Kukruse, Haimre and Tammiku cemeteries and from the early burials in Viru-Nigula churchyard. As artefacts, the cross pendants are more or less stylized Greek crosses. In most cases the pendants were either used as a part of jewellery or worn singularly around the neck area. From the total of 48 analyzed cross-pendants one fourth could be interpreted as locally produced items. Of local origin are probably lead pendants from Pada cemetery, heads of decorative pins worn as cross-shaped pendants from Viru-Nigula and Kaberla cemeteries, and obvious scrap and unfinished pendant crosses. In addition, I would like to draw attention that cross-pendants from inhumation burials do not contain many widespread forms, such as ‘Scandinavian’ and some other widespread Russian origin types, which have been found in hoards, from hill-forts and cremation burials. This observation could confirm that cross pendants from 12th–13th century inhumation burials were communally recognized and some of them were locally manufactured. As the next stage of my investigation I studied different occurrence contexts of cross pendants in cemeteries and burials: the location of burials with

  20. A c-di-GMP effector system controls cell adhesion by inside-out signaling and surface protein cleavage.

    Peter D Newell

    Full Text Available In Pseudomonas fluorescens Pf0-1 the availability of inorganic phosphate (Pi is an environmental signal that controls biofilm formation through a cyclic dimeric GMP (c-di-GMP signaling pathway. In low Pi conditions, a c-di-GMP phosphodiesterase (PDE RapA is expressed, depleting cellular c-di-GMP and causing the loss of a critical outer-membrane adhesin LapA from the cell surface. This response involves an inner membrane protein LapD, which binds c-di-GMP in the cytoplasm and exerts a periplasmic output promoting LapA maintenance on the cell surface. Here we report how LapD differentially controls maintenance and release of LapA: c-di-GMP binding to LapD promotes interaction with and inhibition of the periplasmic protease LapG, which targets the N-terminus of LapA. We identify conserved amino acids in LapA required for cleavage by LapG. Mutating these residues in chromosomal lapA inhibits LapG activity in vivo, leading to retention of the adhesin on the cell surface. Mutations with defined effects on LapD's ability to control LapA localization in vivo show concomitant effects on c-di-GMP-dependent LapG inhibition in vitro. To establish the physiological importance of the LapD-LapG effector system, we track cell attachment and LapA protein localization during Pi starvation. Under this condition, the LapA adhesin is released from the surface of cells and biofilms detach from the substratum. This response requires c-di-GMP depletion by RapA, signaling through LapD, and proteolytic cleavage of LapA by LapG. These data, in combination with the companion study by Navarro et al. presenting a structural analysis of LapD's signaling mechanism, give a detailed description of a complete c-di-GMP control circuit--from environmental signal to molecular output. They describe a novel paradigm in bacterial signal transduction: regulation of a periplasmic enzyme by an inner membrane signaling protein that binds a cytoplasmic second messenger.

  1. Differential regulation of c-di-GMP metabolic enzymes by environmental signals modulates biofilm formation in Yersinia pestis

    Gai-Xian eRen

    2016-06-01

    Full Text Available Cyclic diguanylate (c-di-GMP is essential for Yersinia pestis biofilm formation, which is important for flea-borne blockage-dependent plague transmission. Two diguanylate cyclases (DGCs, HmsT and HmsD and one phosphodiesterase (PDE, HmsP are responsible for the synthesis and degradation of c-di-GMP in Y. pestis. Here, we systematically analyzed the effect of various environmental signals on regulation of the biofilm phenotype, the c-di-GMP levels, and expression of HmsT, HmsD and HmsP in Y. pestis. Biofilm formation was higher in the presence of nonlethal high concentration of CaCl2, MgCl2, CuSO4, sucrose, sodium dodecyl sulfonate, or dithiothreitol, and was lower in the presence of FeCl2 or NaCl. In addition, we found that HmsD plays a major role in biofilm formation in acidic or redox environments. These environmental signals differentially regulated expression of HmsT, HmsP and HmsD, resulting in changes in the intracellular levels of c-di-GMP in Y. pestis. Our results suggest that bacteria can sense various environmental signals, and differentially regulates their DGCs and PDEs to coordinately regulate and adapt metabolism of c-di-GMP and biofilm formation to changing environments.

  2. Measurement of the Ratio GEP/GMP to Large Q2: Interesting New Results from Jefferson Lab

    In two experiments at Jefferson Lab the ratio of the electromagnetic form factors of the proton, GEp=GMp, was obtained in the four-momentum transfer squared range of 0.5 to 5.6 GeV2 with a polarization transfer technique, by measuring Pt and Pl, the transverse and longitudinal recoil proton polarization in ep --> e'p reaction. In one-photon approximation GEp=GMp is proportional to Pt=Pl; the great strength of this method is the tight control of systematics it affords. The ratio mu GEp=GMp was found to decrease linearly with Q2, down to 0.29 at Q2=5.6 GeV2; revealing a definite difference between the spatial distributions of charge and magnetization at short distances. The combined results of the two JLab experiments were surprising as they appeared to contradict the consensus based on Rosenbluth separation results for (GEp)2 and (GMp)2: the ratio mu pGEp=GMp obtained with the Rosenbluth method is ∼1 up to 5 GeV2. This unbridgeable difference between cross section and polarization experiments has been reinforced with two recent JLab Rosenbluth experiments; it appears increasingly difficult to explain it away by methodological or systematic errors. We are currently preparing a third experiment at JLab, this time in Hall C, to extend the Q2-range to 9 GeV2

  3. The effect of a nitric oxide donor on the synthesis of cGMP in Hymenolepis diminuta: a radiometric study.

    Onufriev, Mikhail V; Gulyaeva, Natalia V; Terenina, Nadezhda B; Tolstenkov, Oleg O; Gustafsson, Margaretha K S

    2005-01-01

    The formation of cGMP in homogenates of the adult rat-tapeworm Hymenolepis diminuta was followed with a radiometric assay during 3 h after stimulation with the nitric oxide donor sodium nitroprusside (SNP) and in the presence of isobutylmethylxanthine (IBMX). The level of cGMP was stable in worms incubated with IBMX during the first hour. After 3 h of incubation, the level of cGMP had declined by 27%. Addition of SNP stimulated the formation of cGMP during the first hour of incubation. After 3 h of incubation, a two-fold decline in cGMP formation was observed. The rate of nitric oxide (NO) release by the worm was determined by a spectrophotometric assay for the accumulation of nitrites and nitrates, the stable degradation products of NO, using the Griess reaction. The results are discussed from the perspective of the current concept on the role of the nitrergic mechanisms in the flatworm nervous system. PMID:15614585

  4. 浅析医疗器械 GMP 对质量管理体系资源管理的要求%Opinion on the GMP Requirements on Resource Management within QMS of Medical Device Enterprises

    田少雷

    2013-01-01

      对医疗器械生产质量管理体系中资源管理的重要内容以及GMP的相关要求进行了探讨,并介绍了我国医疗器械生产企业在资源管理方面的常见问题,以供相关企业在建立、完善生产质量管理体系和实施医疗器械GMP时参考。%The important issues and relative requirements of GMP on the management of resources within quality management system (QMS) of medical device enterprises are discussed with the aim of providing reference for relative enterprises to build and improve their QMS and to implement GMP. Otherwise, those typical found problems in GMP inspections are also summarized.

  5. Opinion on the GMP Requirements on Resource Management within QMS of Medical Device Enterprises%浅析医疗器械 GMP 对质量管理体系资源管理的要求

    田少雷

    2013-01-01

    The important issues and relative requirements of GMP on the management of resources within quality management system (QMS) of medical device enterprises are discussed with the aim of providing reference for relative enterprises to build and improve their QMS and to implement GMP. Otherwise, those typical found problems in GMP inspections are also summarized.%  对医疗器械生产质量管理体系中资源管理的重要内容以及GMP的相关要求进行了探讨,并介绍了我国医疗器械生产企业在资源管理方面的常见问题,以供相关企业在建立、完善生产质量管理体系和实施医疗器械GMP时参考。

  6. Thrombin has biphasic effects on the nitric oxide-cGMP pathway in endothelial cells and contributes to experimental pulmonary hypertension.

    Katrin F Nickel

    Full Text Available BACKGROUND: A potential role for coagulation factors in pulmonary arterial hypertension has been recently described, but the mechanism of action is currently not known. Here, we investigated the interactions between thrombin and the nitric oxide-cGMP pathway in pulmonary endothelial cells and experimental pulmonary hypertension. PRINCIPAL FINDINGS: Chronic treatment with the selective thrombin inhibitor melagatran (0.9 mg/kg daily via implanted minipumps reduced right ventricular hypertrophy in the rat monocrotaline model of experimental pulmonary hypertension. In vitro, thrombin was found to have biphasic effects on key regulators of the nitric oxide-cGMP pathway in endothelial cells (HUVECs. Acute thrombin stimulation led to increased expression of the cGMP-elevating factors endothelial nitric oxide synthase (eNOS and soluble guanylate cyclase (sGC subunits, leading to increased cGMP levels. By contrast, prolonged exposition of pulmonary endothelial cells to thrombin revealed a characteristic pattern of differential expression of the key regulators of the nitric oxide-cGMP pathway, in which specifically the factors contributing to cGMP elevation (eNOS and sGC were reduced and the cGMP-hydrolyzing PDE5 was elevated (qPCR and Western blot. In line with the differential expression of key regulators of the nitric oxide-cGMP pathway, a reduction of cGMP by prolonged thrombin stimulation was found. The effects of prolonged thrombin exposure were confirmed in endothelial cells of pulmonary origin (HPAECs and HPMECs. Similar effects could be induced by activation of protease-activated receptor-1 (PAR-1. CONCLUSION: These findings suggest a link between thrombin generation and cGMP depletion in lung endothelial cells through negative regulation of the nitric oxide-cGMP pathway, possibly mediated via PAR-1, which could be of relevance in pulmonary arterial hypertension.

  7. Analysis Application of GMP, SSOP, GTP And The Potential For Business Development

    Bhiaztika Ristyanadi

    2014-12-01

    Full Text Available Establishing crab meat canning industry which has several units production scattered in Madura region. A unit production is assigned to provide canning raw material requirements such as strip crab meat. This study aims to analyze problems of the company to know defense strategy and how to increase productivity. Further more, evaluating quality management system activities aims to improve the system and product  quality to conform to the wishes of consumers. Improvement of product quality conducted an assessment of the quality management system which  applied to unit production using the scoring method. This assessment aims to reduce the risk of production failure,  with prevent infringement of production processes. Assessment of the production aspects including GMP, GTP and SSOP. Then to identify problems and analyze what strategies will be used can used a SWOT analysis. Analysis strategy using SWOT method aims to determine what factors can affect company productivity. Observation result of GMP overall implementation on unit process did not implemented properly. Average total percentage earned was only 50% which means GMP implementation didnt meet eligibility standarts. While the GTP application only obtained between 25% to 50%. That percentage result indicates that the implementation of the GTP did meet the standards. It is also found in the SSOP observation which indicates average percentage result was only 25% to 50%. While the results of the SWOT analysis namely the company improvement strategy which is used to improve productivity. It can be done by increasing the number of partnerships, increasing supply of raw materials and improving product quality. Keywords: Quality Management System, Quality, SWOT

  8. Purine salvage pathways of Bacillus subtilis and effect of guanine on growth of GMP reductase mutants.

    Endo, T; Uratani, B; Freese, E

    1983-07-01

    We have isolated numerous mutants containing mutations in the salvage pathways of purine synthesis. The mutations cause deficiencies in adenine phosphoribosyltransferase (adeF), in hypoxanthine-guanine phosphoribosyltransferase (guaF), in adenine deaminase (adeC), in inosine-guanosine phosphorylase, (guaP), and in GMP reductase (guaC). The physiological properties of mutants containing one or more of these mutations and corresponding enzyme measurements have been used to derive a metabolic chart of the purine salvage pathway of Bacillus subtilis. PMID:6408059

  9. Role of NO-cGMP signalling pathway in mediation of ischemia-reperfusion lung injury

    Egemnazarov, Bakytbek

    2008-01-01

    Ischemia reperfusion (I/R) lung injury is a complex pathological process, which occurs in many clinical situations. Previous studies reported equivocal results about the role of nitric oxide (NO) synthase isoforms in the mediation of the injury and their possible mechanisms of action. Therefore, the aim of our study was to evaluate the role of NO synthase isoforms and NO-cGMP signaling pathway on I/R injury of the lung in an isolated perfused organ model employing rabbits as well as wild type...

  10. Automated system for separation and distribution of 123I according to the GMP requirements

    The radioisotope of 123I is routinely used in nuclear medicine, especially for SPECT imaging. The chemical separation of the produced radioisotope from the irradiated 123Te target is based on the volatile behaviour of the iodine in high temperature. However, in this case, high attention and perfect operations are required from the staff. Therefore we have developed a PC-controlled automated system for dry distillation of 123I from the irradiated target material and for its dispensation to pre-decided users. The main goal was, besides avoiding the error from human handling, to fulfill the regulations of the Good Manufacturing Practice (GMP) and the personal dosimetric aspect

  11. Measurements of the elastic electromagnetic form factor ratio μpGEp/GMp via polarization transfer

    We present measurements of the ratio of the proton elastic electromagnetic form factors, μpGEp/GMp. The Jefferson Lab Hall A Focal Plane Polarimeter was used to determine the longitudinal and transverse components of the recoil proton polarization in ep elastic scattering; the ratio of these polarization components is proportional to the ratio of the two form factors. These data reproduce the observation of Jones et al. [Phys. Rev. Lett. 84, 1398 (2000)], that the form factor ratio decreases significantly from unity above Q2 = 1 GeV2

  12. Complex regulatory network encompassing the Csr, c-di-GMP and motility systems of Salmonella Typhimurium

    Jonas, Kristina; Edwards, Adrianne N.; Ahmad, Irfan; Romeo, Tony; Römling, Ute; Melefors, Öjar

    2009-01-01

    Bacterial survival depends on the ability to switch between sessile and motile life styles in response to changing environmental conditions. In many species, this switch is governed by (3′-5′)-cyclic-diguanosine monophosphate (c-di-GMP) a signalling molecule, which is metabolized by proteins containing GGDEF and/or EAL domains. Salmonella Typhimurium contains 20 such proteins. Here, we show that the RNA-binding protein CsrA regulates the expression of eight genes encoding GGDEF, GGDEF-EAL and...

  13. High Glucose Inhibits the Aspirin-Induced Activation of the Nitric Oxide/cGMP/cGMP-Dependent Protein Kinase Pathway and Does Not Affect the Aspirin-Induced Inhibition of Thromboxane Synthesis in Human Platelets

    Russo, Isabella; Viretto, Michela; Barale, Cristina; Mattiello, Luigi; Doronzo, Gabriella; Pagliarino, Andrea; Cavalot, Franco; Trovati, Mariella; Anfossi, Giovanni

    2012-01-01

    Since hyperglycemia is involved in the “aspirin resistance” occurring in diabetes, we aimed at evaluating whether high glucose interferes with the aspirin-induced inhibition of thromboxane synthesis and/or activation of the nitric oxide (NO)/cGMP/cGMP-dependent protein kinase (PKG) pathway in platelets. For this purpose, in platelets from 60 healthy volunteers incubated for 60 min with 5–25 mmol/L d-glucose or iso-osmolar mannitol, we evaluated the influence of a 30-min incubation with lysine...

  14. Clear cutting (10-13th century) and deep stable economy (18-19th century) as responsible interventions for sand drifting and plaggic deposition in cultural landscapes on aeolian sands (SE-Netherlands).

    van Mourik, Jan; Vera, Hein; Wallinga, Jakob

    2013-04-01

    The landscape in extensive areas in SE-Netherlands is underlain by coversand, deposited during the Late Glacial of the Weichselian. In the Preboreal, aeolian processes reduced soil formation. From the Preboreal to the Atlantic a deciduous climax forest developed. The geomorphology was a coversand landscape, composed of ridges (umbric podzols), coversand plains (gleyic podzols), coversand depressions (histic podzols) and small valleys (gleysols). The area was used by hunting people during the Late Paleolithic and Mesolithic. During the Bronze and Iron Ages the area was populated by people, living from forest grazing, shifting cultivation and trade. The natural deciduous forest gradually degraded into Calluna heath. The deforestation accelerated the soil acidification and affected the hydrology, which is reflected in drying out of ridges and wetting of depressions, promoting the development of histic podzols and even histosols. Aeolian erosion was during this period restricted to local, small scale sand drifting, related to natural hazards as forest fires and hurricanes and shifting cultivation. Sustainable crop productivity on chemically poor sandy substrates required application of organic fertilizers, composed of a mixture of organic litter and animal manure with a very low mineral compound, produced in shallow stables. At least since 1000 AD, heath management was regulated by a series of rules that aimed to protect the valuable heat lands against degradation. During the 11th, 12th and 13th centuries there was an increasing demand for wood and clear cutting transformed the majority of the forests in driftsand landscapes. The most important market was formed by the very wealthy Flemish cities. The exposed soil surface was subjected to wind erosion and sand drifting which endangered the Calluna heath, arable land and even farmhouses. As a consequence, umbric podzols, the natural climax soil under deciduous forests on coversand, degraded into larger scale driftsand

  15. A cyclic GMP-dependent calcium-activated chloride current in smooth-muscle cells from rat mesenteric resistance arteries

    Matchkov, Vladimir; Aalkjær, Christian; Nilsson, Holger

    2004-01-01

    We have previously demonstrated the presence of a cyclic GMP (cGMP)-dependent calcium-activated inward current in vascular smooth-muscle cells, and suggested this to be of importance in synchronizing smooth-muscle contraction. Here we demonstrate the characteristics of this current. Using...... conventional patch-clamp technique, whole-cell currents were evoked in freshly isolated smooth-muscle cells from rat mesenteric resistance arteries by elevation of intracellular calcium with either 10 mM caffeine, 1 microM BAY K8644, 0.4 microM ionomycin, or by high calcium concentration (900 nM) in the....... Under conditions of high calcium in the patch-pipette solution, a current similar to the latter could be identified also in the mesenteric artery smooth-muscle cells. We conclude that smooth-muscle cells from rat mesenteric resistance arteries have a novel cGMP-dependent calcium-activated chloride...

  16. The rhodopsin-guanylyl cyclase of the aquatic fungus Blastocladiella emersonii enables fast optical control of cGMP signaling.

    Scheib, Ulrike; Stehfest, Katja; Gee, Christine E; Körschen, Heinz G; Fudim, Roman; Oertner, Thomas G; Hegemann, Peter

    2015-08-11

    Blastocladiomycota fungi form motile zoospores that are guided by sensory photoreceptors to areas of optimal light conditions. We showed that the microbial rhodopsin of Blastocladiella emersonii is a rhodopsin-guanylyl cyclase (RhGC), a member of a previously uncharacterized rhodopsin class of light-activated enzymes that generate the second messenger cyclic guanosine monophosphate (cGMP). Upon application of a short light flash, recombinant RhGC converted within 8 ms into a signaling state with blue-shifted absorption from which the dark state recovered within 100 ms. When expressed in Xenopus oocytes, Chinese hamster ovary cells, or mammalian neurons, RhGC generated cGMP in response to green light in a light dose-dependent manner on a subsecond time scale. Thus, we propose RhGC as a versatile tool for the optogenetic analysis of cGMP-dependent signaling processes in cell biology and the neurosciences. PMID:26268609

  17. Vibrio cholerae VpsT Regulates Matrix Production and Motility by Directly Sensing Cyclic di-GMP

    Krasteva, P.; Fong, J; Shikuma, N; Beyhan, S; Navarro, M; Yildiz, F; Sondermann, H

    2010-01-01

    Microorganisms can switch from a planktonic, free-swimming life-style to a sessile, colonial state, called a biofilm, which confers resistance to environmental stress. Conversion between the motile and biofilm life-styles has been attributed to increased levels of the prokaryotic second messenger cyclic di-guanosine monophosphate (c-di-GMP), yet the signaling mechanisms mediating such a global switch are poorly understood. Here we show that the transcriptional regulator VpsT from Vibrio cholerae directly senses c-di-GMP to inversely control extracellular matrix production and motility, which identifies VpsT as a master regulator for biofilm formation. Rather than being regulated by phosphorylation, VpsT undergoes a change in oligomerization on c-di-GMP binding.

  18. Role of Ser102 and Ser104 as Regulators of cGMP Hydrolysis by PDE5A

    Carøe Nordgaard, Julie; Kruse, Lars Schack; Gammeltoft, Steen; Kruuse, Christina

    2014-01-01

    BACKGROUND: Phosphodiesterases (PDEs) cleave phosphodiester bonds in cyclic nucleotides and play diverse roles in cell biology. PDE5A is a cytoplasmic phosphodiesterase which specifically degrades cyclic guanosine monophosphate (cGMP), a cell signaling molecule that plays important roles in...... quantified in cell extracts by standard in vitro assay using [3H] cGMP. RESULTS: The intracellular distribution of wild-type, single and double mutant PDE5A was similar and was not altered by exposure to sildenafil, cilostazol, glyceryl trinitrate, calcitonin gene-related peptide (CGRP) or sumatriptan. PDE5...... doublet. INTERPRETATION: Ser102 and Ser104 may influence the conformational flexibility of PDE5A, which may in turn influence phosphorylation status, allosteric regulation by cGMP or other as yet unknown regulatory mechanisms for PDE5A. PDE5A activation could be important in reversal of migraine...

  19. Hypergravity differentially modulates cGMP efflux in human melanocytic cells stimulated by nitric oxide and natriuretic peptides

    Ivanova, K.; Stieber, C.; Lambers, B.; Block, I.; Krieg, R.; Wellmann, A.; Gerzer, R.

    Nitric oxide NO plays a key role in many patho physiologic processes including inflammation and skin cancer The diverse cellular effects of NO are mainly mediated by activation of the soluble guanylyl cyclase sGC isoform that leads to increases in intracellular cGMP levels whereas the membrane-bound isoforms serve as receptors for natriuretic peptides e g ANP In human skin epidermal melanocytes represent the principal cells for skin pigmentation by synthesizing the pigment melanin Melanin acts as a scavenger for free radicals that may arise during metabolic stress as a result of potentially harmful effects of the environment In previous studies we found that long-term exposure to hypergravity stimulated cGMP efflux in normal human melanocytes NHMs and non-metastatic melanoma cells at least partly by an enhanced expression of the multidrug resistance proteins MRP and cGMP transporters MRP4 5 The present study investigated whether hypergravity generated by centrifugal acceleration may modulate the cGMP efflux in NO-stimulated NHMs and melanoma cells MCs with different metastatic potential The NONOates PAPA-NO and DETA-NO were used as direct NO donors for cell stimulation In the presence of 0 1 mM DETA-NO t 1 2 sim 20 h long-term application of hypergravity up to 5 g for 24 h reduced intracellular cGMP levels by stimulating cGMP efflux in NHMs and non-metastatic MCs in comparison to 1 g whereas exposure to 5 g for 6 h in the presence of 0 1 mM PAPA-NO t 1 2 sim 30 min was not effective The hypergravity-stimulated

  20. The Cyclic AMP-Vfr Signaling Pathway in Pseudomonas aeruginosa Is Inhibited by Cyclic Di-GMP

    Almblad, Henrik; Harrison, Joe J; Rybtke, Morten;

    2015-01-01

    a direct result of elevated c-di-GMP content. Overproduction of c-di-GMP causes a decrease in the transcription of virulence factor genes that are regulated by the global virulence regulator Vfr. The low level of Vfr-dependent transcription is caused by a low level of its coactivator, cyclic AMP (c...

  1. Clearance of Pseudomonas aeruginosa Foreign-Body Biofilm Infections through Reduction of the Cyclic Di-GMP Level in the Bacteria

    Christensen, Louise D.; van Gennip, Maria; Rybtke, Morten Theil; Wu, Hong; Chiang, Wen-Chi; Alhede, Morten; Høiby, Niels; Nielsen, Thomas Eiland; Givskov, Michael; Tolker-Nielsen, Tim

    2013-01-01

    be used for biofilm control in vivo. We constructed a Pseudomonas aeruginosa strain in which a reduction in the c-di-GMP level can be achieved via induction of the Escherichia coli YhjH c-di-GMP phosphodiesterase. Initial experiments showed that induction of yhjH expression led to dispersal of the...

  2. The CRP/FNR family protein Bcam1349 is a c-di-GMP effector that regulates biofilm formation in the respiratory pathogen Burkholderia cenocepacia

    Fazli, Mustafa; O'Connell, Aileen; Nilsson, Martin; Niehaus, Karsten; Dow, J Maxwell; Givskov, Michael; Ryan, Robert P; Tolker-Nielsen, Tim

    2011-01-01

    defect with physiological c-di-GMP levels. The bcam1349 gene is predicted to encode a transcriptional regulator of the CRP/FNR superfamily. Analyses of purified Bcam1349 protein and truncations demonstrated that it binds c-di-GMP in vitro. The Bcam1349 protein was shown to regulate the production of a...

  3. Distribution of cyclic-GMP-dependent protein kinase in various rat tissues and cell lines determined by a sensitive and specific radioimmunoassay

    Cyclic-GMP-dependent protein kinase purified from bovine lung was radioiodinated by the Bolton-Hunter procedure yielding a specific radioactivity of 2200 Gi/mmol of enzyme. Using a specific precipitating rabbit antiserum to the cyclic-GMP-dependent protein kinase, a sensitive radioimmunoassay was developed which can detect 200 pg (1.33 fmol) of cyclic-GMP-dependent protein kinase. Immunoreactivity like that of cyclic-GMP-dependent protein kinase was detectable in extracts of all rat tissues tested, in extracts of cultured rat brain and heart cells, and in extracts of rat glioma (C6) and neuroblastoma x glioma hybrid cells. In extracts of several tissues and cell lines the presence of cyclic-GMP-dependent protein kinase was also demonstrated by a photoaffinity-labeling procedure using 8-azidoinosine 3',5'-[32P]monophosphate. The results suggest that cyclic-GMP-dependent protein kinase is ubiquitously distributed although its level varies significantly from tissue to tissue and cell type to cell type. The results also support the hypothesis that cyclic-GMP-dependent protein kinase is involved in mediating some of the intracellular effects of those hormones, neutrotransmitters and drugs which regulate the intracellular level of cyclic GMP. (orig.)

  4. Regulation of a Drosophila cGMP-PDE by prenylation and interaction with a prenyl-binding protein

    Day, Jonathan P.; Cleghon, Vaughn; Houslay, Miles Douglas; Davies, Shireen A.

    2008-01-01

    Abstract Post-translational modification by isoprenylation is a pivotal process for the correct functioning of many signalling proteins. The Drosophila melanogaster cGMP-specific phosphodiesterase, DmPDE5/6, possesses a CaaX-box prenylation signal-motif, as do several novel cGMP-PDEs from insect and echinoid species. DmPDE5/6 is prenylated in vivo at Cys1128 and is localised to the plasma membrane when expressed in Drosophila S2 cells. Site-directed mutagenesis of the prenylated c...

  5. Derivation of Xeno-Free and GMP-Grade Human Embryonic Stem Cells – Platforms for Future Clinical Applications

    Tannenbaum, Shelly E.; Tako Turetsky, Tikva; Singer, Orna; Aizenman, Einat; Kirshberg, Sophie; Ilouz, Nili; Gil, Yaniv; Berman-Zaken, Yael; Perlman, Temima Schnitzer; Geva, Nitshia; Levy, Ora; Arbell, Daniel; Simon, Alex; Ben-Meir, Assaf; Shufaro, Yoel

    2012-01-01

    Clinically compliant human embryonic stem cells (hESCs) should be developed in adherence to ethical standards, without risk of contamination by adventitious agents. Here we developed for the first time animal-component free and good manufacturing practice (GMP)-compliant hESCs. After vendor and raw material qualification, we derived xeno-free, GMP-grade feeders from umbilical cord tissue, and utilized them within a novel, xeno-free hESC culture system. We derived and characterized three hESC ...

  6. Cavernosum smooth muscle relaxation induced by Schisandrol A via the NO-cGMP signaling pathway.

    Liu, W; Choi, B R; Bak, Y O; Zhang, L T; Zhou, L X; Huang, Y R; Zhao, C; Park, J K

    2016-01-01

    To evaluate the effect of Schisandrol A on rabbit corpus cavernosum smooth muscle and elucidate the potential mechanism. Penises were obtained from healthy male New Zealand White rabbits (2.5-3.0 kg). The pre-contracted penis with phenylephrine (Phe, 10 µM) was treated with accumulative concentrations of Schisandrol A (10-7, 10-6, 10-5 and 10-4 M). The change in intracavernosum pressure (ICP) and tension was recorded, cyclic nucleotides in the cavernosum tissue were measured by radioimmunoassay, mRNA level and expression of endothelial nitric oxide synthase (eNOS) and neuronal NOS (nNOS) were measured by real time PCR and western blot respectively. The corpus cavernosum smooth muscle relaxation induced by Schisandrol A was in a dose-dependent manner. Pre-treatment with NOS inhibitor (Nω nitro-L-arginine-methyl ester, L-NAME) or guanylyl cyclase inhibitor (1H-(1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one, ODQ) significantly diminished the relaxation. The cyclic guanosine monophosphate (cGMP) level was significantly increased in the cavernosum tissue. Real time PCR and western blot showed the mRNA level and expression of eNOS and nNOS was also upregulated. Schisandrol A relaxes the cavernosum smooth muscle by activating NO-cGMP signaling pathway. It may be a new promising treatment for erectile dysfunction and cardiovascular disease. PMID:27064883

  7. Altered cGMP dynamics at the plasma membrane contribute to diarrhea in ulcerative colitis.

    Arora, Kavisha; Sinha, Chandrima; Zhang, Weiqiang; Moon, Chang Suk; Ren, Aixia; Yarlagadda, Sunitha; Dostmann, Wolfgang R; Adebiyi, Adebowale; Haberman, Yael; Denson, Lee A; Wang, Xusheng; Naren, Anjaparavanda P

    2015-10-01

    Ulcerative colitis (UC) belongs to inflammatory bowel disorders, a group of gastrointestinal disorders that can produce serious recurring diarrhea in affected patients. The mechanism for UC- and inflammatory bowel disorder-associated diarrhea is not well understood. The cystic fibrosis transmembrane-conductance regulator (CFTR) chloride channel plays an important role in fluid and water transport across the intestinal mucosa. CFTR channel function is regulated in a compartmentalized manner through the formation of CFTR-containing macromolecular complexes at the plasma membrane. In this study, we demonstrate the involvement of a novel macromolecular signaling pathway that causes diarrhea in UC. We found that a nitric oxide-producing enzyme, inducible nitric oxide synthase (iNOS), is overexpressed under the plasma membrane and generates compartmentalized cGMP in gut epithelia in UC. The scaffolding protein Na(+)/H(+) exchanger regulatory factor 2 (NHERF2) bridges iNOS with CFTR, forming CFTR-NHERF2-iNOS macromolecular complexes that potentiate CFTR channel function via the nitric oxide-cGMP pathway under inflammatory conditions both in vitro and in vivo. Potential disruption of these complexes in Nherf2(-/-) mice may render them more resistant to CFTR-mediated secretory diarrhea than Nherf2(+/+) mice in murine colitis models. Our study provides insight into the mechanism of pathophysiologic occurrence of diarrhea in UC and suggests that targeting CFTR and CFTR-containing macromolecular complexes will ameliorate diarrheal symptoms and improve conditions associated with inflammatory bowel disorders. PMID:26261085

  8. Amyloid-β impairs, and ibuprofen restores, the cGMP pathway, synaptic expression of AMPA receptors and long-term potentiation in the hippocampus.

    Monfort, Pilar; Felipo, Vicente

    2010-01-01

    Amyloid-β (Aβ) rapidly impairs hippocampal long-term potentiation (LTP) and cognitive function in rats. We hypothesized that: a) Aβ-induced impairment of LTP would be due to impairment of the nitric oxide (NO)-cGMP pathway and AMPA receptor translocation; and b) treatment with the anti-inflammatory drug ibuprofen would restore the NO-cGMP pathway and LTP. The aims of this work were to assess whether ibuprofen prevents and/or rescues Aβ-induced LTP impairments in hippocampal slices and to analyze the role of the altered NO-cGMP-protein kinase G pathway and AMPA receptor phosphorylation and synaptic expression in the mechanisms by which Aβ impairs and ibuprofen restores LTP. Aβ impairs tetanus-induced activation of guanylate cyclase and cGMP increase, preventing protein kinase G activation, phosphorylation of GluR1 in Ser845 and AMPA receptors translocation to synaptic membranes, which is responsible for LTP impairment by Aβ. Ibuprofen prevents LTP impairment by Aβ by restoring guanylate cyclase activation and increase in cGMP and, subsequently, activation of protein kinase G, phosphorylation of GluR1 in Ser845 and synaptic expression of AMPA receptors. Restoration of cGMP levels is enough to restore all this process as indicated by the fact that the cGMP analog 8-Br-cGMP also normalizes the function of this pathway and restores LTP in the presence of Aβ. These results indicate that Aβ impairs LTP by impairing the NO-cGMP pathway and that ibuprofen restores LTP by restoring this pathway. These data suggest that restoring cGMP levels may have therapeutic utility to improve cognitive function impaired by Aβ. PMID:20858955

  9. El cuerpo como materia educativa en la prosa castellana de los siglos 13 y 14 - The body as educational content in late medieval spanish prose: 13th and 14th centuries

    Miguel Vicente Pedraz, España

    2012-07-01

    Full Text Available O CORPO COMO CONTEÚDO DA EDUCAÇÃO EM PROSA ESPANHOLA DOS SÉCULOS 13 E 14 ResumoO artigo propõe a seguinte hipótese: a gênese medieval da educação física remete, sobretudo, à emergência da sensibilidade cortesã, manifestada na aparência distinguida e no autodomínio gestual. A partir dessa perspectiva, as destrezas corporais próprias da atividade bélica ou da prática lúdica cavalheiresca constituem um aspecto de menor relevância; somente uma expressão dessa sensibilidade, uma vez que o fundamental é que manifesta o relativo declínio da idéia de estirpe e dos atributos do sangue como dispositivso determinantes da excelência, processo que abriria espaço a um crescente interesse educativo pela instrução do corpo para além do exercício ou do mero endurecimento físico. A partir da perspectiva de Norbert Elias, exposta em O processo civilizatório, se mostram alguns exemplos literários da pujança dessa nova sensibilidade na literatura castelhana da baixa Idade Média.Palvras-chave: educação do príncipe, corpo, cortesía, literatura medieval castelhana.  THE BODY AS EDUCATIONAL CONTENT IN LATE MEDIEVAL SPANISH PROSE: 13TH AND 14TH CENTURIES AbstractThis paper advances the following hypothesis: physical education in the Middle Ages originates mainly in the emergence of a new sensibility, exhibited in control of gestures, posture and speech. Thus, the physical strength and bodily skills required of a knight in order to face combat represent an expression, among many others, of this new chivalrous sensibility, which would eventually lead to the decline of the concepts of lineage and blood as supreme marks of nobility and excellence, as well as to a growing educational interest in the instruction of the body, beyond mere physical exercise and training. Following the theory proposed by Norbert Elias in his Process of civilization, this paper aims to offer a selection of literary examples of this new sensibility

  10. Activity of cGMP-Dependent Protein Kinase (PKG) Affects Sucrose Responsiveness and Habituation in "Drosophila melanogaster"

    Scheiner, Ricarda; Sokolowski, Marla B.; Erber, Joachim

    2004-01-01

    The cGMP-dependent protein kinase (PKG) has many cellular functions in vertebrates and insects that affect complex behaviors such as locomotion and foraging. The "foraging" ("for") gene encodes a PKG in "Drosophila melanogaster." Here, we demonstrate a function for the "for" gene in sensory responsiveness and nonassociative learning. Larvae of the…

  11. Chemotaxis in Dictyostelium discoideum : Effect of Concanavalin A on Chemoattractant Mediated Cyclic GMP Accumulation and Light Scattering Decrease

    Mato, José M.; Haastert, Peter J.M. van; Krens, Frans A.; Konijn, Theo M.

    1978-01-01

    In cells of the cellular slime mold Dictyostelium discoideum concanavalin A (Con A), at a concentration of 100 µg per ml, inhibits folic acid and cyclic AMP induced decrease in light scattering. Con A has no effect on folic acid mediated cyclic GMP accumulation and increases cyclic AMP mediated cycl

  12. Crystallization and preliminary X-ray diffraction studies of Xanthomonas campestris PNPase in the presence of c-di-GMP

    Crystals of S1-domain-truncated X. campestris PNPase were grown in the presence of c-di-GMP and diffracted to a resolution of 2.00 Å. Bacterial polynucleotide phosphorylase (PNPase) is a 3′–5′ processive exoribonuclease that participates in mRNA turnover and quality control of rRNA precursors in many bacterial species. It also associates with the RNase E scaffold and other components to form a multi-enzyme RNA degradasome machinery that performs a wider regulatory role in degradation, quality control and maturation of mRNA and noncoding RNA. Several crystal structures of bacterial PNPases, as well as some biological activity studies, have been published. However, how the enzymatic activity of PNPase is regulated is less well understood. Recently, Escherichia coli PNPase was found to be a direct c-di-GMP binding target, raising the possibility that c-di-GMP may participate in the regulation of RNA processing. Here, the successful cloning, purification and crystallization of S1-domain-truncated Xanthomonas campestris PNPase (XcPNPaseΔS1) in the presence of c-di-GMP are reported. The crystals belonged to the monoclinic space group C2, with unit-cell parameters a = 132.76, b = 128.38, c = 133.01 Å, γ = 93.3°, and diffracted to a resolution of 2.00 Å

  13. Effects of Na+ on ultraviolet light-induced photorelaxation and c-GMP levels in rabbit aorta

    Isolated strips of rabbit aorta in a state of drug-induced contraction relax reversibly when irradiated with ultraviolet light. The authors previously found that the magnitude of the photorelaxation progressively diminished as the extracellular Na+ ion concentration was reduced from 145 mM to 85 mM. At 85 mM Na+, there was minimal photorelaxation, even though the preparation continued to respond to vasoconstricting agents. The reduction in photosensitivity is not an osmotic effect because restoration of osmolarity did not restore photosensitivity. Neither the mechanism underlaying photorelaxation nor its modification by Na+ is precisely known. In order to examine these further the authors measured cyclic GMP levels in the absence and presence of UV light at both normal and reduced Na+ levels. At 145 mM Na+, irradiation resulted in an increase of cGMP from 0.299 to 0.717 fmole/μg protein. At 85 mM Na+, the corresponding levels were 0.541 and 1.24 fmole/μg protein. Thus, cGMP levels increase (approximately double) with UV irradiation at both reduced and normal Na+ concentrations even though there is little or no photorelaxation in the reduced Na+ environment. The reduction in Na+ may uncouple a link between cGMP elevation and cytoplasmic calcium in the aortic cell

  14. Cyclic di-GMP contributes to adaption and virulence of Bacillus thuringiensis through a riboswitch-regulated collagen adhesion protein.

    Tang, Qing; Yin, Kang; Qian, Hongliang; Zhao, Youwen; Wang, Wen; Chou, Shan-Ho; Fu, Yang; He, Jin

    2016-01-01

    Cyclic di-GMP is a ubiquitous second messenger that regulates diverse cellular processes in bacteria by binding to various protein or riboswitch effectors. In Bacillus thuringiensis BMB171, a c-di-GMP riboswitch termed Bc2 RNA resides in the 5'-untranslated region (5'-UTR) of an mRNA that encodes a collagen adhesion protein (Cap). The expression of cap was strongly repressed in parent strain BMB171 because of the presence of Bc2 RNA but was significantly promoted in the Bc2 RNA markerless deletion mutant. Bc2 RNA acts as a genetic "on" switch, which forms an anti-terminator structure to promote cap read-through transcription upon c-di-GMP binding. As a result, cap transcription was de-repressed under high c-di-GMP levels. Therefore, Bc2 RNA regulates cap expression using a repression/de-repression model. Bc2 RNA-regulated Cap was also found to be tightly associated with motility, aggregation, exopolysaccharide secretion, biofilm formation, and virulence of B. thuringiensis BMB171 against its host insect Helicoverpa armigera. PMID:27381437

  15. Application of halophilic nuclease H of Micrococcus varians subsp. halophilus to commercial production of flavoring agent 5'-GMP.

    Kamekura, M; Hamakawa, T; Onishi, H.

    1982-01-01

    RNA was degraded at 60 degrees C for 24 h by halophilic nuclease H in supernatants from broth cultures of Micrococcus varians subsp. halophilus containing 12% NaCl. Since contaminating 5'-nucleotidase exhibited almost no activity under these conditions, the 5'-GMP formed could be recovered from the reaction mixture, and the yield was 805 mg from 5 g of RNA.

  16. Clinical grade Treg: GMP isolation, improvement of purity by CD127 Depletion, Treg expansion, and Treg cryopreservation.

    Peters, J.H.; Preijers, F.W.M.B.; Woestenenk, R.; Hilbrands, L.B.; Koenen, H.J.P.M.; Joosten, I.

    2008-01-01

    BACKGROUND: Treg based immunotherapy is of great interest to facilitate tolerance in autoimmunity and transplantation. For clinical trials, it is essential to have a clinical grade Treg isolation protocol in accordance with Good Manufacturing Practice (GMP) guidelines. To obtain sufficient Treg for

  17. Crystal structures of Apo and GMP bound hypoxanthine-guanine phosphoribosyltransferase from Legionella pneumophila and the implications in gouty arthritis.

    Zhang, Nannan; Gong, Xiaojian; Lu, Min; Chen, Xiaofang; Qin, Ximing; Ge, Honghua

    2016-06-01

    Hypoxanthine-guanine phosphoribosyltransferase (HGPRT) (EC 2.4.2.8) reversibly catalyzes the transfer of the 5-phophoribosyl group from 5-phosphoribosyl-alpha-1-pyrophosphate (PRPP) to hypoxanthine or guanine to form inosine monophosphate (IMP) or guanosine monophosphate (GMP) in the purine salvage pathway. To investigate the catalytic mechanism of this enzyme in the intracellular pathogen Legionella pneumophila, we determined the crystal structures of the L. pneumophila HGPRT (LpHGPRT) both in its apo-form and in complex with GMP. The structures reveal that LpHGPRT comprises a core domain and a hood domain which are packed together to create a cavity for GMP-binding and the enzymatic catalysis. The binding of GMP induces conformational changes of the stable loop II. This new binding site is closely related to the Gout arthritis-linked human HGPRT mutation site (Ser103Arg). Finally, these structures of LpHGPRT provide insights into the catalytic mechanism of HGPRT. PMID:26968365

  18. Cytoplasmic and membranous CD24 marker expression has indirect correlation with cAMP/cGMP ratio

    Hossein Goudarzi

    2014-02-01

    Full Text Available Background- CD24 is a cell adhesion molecule that has been implicated in metastatic tumor progression cells. Our aim was clarify correlation between CD24 expression and cAMP:cGMP ratio in murine colorectal cancer cell line (CT26 after using cholera toxin.Materials and Methods- The CT26 cells were cultured in microtubes for assaying cAMP and cGMP; also the cells were cultured in flasks for assaying cytoplasmic and membranous CD24 expression. The Real-Time PCR was done for cDNA that was synthesized from CT26 cells’ mRNA. Also, expression CD24 marker of cells was determined by Anti-CD24 antibody and Goat Anti-Rabbit IgG-FITC (flow cytometry.Results- The cholera toxin grew cAMP:cGMP ratio and it influenced cytoplasmic and membrane CD24 expression.Conclusion- There are indirect correlation between cAMP:cGMP ratio and CD24 expression.

  19. Main flood peaks in the medieval Carpathian Basin (1000-1500): Annual and decadal overview

    Kiss, Andrea

    2013-04-01

    The analysis of over 140 reported floods is mainly based on contemporary legal evidence (charters), partly on other types of contemporary documentary evidence. Majority of sources contains data on individual flood events (i.e. occurrence, seasonality, magnitude). Concerning main flood peaks, evidence on annual and multi-annual (decadal, multi-decadal) level is also available. Despite data increase in the 13th century, only in the 14th-15th centuries documentation is representative enough to draw further conclusions. Apart from secondary flood peaks (probably in the mid-13th century and the turn of the 13th-14th centuries), three main periods with high flood frequencies are detected: 1330s-1350s, 1390s-1430s, and the late 1480s-1490s (continuing in the early 16th century). The first major flood peak was primarily reported in the eastern Carpathian Basin (the Tisa catchment), and can be characterised by a number of high-intensity flood events (with 1342-1343 in centre). During the second major, prolonged flood peak of 1390s-1430s, and that of the third, late 15th century one the importance of floods occurred on the Danube and in the Danube catchment area has to be as well highlighted. Moreover, in the first half of the 15th century long-term hydrological problems (prolonged high water-level and high flood frequency problems) can be identified. In some cases high flood-frequency periods were accompanied by documented hydromorphological impacts and some impacts on society can be also detected. Results show good agreement with the decadal precipitation reconstruction based on speleothem investigations carried out in North-Hungary.

  20. Bench to Bedside: Stability Studies of GMP Produced Trastuzumab-TCMC in Support of a Clinical Trial

    Diane E. Milenic

    2015-07-01

    Full Text Available The first-in-human phase 1 clinical radioimmunotherapy (RIT trial with 212Pb-1,4,7,10-tetraaza-1,4,7,10-tetra-(2-carbamoylmethyl-cyclododecane-trastuzumab (212Pb-TCMC-trastuzumab was completed in October 2014 as a joint effort at the University of Alabama (UAB and the University of California San Diego Moores Cancer Center. The preliminary reports indicate that after five dose-levels of intraperitoneally administered 212Pb-TCMC-trastuzumab, patients with carcinomatosis experienced minimal agent-related toxicity. This report presents the data accumulated to date on the stability of the clinical grade, produced according to current good manufacturing practices (cGMP, TCMC-trastuzumab conducted in support of that clinical trial. Of the eleven tests performed with the cGMP TCMC-trastuzumab all but one remained within specifications throughout the 5 year testing period. The protein concentration varied by 0.01 mg/mL at 48 months. Two other assays, ion-exchange high performance liquid chromatography (IEX-HPLC and a competitive radioimmunoassay (RIA indicated that the cGMP TCMC-trastuzumab integrity may be changing, although the change thus far is within specifications. Subsequent stability testing will confirm if a trend has truly developed. The cGMP TCMC-trastuzumab was also evaluated for tolerance to higher temperatures and the potential of storage at −80 °C. The immunoconjugate proved stable when subjected to the lower temperatures and to multiple freeze-thaw cycles. The size exclusion (SE HPLC analysis of the 203Pb-TCMC-trastuzumab was the only indicator that cGMP TCMC-trastuzumab may be sensitive to storage at 37 °C for 3 months.

  1. Physiological and Molecular Effects of the Cyclic Nucleotides cAMP and cGMP on Arabidopsis thaliana

    Herrera, Natalia M.

    2012-12-01

    The cyclic nucleotide monophosphates (CNs), cAMP and cGMP, are second messengers that participate in the regulation of development, metabolism and adaptive responses. In plants, CNs are associated with the control of pathogen responses, pollen tube orientation, abiotic stress response, membrane transport regulation, stomatal movement and light perception. In this study, we hypothesize that cAMP and cGMP promote changes in the transcription level of genes related to photosynthesis, high light and membrane transport in Arabidopsis thaliana leaves and, that these changes at the molecular level can have functional biological consequences. For this reason we tested if CNs modulate the photosynthetic rate, responses to high light and root ion transport. Real time quantitative PCR was used to assess transcription levels of selected genes and infrared gas analyzers coupled to fluorescence sensors were used to measure the photosynthetic parameters. We present evidence that both cAMP and cGMP modulate foliar mRNA levels early after stimulation. The two CNs trigger different responses indicating that the signals have specificity. A comparison of proteomic and transcriptional changes suggest that both transcriptional and post-transcriptional mechanisms are modulated by CNs. cGMP up-regulates the mRNA levels of components of the photosynthesis and carbon metabolism. However, neither cAMP nor cGMP trigger differences in the rate of carbon assimilation, maximum efficiency of the photosystem II (PSII), or PSII operating efficiency. It was also demonstrated that CN regulate the expression of its own targets, the cyclic nucleotide gated channels - CNGC. Further studies are needed to identify the components of the signaling transduction pathway that mediate cellular changes and their respective regulatory and/or signaling roles.

  2. cGMP-phosphodiesterase inhibition enhances photic responses and synchronization of the biological circadian clock in rodents.

    Santiago A Plano

    Full Text Available The master circadian clock in mammals is located in the hypothalamic suprachiasmatic nuclei (SCN and is synchronized by several environmental stimuli, mainly the light-dark (LD cycle. Light pulses in the late subjective night induce phase advances in locomotor circadian rhythms and the expression of clock genes (such as Per1-2. The mechanism responsible for light-induced phase advances involves the activation of guanylyl cyclase (GC, cGMP and its related protein kinase (PKG. Pharmacological manipulation of cGMP by phosphodiesterase (PDE inhibition (e.g., sildenafil increases low-intensity light-induced circadian responses, which could reflect the ability of the cGMP-dependent pathway to directly affect the photic sensitivity of the master circadian clock within the SCN. Indeed, sildenafil is also able to increase the phase-shifting effect of saturating (1200 lux light pulses leading to phase advances of about 9 hours, as well as in C57 a mouse strain that shows reduced phase advances. In addition, sildenafil was effective in both male and female hamsters, as well as after oral administration. Other PDE inhibitors (such as vardenafil and tadalafil also increased light-induced phase advances of locomotor activity rhythms and accelerated reentrainment after a phase advance in the LD cycle. Pharmacological inhibition of the main downstream target of cGMP, PKG, blocked light-induced expression of Per1. Our results indicate that the cGMP-dependent pathway can directly modulate the light-induced expression of clock-genes within the SCN and the magnitude of light-induced phase advances of overt rhythms, and provide promising tools to design treatments for human circadian disruptions.

  3. The Role of cGMP on Adenosine A1 Receptor-mediated Inhibition of Synaptic Transmission at the Hippocampus

    Pinto, Isa; Serpa, André; Sebastião, Ana M.; Cascalheira, José F.

    2016-01-01

    Both adenosine A1 receptor and cGMP inhibit synaptic transmission at the hippocampus and recently it was found that A1 receptor increased cGMP levels in hippocampus, but the role of cGMP on A1 receptor-mediated inhibition of synaptic transmission remains to be established. In the present work we investigated if blocking the NOS/sGC/cGMP/PKG pathway using nitric oxide synthase (NOS), protein kinase G (PKG), and soluble guanylyl cyclase (sGC) inhibitors modify the A1 receptor effect on synaptic transmission. Neurotransmission was evaluated by measuring the slope of field excitatory postsynaptic potentials (fEPSPs) evoked by electrical stimulation at hippocampal slices. N6-cyclopentyladenosine (CPA, 15 nM), a selective A1 receptor agonist, reversibly decreased the fEPSPs by 54 ± 5%. Incubation of the slices with an inhibitor of NOS (L-NAME, 200 μM) decreased the CPA effect on fEPSPs by 57 ± 9% in female rats. In males, ODQ (10 μM), an sGC inhibitor, decreased the CPA inhibitory effect on fEPSPs by 23 ± 6%, but only when adenosine deaminase (ADA,1 U/ml) was present; similar results were found in females, where ODQ decreased CPA-induced inhibition of fEPSP slope by 23 ± 7%. In male rats, the presence of the PKG inhibitor (KT5823, 1 nM) decreased the CPA effect by 45.0 ± 9%; similar results were obtained in females, where KT5823 caused a 32 ± 9% decrease on the CPA effect. In conclusion, the results suggest that the inhibitory action of adenosine A1 receptors on synaptic transmission at hippocampus is, in part, mediated by the NOS/sGC/cGMP/PKG pathway.

  4. GMP production of [18F]FDOPA and issues concerning its quality analyses as in USP 'Fluorodopa F 18 Injection'

    Lately, 6-[18F]fluoro-L-DOPA (FDOPA) has found increase in its clinical demand for whole-body positron emission tomography (PET) scans, and two key issues in fulfilling this demand are the difficulties in producing FDOPA under the recently imposed PET drug good manufacturing practice (GMP) regulations and in providing it in the quality meeting the terms of major compendia. This paper describes the approaches for the GMP production of FDOPA and for the product testing to meet the standard of United States Pharmacopeia (USP) 'Fluorodopa F 18 Injection.' FDOPA was produced by the carrier-added electrophilic aromatic substitution reaction in the facility complying Pharmaceutical Inspection Cooperation Scheme clean room standard. The special aseptic handling technique was applied to minimize the bioburden. The product quality control followed all testing items and procedures, including three different settings of high performance liquid chromatography (HPLC). The process yielded FDOPA average 2.60±0.26 GBq (N=22) in every batch. All qualities of the product were within the specifications described in the USP 'Fluorodopa F 18 Injection.' The entire production was audited by the government authority and certified to comply with the latest PET drug GMP regulation. Our efforts in producing FDOPA following all aspects of GMP requirements have resulted in a product with the USP quality and certified as GMP complied. The routine production yields enough doses for three to four whole-body scans in each batch. The issues discussed in the report provide good reference for producers planning in routine production for PET drugs that are not commonly produced or with complicated compendial quality control tests. (author)

  5. 3',5'-Cyclic diguanylic acid (c-di-GMP) inhibits basal and growth factor-stimulated human colon cancer cell proliferation

    The novel cyclic dinucleotide, 3',5'-cyclic diguanylic acid, cGpGp (c-di-GMP), is a naturally occurring small molecule that regulates important signaling mechanisms in prokaryotes. Recently, we showed that c-di-GMP has 'drug-like' properties and that c-di-GMP treatment might be a useful antimicrobial approach to attenuate the virulence and pathogenesis of Staphylococcus aureus and prevent or treat infection. In the present communication, we report that c-di-GMP (≤50 μM) has striking properties regarding inhibition of cancer cell proliferation in vitro. c-di-GMP inhibits both basal and growth factor (acetylcholine and epidermal growth factor)-induced cell proliferation of human colon cancer (H508) cells. Toxicity studies revealed that exposure of normal rat kidney cells and human neuroblastoma cells to c-di-GMP at biologically relevant doses showed no lethal cytotoxicity. Cyclic dinucleotides, such as c-di-GMP, represent an attractive and novel 'drug-platform technology' that can be used not only to develop new antimicrobial agents, but also to develop novel therapeutic agents to prevent or treat cancer

  6. Jefferson Lab Hall A Beamline Instrumentation and Calibration for GMP experiment

    Gautam, Thir Narayan

    2015-10-01

    The nucleon electromagnetic form factors characterize the distributions of electric charge and magnetization current inside the nucleon and thus reflect the internal structure determined by Quantum Chromodynamics. The GMp experiment is a first experiment run in Hall A at Jefferson Lab after the upgrade to double the beam energy with the goal to precisely measure electron-proton elastic cross section in the Q2 range of 7 to 17 GeV2 with an accuracy of better than 2%; several time better than existing data at the highest Q2. In order to achieve this accuracy, a determination of the accumulated beam charge of better than 0.5% is required. The new 12 GeV beamline was commissioned during the spring of 2015, with the main instrumentation consisting of beam charge and position monitors. In this talk, the procedures and the results of the calibrations of these beamline components will be presented.

  7. GMP production of 68Ga-labelled DOTA-NOC on IBA Synthera

    68Ga labelled peptides are increasingly being used clinically due to widespread availability of 68Ge/68Ga generators and their recognised importance in the diagnosis of neuroendocrine tumours. A simple and reproducible GMP-compliant process for labelling peptides with generator-produced 68Ga is crucial to meet current pharmaceutical requirements. Here, we describe the development of a cassette-based 68Ga-radiolabelling process using an automated commercial synthesizer coupled with a standard 68Ge/68Ga generator. The radiolabeled somatostatin analogue [68Ga]DOTA-NOC was successfully synthesized in about 30 min with 80.3 - 3.20 % of decay corrected radiochemical yield. The process is easily adaptable to the labelling of other peptides such as DOTA-TOC or DOTA-TATE for diagnosis or therapy. (author)

  8. Selective binding of 2'-F-c-di-GMP to Ct-E88 and Cb-E43, new class I riboswitches from Clostridium tetani and Clostridium botulinum respectively.

    Luo, Yiling; Zhou, Jie; Wang, Jingxin; Dayie, T Kwaku; Sintim, Herman O

    2013-06-01

    C-di-GMP is a second messenger in bacteria and partly regulates bacterial physiology by binding to class I and II riboswitches. Four class I c-di-GMP riboswitch aptamer candidates, Ct-E88, Cb-17B, Cb-E43 and Cd-630 RNAs, selected from a GEMM RNA sequence motif in the Rfam database, were expressed and experimentally verified to bind to c-di-GMP. The two newly characterized c-di-GMP riboswitches, Ct-E88 and Cb-E43, bound c-di-GMP with nanomolar Kd whereas the affinities of Cb-17B and Cd-630 for c-di-GMP were at least a 100-fold weaker. Interestingly, whereas the three riboswitches (Vc2, Et-E88 and Cb-E43) bound c-di-GMP with similar Kd values, 2'-modified analogs of c-di-GMP differentially bound to these three class I aptamers. For example, 2'-F-c-di-GMP bound Vc2 with a Kd value of 102 nM whereas the Kd value of 2'-F-c-di-GMP-Ct-E88 is 43 μM (422× higher than that for Vc2 RNA), revealing that there are differences in the binding sites of functional class I c-di-GMP riboswitches. PMID:23559271

  9. C-di-GMP hydrolysis by Pseudomonas aeruginosa HD-GYP phosphodiesterases: analysis of the reaction mechanism and novel roles for pGpG.

    Valentina Stelitano

    Full Text Available In biofilms, the bacterial community optimizes the strategies to sense the environment and to communicate from cell to cell. A key player in the development of a bacterial biofilm is the second messenger c-di-GMP, whose intracellular levels are modulated by the opposite activity of diguanylate cyclases and phosphodiesterases. Given the huge impact of bacterial biofilms on human health, understanding the molecular details of c-di-GMP metabolism represents a critical step in the development of novel therapeutic approaches against biofilms. In this study, we present a detailed biochemical characterization of two c-di-GMP phosphodiesterases of the HD-GYP subtype from the human pathogen Pseudomonas aeruginosa, namely PA4781 and PA4108. Upstream of the catalytic HD-GYP domain, PA4781 contains a REC domain typical of two-component systems, while PA4108 contains an uncharacterized domain of unknown function. Our findings shed light on the activity and catalytic mechanism of these phosphodiesterases. We show that both enzymes hydrolyse c-di-GMP in a two-step reaction via the linear intermediate pGpG and that they produce GMP in vitro at a surprisingly low rate. In addition, our data indicate that the non-phosphorylated REC domain of PA4781 prevents accessibility of c-di-GMP to the active site. Both PA4108 and phosphorylated PA4781 are also capable to use pGpG as an alternative substrate and to hydrolyse it into GMP; the affinity of PA4781 for pGpG is one order of magnitude higher than that for c-di-GMP. These results suggest that these enzymes may not work (primarily as genuine phosphodiesterases. Moreover, the unexpected affinity of PA4781 for pGpG may indicate that pGpG could also act as a signal molecule in its own right, thus further widening the c-di-GMP-related signalling scenario.

  10. Hyperammonemia inhibits the natriuretic peptide receptor 2 (NPR-2)-mediated cyclic GMP synthesis in the astrocytic compartment of rat cerebral cortex slices.

    Zielińska, Magdalena; Fresko, Inez; Konopacka, Agnieszka; Felipo, Vicente; Albrecht, Jan

    2007-11-01

    The decrease of cyclic GMP (cGMP) level in the brain, contributing to cognitive and memory deficit in hyperammonemia (HA), has been attributed to the interference of ammonia with the NMDA/nitric oxide/soluble guanylate cyclase (GC)/cGMP pathway in neurons. The present study tested the hypotheses that (a) HA also affects cGMP synthesis elicited by stimulation of the natriuretic peptide receptor 2 (NPR-2) with its natural ligand, C-type natriuretic peptide (CNP) and (b) the latter effect may involve astrocytes, the ammonia-sensitive cells. In the cerebral cortical slices of control rats, CNP stimulated cGMP synthesis in a degree comparable to the NO donor, S-nitroso-N-acetylpenicillamine (SNAP) used at an optimal concentration. Fluoroacetate (FA), a metabolic inhibitor specifically affecting astrocytic mitochondria, inhibited the CNP-dependent cGMP synthesis by about 50%. Ammonium acetate-induced HA decreased by 68% the CNP-dependent cGMP generation in slices incubated in the absence of FA. In slices incubated in the presence of FA, cGMP synthesis in slices derived from HA rats did not differ from that in control slices. The results indicate that HA inhibits CNP-dependent cGMP synthesis in the FA-vulnerable, astrocytic compartment, but not in the FA-resistant compartment(s) of the brain. HA did not affect the expression of NPR-2 mRNA in the cerebral cortex tissue as tested using real-time PCR, indicating that the effect of ammonia involves as yet unidentified events occurring posttranscriptionally. Deregulation of NPR-2 function in astrocytes by ammonia may contribute to neurophysiological symptoms of HA. PMID:17629948

  11. Structural and functional characteristics of cGMP-dependent methionine oxidation in Arabidopsis thaliana proteins

    Marondedze, Claudius

    2013-01-05

    Background: Increasing structural and biochemical evidence suggests that post-translational methionine oxidation of proteins is not just a result of cellular damage but may provide the cell with information on the cellular oxidative status. In addition, oxidation of methionine residues in key regulatory proteins, such as calmodulin, does influence cellular homeostasis. Previous findings also indicate that oxidation of methionine residues in signaling molecules may have a role in stress responses since these specific structural modifications can in turn change biological activities of proteins. Findings. Here we use tandem mass spectrometry-based proteomics to show that treatment of Arabidopsis thaliana cells with a non-oxidative signaling molecule, the cell-permeant second messenger analogue, 8-bromo-3,5-cyclic guanosine monophosphate (8-Br-cGMP), results in a time-dependent increase in the content of oxidised methionine residues. Interestingly, the group of proteins affected by cGMP-dependent methionine oxidation is functionally enriched for stress response proteins. Furthermore, we also noted distinct signatures in the frequency of amino acids flanking oxidised and un-oxidised methionine residues on both the C- and N-terminus. Conclusions: Given both a structural and functional bias in methionine oxidation events in response to a signaling molecule, we propose that these are indicative of a specific role of such post-translational modifications in the direct or indirect regulation of cellular responses. The mechanisms that determine the specificity of the modifications remain to be elucidated. 2013 Marondedze et al.; licensee BioMed Central Ltd.

  12. Direct photolabeling of the cGMP-stimulated cyclic nucleotide phosphodiesterase

    cGMP-stimulated phosphodiesterase (PDE) has been directly photolabeled with [32P]cGMP using UV light. Sequence analysis of peptide fragments obtained from partial proteolysis or cyanogen bromide cleavage indicate that two different domains are labeled. One site, on a Mr = 36,000 chymotryptic fragment located near the COOH terminus, has characteristics consistent with it being close to or part of the catalytic site of the enzyme. This peptide contains a region of sequence that is highly conserved in all mammalian cyclic nucleotide PDEs and has been postulated to contain the catalytic domain of the enzyme. The other site, on a Mr = 28,000 cyanogen bromide cleavage fragment located near the middle of the molecule, probably makes up part of the allosteric site of the molecule. Labeling of the enzyme is concentration dependent and Scatchard analysis of labeling yields a biphasic plot with apparent half labeling concentrations of about 1 and 30 microM consistent with two types of sites being labeled. Limited proteolysis of the PDE by chymotrypsin yields five prominent fragments that separate by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) at Mr = 60,000, 57,000, 36,000, 21,000, and 17,000. Both the Mr = 60,000 and 57,000 apparently have blocked NH2 termini suggesting that the Mr = 57,000 fragment is a subfragment of the Mr = 60,000 fragment. Primary sequence analysis indicates that both the Mr = 21,000 and 17,000 fragments are subfragments of the Mr = 36,000 fragment. Autoradiographs of photolabeled then partially proteolyzed enzyme show labeled bands at Mr = 60,000, 57,000, and 36,000. Addition of 5 microM cAMP prior to photolabeling eliminates photolabeling of the Mr = 36,000 fragment but not the Mr = 60,000 or 57,000 fragments

  13. Development of a fully automated and GMP compliant [11C]-radiosynthesis module

    Full text: The automation of routinely produced and GMP compliant [11C]radiotracers is an important process for any PET centre. Automation allows for highly reproducible synthesis with large amount of radioactivity while minimising exposure to the radiochemist. GMP compliance is required to meet the upcoming regulations for PET radio-pharmaceutical production. Based on these requirements, we have designed at the ARMC PET Centre, a reliable and versatile chemistry module for the production of [11C]-radiotracers. This aim has been achieved by using disposable manifolds and tubing in conjunction with newly prepared reaction vessels. Cross-contamination by subsequent productions of different radiophammaceuticals is then fully prevented. Automation of the synthesis sequences is performed by a PLC Siemens (CPU216), and the programs allows for the production of different radiotracers according to their specific reaction parameters such as temperature and reaction time. Relevant digital and analog parameters are measured throughout the synthesis and fed back to the PLC to allow clear monitoring of the process by the user. Our new radiosynthesis module is based on the conventional production of [11C]-methyliodide, followed by its reaction with the relevant precursor at room temperature using the loop technique. The crude labelled product is then automatically injected into the HPLC column. After collection of the pure product, the mobile phase is discarded, and the final product eluted with ethanol and formulated with a physiological solution using the 'SepPak technique'. The synthesis time is about 30 minutes including purification and formulation of the product. The [11C]-radiosynthesis prototype module is currently under evaluation. Copyright (2002) The Australian and New Zealand Society of Nuclear Medicine Inc

  14. Chemical and functional properties of glycomacropeptide (GMP) and its role in the detection of cheese whey adulteration in milk: a review

    7; ,; Sharma, Rajan; Rajput, Yudhishthir; Mann, Bimlesh

    2013-01-01

    International audience Glycomacropeptide (GMP) is a C-terminal part (f 106–169) of kappa-casein which is released in whey during cheese making by the action of chymosin. GMP being a biologically active component has gained much attention in the past decade. It also has unique chemical and functional properties. Many of the biological properties have been ascribed to the carbohydrate moieties attached to the peptide. The unique set of amino acids in GMP makes it a sought-after ingredient wi...

  15. Angiotensin II increases phosphodiesterase 5A expression in vascular smooth muscle cells: A mechanism by which angiotensin II antagonizes cGMP signaling

    Kim, Dongsoo; Aizawa, Toru; Wei, Heng; Pi, Xinchun; Rybalkin, Sergei D.; Berk, Bradford C.; Yan, Chen

    2004-01-01

    Angiotensin II (Ang II) and nitric oxide (NO)/natriuretic peptide (NP) signaling pathways mutually regulate each other. Imbalance of Ang II and NO/NP has been implicated in the pathophysiology of many vascular diseases. cGMP functions as a key mediator in the interaction between Ang II and NO/NP. Cyclic nucleotide phosphodiesterase 5A (PDE5A) is important in modulating cGMP signaling by hydrolyzing cGMP in vascular smooth muscle cells (VSMC). Therefore, we examined whether Ang II negatively m...

  16. In vitro and in vivo generation and characterization of Pseudomonas aeruginosa biofilm-dispersed cells via c-di-GMP manipulation

    Chua, Song Lin; Hultqvist, Louise D; Yuan, Mingjun; Rybtke, Morten; Nielsen, Thomas E; Givskov, Michael; Tolker-Nielsen, Tim; Yang, Liang

    2015-01-01

    Bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP) is a global secondary bacterial messenger that controls the formation of drug-resistant multicellular biofilms. Lowering the intracellular c-di-GMP content can disperse biofilms, and it is proposed as a biofilm eradication strategy...... biofilms by reducing the intracellular c-di-GMP content through modulation of phosphodiesterases (PDEs). Unlike conventional protocols that demonstrate biofilm dispersal by biomass quantification, our protocols enable physiological characterization of the dispersed cells. Biomarkers of dispersed cells are...

  17. Similar expression patterns of bestrophin-4 and cGMP dependent Ca2+-activated chloride channel activity in the vasculature

    Bouzinova, Elena V.; Larsen, Per; Matchkov, Vladimir; Nilsson, Holger; Aalkjær, Christian

    2008-01-01

    (abstract by Matchkov et. al) that siRNA mediated downregulation of bestrophin-4 is associated with the disappearance of a recently demonstrated2 cGMP-dependent Ca2+-activated Cl- current in vascular smooth muscle cells (SMCs). Here we study the distribution of bestrophin-4-and cGMP dependent Cl- channel...... expressed epitope) Western blot detected a ~65 kDa band in cell lysates from rat mesenteric small arteries and aorta, which was not seen in pulmonary arteries and when preincubated with the immunizing peptide. The distribution of bestrophin-4 mRNA and protein has a pattern similar to the cGMP-dependent Cl......- current in SMCs of different origins. Immunohistochemistry identified bestrophin-4 both in endothelial and SMCs of the vascular tree in the brain, heart, kidney and mesentery, but not in the lungs. We suggest that bestrophin-4 is important for the cGMP dependent, Ca2+ activated Cl- conductance in many...

  18. Discussion on how to implement GMP and HACCP in pickled vegetables factories%酱腌菜加工厂如何实施GMP和HACCP

    沈子林; 管有根; 翁本德

    2002-01-01

    食品良好生产规范(GMP)是食品生产全过程中保证食品具有高度安全性的良好生产管理系统,是国际上普遍采用的先进管理办法.我国酱腌菜加工企业也应实施建立GMP,同时要进行HACCP应用.

  19. CSF concentrations of cAMP and cGMP are lower in patients with Creutzfeldt-Jakob disease but not Parkinson's disease and amyotrophic lateral sclerosis.

    Patrick Oeckl

    Full Text Available BACKGROUND: The cyclic nucleotides cyclic adenosine-3',5'-monophosphate (cAMP and cyclic guanosine-3',5'-monophosphate (cGMP are important second messengers and are potential biomarkers for Parkinson's disease (PD, amyotrophic lateral sclerosis (ALS and Creutzfeldt-Jakob disease (CJD. METHODOLOGY/PRINCIPAL FINDINGS: Here, we investigated by liquid chromatography/tandem mass spectrometry (LC-MS/MS the cerebrospinal fluid (CSF concentrations of cAMP and cGMP of 82 patients and evaluated their diagnostic potency as biomarkers. For comparison with a well-accepted biomarker, we measured tau concentrations in CSF of CJD and control patients. CJD patients (n = 15 had lower cAMP (-70% and cGMP (-55% concentrations in CSF compared with controls (n = 11. There was no difference in PD, PD dementia (PDD and ALS cases. Receiver operating characteristic (ROC curve analyses confirmed cAMP and cGMP as valuable diagnostic markers for CJD indicated by the area under the curve (AUC of 0.86 (cAMP and 0.85 (cGMP. We calculated a sensitivity of 100% and specificity of 64% for cAMP and a sensitivity of 67% and specificity of 100% for cGMP. The combination of both nucleotides increased the sensitivity to 80% and specificity to 91% for the term cAMPxcGMP (AUC 0.92 and to 93% and 100% for the ratio tau/cAMP (AUC 0.99. CONCLUSIONS/SIGNIFICANCE: We conclude that the CSF determination of cAMP and cGMP may easily be included in the diagnosis of CJD and could be helpful in monitoring disease progression as well as in therapy control.

  20. DNA-binding properties of a cGMP-binding CRP homologue that controls development of metabolically dormant cysts of Rhodospirillum centenum.

    Roychowdhury, Sugata; Dong, Qian; Bauer, Carl E

    2015-11-01

    Rhodospirillum centenum utilizes 3',5'-cyclic guanosine monophosphate (cGMP) as a messenger to regulate development of desiccation-resistant cysts. In this study, we demonstrated that gcyA, gcyB and gcyC, coding for putative subunits of a guanylyl cyclase, increase expression from 8- to 500-fold when cells transition from vegetative to cyst phases of growth. This induction did not occur in a strain that is defective in cGMP synthesis or in a strain that contains a deletion of cgrA that codes for a cGMP-binding homologue of Escherichia coli catabolite repressor protein (CRP). We also demonstrated that cgrA auto-induces its own expression in the presence of cGMP, indicating that a feed-forward loop is used to ramp up cGMP production as cells undergo encystment. Inspection of an intragenic region upstream of gcyB revealed a sequence that is identical to the CRP consensus sequence from E. coli. DNase I and fluorescence anisotropy analyses demonstrated that CgrA bound to this target sequence at a protein : cGMP ratio of 1 : 2 with Kd ∼61 nM. This was in contrast to CgrA in the presence of cAMP, which exhibited Kd ∼1795 nM. CgrA thus constitutes a novel variant of CRP that utilizes cGMP to regulate production of cGMP synthase for the control of cyst development. PMID:26362215

  1. Fully automated radiosynthesis of both enantiomers of [18F]Flubatine under GMP conditions for human application

    A fully automatized radiosynthesis of (+)- and (−)-[18F]Flubatine ((+)- and (−)NCFHEB) by means of a commercially available synthesis module (TRACERlab FX FN) under GMP conditions is reported. Radiochemical yields of 30% within an overall synthesis time of 40 min were achieved in more than 70 individual syntheses. Specific activities were approximately 3000 GBq/μmol and radiochemical purity was determined to be at least 97%. - Highlights: • A fully automatized radiosynthesis of Flubatine under GMP conditions is reported. • Radiochemical yields of 30% in 40 min were achieved in more than 70 syntheses. • Specific activitiy of 3000 GBq/μmol and radiochemical purity of more than 97% were achieved

  2. An Overgrowth Disorder Associated with Excessive Production of cGMP Due to a Gain-of-Function Mutation of the Natriuretic Peptide Receptor 2 Gene

    Miura, Kohji; Namba, Noriyuki; Fujiwara, Makoto; Ohata, Yasuhisa; Ishida, Hidekazu; Kitaoka, Taichi; Kubota, Takuo; Hirai, Haruhiko; Higuchi, Chikahisa; Tsumaki, Noriyuki; Yoshikawa, Hideki; Sakai, Norio; Michigami, Toshimi; Ozono, Keiichi

    2012-01-01

    We describe a three-generation family with tall stature, scoliosis and macrodactyly of the great toes and a heterozygous p.Val883Met mutation in Npr2, the gene that encodes the CNP receptor NPR2 (natriuretic peptide receptor 2). When expressed in HEK293A cells, the mutant Npr2 cDNA generated intracellular cGMP (cyclic guanosine monophosphate) in the absence of CNP ligand. In the presence of CNP, cGMP production was greater in cells that had been transfected with the mutant Npr2 cDNA compared to wild-type cDNA. Transgenic mice in which the mutant Npr2 was expressed in chondrocytes driven by the promoter and intronic enhancer of the Col11a2 gene exhibited an enhanced production of cGMP in cartilage, leading to a similar phenotype to that observed in the patients. In addition, blood cGMP concentrations were elevated in the patients. These results indicate that p.Val883Met is a constitutive active gain-of-function mutation and elevated levels of cGMP in growth plates lead to the elongation of long bones. Our findings reveal a critical role for NPR2 in skeletal growth in both humans and mice, and may provide a potential target for prevention and treatment of diseases caused by impaired production of cGMP. PMID:22870295

  3. An overgrowth disorder associated with excessive production of cGMP due to a gain-of-function mutation of the natriuretic peptide receptor 2 gene.

    Kohji Miura

    Full Text Available We describe a three-generation family with tall stature, scoliosis and macrodactyly of the great toes and a heterozygous p.Val883Met mutation in Npr2, the gene that encodes the CNP receptor NPR2 (natriuretic peptide receptor 2. When expressed in HEK293A cells, the mutant Npr2 cDNA generated intracellular cGMP (cyclic guanosine monophosphate in the absence of CNP ligand. In the presence of CNP, cGMP production was greater in cells that had been transfected with the mutant Npr2 cDNA compared to wild-type cDNA. Transgenic mice in which the mutant Npr2 was expressed in chondrocytes driven by the promoter and intronic enhancer of the Col11a2 gene exhibited an enhanced production of cGMP in cartilage, leading to a similar phenotype to that observed in the patients. In addition, blood cGMP concentrations were elevated in the patients. These results indicate that p.Val883Met is a constitutive active gain-of-function mutation and elevated levels of cGMP in growth plates lead to the elongation of long bones. Our findings reveal a critical role for NPR2 in skeletal growth in both humans and mice, and may provide a potential target for prevention and treatment of diseases caused by impaired production of cGMP.

  4. Clinical application of combined detection of serum hs-CRP, GMP-140 and cTnI in patients with coronary heart diseases

    Objective: To explore the clinical significance of changes of serum hs-CRP, GMP-140 and cTnI levels in patients with coronary heart diseases. Methods: Serum GMP-140 (with RIA), cTnI (with ELISA) and hs-CRP (with immuno turbidity method) levels were determined in 91 patients with coronary heart diseases (42 SAP, 34UAP, 15AMI) and 35 controls. Results: Serum hs-CRP, GMP-140, cTnI levels in patients with coronary heart diseases were significantly higher than those in controls (P <0.01). Among the patients with of coronary heart diseases, the magnitude of changes of the levels of serum hs-CRP, GMP-140 and cTnI levels in AMI and UAP groups were significantly larger than those in SAP group (P < 0.05). Serum hs-CRP levels were positively correlated with serum GMP-140 and cTnI levels (r = 0.6214, 0.6023, P < 0.01). Conclusion: Serum hs-CRP, GMP140 and cTnI levels were closely related to the diseases process of coronary heart diseases and were of great clinical importance for assessment of the disease and outcome prediction. (authors)

  5. The NO/cGMP pathway inhibits transient cAMP signals through the activation of PDE2 in striatal neurons

    Marina ePolito

    2013-11-01

    Full Text Available The NO-cGMP signaling plays an important role in the regulation of striatal function although the mechanisms of action of cGMP specifically in medium spiny neurons (MSNs remain unclear. Using genetically encoded fluorescent biosensors, including a novel Epac-based sensor (EPAC-SH150 with increased sensitivity for cAMP, we analyze the cGMP response to NO and whether it affected cAMP/PKA signaling in MSNs. The Cygnet2 sensor for cGMP reported large responses to NO donors in both striatonigral and striatopallidal MSNs, and this cGMP signal was controlled partially by PDE2. At the level of cAMP brief forskolin stimulations produced transient cAMP signals which differed between D1 and D2 medium spiny neurons. NO inhibited these cAMP transients through cGMP-dependent PDE2 activation, an effect that was translated and magnified downstream of cAMP, at the level of PKA. PDE2 thus appears as a critical effector of NO which modulates the post-synaptic response of MSNs to dopaminergic transmission.

  6. Clinical significance of determination of plasma ET-1, GMP-140 and serum cTnl levels in patients with coronary heart disease

    Objective: To explore the clinical significance of changes of plasma ET-1, GMP-140 and serum cTnI levels in patients with CHD. Methods: Plasma ET-1, GMP-140 levels (with RIA) and serum cTnI levels (with ELISA) were determined in 96 patients with CHD (36 SAP, 41UAP, 19AMI) and 35 controls. Results: Plasma ET-1, GMP-140 and serum cTnI levels in patients with CHD were significantly higher than those in controls (P<0.01). Among the CHD patients, magnitidue of changes of the levels of plasma ET-1, GMP-140 and serum cTnI in AMI and UAP groups were significantly larger than those in SAP groups (P<0.05). Serum cTnI levels were positively correlated with plasma ET-1, GMP-140 levels. Conclusion: The plasma ET-1, GMP-140 and serum cTnI levels were closely related to the diseases process of CHD and were of great clinical importance. (authors)

  7. C-di-GMP regulates Pseudomonas aeruginosa stress response to tellurite during both planktonic and biofilm modes of growth

    Chua, Song Lin; Sivakumar, Krishnakumar; Rybtke, Morten Levin; Yuan, Mingjun; Andersen, Jens Bo; Nielsen, Thomas E.; Givskov, Michael; Tolker-Nielsen, Tim; Cao, Bin; Kjelleberg, Staffan; Yang, Liang

    2015-01-01

    tellurite (TeO3(2-)) exposure induced the intracellular content of the secondary messenger cyclic di-GMP (c-di-GMP) of Pseudomonas aeruginosa. Two diguanylate cyclases (DGCs), SadC and SiaD, were responsible for the increased intracellular content of c-di-GMP. Enhanced c-di-GMP levels by TeO3(2-) further...... increased P. aeruginosa biofilm formation and resistance to TeO3(2-). P. aeruginosa ΔsadCΔsiaD and PAO1/p(lac)-yhjH mutants with low intracellular c-di-GMP content were more sensitive to TeO3(2-) exposure and had low relative fitness compared to the wild-type PAO1 planktonic and biofilm cultures exposed to...... TeO3(2-). Our study provided evidence that c-di-GMP level can play an important role in mediating stress response in microbial communities during both planktonic and biofilm modes of growth....

  8. An efficient and aseptic preparation of 'sodium fluoride (18F) injection' in a GMP compliant facility

    'Sodium fluoride (18F) injection' is an isotonic NaCl solution containing [18F]NaF to be used as bone imaging agent. Although its new drug application (NDA) was approved by the United States (US) Food and Drug Administration (FDA) in 1972, it has not been commercially available since 1975 due to mostly the popularity of 99mTc-methylene diphosphonate (MDP). Recently, advances in positron emission tomography (PET)/CT technology and the often interrupted 99mTc supply have led to the renewed interest in the use of [18F]NaF to detect bone metastases in cancer patients. This report introduces an efficient, low-cost and aseptic preparation of 'Sodium fluoride (18F) injection' for PET scan. 18F-fluoride in target water from cyclotron was adsorbed onto four different forms of anion-exchange resins then desorbed by isotonic NaCl solution into the product vial. One of the resins that yielded the product at the suitable pH was used for the aseptic preparation. The components for this setup, including stopcocks, extension tubes, etc., were all single-use, individually packed and sterile. The process was done in a lead-line isolator maintained in grade A (PIC/S) aseptic condition. The quality of the obtained 'Sodium fluoride (18F) injection' was analyzed according to its monograph in the European Pharmacopoeia (EP). The resin in the chloride form yielded the product of pH 6.7 and was chosen for the subsequent preparation. The radiochemical yield was quantitative. The product met all criteria specified in EP, including biological, physical and chemical specifications. This method is an efficient, space-saving and extremely low-cost operation that easily performed in an aseptic environment meeting Good Manufacturing Practice (GMP) standard. The quality of the 'Sodium fluoride (18F) injection' so yielded meets EP specifications. This setup provides hospital with facility meeting GMP standard a cost effective and efficient method for ''Sodium fluoride (18F) injection'' production

  9. Role of Ser102 and Ser104 as Regulators of cGMP Hydrolysis by PDE5A

    Carøe Nordgaard, Julie; Kruse, Lars Schack; Gammeltoft, Steen; Kruuse, Christina Rostrup

    2014-01-01

    Background Phosphodiesterases (PDEs) cleave phosphodiester bonds in cyclic nucleotides and play diverse roles in cell biology. PDE5A is a cytoplasmic phosphodiesterase which specifically degrades cyclic guanosine monophosphate (cGMP), a cell signaling molecule that plays important roles in neuronal signaling and vascular smooth muscle contraction. Inhibition of PDE5A induces headache resembling migraine headaches. Aim To test the hypothesis that Ser102 and Ser104 in PDE5A and/or their phospho...

  10. High levels of cyclic-di-GMP in plant-associated Pseudomonas correlate with evasion of plant immunity.

    Pfeilmeier, Sebastian; Saur, Isabel Marie-Luise; Rathjen, John Paul; Zipfel, Cyril; Malone, Jacob George

    2016-05-01

    The plant innate immune system employs plasma membrane-localized receptors that specifically perceive pathogen/microbe-associated molecular patterns (PAMPs/MAMPs). This induces a defence response called pattern-triggered immunity (PTI) to fend off pathogen attack. Commensal bacteria are also exposed to potential immune recognition and must employ strategies to evade and/or suppress PTI to successfully colonize the plant. During plant infection, the flagellum has an ambiguous role, acting as both a virulence factor and also as a potent immunogen as a result of the recognition of its main building block, flagellin, by the plant pattern recognition receptors (PRRs), including FLAGELLIN SENSING2 (FLS2). Therefore, strict control of flagella synthesis is especially important for plant-associated bacteria. Here, we show that cyclic-di-GMP [bis-(3'-5')-cyclic di-guanosine monophosphate], a central regulator of bacterial lifestyle, is involved in the evasion of PTI. Elevated cyclic-di-GMP levels in the pathogen Pseudomonas syringae pv. tomato (Pto) DC3000, the opportunist P. aeruginosa PAO1 and the commensal P. protegens Pf-5 inhibit flagellin synthesis and help the bacteria to evade FLS2-mediated signalling in Nicotiana benthamiana and Arabidopsis thaliana. Despite this, high cellular cyclic-di-GMP concentrations were shown to drastically reduce the virulence of Pto DC3000 during plant infection. We propose that this is a result of reduced flagellar motility and/or additional pleiotropic effects of cyclic-di-GMP signalling on bacterial behaviour. PMID:26202381

  11. Effects of low dose radiation on cAMP ,cGMP and Fos protein expressions in mouse hypothalamus

    Objective: In the present study the authors observed the effects of low dose radiation (LDR) on cAMP, cGMP and Fos expressions in mouse hypothalamus. Methods: The changes of cAMP and cGMP contents as well as Fos protein expression in hypothalamus were determined using competitive protein binding assay, radioimmunoassay and immuno-histochemistry, respectively. Results: The cAMP content in the hypothalamus increased rapidly after irradiation with 75 mGy X-rays. It reached its peak value at 15 min, and then returned to nearly normal value at 1 h, followed by small fluctuations. The changes of cGMP content were basically opposite to those of cAMP. The Fos protein was positive in a variety of neurons of hypothalamus after irradiation, and it reached the peak value at 8 h. Then it gradually returned to the control level 48 h after irradiation. Conclusion: These findings suggest that LDR could influence the neuronal signal transduction in mouse hypothalamus with modulating functions on the immune system

  12. GMP synthase is essential for viability and infectivity of Trypanosoma brucei despite a redundant purine salvage pathway.

    Li, Qiong; Leija, Christopher; Rijo-Ferreira, Filipa; Chen, Jun; Cestari, Igor; Stuart, Kenneth; Tu, Benjamin P; Phillips, Margaret A

    2015-09-01

    The causative agent of human African trypanosomiasis, Trypanosoma brucei, lacks de novo purine biosynthesis and depends on purine salvage from the host. The purine salvage pathway is redundant and contains two routes to guanosine-5'-monophosphate (GMP) formation: conversion from xanthosine-5'-monophosphate (XMP) by GMP synthase (GMPS) or direct salvage of guanine by hypoxanthine-guanine phosphoribosyltransferase (HGPRT). We show recombinant T. brucei GMPS efficiently catalyzes GMP formation. Genetic knockout of GMPS in bloodstream parasites led to depletion of guanine nucleotide pools and was lethal. Growth of gmps null cells was only rescued by supraphysiological guanine concentrations (100 μM) or by expression of an extrachromosomal copy of GMPS. Hypoxanthine was a competitive inhibitor of guanine rescue, consistent with a common uptake/metabolic conversion mechanism. In mice, gmps null parasites were unable to establish an infection demonstrating that GMPS is essential for virulence and that plasma guanine is insufficient to support parasite purine requirements. These data validate GMPS as a potential therapeutic target for treatment of human African trypanosomiasis. The ability to strategically inhibit key metabolic enzymes in the purine pathway unexpectedly bypasses its functional redundancy by exploiting both the nature of pathway flux and the limited nutrient environment of the parasite's extracellular niche. PMID:26043892

  13. The structure of substrate-free microbial ribonuclease binase and of its complexes with 3'GMP and sulfate ions.

    Polyakov, K M; Lebedev, A A; Okorokov, A L; Panov, K I; Schulga, A A; Pavlovsky, A G; Karpeisky, M Ya; Dodson, G G

    2002-05-01

    The structures of Bacillus intermedius ribonuclease (binase), an extracellular 109-residue enzyme, and its complexes with 3'GMP and sulfate ions were solved at 1.65 and 2.0 A, respectively. The structures were refined using REFMAC. The crystal of free binase belongs to the space group C2, whereas the crystals of complexes belong to the space group P2(1)2(1)2(1). In both crystal lattices the asymmetric unit contains two molecules which form an identical dimer. The structure of the dimer is such that only one of its subunits can bind the nucleotide in the 3'GMP-binase complex, where the guanyl base is located in the recognition loop of the enzyme. In both binase complex structures the phosphate group of 3'GMP or one of the sulfate ions make an electrostatic interaction with the binase molecule at the catalytic site. A second phosphate-binding site was found in the structures of the complexes at the cleft formed by the loop 34-39, the main chain of Arg82 and the side chain of Trp34. Comparison of the complex and unliganded enzyme crystal structures shows that there are some small but distinct differences in the specificity loop (56-62) and in the loops 34-39 and 99-104 associated with the binding of the nucleotide and sulfate ions. PMID:11976484

  14. cGMP production of patient-specific iPSCs and photoreceptor precursor cells to treat retinal degenerative blindness.

    Wiley, Luke A; Burnight, Erin R; DeLuca, Adam P; Anfinson, Kristin R; Cranston, Cathryn M; Kaalberg, Emily E; Penticoff, Jessica A; Affatigato, Louisa M; Mullins, Robert F; Stone, Edwin M; Tucker, Budd A

    2016-01-01

    Immunologically-matched, induced pluripotent stem cell (iPSC)-derived photoreceptor precursor cells have the potential to restore vision to patients with retinal degenerative diseases like retinitis pigmentosa. The purpose of this study was to develop clinically-compatible methods for manufacturing photoreceptor precursor cells from adult skin in a non-profit cGMP environment. Biopsies were obtained from 35 adult patients with inherited retinal degeneration and fibroblast lines were established under ISO class 5 cGMP conditions. Patient-specific iPSCs were then generated, clonally expanded and validated. Post-mitotic photoreceptor precursor cells were generated using a stepwise cGMP-compliant 3D differentiation protocol. The recapitulation of the enhanced S-cone phenotype in retinal organoids generated from a patient with NR2E3 mutations demonstrated the fidelity of these protocols. Transplantation into immune compromised animals revealed no evidence of abnormal proliferation or tumor formation. These studies will enable clinical trials to test the safety and efficiency of patient-specific photoreceptor cell replacement in humans. PMID:27471043

  15. Role of cyclic GMP in cells with the properties of smooth muscle cultured from the rat myometrium

    Cells growing in culture with previously described properties of rat uterine smooth muscle accumulated 45Ca2+ from the medium. Ca2+ uptake by these cells was stimulated by the addition to the medium of 8-bromo-cGMP but not by 8-bromo-cAMP. Ca2+ uptake was also stimulated by carbachol and by the nitro-vasodilator nitroprusside. Although cholinergic agonists have been shown previously to stimulate contraction but not cGMP synthesis in the rat myometrium, both carbachol and nitroprusside stimulated cGMP production by the cultured cells. These results suggested the cells had cholinergic receptor-medicated functions that reflected some neurotransmitter-sensitive properties of uterine smooth muscle in situ. When determined by a specific radioligand binding assay, subcellular fractions of the cultured cells bound muscarinic cholinergic agonists and antagonists with affinities expected of the muscarinic receptor. The cells were also sensitive to the β-adrenergic catecholamine agonist isoproterenol, which stimulated cAMP production but not Ca2+ uptake. Carbachol failed to inhibit isoproterenol-dependent cAMP production, which is an important property of the cholinergic receptor in uterine smooth muscle in situ. These results suggest some but not all acetylcholine-sensitive properties of uterine smooth muscle may be retained in cell culture

  16. Homologs of the LapD-LapG c-di-GMP Effector System Control Biofilm Formation by Bordetella bronchiseptica.

    Nicolás Ambrosis

    Full Text Available Biofilm formation is important for infection by many pathogens. Bordetella bronchiseptica causes respiratory tract infections in mammals and forms biofilm structures in nasal epithelium of infected mice. We previously demonstrated that cyclic di-GMP is involved in biofilm formation in B. bronchiseptica. In the present work, based on their previously reported function in Pseudomonas fluorescens, we identified three genes in the B. bronchiseptica genome likely involved in c-di-GMP-dependent biofilm formation: brtA, lapD and lapG. Genetic analysis confirmed a role for BrtA, LapD and LapG in biofilm formation using microtiter plate assays, as well as scanning electron and fluorescent microscopy to analyze the phenotypes of mutants lacking these proteins. In vitro and in vivo studies showed that the protease LapG of B. bronchiseptica cleaves the N-terminal domain of BrtA, as well as the LapA protein of P. fluorescens, indicating functional conservation between these species. Furthermore, while BrtA and LapG appear to have little or no impact on colonization in a mouse model of infection, a B. bronchiseptica strain lacking the LapG protease has a significantly higher rate of inducing a severe disease outcome compared to the wild type. These findings support a role for c-di-GMP acting through BrtA/LapD/LapG to modulate biofilm formation, as well as impact pathogenesis, by B. bronchiseptica.

  17. Homologs of the LapD-LapG c-di-GMP Effector System Control Biofilm Formation by Bordetella bronchiseptica

    Ambrosis, Nicolás; Boyd, Chelsea D.; O´Toole, George A.; Fernández, Julieta; Sisti, Federico

    2016-01-01

    Biofilm formation is important for infection by many pathogens. Bordetella bronchiseptica causes respiratory tract infections in mammals and forms biofilm structures in nasal epithelium of infected mice. We previously demonstrated that cyclic di-GMP is involved in biofilm formation in B. bronchiseptica. In the present work, based on their previously reported function in Pseudomonas fluorescens, we identified three genes in the B. bronchiseptica genome likely involved in c-di-GMP-dependent biofilm formation: brtA, lapD and lapG. Genetic analysis confirmed a role for BrtA, LapD and LapG in biofilm formation using microtiter plate assays, as well as scanning electron and fluorescent microscopy to analyze the phenotypes of mutants lacking these proteins. In vitro and in vivo studies showed that the protease LapG of B. bronchiseptica cleaves the N-terminal domain of BrtA, as well as the LapA protein of P. fluorescens, indicating functional conservation between these species. Furthermore, while BrtA and LapG appear to have little or no impact on colonization in a mouse model of infection, a B. bronchiseptica strain lacking the LapG protease has a significantly higher rate of inducing a severe disease outcome compared to the wild type. These findings support a role for c-di-GMP acting through BrtA/LapD/LapG to modulate biofilm formation, as well as impact pathogenesis, by B. bronchiseptica PMID:27380521

  18. Procedures development and methodology of control for application of good manufacture practices (GMP) on human blood irradiation

    The irradiation of human blood is used to avoid the TA-GVHD (transfusion-associated graft-versus-host-disease), a rare but devastating adverse effect of leukocytes present in blood components for a immunocompetent transfusion recipients. Usually this irradiation practice is performed to a physical elimination of leukocytes. The implementation of the procedures will assure that the properly dose in a range of 25 Gy to 50 Gy will be delivered to the blood in the bag collected in a blood tissue bank. The studies of the procedures in order to establish a GMP (Good Manufacturing Practices) were developed under the guidelines of the standard ISO 11137 - Sterilization of health care products - Requirements for validation and routine control - Radiation sterilization. In this work, two dosimetric systems were used for dose mapping during the studies of irradiator qualification, loading pattern, irradiation process validation and auditing. The CaS04:Dy dosimeter presented difficulties concerning to uncertainty on dose measurement, stability, traceability and calibration. The PMMA and Gafchromic dosimetric systems have shown a better performance and were adopted on studies of irradiators qualification that are necessary to implementation of GMP. The irradiation tests have been done in a Gammacell 220 irradiator. The developed procedures can be adapted for different kinds of gamma irradiators, allowing implanting a quality assurance program and a GMP for blood irradiation. (author)

  19. ADOPTION OF GOOD MANAGEMENT PRACTICE (GMP IN SMALL AND MEDIUM SCALE VANNAMEI SHRIMP FARMS ON THE NORTHERN SHORE EAST JAVA

    Paula Florina

    2012-09-01

    Full Text Available The objectives of this research were to describe the value chain of Vannamei shrimp industry on the northern shore of East Java, to identify the current management practice that is being implemented by small to medium scale shrimp farmers, to analyze the level of adoption of Good Management Practice by small to medium scale shrimp farmers and to identify the challenges that prevent small to medium scale farmers from adopting Good Management Practice. This research utilized descriptive qualitative method using multiple-case study. Data collection was conducted through interviews with experts and various actors in shrimp industry and interviews and observations in shrimp farms. From this research it can be concluded that the farming activity gives the largest contribution in final product value, that the current management practice in observed intensive Vannamei shrimp farms is in line with recommendations of various institutions with the exception of waste management, that the observed intensive Vannamei shrimp farms have high adoption level of Good Management Practice (GMP for various stages of shrimp farming activity with the exception of waste management, and that there is a tendency that farmers follow the good practices that have direct impact to the success of the production, and that the main challenges of GMP adoption are the lack of a comprehensive GMP manual, the lack of structured technology dissemination, and the lack of incentive for the farmers to adopt waste management practices.Keywords: Litopenaeus Vannamei, Shrimp Farm, Good Management Practice, Value Chain, Multiple-Case Study

  20. “十三五”时期的新挑战、新机遇与新战略*--以安徽为例%The New Challenge, New Chance and New Strategy During the 13th Five-year Plan Period--A Case Study on Anhui Province’s Development Situation

    蒋晓岚; 孔令刚

    2015-01-01

    “十三五”时期是我国实现转型升级,顺利走完工业化发展阶段,打造经济升级版取得实效的关键时期,在“十三五”时期,我国将面临更为复杂多变的形势。以安徽为例,基于对“十三五”时期机遇、挑战的分析“,十三五”时期安徽将进入工业化后期和城镇化加速发展期,因此“,十三五”期间要重点实施深度城镇化战略、立体工业化战略和一、二、三产业联动融合发展战略,构建新的发展动力机制,加快推进四化协调发展下的新型工业化和新型城镇化进程。%The 13th Five-year Plan Period is the key time to realize industry transformation and upgrading, to finish the industrialization phrase favorably, to build economy upgraded version. During this period, we will face much more complicated and multivariate situation. Take Anhui as an example, this article concludes that Anhui province will reach the late stage of industrialization and the rapid developing stage of urbanization base on the analysis of the chance and challenge in the 13th five year plan period. During the 13th five year plan period, Anhui province should pay more attention to exerting profound urbanization strategy, the three-dimensional industrialization strategy and three industries interactive integration development strategy. Furthermore, Anhui province should build the new development motive mechanism, accelerate forcing the new industrialization and new urbanization under the coordinated development of the four modernizations.

  1. Crystallization studies of the murine c-di-GMP sensor protein STING

    Crystals of native and selenomethionine-labelled STING138–344 protein from M. musculus diffracted to resolutions of 2.39 and 2.2 Å, respectively The innate immune response is the first defence system against pathogenic microorganisms, and cytosolic detection of pathogen-derived DNA is believed to be one of the major mechanisms of interferon production. Recently, the mammalian ER membrane protein STING (stimulator of IFNgenes; also known as MITA, ERIS, MPYS and TMEM173) has been found to be the master regulator linking the detection of cytosolic DNA to TANK-binding kinase 1 (TBK1) and its downstream transcription factor IFN regulatory factor 3 (IRF3). In addition, STING itself was soon discovered to be a direct sensor of bacterial cyclic dinucleotides such as c-di-GMP or c-di-AMP. However, structural studies of apo STING and its complexes with these cyclic dinucleotides and with other cognate binding proteins are essential in order to fully understand the roles played by STING in these crucial signalling pathways. In this manuscript, the successful crystallization of the C-terminal domain of murine STING (STING-CTD; residues 138–344) is reported. Native and SeMet-labelled crystals were obtained and diffracted to moderate resolutions of 2.39 and 2.2 Å, respectively

  2. Automated radiosynthesis of GMP quality [18F]HX4 for PET imaging of hypoxia

    Introduction: [18F]HX4 is a 2-nitroimidazole based investigational radiotracer for imaging hypoxia. Methods: A two-step, one-pot synthetic procedure was developed on a GE Tracerlab MX-FDG with a disposable cassette. Nucleophilic substitution of a nosylate group with [18F]fluoride was followed by solvent evaporation and acidic removal of the acetyl protecting group. HPLC purification in a bio-compatible solvent mixture was developed. Results and conclusions: Using starting activities of 80–110 GBq [18F]fluoride, GMP compliant [18F]HX4 was produced in non-decay corrected radiochemical yields of 12 ± 3% (n = 9) in 55 min including HPLC purification. No reformulation steps were required. The mean specific activity of the final product was 2450 GBq/μmol. Modifications to the process and final formulation were included to prevent decomposition of the product, and these changes resulted in an improved stability of the formulated [18F]HX4, with a shelf-life of at least 8 h post-synthesis. The product consistently passed all required quality control tests to determine that the [18F]HX4 was suitable for clinical use. Using a 90 minute target bombardment, and 80–110 GBq starting [18F]fluoride, the method produced multiple patient doses

  3. Diurnal cycles in serotonin acetyltransferase activity and cyclic GMP content of cultured chick pineal glands.

    Wainwright, S D

    1980-06-12

    Levels of serotonin N-acetyltransferase (NAT: acetul CoA:arylamine N-acetyltransferase; EC 2.1.1.5.) activity in the chick pineal gland exhibit a marked diurnal variation in birds kept under a diurnal cycle of ilumination. Activity begins to rise rapidly at the start of the dark phase of the cycle and reaches maximum levels at mid-dark phase about 25-fold greater than the minimum basal level at mid-light phase. Thereafter, the level of activity declines to the basal level about the start of the light phase. This diurnal cycle in chick pineal NAT activity found in vivo has recently been reproduced in vitro with intact glands incubated in organ culture. The mechanism of the 'biological clock' which regulates these variations in level of chick pineal NAT activity is unknown. However, I now report that chick pineal glands cultured under a diurnal cycle of illumination exhibit a diurnal cycle in content of cyclic GMP which roughly parallels the cycles in NAT activity. In contrast, there was no correlation between variations in pineal content of cyclic AMP and in level of NAT activity. PMID:6250035

  4. Microneedle characterisation: the need for universal acceptance criteria and GMP specifications when moving towards commercialisation.

    Lutton, Rebecca E M; Moore, Jessica; Larrañeta, Eneko; Ligett, Stephen; Woolfson, A David; Donnelly, Ryan F

    2015-08-01

    With interest in microneedles as a novel drug transdermal delivery system increasing rapidly since the late 1990s (Margetts and Sawyer Contin Educ Anaesthesia Crit Care Pain. 7(5):171-76, 2007), a diverse range of microneedle systems have been fabricated with varying designs and dimensions. However, there are still very few commercially available microneedle products. One major issue regarding microneedle manufacture on an industrial scale is the lack of specific quality standards for this novel dosage form in the context of Good Manufacturing Practice (GMP). A range of mechanical characterisation tests and microneedle insertion analysis techniques are used by researchers working on microneedle systems to assess the safety and performance profiles of their various designs. The lack of standardised tests and equipment used to demonstrate microneedle mechanical properties and insertion capability makes it difficult to directly compare the in use performance of candidate systems. This review highlights the mechanical tests and insertion analytical techniques used by various groups to characterise microneedles. This in turn exposes the urgent need for consistency across the range of microneedle systems in order to promote innovation and the successful commercialisation of microneedle products. PMID:26022578

  5. Annual Interviews

    Human Resources Department

    2005-01-01

    Annex II, page 1, Section 3 of the Administrative Circular no. 26 (Rev. 5) states that "The annual interview shall usually take place between 15 November of the reference year and 15 February of the following year." Following the meeting of the Executive Board on 7 December 2004 and the meeting of the Standing Concertation Committee on 19 January 2005, it has been decided, for the advancement exercise of 2005, to extend this period until 15 March 2005. Human Resources Department Tel. 73566

  6. Diguanylate cyclase null mutant reveals that C-Di-GMP pathway regulates the motility and adherence of the extremophile bacterium Acidithiobacillus caldus.

    Castro, Matías; Deane, Shelly M; Ruiz, Lina; Rawlings, Douglas E; Guiliani, Nicolas

    2015-01-01

    An understanding of biofilm formation is relevant to the design of biological strategies to improve the efficiency of the bioleaching process and to prevent environmental damages caused by acid mine/rock drainage. For this reason, our laboratory is focused on the characterization of the molecular mechanisms involved in biofilm formation in different biomining bacteria. In many bacteria, the intracellular levels of c-di-GMP molecules regulate the transition from the motile planktonic state to sessile community-based behaviors, such as biofilm development, through different kinds of effectors. Thus, we recently started a study of the c-di-GMP pathway in several biomining bacteria including Acidithiobacillus caldus. C-di-GMP molecules are synthesized by diguanylate cyclases (DGCs) and degraded by phosphodiesterases (PDEs). We previously reported the existence of intermediates involved in c-di-GMP pathway from different Acidithiobacillus species. Here, we report our work related to At. caldus ATCC 51756. We identified several putative-ORFs encoding DGC and PDE and effector proteins. By using total RNA extracted from At. caldus cells and RT-PCR, we demonstrated that these genes are expressed. We also demonstrated the presence of c-di-GMP by mass spectrometry and showed that genes for several of the DGC enzymes were functional by heterologous genetic complementation in Salmonella enterica serovar Typhimurium mutants. Moreover, we developed a DGC defective mutant strain (Δc1319) that strongly indicated that the c-di-GMP pathway regulates the swarming motility and adherence to sulfur surfaces by At. caldus. Together, our results revealed that At. caldus possesses a functional c-di-GMP pathway which could be significant for ores colonization during the bioleaching process. PMID:25689133

  7. Ammonia inhibits the C-type natriuretic peptide-dependent cyclic GMP synthesis and calcium accumulation in a rat brain endothelial cell line.

    Konopacka, Agnieszka; Zielińska, Magdalena; Albrecht, Jan

    2008-05-01

    Recently we reported a decrease of C-type natriuretic peptide (CNP)-dependent, natriuretic peptide receptor 2 (NPR2)-mediated cyclic GMP (cGMP) synthesis in a non-neuronal compartment of cerebral cortical slices of hyperammonemic rats [Zielińska, M., Fresko, I., Konopacka, A., Felipo, V., Albrecht, J., 2007. Hyperammonemia inhibits the natriuretic peptide receptor 2 (NPR2)-mediated cyclic GMP synthesis in the astrocytic compartment of rat cerebral cortex slices. Neurotoxicology 28, 1260-1263]. Here we accounted for the possible involvement of cerebral capillary endothelial cells in this response by measuring the effect of ammonia on the CNP-mediated cGMP formation and intracellular calcium ([Ca2+]i) accumulation in a rat cerebral endothelial cell line (RBE-4). We first established that stimulation of cGMP synthesis in RBE-4 cells was coupled to protein kinase G (PKG)-mediated Ca2+ influx from the medium which was inhibited by an L-type channel blocker nimodipine. Ammonia treatment (1h, 5mM NH4Cl) evoked a substantial decrease of CNP-stimulated cGMP synthesis which was related to a decreased binding of CNP to NPR2 receptors, and depressed the CNP-dependent [Ca2+]i accumulation in these cells. Ammonia also abolished the CNP-dependent Ca2+ accumulation in the absence of Na+. In cells incubated with ammonia in the absence of Ca2+ a slight CNP-dependent increase of [Ca2+]i was observed, most likely representing Ca2+ release from intracellular stores. Depression of CNP-dependent cGMP-mediated [Ca2+]i accumulation may contribute to cerebral vascular endothelial dysfunction associated with hyperammonemia or hepatic encephalopathy. PMID:18222015

  8. Diguanylate cyclase null mutant reveals that C-Di-GMP pathway regulates the motility and adherence of the extremophile bacterium Acidithiobacillus caldus.

    Matías Castro

    Full Text Available An understanding of biofilm formation is relevant to the design of biological strategies to improve the efficiency of the bioleaching process and to prevent environmental damages caused by acid mine/rock drainage. For this reason, our laboratory is focused on the characterization of the molecular mechanisms involved in biofilm formation in different biomining bacteria. In many bacteria, the intracellular levels of c-di-GMP molecules regulate the transition from the motile planktonic state to sessile community-based behaviors, such as biofilm development, through different kinds of effectors. Thus, we recently started a study of the c-di-GMP pathway in several biomining bacteria including Acidithiobacillus caldus. C-di-GMP molecules are synthesized by diguanylate cyclases (DGCs and degraded by phosphodiesterases (PDEs. We previously reported the existence of intermediates involved in c-di-GMP pathway from different Acidithiobacillus species. Here, we report our work related to At. caldus ATCC 51756. We identified several putative-ORFs encoding DGC and PDE and effector proteins. By using total RNA extracted from At. caldus cells and RT-PCR, we demonstrated that these genes are expressed. We also demonstrated the presence of c-di-GMP by mass spectrometry and showed that genes for several of the DGC enzymes were functional by heterologous genetic complementation in Salmonella enterica serovar Typhimurium mutants. Moreover, we developed a DGC defective mutant strain (Δc1319 that strongly indicated that the c-di-GMP pathway regulates the swarming motility and adherence to sulfur surfaces by At. caldus. Together, our results revealed that At. caldus possesses a functional c-di-GMP pathway which could be significant for ores colonization during the bioleaching process.

  9. Characterization of the cGMP-dependent protein kinase SmcGK1 of Schistosoma mansoni

    Silke Leutner

    2011-06-01

    Full Text Available Schistosomes are trematode parasites and of worldwide medical importance for humans and animals. Growth and development of these parasites require a specific host environment, but also permanent communication processes between the two genders. Accumulating molecular evidence indicates that the responsible interactions are mediated by signal transduction processes. Conserved signaling molecules were identified, and first approaches made for their characterization. However, no representative of the conserved family of cGMP-dependent protein kinases (cGKs has been described in this parasite yet. Within the Schistosoma mansoni genome data-set we identified cGK homologs, of which one was investigated in more detail in this study. We present the cloning of SmcGK1, whose sequence shows homology to cGKs of higher eukaryotes. SmcGK1 was found to be gender-independently transcribed in adult schistosomes. The occurrence of SmcGK1 sense and antisense transcripts suggests that the expression of this gene is controlled at the post-transcriptional level. In situ hybridization experiments demonstrated a gonad-preferential expression profile in both genders indicating a role of SmcGK1, at least during sexual development of schistosomes. Using a cGK-specific inhibitor to treat adult schistosomes in vitro finally resulted in a multifaceted phenotype including slow motion, oocyte congestion, and reduced egg production.Esquistossomos são parasitas trematodos de importância médica em todo o mundo para o homem e os animais. O crescimento e o desenvolvimento destes parasitas requerem um ambiente específico do hospedeiro, mas também um processo de comunicação permanente entre parasitas dos dois sexos. Evidência molecular tem se acumulado e indica que as interações são mediadas por processos de transdução de sinal. Moléculas sinalizadoras conservadas foram identificadas, e as primeiras abordagens têm sido feitas para sua caracterização. Contudo, não foi

  10. GMP-compliant isolation and large-scale expansion of bone marrow-derived MSC.

    Natalie Fekete

    Full Text Available BACKGROUND: Mesenchymal stromal cells (MSC have gained importance in tissue repair, tissue engineering and in immunosupressive therapy during the last years. Due to the limited availability of MSC in the bone marrow, ex vivo amplification prior to clinical application is requisite to obtain therapeutic applicable cell doses. Translation of preclinical into clinical-grade large-scale MSC expansion necessitates precise definition and standardization of all procedural parameters including cell seeding density, culture medium and cultivation devices. While xenogeneic additives such as fetal calf serum are still widely used for cell culture, its use in the clinical context is associated with many risks, such as prion and viral transmission or adverse immunological reactions against xenogeneic components. METHODS AND FINDINGS: We established animal-free expansion protocols using platelet lysate as medium supplement and thereby could confirm its safety and feasibility for large-scale MSC isolation and expansion. Five different GMP-compliant standardized protocols designed for the safe, reliable, efficient and economical isolation and expansion of MSC was performed and MSC obtained were analyzed for differentiation capacity by qPCR and histochemistry. Expression of standard MSC markers as defined by the International Society for Cellular Therapy as well as expression of additional MSC markers and of various chemokine and cytokine receptors was analysed by flow cytometry. Changes of metabolic markers and cytokines in the medium were addressed using the LUMINEX platform. CONCLUSIONS: The five different systems for isolation and expansion of MSC described in this study are all suitable to produce at least 100 millions of MSC, which is commonly regarded as a single clinical dose. Final products are equal according to the minimal criteria for MSC defined by the ISCT. We showed that chemokine and integrin receptors analyzed had the same expression pattern

  11. The impact of cGMP compliance on consumer confidence in dietary supplement products.

    Crowley, Richard; FitzGerald, Libby Harvey

    2006-04-01

    The FDA estimates that US citizens spend more than $ 8.5 billion a year on dietary supplements and world wide the market is estimated at more than $ 60 billion. However, although a majority of consumers express confidence in the safety of these products, 74% believe the government should be more involved in ensuring that these products are safe and efficacious. Recent regulatory initiatives such as the imminent adoption of cGMPs for dietary supplements in the US, implementation of cGMPs in Canada and the recent EU dietary supplement initiative represent legislative and industry response to public clamor for more comprehensive oversight of dietary supplements. Regardless of mandated practices, the majority of dietary supplement manufacturers have done an excellent job of protecting the safety and quality of their products. The promulgation of these cGMPs will help ensure consumers that equal standards are followed throughout the industry. For some companies with established processes based on existing food or pharmaceutical cGMP regulations, the transition will be relatively painless while, for many, it will represent a significant increase in the level of documentation and testing. However, consumers deserve and demand that products meet standards for safety and quality and the implementation of cGMPs for these products are an important first step. Although the cGMPs are designed to ensure products are safe from a standpoint of identity, purity, quality, strength and composition, they do not address preclinical or clinical testing of ingredients for safety or efficacy. This would involve ingredients meeting the requirements of Generally Recognized as Safe (GRAS) status or going through the New Dietary Ingredient (NDI) process. PMID:16469425

  12. The impact of cGMP compliance on consumer confidence in dietary supplement products

    The FDA estimates that US citizens spend more than $ 8.5 billion a year on dietary supplements and world wide the market is estimated at more than $ 60 billion. However, although a majority of consumers express confidence in the safety of these products, 74% believe the government should be more involved in ensuring that these products are safe and efficacious. Recent regulatory initiatives such as the imminent adoption of cGMPs for dietary supplements in the US, implementation of cGMPs in Canada and the recent EU dietary supplement initiative represent legislative and industry response to public clamor for more comprehensive oversight of dietary supplements. Regardless of mandated practices, the majority of dietary supplement manufacturers have done an excellent job of protecting the safety and quality of their products. The promulgation of these cGMPs will help ensure consumers that equal standards are followed throughout the industry. For some companies with established processes based on existing food or pharmaceutical cGMP regulations, the transition will be relatively painless while, for many, it will represent a significant increase in the level of documentation and testing. However, consumers deserve and demand that products meet standards for safety and quality and the implementation of cGMPs for these products are an important first step. Although the cGMPs are designed to ensure products are safe from a standpoint of identity, purity, quality, strength and composition, they do not address preclinical or clinical testing of ingredients for safety or efficacy. This would involve ingredients meeting the requirements of Generally Recognized as Safe (GRAS) status or going through the New Dietary Ingredient (NDI) process

  13. Annual budget

    This paper reports that all businesses, including individuals, should have a budget - that is, an estimation of income and expense over an annual cycle. For companies, the budget is generally prepared and approved about one quarter before the start of the company's fiscal year and is updated and revised each quarter during the year. Although budgeting is a task dreaded by most exploration managers, it is usually the vehicle by which drilling prospects, the heart of any exploration program, are sold to the final decision makers. The budgeting process should be viewed as an opportunity rather than as a chore to be completed as quickly as possible

  14. Time-dependent inhibitory effects of cGMP-analogues on thrombin-induced platelet-derived microparticles formation, platelet aggregation, and P-selectin expression

    Nygaard, Gyrid [Proteomic Unit at University of Bergen (PROBE), University of Bergen, Bergen (Norway); Department of Biomedicine, University of Bergen, Bergen (Norway); Herfindal, Lars; Kopperud, Reidun [Department of Biomedicine, University of Bergen, Bergen (Norway); Aragay, Anna M. [Department of Biomedicine, University of Bergen, Bergen (Norway); Molecular Biology Institute of Barcelona (IBMB, CSIC), Barcelona (Spain); Holmsen, Holm; Døskeland, Stein Ove; Kleppe, Rune [Department of Biomedicine, University of Bergen, Bergen (Norway); Selheim, Frode, E-mail: Frode.Selheim@biomed.uib.no [Proteomic Unit at University of Bergen (PROBE), University of Bergen, Bergen (Norway); Department of Biomedicine, University of Bergen, Bergen (Norway)

    2014-07-04

    Highlights: • We investigated the impact of cyclic nucleotide analogues on platelet activation. • Different time dependence were found for inhibition of platelet activation. • Additive effect was found using PKA- and PKG-activating analogues. • Our results may explain some of the discrepancies reported for cNMP signalling. - Abstract: In platelets, nitric oxide (NO) activates cGMP/PKG signalling, whereas prostaglandins and adenosine signal through cAMP/PKA. Cyclic nucleotide signalling has been considered to play an inhibitory role in platelets. However, an early stimulatory effect of NO and cGMP-PKG signalling in low dose agonist-induced platelet activation have recently been suggested. Here, we investigated whether different experimental conditions could explain some of the discrepancy reported for platelet cGMP-PKG-signalling. We treated gel-filtered human platelets with cGMP and cAMP analogues, and used flow cytometric assays to detect low dose thrombin-induced formation of small platelet aggregates, single platelet disappearance (SPD), platelet-derived microparticles (PMP) and thrombin receptor agonist peptide (TRAP)-induced P-selectin expression. All four agonist-induced platelet activation phases were blocked when platelets were costimulated with the PKG activators 8-Br-PET-cGMP or 8-pCPT-cGMP and low-doses of thrombin or TRAP. However, extended incubation with 8-Br-PET-cGMP decreased its inhibition of TRAP-induced P-selectin expression in a time-dependent manner. This effect did not involve desensitisation of PKG or PKA activity, measured as site-specific VASP phosphorylation. Moreover, PKG activators in combination with the PKA activator Sp-5,6-DCL-cBIMPS revealed additive inhibitory effect on TRAP-induced P-selectin expression. Taken together, we found no evidence for a stimulatory role of cGMP/PKG in platelets activation and conclude rather that cGMP/PKG signalling has an important inhibitory function in human platelet activation.

  15. Radiological safety and GMP in the bulk batch manufacturing, formulation and dispensing of radiopharmaceuticals

    Full text: Radiopharmaceutical Program of BRIT is involved in the manufacturing of ready to use radiopharmaceuticals for therapy for the last three decades. Ready to use Radiopharmaceutical products include, 131I-Sodium iodide solution and capsules for thyroid carcinoma and metastatic lesions, 131I MIBG injection for diagnosis and therapy of adrenal medullae tumors and their mets, 153Sm-EDTMP injection and 32P- Sodium orthophosphate injection for bone pain palliative treatment. BRIT's radiopharmaceutical production facility is a radioisotope laboratory classified as Type-III facility for handling radioisotopes of Group-II, Group-III and Group-IV approved by AERB. This facility meets all the radiological safety requirements as per AERB guidelines. Production of above mentioned radiopharmaceuticals is carried out in Production Plants (PP), β, γ Glove boxes (GB) and Fume hoods (FH). Typical production procedure involves bulk processing, formulation, sterilization and dispensing of doses. Production Plants (PP) are exclusively designed facilities to carry out the production in a radiologically safe manner, at the same time maintaining aseptic conditions required for injectables as per the current GMP. Each production plant has a leak tight Isolator box, made up of high quality SS which has provisions for remote handling devices like, Tongs, dispensing systems, service points for vacuum, gas, compressed air, water and electric power. This Isolator box is shielded from all sides by required amount of lead (2 inch or 4 inch), and has an access port called 'Transport Port Box' with double door transport lock and is equipped with a trolley. Two filter unit systems are fitted at the top of the plant and each unit comprises of activated Charcoal filter and HEPA filter in tandem, this in turn is connected to special exhaust meant for radioactive gases. Similarly, the designs of β, γ Glove boxes (GB) and Fume hoods (FH) also incorporate all radiological safety features to

  16. Free and total GMP (glycomacropeptide contents of milk during bovine lactation Variação dos teores de GMP (glicomacropeptídeo livre e total no leite bovino durante a lactação

    Andréa Maria Furlanetti

    2003-12-01

    Full Text Available Individual milk samples taken every two weeks from parturition to the end of lactation from 34 animals of three different herds and breeds were analyzed for free-GMP. A milk pool of each herd was analyzed for free and total GMP (released from k-casein by the action of rennin and the data were correlated with sanitary conditions of animal and udder, phase of lactation and milk production. Most udder problems were concentrated near parturition, with few and spaced occurrences of clinical mastitis. The Californian Mastitis Test (CMT results showed oscillations compatible with the phases of lactation period and environmental conditions. The widest variations in free-GMP occurred as a function of lactation period and as a consequence of clinical or subclinical mastitis. Higher levels were observed at the beginning of lactation (5.87mg L-1 of sialic acid, becoming normal with mean values of about 3.30mg L-1 at the end of the second month, and increasing again during the final third of lactation. On average, the same trends were observed for total GMP released by commercial rennet, beginning with slightly high values (35.59mg L-1, becoming normal by the sixth month with values close to 27.15mg L-1, and rising gradually up to the end of lactation, with 58.35mg L-1 of sialic acid. These results prove to be useful for the correct interpretation of tests applied to milk selection with respect to proteolytic status or even to restrain frauds by the addition of whey to milk.Amostras quinzenais, desde o parto até o final do período de lactação, obtidas de 34 vacas de três diferentes raças e propriedades, foram analisadas quanto à presença de GMP livre. Um "pool" das amostras quinzenais de cada rebanho foi analisada tanto para o conteúdo de GMP livre quanto para o GMP total (liberado da k-caseína pela ação da renina, correlacionando-os com as condições sanitárias do animal e do úbere, à fase da lactação e à produção de leite. A maioria

  17. Regulation of Motility and Phenazine Pigment Production by FliA Is Cyclic-di-GMP Dependent in Pseudomonas aeruginosa PAO1

    Lo, Yi-Ling; Shen, Lunda; Chang, Chih-Hsuan; Bhuwan, Manish; Chiu, Cheng-Hsun; Chang, Hwan-You

    2016-01-01

    The transcription factor FliA, also called sigma 28, is a major regulator of bacterial flagellar biosynthesis genes. Growing evidence suggest that in addition to motility, FliA is involved in controlling numerous bacterial behaviors, even though the underlying regulatory mechanism remains unclear. By using a transcriptional fusion to gfp that responds to cyclic (c)-di-GMP, this study revealed a higher c-di-GMP concentration in the fliA deletion mutant of Pseudomonas aeruginosa than in its wild-type strain PAO1. A comparative analysis of transcriptome profiles of P. aeruginosa PAO1 and its fliA deletion mutant revealed an altered expression of several c-di-GMP-modulating enzyme-encoding genes in the fliA deletion mutant. Moreover, the downregulation of PA4367 (bifA), a Glu-Ala-Leu motif-containing phosphodiesterase, in the fliA deletion mutant was confirmed using the β-glucuronidase reporter gene assay. FliA also altered pyocyanin and pyorubin production by modulating the c-di-GMP concentration. Complementing the fliA mutant strain with bifA restored the motility defect and pigment overproduction of the fliA mutant. Our results indicate that in addition to regulating flagellar gene transcription, FliA can modulate the c-di-GMP concentration to regulate the swarming motility and phenazine pigment production in P. aeruginosa. PMID:27175902

  18. Hypercapnic vasodilatation in isolated rat basilar arteries is exerted via low pH and does not involve nitric oxide synthase stimulation or cyclic GMP production

    You, J P; Wang, Qian; Zhang, W; Jansen-Olesen, I; Paulson, O B; Lassen, N A; Edvinsson, L

    1994-01-01

    relaxation by 54% and 70%, respectively. The effect of L-NOARG was completely reversed by L-arginine. Blockade of nerve excitation with tetrodotoxin (TTX) had no affect on the 15% CO2 elicited vasodilatation. Measurements of cGMP in vessel segments showed no significant increase in cGMP content in response...... to hypercapnia. L-NOARG and MB, but not TTX, significantly reduced the basal cGMP content in cerebral vessels. Adding 1.5% halothane to the incubation medium did not result in a significant increase in cGMP content. Lowering the pH by cumulative application of 0.12 M HCl resulted in relaxation...... that high CO2 elicits vasodilatation of isolated rat basilar arteries by a mechanism independent of nitric oxide synthase (NOS) activity. The markedly reduced basal cGMP levels in cerebral vessels by L-NOARG and MB suggest that there exists a basal NO formation in the cerebral vessel wall....

  19. Hyperammonemia alters the modulation by different neurosteroids of the glutamate-nitric oxide-cyclic GMP pathway through NMDA- GABAA - or sigma receptors in cerebellum in vivo.

    González-Usano, Alba; Cauli, Omar; Agustí, Ana; Felipo, Vicente

    2013-04-01

    Several neurosteroids modulate the glutamate-nitric oxide (NO)-cGMP pathway in cerebellum through modulation of NMDA- GABAA - or sigma receptors. Hyperammonemia alters the concentration of several neurosteroids and impairs the glutamate-NO-cGMP pathway, leading to impaired learning ability. This work aimed to assess whether chronic hyperammonemia alters the modulation by different neurosteroids of GABAA, NMDA, and/or sigma receptors and of the glutamate-NO-cGMP pathway in cerebellum. Neurosteroids were administered through microdialysis probes, and extracellular cGMP and citrulline were measured. Then NMDA was administered to assess the effects on the glutamate-NO-cGMP pathway activation. Hyperammonemia completely modifies the effects of pregnanolone and pregnenolone. Pregnanolone acts as a GABAA receptor agonist in controls, but as an NMDA receptor antagonist in hyperammonemic rats. Pregnenolone does not induce any effect in controls, but acts as a sigma receptor agonist in hyperammonemic rats. Hyperammonemia potentiates the actions of tetrahydrodeoxy-corticosterone (THDOC) as a GABAA receptor agonist, allopregnanolone as an NMDA receptor antagonist, and pregnenolone sulfate as an NMDA receptor activation enhancer. Neurosteroids that reduce the pathway (pregnanolone, THDOC, allopregnanolone, DHEAS) may contribute to cognitive impairment in hyperammonemia and hepatic encephalopathy. Pregnenolone would impair cognitive function in hyperammonemia. Neurosteroids that restore the pathway in hyperammonemia (pregnenolone sulfate) could restore cognitive function in hyperammonemia and encephalopathy. PMID:23227932

  20. Participation of the NO/cGMP/K+ATP pathway in the antinociception induced by Walker tumor bearing in rats

    Implantation of Walker 256 tumor decreases acute systemic inflammation in rats. Inflammatory hyperalgesia is one of the most important events of acute inflammation. The L-arginine/NO/cGMP/K+ATP pathway has been proposed as the mechanism of peripheral antinociception mediated by several drugs and physical exercise. The objective of this study was to investigate a possible involvement of the NO/cGMP/K+ATP pathway in antinociception induced in Walker 256 tumor-bearing male Wistar rats (180-220 g). The groups consisted of 5-6 animals. Mechanical inflammatory hypernociception was evaluated using an electronic version of the von Frey test. Walker tumor (4th and 7th day post-implantation) reduced prostaglandin E2- (PGE2, 400 ng/paw; 50 µL; intraplantar injection) and carrageenan-induced hypernociception (500 µg/paw; 100 µL; intraplantar injection). Walker tumor-induced analgesia was reversed (99.3% for carrageenan and 77.2% for PGE2) by a selective inhibitor of nitric oxide synthase (L-NAME; 90 mg/kg, ip) and L-arginine (200 mg/kg, ip), which prevented (80% for carrageenan and 65% for PGE2) the effect of L-NAME. Treatment with the soluble guanylyl cyclase inhibitor ODQ (100% for carrageenan and 95% for PGE2; 8 µg/paw) and the ATP-sensitive K+ channel (KATP) blocker glibenclamide (87.5% for carrageenan and 100% for PGE2; 160 µg/paw) reversed the antinociceptive effect of tumor bearing in a statistically significant manner (P < 0.05). The present study confirmed an intrinsic peripheral antinociceptive effect of Walker tumor bearing in rats. This antinociceptive effect seemed to be mediated by activation of the NO/cGMP pathway followed by the opening of KATP channels

  1. Studies of the effect of glyceryl trinitrate and cyclic GMP on calcium turnover in bovine mesenteric artery

    It was recently observed that the relaxation induced by glyceryl trinitrate (GTN) showed a biphasic concentration-response curve; a high-sensitivity component represented by concentrations 1 nM. The effect of two glyceryl trinitrate concentrations (0.1 nM and 1 μM) were tested on the uptake of 45Ca2+ to tissue pieces of bovine mesenteric arteries (BMA) as well as on the uptake of 45Ca2+ to a microsomal preparation of BMA. The effect of GTN and 8-Br-cGMP was also studied on the IP3-induced release of Ca2+ from the microsomal preparation preloaded with 45Ca2+. The influence of IP3 and GTN on the activity of Ca2+-ATPase in the microsomal preparation was tested as well. The phenylephrine-stimulated uptake of Ca2+ to tissue pieces of BMA was significantly reduced by the high GTN-concentration (1 μM) but not by the lower concentration. The uptake of Ca2+ to the microsomal preparation was significantly stimulated by the two GTN-concentrations tested, as well as by 8-Br-cGMP (0.1 mM). The calcium release induced by IP3 (1 μM) from the microsomal preparation was inhibited by both the low and the high GTN-concentration and by 8-Br-cGMP (0.1 mM). The Ca2+-ATPase activity was stimulated by both GTN-concentrations tested while it was inhibitedby IP3. It is concluded that GTN is able to induce a reduction of the free intracellular Ca2+ by several mechanisms, which are of importance for the relaxation represented by the high-affinity component. The low-affinity component in addition reduces the inflow of Ca2+ over the plasma membrane. (author)

  2. Lentivirus-induced 'Smart' dendritic cells: Pharmacodynamics and GMP-compliant production for immunotherapy against TRP2-positive melanoma.

    Sundarasetty, B S; Chan, L; Darling, D; Giunti, G; Farzaneh, F; Schenck, F; Naundorf, S; Kuehlcke, K; Ruggiero, E; Schmidt, M; von Kalle, C; Rothe, M; Hoon, D S B; Gerasch, L; Figueiredo, C; Koehl, U; Blasczyk, R; Gutzmer, R; Stripecke, R

    2015-09-01

    Monocyte-derived conventional dendritic cells (ConvDCs) loaded with melanoma antigens showed modest responses in clinical trials. Efficacy studies were hampered by difficulties in ConvDC manufacturing and low potency. Overcoming these issues, we demonstrated higher potency of lentiviral vector (LV)-programmed DCs. Monocytes were directly induced to self-differentiate into DCs (SmartDC-TRP2) upon transduction with a tricistronic LV encoding for cytokines (granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4)) and a melanoma antigen (tyrosinase-related protein 2 (TRP2)). Here, SmartDC-TRP2 generated with monocytes from five advanced melanoma patients were tested in autologous DC:T cell stimulation assays, validating the activation of functional TRP2-specific cytotoxic T lymphocytes (CTLs) for all patients. We described methods compliant to good manufacturing practices (GMP) to produce LV and SmartDC-TRP2. Feasibility of monocyte transduction in a bag system and cryopreservation following a 24-h standard operating procedure were achieved. After thawing, 50% of the initial monocyte input was recovered and SmartDC-TRP2 self-differentiated in vitro, showing uniform expression of DC markers, detectable LV copies and a polyclonal LV integration pattern not biased to oncogenic loci. GMP-grade SmartDC-TRP2 expanded TRP2-specific autologous CTLs in vitro. These results demonstrated a simpler GMP-compliant method of manufacturing an effective individualized DC vaccine. Such DC vaccine, when in combination with checkpoint inhibition therapies, might provide higher specificity against melanoma. PMID:25965393

  3. The Role of Aquaporin 1 Activated by cGMP in Myocardial Edema Caused by Cardiopulmonary Bypass in Sheep

    Fang-bao Ding

    2013-11-01

    Full Text Available Background/Aims: Most cardiac procedures involve the use of cardiopulmonary bypass (CPB, which pumps oxygenated blood to the body while the heart and lungs are isolated. CPB can cause profound alterations V in the homeostasis of physiological fluids, which often results in myocardial edema. In our study, we used sheep CPB model of in vivo and in vitro to assess the relationship between cGMP and AQP1 during CPB. Methods: ODQ, a specific inhibitor of soluble guanylate cyclase (sGC, was used to treat the CPB animals or cardiomyocytes. Left ventricular function of each group was determined by pressure-volume system. Water content of myocardial tissue was assessed by dry-wet weight, and cardiomyocytes water permeability was also calculated. The concentration of cGMP was determined by Radioimmunoassay (RIA. mRNA and protein expression of AQP1 were detected by real-time PCR and western blot, respectively. Results: The relative expression level of AQP1 mRNA and protein at each time point (0, 6, 12, 24 or 48 h after CPB was significantly increased (1.18-fold at 12 h, 1.77-fold at 24 h and 2.18-fold at 48h compared with each sham group, the protein expression of AQP1 also showed a rising trend after CPB. The degree of myocardial edema (75.1% at 12 h, 79.3% at 24 h and 81.0% at 48h increased following the CPB surgery. The mRNA expression level of AQP1 was significantly decreased by 39.7% (pin vitro experiments showed the same changing trends as in vivo. Conclusion: cGMP pathway controls water channels and then affects water intake during CPB through an AQP1-mediated pathway.

  4. Food GMP Certification System in Taiwan%我国台湾的食品GMP认证制度

    一泓

    2002-01-01

    @@ 1食品GMP概况 1.1食品GMP的概念 GMP是英文Good Manufacturing Practice的缩写,中文的意思是"良好作业规范",或是"优良制造标准",是一种特别注重制造过程中产品质量与卫生安全的自主性管理制度.当它用于食品工业管理时,称作食品GMP.

  5. Receptor-mediated rapid action of 1 alpha,25-dihydroxycholecalciferol: increase of intracellular cGMP in human skin fibroblasts.

    Barsony, J; Marx, S. J.

    1988-01-01

    The intracellular cGMP concentration in normal human cultured fibroblasts was increased 2- to 3-fold by 1 alpha,25-dihydroxycholecalciferol [1 alpha,25-(OH)2D3] in a dose-dependent manner between 0.01 nM and 1 microM. The response was detectable within 1 min, reached a maximum (225% +/- 8% of baseline) at 6-8 min, and was no longer detectable at 30 min. The half-maximal effect of 1 alpha,25-(OH)2D3 was at 1.8 nM, and 24,25-dihydroxycholecalciferol showed an estimated EC50 100-fold higher. 1 b...

  6. Development and Application of GMP Management Information System%兽药GMP信息管理系统的开发与应用

    罗舜庭; 童伟; 谭志坚; 徐桓; 宋艳丽

    2012-01-01

    This paper introduced good manufacture practice MIS of veterinary drug based on the management of GMP and the products' procedures, presented the development of the GMP MIS and its applications in veterinary drugs enterprises and the system' s advantages. Finally this paper indicated the meaning and the prospect of the GMP informatization system in the veterinary drugs enterprises.%介绍了基于兽药GMP以生产质量管理为核心的兽药GMP信息管理系统的研发背景、系统特点以及其在兽药生产企业的应用情况,并对其在兽药生产企业信息化过程中的意义和前景进行了阐述,以期为兽药生产管理的信息化建设提供参考。

  7. Analysis of guanosine-5'-monophosphate at nM level based on the co-luminescence effect of Tb-Gd-GMP system

    This paper first found the co-luminescence effect of guanosine-5'-monophosphate (GMP) system. Experiment showed that La3+, Gd3+, Sc3+ and Y3+ all could enhance the luminescence of Tb-GMP system, among which Gd3+ has the greatest enhancement. Under the optimum conditions, the enhanced intensity of the system is in proportion to the concentration of GMP in the range from 3x10-8 to 3x10-5 mol/l. The detection limit is 3.9x10-9 mol/l. In the study of mechanism, we propose that both Tb and Gd complexes can form bigger netlike compound by the link of the oxygen bridge, through which the energy absorbed by Gd complex may transfer to Tb3+ in Tb complex

  8. GMP-compliant automated synthesis of [18F]AV-45 (Florbetapir F 18) for imaging β-amyloid plaques in human brain

    We report herein the Good Manufacturing Practice (GMP)-compliant automated synthesis of 18F-labeled styrylpyridine, AV-45 (Florbetapir), a novel tracer for positron emission tomography (PET) imaging of β-amyloid (Aβ) plaques in the brain of Alzheimer's disease patients. [18F]AV-45 was prepared in 105 min using a tosylate precursor with Sumitomo modules for radiosynthesis under GMP-compliant conditions. The overall yield was 25.4±7.7% with a final radiochemical purity of 95.3±2.2% (n=19). The specific activity of [18F]AV-45 reached as high as 470±135 TBq/mmol (n=19). The present studies show that [18F]AV-45 can be manufactured under GMP-compliant conditions and could be widely available for routine clinical use.

  9. Potentiation of cGMP signaling increases oxygen delivery and oxidative metabolism in contracting skeletal muscle of older but not young humans

    Nyberg, Michael; Piil, Peter; Egelund, Jon;

    2015-01-01

    Aging is associated with progressive loss of cardiovascular and skeletal muscle function. The impairment in physical capacity with advancing age could be related to an insufficient peripheral O2 delivery to the exercising muscles. Furthermore, the mechanisms underlying an impaired blood flow...... regulation remain unresolved. Cyclic guanosine monophosphate (cGMP) is one of the main second messengers that mediate smooth muscle vasodilation and alterations in cGMP signaling could, therefore, be one mechanism by which skeletal muscle perfusion is impaired with advancing age. The current study aimed to...... evaluate the effect of inhibiting the main enzyme involved in cGMP degradation, phosphodiesterase 5 (PDE5), on blood flow and O2 delivery in contracting skeletal muscle of young and older humans. A group of young (23 ± 1 years) and a group of older (72 ± 2 years) male human subjects performed submaximal...

  10. Annual report

    This is the thirty-ninth annual report of the Atomic Energy Control Board. The period covered by this report is the year ending March 31, 1986. The Atomic Energy Control Board (AECB) was established in 1946, by the Atomic Energy Control Act (AEC Act), (Revised Statues of Canada (R.S.C.) 1970 cA19). It is a departmental corporation (Schedule B) within the meaning and purpose of the Financial Administration Act. The AECB controls the development, application and use of atomic energy in Canada, and participates on behalf of Canada in international measures of control. The AECB is also repsonsible for the administration of the Nuclear Liability Act, (R.S.C. 1970 c29 1st Supp) as amended, including the designation of nuclear installations and the prescription of basic insurance to be carried by the operators of such nuclear installations. The AECB reports to Parliament through a designated Minister, currently the Minister of Energy, Mines and Resources

  11. Role of NO-cGMP pathway in ovine cervical relaxation induced by Erythroxylum caatingae Plowman.

    Santos, K C; Monte, A P O; Lima, J T; Ribeiro, L A A; Palheta Junior, R C

    2016-01-01

    Erythroxylum caatingae Plowman has a myorelaxing effect on smooth muscle tissue. We investigated the effect of the crude ethanolic extract of E. caatingae Plowman (Ec-EtOH) on the contractility of the ovine cervix. In an isometric system, circular strips were subjected to 90mM potassium (K(+)) or 30μM carbamylcholine (CCh)-induced contraction. We then exposed the tissue to cumulative concentrations of Ec-EtOH (1-729 μg/ml). In other bath solutions, the tissues were exposed to l-NG-nitroarginine methyl ester (l-NAME; 100μM), l-NAME (100μM)+l-arginine (300μM), 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, ODQ; 5μM), 4-aminopyridine (4-AP; 3mM), tetraethylammonium (TEA; 0.3mM), glybenclamide (1μM), atosiban (10μM) or verapamil (3μM), followed by the addition of Ec-EtOH (1-729 μg/ml). We also evaluated the effect of cervical Ec-EtOH infusion (2mg) on cervical contractility in vivo. Ec-EtOH decreased cervical contractility induced by K(+) or CCh, and 729 μg/ml Ec-EtOH decreased 85.4±5.1% the amplitude of basal contractility in vitro, with an EC50 of 17.9±3.7 μg/ml. This effect of Ec-EtOH was prevented by l-NAME or ODQ. l-arginine impaired the blunting effect of l-NAME on cervical relaxation caused by Ec-EtOH. However, the potassium channel blockers 4-AP, TEA, and glybenclamide did not modify this myorelaxation triggered by Ec-EtOH. Ec-EtOH also decreased acetylcholine-induced contractions in tissue preincubated with verapamil. In addition, Ec-EtOH decreased ovine cervical contractions in vivo. Thus, Ec-EtOH had a relaxant effect on ovine cervical contractions. This may involve the nitric oxide signal, mediated by cGMP cellular transduction, and be related to intracellular calcium sequestration. PMID:26619941

  12. 13th AINSE radiation biology conference: conference handbook

    The forty one papers presented at this conference covered the areas of radiation induced lesions, apoptosis, genetics and radiobiological consequences of low level radiation exposure, clinical applications of radiation, mammalian cells radiosensitivity and radiation-activated proteins

  13. MANAGEMENT BOARD MEETING OF 13th MARCH 2003

    2003-01-01

    Status of Earned Value Management (EVM) J. Ferguson, the EVM Project Leader, reported that the concerted EVM data collection phase that had begun in January was nearing completion, with the last of the 23 groups concerned having begun the data collection process and data corresponding to over 85% of the overall LHC cost-to-completion figure now incorporated in the system. The project team was working closely with the groups that had yet to submit their full data to ensure that the exercise was completed by Easter. The period leading up to the June meetings of the Council and its committees, at which the first EVM report was due to be presented, would then be devoted to tuning the data and ensuring consistent reporting across all the groups concerned. Preparation of the March Meetings of the Council and its Committees The Director-General stated that, in addition to regular items of business, such as the approval of the 2003 Budget at 2003 prices, contract adjudications and a progress report on technology tra...

  14. Prague Overview Week, September 13th-20th, 2003

    Smith, K

    The week already began on Saturday for a number of system and sub-system meetings, but got into full swing on the Sunday, with a marathon session on how to tackle the challenging task of commissioning the full ATLAS spectrometer. The seeds of lots of ideas have already germinated and should begin to flower soon. Simulation studies of cosmic ray fluxes (and even early experimental measurements in Pit 1) have shown useful rates for the pre-beam phase of commissioning, and single beam measurements will also offer useful opportunities, in particular for trigger timing. Commissioning is clearly a topic which will continue to grow in importance during the next few years. On Monday we were welcomed by our Czech hosts to the first "official" session of the Overview Week, held in the Department of Mechanical Engineering of the Czech Technical University, (CTU), the oldest Technical University in Europe. ATLAS (Prague) includes groups from the CTU, the even longer established Charles University and the Czech Acad...

  15. 13th European Conference on Mathematics for Industry

    Mattheij, RMM; Peletier, MA

    2006-01-01

    ECMI has a brand name in Industrial Mathematics and organises successful biannual conferences. This time, the conference on Industrial Mathematics held in Eindhoven in June 2004 Mathematics focused on Aerospace, Electronic Industry, Chemical Technology, Life Sciences, Materials, Geophysics, Financial Mathematics and Water flow. The majority of the invited talks on these topics can be found in these proceedings. Apart form these lectures, a large number of contributed papers and minisymposium papers are included here. They give an interesting and impressive overview of the important place mathematics has achieved in solving all kinds of problems met in industry, and commerce in particular.

  16. The 13th Tihany Symposium on Radiation Chemistry

    Wojnárovits, László; Takács, Erzsébet

    2016-07-01

    The Symposium was held in Balatonalmádi, a beautiful city by the Lake Balaton, Hungary, between August 29 and September 3, 2015. This time - to meet the expectations of many colleagues and friends - a place close to the village Tihany was selected, where the first Tihany Symposium was organized in 1962. The participants beside the excellent lectures could also enjoy the panorama of the Lake from the rooms and from the terrace of Hotel Ramada. The number of participants was close to 170 from about 33 countries. The highest number of participants arrived from Poland (14), followed by France (11), Turkey (9) and China (9). The Symposium had 6-6 colleagues from Brazil, Israel, and Romania. Beside China, Asia was represented by a few scientists from the Republic of Korea, Malaysia, Thailand, Philippines and from Saudi Arabia.

  17. 13th workshop on what comes beyond the standard models

    Nielsen, Holger Bech; Lukman, Dragan

    2010-01-01

    1. Noncommutativity and Topology within Lattice Field Theories 2. The Construction of Quantum Field Operators 3. The Bargmann-Wigner Formalism for Spin 2 Fields 4. New Light on Dark Matter from the LHC 5. Extra Dimensional Metric Reversal Symmetry and its Prospect... 6. Masses and Mixing Matrices of Families within SU(3) Flavor Symmetry ... 7. Dark Atoms of the Universe: OHe Nuclear Physics, 8. Can the Matter-Antimatter Asymmetry be Easier to Understand Within the "Spin-charge-family-theory", .. 9. Mass Matrices of Twice Four Families of Quarks and Leptons, ...in the "Spin-charge-family-theory" 10. Bohmian Quantum Mechanics or What Comes Before the Standard Model 11. Backward Causation in Complex Action Model ... 12. Is the Prediction of the "Spin-charge-family-theory" in Disagreement with the XENON100..? 13. Masses and Mixing Matrices of Families of Quarks and Leptons Within the "Spin-charge-family-theory" 14. Can the Stable Fifth Family of the "Spin-charge-family-theory" ...Form the Fifth Antibaryon Cluster...

  18. Proceedings of the 13th International Stellarator Workshop

    These proceedings include submitted abstracts and papers summarising the work as presented by participants in either oral or poster sessions and the highlights of the Workshop presented by Professor Jeffrey Harris. He stressed that while notable progress has been made in stellarator research overall, the world stellarator program faces important challenges in the coming years. Lead times for the construction of the next generation of devices are approximately 5 years, which makes maintaining continuity in research programs more difficult. The development of a compelling reactor vision based on the stellarator is an important task, especially in the context of renewed activity on the ITER tokamak project. The sharing of ideas, techniques, and results via international collaboration is more important than ever. Individual presentations have been separately indexed

  19. EDS'09: 13th International Conference on Elastic & Diffractive Scattering

    CERN. Geneva

    2009-01-01

    The series of International Conferences on Elastic and Diffractive Scattering was founded in 1985 in the picturesque old French town of Blois, famous for its XIV - XVIIth century château, inside of which the first meeting took place. Since then, meetings have been organised every two years in different places of the world: New York (1987), Evanston (1989), Isola d'Elba (1991), Providence (1993), Blois (1995), Seoul (1997), Protvino (1999), Prague (2001), Helsinki (2003), Blois (2005) and Hamburg (2007). The conference will focus on the most recent experimental and theoretical results in particle physics with an emphasis on Quantum Chromodynamics (QCD). http://cern.ch/eds09/ The conference agenda is now full. No further contributions can be accepted.

  20. 13th International Mass Spectrometry Conference. Book of Abstracts

    The collection contains abstracts of several hundred papers presented at the international conference on new research and development results and applications of mass spectrometry. Abstracts falling into the INIS scope were indexed separately in the INIS database. (Roboz, P.)

  1. 13th Conference of the Canadian Number Theory Association

    Alaca, Şaban; Williams, Kenneth

    2015-01-01

    The theory of numbers continues to occupy a central place in modern mathematics because of both its long history over many centuries as well as its many diverse applications to other fields such as discrete mathematics, cryptography, and coding theory. The proof by Andrew Wiles (with Richard Taylor) of Fermat’s last theorem published in 1995 illustrates the high level of difficulty of problems encountered in number-theoretic research as well as the usefulness of the new ideas arising from its proof. The thirteenth conference of the Canadian Number Theory Association was held at Carleton University, Ottawa, Ontario, Canada from June 16 to 20, 2014. Ninety-nine talks were presented at the conference on the theme of advances in the theory of numbers. Topics of the talks reflected the diversity of current trends and activities in modern number theory. These topics included modular forms, hypergeometric functions, elliptic curves, distribution of prime numbers, diophantine equations, L-functions, Diophantine app...

  2. 13th Forum for Specification and Design Languages (FDL) conference

    Morawiec, Adam; System Specification and Design Languages : Selected Contributions from FDL 2010

    2012-01-01

    This book brings together a selection of the best papers from the thirteenth edition of the Forum on specification and Design Languages Conference (FDL), which was held in Southampton, UK in September 2010.  FDL is a well established international forum devoted to dissemination of research results, practical experiences and new ideas in the application of specification, design and verification languages to the design, modelling and verification of integrated circuits, complex hardware/software embedded systems, and mixed-technology systems. Covers design verification, automatic synthesis and mechanized debug aids; Includes language-based modeling and design techniques for embedded systems; Covers design, modeling and verification of mixed physical domain and mixed signal systems that include significant analog parts in electrical and non-electrical domains; Includes formal and semi-formal system level design methods for complex embedded systems based on the Unified Modelling Language (UML) and Model Driven E...

  3. The absence of the Pseudomonas aeruginosa OprF protein leads to increased biofilm formation through variation in c-di-GMP level

    Emeline eBouffartigues

    2015-06-01

    Full Text Available OprF is the major outer membrane porin in bacteria belonging to the Pseudomonas genus. In previous studies, we have shown that OprF is required for full virulence expression of the opportunistic pathogen Pseudomonas aeruginosa. Here, we describe molecular insights on the nature of this relationship and report that the absence of OprF leads to increased biofilm formation and production of the Pel exopolysaccharide. Accordingly, the level of c-di-GMP, a key second messenger in biofilm control, is elevated in an oprF mutant. By decreasing c-di-GMP levels in this mutant, both biofilm formation and pel gene expression phenotypes were restored to wild-type levels. We further investigated the impact on two small RNAs, which are associated with the biofilm lifestyle, and found that expression of rsmZ but not of rsmY was increased in the oprF mutant and this occurs in a c-di-GMP-dependent manner. Finally, the extracytoplasmic function (ECF sigma factors AlgU and SigX displayed higher activity levels in the oprF mutant. Two genes of the SigX regulon involved in c-di-GMP metabolism, PA1181 and adcA (PA4843, were up-regulated in the oprF mutant, partly explaining the increased c-di-GMP level. We hypothesized that the absence of OprF leads to a cell envelope stress that activates SigX and results in a c-di-GMP elevated level due to higher expression of adcA and PA1181. The c-di-GMP level can in turn stimulate Pel synthesis via increased rsmZ sRNA levels and pel mRNA, thus affecting Pel-dependent phenotypes such as cell aggregation and biofilm formation. This work highlights the connection between OprF and c-di-GMP regulatory networks, likely via SigX (ECF, on the regulation of biofilm phenotypes.

  4. The search for mutations in the gene for the beta subunit of the cGMP phosphodiesterase (PDEB) in patients with autosomal recessive retinitis pigmentosa

    Riess, O; Noerremoelle, A; Weber, B; Musarella, M A; Hayden, M R

    1992-01-01

    including 196 bp of the 5' region of the PDEB gene have been assessed for mutations by using single-strand conformational polymorphism analysis in 14 patients from 13 unrelated families with autosomal recessive retinitis pigmentosa (ARRP). No disease-causing mutations were found in this group of affected......The finding of a mutation in the beta subunit of the cyclic GMP (cGMP) phosphodiesterase gene causing retinal degeneration in mice (the Pdeb gene) prompted a search for disease-causing mutations in the human phosphodiesterase gene (PDEB gene) in patients with retinitis pigmentosa. All 22 exons...

  5. cGMP-dependent protein kinase Iα associates with the antidepressant-sensitive serotonin transporter and dictates rapid modulation of serotonin uptake

    Steiner Jennifer A

    2009-08-01

    Full Text Available Abstract Background The Na+/Cl--dependent serotonin (5-hydroxytryptamine, 5-HT transporter (SERT is a critical element in neuronal 5-HT signaling, being responsible for the efficient elimination of 5-HT after release. SERTs are not only targets for exogenous addictive and therapeutic agents but also can be modulated by endogenous, receptor-linked signaling pathways. We have shown that neuronal A3 adenosine receptor activation leads to enhanced presynaptic 5-HT transport in vitro and an increased rate of SERT-mediated 5-HT clearance in vivo. SERT stimulation by A3 adenosine receptors derives from an elevation of cGMP and subsequent activation of both cGMP-dependent protein kinase (PKG and p38 mitogen-activated protein kinase. PKG activators such as 8-Br-cGMP are known to lead to transporter phosphorylation, though how this modification supports SERT regulation is unclear. Results In this report, we explore the kinase isoform specificity underlying the rapid stimulation of SERT activity by PKG activators. Using immortalized, rat serotonergic raphe neurons (RN46A previously shown to support 8-Br-cGMP stimulation of SERT surface trafficking, we document expression of PKGI, and to a lower extent, PKGII. Quantitative analysis of staining profiles using permeabilized or nonpermeabilized conditions reveals that SERT colocalizes with PKGI in both intracellular and cell surface domains of RN46A cell bodies, and exhibits a more restricted, intracellular pattern of colocalization in neuritic processes. In the same cells, SERT demonstrates a lack of colocalization with PKGII in either intracellular or surface membranes. In keeping with the ability of the membrane permeant kinase inhibitor DT-2 to block 8-Br-cGMP stimulation of SERT, we found that DT-2 treatment eliminated cGMP-dependent kinase activity in PKGI-immunoreactive extracts resolved by liquid chromatography. Similarly, treatment of SERT-transfected HeLa cells with small interfering RNAs targeting

  6. Activation of a cGMP-sensitive calcium-dependent chloride channel may cause transition from calcium waves to whole cell oscillations in smooth muscle cells

    Jacobsen, Jens Christian Brings; Aalkjær, Christian; Nilsson, Holger; Matchkov, Vladimir V; Freiberg, Jacob; Holstein-Rathlou, N.-H.

    2007-01-01

    waves sweeping through the cytoplasm when the sarcoplasmic reticulum (SR) is stimulated to release calcium. A rise in cGMP leads to the experimentally observed transition from waves to whole cell calcium oscillations. At the same time, membrane potential starts to oscillate and the frequency...... approximately doubles. In this transition, the simulated results point to a key role for a recently discovered cGMP-sensitive calcium-dependent chloride channel. This channel depolarizes the membrane in response to calcium released from the SR. In turn, depolarization causes a uniform opening of L-type calcium...

  7. Metabotropic glutamate receptor 5 modulates the nitric oxide-cGMP pathway in cerebellum in vivo through activation of AMPA receptors.

    Boix, Jordi; Llansola, Marta; Cabrera-Pastor, Andrea; Felipo, Vicente

    2011-04-01

    Metabotropic glutamate receptors (mGluRs) modulate important processes in cerebellum including long-term depression, which also requires formation of nitric oxide (NO) and cGMP. Some reports suggest that mGluRs could modulate the NO-cGMP pathway in cerebellum. However this modulation has not been studied in detail. The aim of this work was to assess by microdialysis in freely moving rats whether activation of mGluR5 modulates the NO-cGMP pathway in cerebellum in vivo and to analyze the underlying mechanisms. We show that mGluR5 activation increases extracellular glutamate, citrulline and cGMP in cerebellum. Blocking NMDA receptors with MK-801 does not prevent any of these effects, indicating that NMDA receptors activation is not required. However in the presence of MK-801 the effects are more transient, returning faster to basal levels. Blocking AMPA receptors prevents the increase in citrulline and cGMP induced by mGluR5 activation, but not the increase in glutamate. The release of glutamate is prevented by tetrodotoxin but not by fluoroacetate, indicating that glutamate is released from neurons and not from astrocytes. Activation of AMPA receptors increases citrulline and cGMP. These data indicate that activation of mGluR5 induces an increase of extracellular glutamate which activates AMPA receptors, leading to activation of nitric oxide synthase and increased NO, which activates guanylate cyclase, increasing cGMP. The response mediated by AMPA receptors desensitize rapidly. Activation of AMPA receptors also induces a mild depolarization, allowing activation of NMDA receptors which prolongs the duration of the effect initiated by activation of AMPA receptors. These data support that the three types of glutamate receptors: mGluR5, AMPA and NMDA cooperate in the modulation of the grade and duration of activation of the NO-cGMP pathway in cerebellum in vivo. This pathway would modulate cerebellar processes such as long-term depression. PMID:21300123

  8. The function of the glutamate–nitric oxide–cGMP pathway in brain in vivo and learning ability decrease in parallel in mature compared with young rats

    Piedrafita, Blanca; Cauli, Omar; Montoliu, Carmina; Felipo, Vicente

    2007-01-01

    Aging is associated with cognitive impairment, but the underlying mechanisms remain unclear. We have recently reported that the ability of rats to learn a Y-maze conditional discrimination task depends on the function of the glutamate–nitric oxide–cGMP pathway in brain. The aims of the present work were to assess whether the ability of rats to learn this task decreases with age and whether this reduction is associated with a decreased function of the glutamate–nitric oxide–cGMP pathway in bra...

  9. Hyperammonemia alters glycinergic neurotransmission and modulation of the glutamate-nitric oxide-cGMP pathway by extracellular glycine in cerebellum in vivo.

    Cabrera-Pastor, Andrea; Taoro-Gonzalez, Lucas; Felipo, Vicente

    2016-05-01

    The glutamate-nitric oxide (NO)-cGMP pathway modulates some forms of learning. How glycine modulates this pathway is unclear. Glycine could modulate the pathway biphasically, enhancing its function through NMDA receptor activation or reducing it through glycine receptor activation. Chronic hyperammonemia impairs the glutamate-NO-cGMP pathway in the cerebellum and induces cognitive impairment. The possible alterations in hyperammonemia of glycinergic neurotransmission and of glutamate-NO-cGMP pathway modulation by glycine remain unknown. The aims were to assess, by in vivo microdialysis in cerebellum: (i) the effects of different glycine concentrations, administered through the microdialysis probe, on the glutamate-NO-cGMP pathway function; (ii) the effects of tonic glycine receptors activation on the pathway function, by blocking them with strychnine; (iii) whether hyperammonemia alters the pathway modulation by glycine; (iv) and whether hyperammonemia alters extracellular glycine concentration and/or glycine receptor membrane expression. In control rats, low glycine levels reduce the pathway function, likely by activating glycine receptors, while 20 μM glycine enhances the pathway function, likely by enhancing NMDA receptor activation. In hyperammonemic rats, glycine did not reduce the pathway function, but enhanced it when administered at 1-20 μM. Hyperammonemia reduces extracellular glycine concentration by approximately 50% and glycine receptor membrane expression. However, tonic glycine receptor activation seems to be enhanced in hyperammonemic rats, as indicated by the larger increase in extracellular cGMP induced by strychnine. These data show that glycine modulates the glutamate-NO-cGMP pathway biphasically and that hyperammonemia strongly alters glycinergic neurotransmission and modulation by glycine of the glutamate-NO-cGMP pathway. These alterations may contribute to the cerebellar aspects of cognitive alterations in hyperammonemia. The findings

  10. Polychlorinated biphenyls PCB 52, PCB 180, and PCB 138 impair the glutamate-nitric oxide-cGMP pathway in cerebellar neurons in culture by different mechanisms.

    Llansola, Marta; Montoliu, Carmina; Boix, Jordi; Felipo, Vicente

    2010-04-19

    Polychlorinated biphenyls (PCBs) are persistent organic pollutants that accumulate in the food chain and are present in human blood and milk. Children born to mothers exposed to PCBs show cognitive deficits, which are reproduced in rats perinatally exposed to PCBs. It has been proposed that PCB-induced cognitive impairment is due to impairment of the glutamate-nitric oxide (NO)-cGMP pathway. The aim of the present work was to assess whether chronic exposure to the nondioxin-like PCB52, PCB138, or PCB180 alters the function of this pathway in primary cultures of rat cerebellar neurons and to assess whether different PCBs have similar or different mechanisms of action. PCB180 and PCB138 impair the function of the glutamate-NO-cGMP pathway at nanomolar concentrations, and PCB52 impairs the function of the glutamate-NO-cGMP pathway at micromolar concentrations. The mechanisms by which different PCBs impair the function of the glutamate-NO-cGMP pathway are different. Each PCB affects the pathway at more than one step but with different potency and, for some steps, in opposite ways. Exposure to the PCBs alters the basal concentrations of intracellular calcium, NO, and cGMP. The three PCBs increase NO; however, PCB52 and PCB138 increase basal cGMP, while PCB180 decreases it. PCB52 and PCB138 decrease the activation of soluble guanylate cyclase by NO, and PCB180 increases it. Long-term exposure to PCB52, PCB180, or PCB138 reduces the activation of NO synthase and the whole glutamate-NO-cGMP pathway in response to activation of N-methyl-d-aspartate receptors. The EC(50) was 300 nM for PCB52 and 2 nM for PCB138 or PCB180. These results show that chronic exposure to nondioxin like PCBs impairs the function of the glutamate-NO-cGMP pathway in cerebellar neurons by different mechanisms and with different potencies. Impaired function of this pathway would contribute to the cognitive alterations induced by perinatal exposure to PCBs in humans. PMID:20297801

  11. Design and package of a {sup 14}CO{sub 2} field analyzer The Global Monitor Platform (GMP)

    Bright, Michelle; Marino, Bruno D.V.; Gronniger, Glen

    2011-08-01

    Carbon Capture and Sequestration (CCS) is widely accepted as a means to reduce and eliminate the fossil fuel CO{sub 2} (ff- CO{sub 2}) emissions from coal fired power plants. Success of CCS depends on near zero leakage rates over decadal time scales. Currently no commercial methods to determine leakage of ff-CO{sub 2} are available. The Global Monitor Platform (GMP) field analyzer provides high precision analysis of CO{sub 2} isotopes [12C (99%), 13C (<1%), 14C (1.2x10-10 %)] that can differentiate between fossil and biogenic CO{sub 2} emissions. Fossil fuels contain no {sup 14}C; their combustion should lower atmospheric amounts on local to global scales. There is a clear mandate for monitoring, verification and accounting (MVA) of CCS systems nationally and globally to verify CCS integrity, treaty verification (Kyoto Protocol) and to characterize the nuclear fuel cycle. Planetary Emissions Management (PEM), working with the National Secure Manufacturing Center (NSMC), has the goal of designing, ruggedizing and packaging the GMP for field deployment. The system will conduct atmosphere monitoring then adapt the system to monitor water and soil evaluations. Measuring {sup 14}CO{sub 2} in real time will provide quantitative concentration data for ff-CO{sub 2} in the atmosphere and CCS leakage detection. Initial results will be discussed along with design changes for improved detection sensitivity and manufacturability.

  12. Effects of Tang Niao Ning decoction on ET-1, GMP-140 and TXB2 in diabetic rats

    To investigate the effects of Tang Niao Ning decoction on fasting blood glucose (FBG), total cholesterol (TC), triglyceride (TG), endothelin (ET-1), platelet granule membrane protein (GMP-140), thromboxane B2 (TXB2), 6-keto-prostaglandin F1α (6-keto-PGF1α), the selected female Wistar rats were modeled by streptozotocin (STZ) and high fat diets, and were randomly divided into two groups as Tang Niao Ning and model group, while the normal control group was set up. The Tang Niao Ning group started to supply decoctions after these rats had been fed one week with high fat diets. The rats were killed after six weeks. Above items were examed. The results showed that the model group displayed marked difference when the experiment finished compared with the normal group. The contents of fasting blood glucose and blood lipid were increased, accompanied with the phenomena of hoisting ET-1, enhancing GMP-140, increasing TXB2 and droping 6-Keto-PGF1α. After treatment in time with Tang Niao Ning decoction, all kinds of items were adjusted to normal level gradually. It suggested that the treatment with Tang Niao Ning decoction might decrease FBG and blood lipid. In some ways, the decoction could improve the signs of blood stasis. The regulation mechanisms of the decoction was characteristic of wholism through many channels

  13. The phytosulfokine (PSK) receptor is capable of guanylate cyclase activity and enabling cyclic GMP-dependent signaling in plants

    Kwezi, Lusisizwe

    2011-04-19

    Phytosulfokines (PSKs) are sulfated pentapeptides that stimulate plant growth and differentiation mediated by the PSK receptor (PSKR1), which is a leucine-rich repeat receptor-like kinase. We identified a putative guanylate cyclase (GC) catalytic center in PSKR1 that is embedded within the kinase domain and hypothesized that the GC works in conjunction with the kinase in downstream PSK signaling. We expressed the recombinant complete kinase (cytoplasmic) domain of AtPSKR1 and show that it has serine/threonine kinase activity using the Ser/Thr peptide 1 as a substrate with an approximate Km of 7.5 μM and Vmax of 1800 nmol min-1 mg-1 of protein. This same recombinant protein also has GC activity in vitro that is dependent on the presence of either Mg2+ or Mn2+. Overexpression of the full-length AtPSKR1 receptor in Arabidopsis leaf protoplasts raised the endogenous basal cGMP levels over 20-fold, indicating that the receptor has GC activity in vivo. In addition, PSK-α itself, but not the non-sulfated backbone, induces rapid increases in cGMP levels in protoplasts. Together these results indicate that the PSKR1 contains dual GC and kinase catalytic activities that operate in vivo and that this receptor constitutes a novel class of enzymes with overlapping catalytic domains. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

  14. Der Stadtwall in den Gärten: Neue Gedanken zu den historischen Hintergründen der Verlegung der Stadt Hitzacker um die Mitte des 13. Jahrhunderts (The Town Ramparts in the Garden: New Thoughts About the Historical Background of the Relocation of the Town of Hitzacker in the Mid-13th Century

    Thomas Küntzel

    2010-08-01

    Full Text Available This Essay tries to explain the historical context of the „Landgraben“, an impressive ditch and bank west to the city centre of Hitzacker. It first deals with the development of the settlement, of wich the castle-mound of the “Weinberg” already gains importance in the 9th century. This was underlined by the excavations in the area of the Lanke-gardens and the street line south of Hitzacker, proving the rise of the settlement in the 10th/11th centuries and the flourishing in the 12th century. Obviously in the first half of the 13th century the area in the south of Hitzacker was abandoned, probably in connection with the construction of the “Landgraben” (ca. 1230?. Yet the exact date and historical ascription is uncertain, especially wether the Welfen dukes or the princes of Ascania built it – one even can suppose the builing of the “Landgraben” (or parts of it in the beginning of the 14th century.

  15. Crystallization and preliminary X-ray analysis of the flagellar motor ‘brake’ molecule YcgR with c-di-GMP from Escherichia coli

    The flagellar motor ‘brake’ protein YcgR from E. coli was crystallized with c-di-GMP. The crystals diffracted to 2.3 Å resolution and belonged to space group R3:H, with unit-cell parameters a = b = 93.96, c = 109.61 Å. In Escherichia coli and Salmonella enterica, bis-(3′-5′)-cyclic dimeric guanosine monophosphate (c-di-GMP), a ubiquitous bacterial second-messenger molecule that participates in many cellular processes, can regulate flagellar motor speed and reduce cell swimming velocity by binding to the PilZ-containing protein YcgR. Here, the crystallization and preliminary X-ray crystallographic analysis of YcgR with c-di-GMP are reported. The crystals diffracted to 2.3 Å resolution and belonged to space group R3:H, with unit-cell parameters a = b = 93.96, c = 109.61 Å. The asymmetric unit appeared to contain one subunit with a Matthews coefficient of 3.21 Å3 Da−1. The results reported here provide a sound basis for solving the crystal structure of YcgR with c-di-GMP and revealing its structure–function relationship based on the three-dimensional structure

  16. Effects of Kaempferia parviflora Wall. Ex. Baker and sildenafil citrate on cGMP level, cardiac function, and intracellular Ca2+ regulation in rat hearts.

    Weerateerangkul, Punate; Palee, Siripong; Chinda, Kroekkiat; Chattipakorn, Siriporn C; Chattipakorn, Nipon

    2012-09-01

    Although Kaempferia parviflora extract (KPE) and its flavonoids have positive effects on the nitric oxide (NO) signaling pathway, its mechanisms on the heart are still unclear. Because our previous studies demonstrated that KPE decreased defibrillation efficacy in swine similar to that of sildenafil citrate, the phosphodiesterase-5 inhibitor, it is possible that KPE may affect the cardiac NO signaling pathway. In the present study, the effects of KPE and sildenafil citrate on cyclic guanosine monophosphate (cGMP) level, modulation of cardiac function, and Ca transients in ventricular myocytes were investigated. In a rat model, cardiac cGMP level, cardiac function, and Ca transients were measured before and after treatment with KPE and sildenafil citrate. KPE significantly increased the cGMP level and decreased cardiac function and Ca transient. These effects were similar to those found in the sildenafil citrate-treated group. Furthermore, the nonspecific NOS inhibitor could abolish the effects of KPE and sildenafil citrate on Ca transient. KPE has positive effect on NO signaling in the heart, resulting in an increased cGMP level, similar to that of sildenafil citrate. This effect was found to influence the physiology of normal heart via the attenuation of cardiac function and the reduction of Ca transient in ventricular myocytes. PMID:22691878

  17. Studies on the metal-nucleotide (Co-5'IMP, Co-5'GMP) interaction via Moessbauer emission spectroscopy and magnetic susceptibility measurements

    By means of 57Co Moessbauer emission spectroscopy and magnetic susceptibility measurements, the local symmetries and electronic structure of the Co(II) ions in the Co-5'IMP and Co-5'GMP complexes in the hydrated and dehydrated forms have been determined. (Auth.)

  18. Comparative effects of vinpocetine and 8-Br-cyclic GMP on the contraction and 45Ca-fluxes in the rabbit aorta

    Vinpocetine is a highly specific inhibitor of calmodulin-dependent phosphodiesterase (CaM-PDE) with an IC50 of 19 microM and produces a significant accumulation of cyclic GMP but not cyclic AMP in rabbit aorta. In isolated rabbit aortic strips, vinpocetine (0.01 and 0.1 mM) inhibited the contraction and 45Ca uptake due to both phenylephrine (1 microM) and KCl (40 mM), whereas 8-Br-cyclic GMP (0.1-1mM) selectively impaired phenylephrine-induced responses. Furthermore, the KCl-stimulated 45Ca efflux in normal Ca2+ buffer, which reflects elevated cytosolic Ca2+, was greatly diminished by vinpocetine but not by 8-Br-cyclic GMP. However, phenylephrine-induced 45Ca efflux and contraction in Ca2+-free buffer, which reflect Ca2+ release from intracellular sites, were similarly inhibited by both vinpocetine and 8-Br-cyclic GMP. The results suggest that vinpocetine may effect vasodilatation through blockade of the slow channel and selective inhibition of CaM-PDE in the vascular smooth muscle

  19. The Function of the Glutamate-Nitric Oxide-cGMP Pathway in Brain in Vivo and Learning Ability Decrease in Parallel in Mature Compared with Young Rats

    Piedrafita, Blanca; Cauli, Omar; Montoliu, Carmina; Felipo, Vicente

    2007-01-01

    Aging is associated with cognitive impairment, but the underlying mechanisms remain unclear. We have recently reported that the ability of rats to learn a Y-maze conditional discrimination task depends on the function of the glutamate-nitric oxide-cGMP pathway in brain. The aims of the present work were to assess whether the ability of rats to…

  20. The function of the glutamate–nitric oxide–cGMP pathway in brain in vivo and learning ability decrease in parallel in mature compared with young rats

    Piedrafita, Blanca; Cauli, Omar; Montoliu, Carmina; Felipo, Vicente

    2007-01-01

    Aging is associated with cognitive impairment, but the underlying mechanisms remain unclear. We have recently reported that the ability of rats to learn a Y-maze conditional discrimination task depends on the function of the glutamate–nitric oxide–cGMP pathway in brain. The aims of the present work were to assess whether the ability of rats to learn this task decreases with age and whether this reduction is associated with a decreased function of the glutamate–nitric oxide–cGMP pathway in brain in vivo, as analyzed by microdialysis in freely moving rats. We show that 7-mo-old rats need significantly more (192 ± 64%) trials than do 3-mo-old rats to learn the Y-maze task. Moreover, the function of the glutamate–nitric oxide–cGMP pathway is reduced by 60 ± 23% in 7-mo-old rats compared with 3-mo-old rats. The results reported support the idea that the reduction in the ability to learn the Y-maze task (and likely other types of learning) of mature compared with young rats would be a consequence of reduced function of the glutamate–nitric oxide–cGMP pathway. PMID:17412964

  1. Pregnenolone sulfate restores the glutamate-nitric-oxide-cGMP pathway and extracellular GABA in cerebellum and learning and motor coordination in hyperammonemic rats.

    Gonzalez-Usano, Alba; Cauli, Omar; Agusti, Ana; Felipo, Vicente

    2014-02-19

    Around 40% of cirrhotic patients show minimal hepatic encephalopathy (MHE), with mild cognitive impairment which reduces their quality of life and life span. Treatment of MHE is unsatisfactory, and there are no specific treatments for the neurological alterations in MHE. Hyperammonemia is the main contributor to neurological alterations in MHE. New agents acting on molecular targets involved in brain mechanisms leading to neurological alterations are needed to treat MHE. Chronic hyperammonemia impairs learning of a Y-maze task by impairing the glutamate-nitric-oxide (NO)-cGMP pathway in cerebellum, in part by enhancing GABA(A) receptor activation, which also induces motor in-coordination. Acute pregnenolone sulfate (PregS) restores the glutamate-NO-cGMP pathway in hyperammonemic rats. This work aimed to assess whether chronic treatment of hyperammonemic rats with PregS restores (1) motor coordination; (2) extracellular GABA in cerebellum; (3) learning of the Y-maze task; (4) the glutamate-NO-cGMP pathway in cerebellum. Chronic intracerebral administration of PregS normalizes motor coordination likely due to extracellular GABA reduction. PregS restores learning ability by restoring the glutamate-NO-cGMP pathway, likely due to both enhanced NMDA receptor activation and reduced GABA(A) receptor activation. Similar treatments would improve cognitive and motor alterations in patients with MHE. PMID:24256194

  2. Experience towards compliance with current good manufacturing practice (cGMP) requirements in manufacturing of Tc-99m generator at Nuclear Malaysia

    Manufacturing of Tc-99m generator need to be carried out in compliance with the current Good Manufacturing Practice (cGMP) requirements of the Pharmaceutical Inspection Convention and Pharmaceutical Inspection Co-operation Scheme, PIC/S. The objective of this paper is to present our experience to get a licence to manufacture Tc-99m generator that comply with the cGMP requirements. The existing clean room has been renovated and qualified together with heat-ventilation air-conditioning (HVAC) system. HVAC system is now at stage three of the qualification process. The hot cell and laminar air-flow units have been qualified. The critical equipment including Oven and Autoclave had been qualified for its applications. The water for injection (WFI) system has just completed its stage one qualification. Production operators have been trained for the competency in cGMP requirements. In conclusion, manufacturing of Tc-99m generator at Nuclear Malaysia is carried out in the manner of cGMP requirements of PIC/S. (Author)

  3. YfiBNR mediates cyclic di-GMP dependent small colony variant formation and persistence in Pseudomonas aeruginosa.

    Jacob G Malone

    2010-03-01

    Full Text Available During long-term cystic fibrosis lung infections, Pseudomonas aeruginosa undergoes genetic adaptation resulting in progressively increased persistence and the generation of adaptive colony morphotypes. This includes small colony variants (SCVs, auto-aggregative, hyper-adherent cells whose appearance correlates with poor lung function and persistence of infection. The SCV morphotype is strongly linked to elevated levels of cyclic-di-GMP, a ubiquitous bacterial second messenger that regulates the transition between motile and sessile, cooperative lifestyles. A genetic screen in PA01 for SCV-related loci identified the yfiBNR operon, encoding a tripartite signaling module that regulates c-di-GMP levels in P. aeruginosa. Subsequent analysis determined that YfiN is a membrane-integral diguanylate cyclase whose activity is tightly controlled by YfiR, a small periplasmic protein, and the OmpA/Pal-like outer-membrane lipoprotein YfiB. Exopolysaccharide synthesis was identified as the principal downstream target for YfiBNR, with increased production of Pel and Psl exopolysaccharides responsible for many characteristic SCV behaviors. An yfi-dependent SCV was isolated from the sputum of a CF patient. Consequently, the effect of the SCV morphology on persistence of infection was analyzed in vitro and in vivo using the YfiN-mediated SCV as a representative strain. The SCV strain exhibited strong, exopolysaccharide-dependent resistance to nematode scavenging and macrophage phagocytosis. Furthermore, the SCV strain effectively persisted over many weeks in mouse infection models, despite exhibiting a marked fitness disadvantage in vitro. Exposure to sub-inhibitory concentrations of antibiotics significantly decreased both the number of suppressors arising, and the relative fitness disadvantage of the SCV mutant in vitro, suggesting that the SCV persistence phenotype may play a more important role during antimicrobial chemotherapy. This study establishes Yfi

  4. MrkH, a novel c-di-GMP-dependent transcriptional activator, controls Klebsiella pneumoniae biofilm formation by regulating type 3 fimbriae expression.

    Jonathan J Wilksch

    2011-08-01

    Full Text Available Klebsiella pneumoniae causes significant morbidity and mortality worldwide, particularly amongst hospitalized individuals. The principle mechanism for pathogenesis in hospital environments involves the formation of biofilms, primarily on implanted medical devices. In this study, we constructed a transposon mutant library in a clinical isolate, K. pneumoniae AJ218, to identify the genes and pathways implicated in biofilm formation. Three mutants severely defective in biofilm formation contained insertions within the mrkABCDF genes encoding the main structural subunit and assembly machinery for type 3 fimbriae. Two other mutants carried insertions within the yfiN and mrkJ genes, which encode GGDEF domain- and EAL domain-containing c-di-GMP turnover enzymes, respectively. The remaining two isolates contained insertions that inactivated the mrkH and mrkI genes, which encode for novel proteins with a c-di-GMP-binding PilZ domain and a LuxR-type transcriptional regulator, respectively. Biochemical and functional assays indicated that the effects of these factors on biofilm formation accompany concomitant changes in type 3 fimbriae expression. We mapped the transcriptional start site of mrkA, demonstrated that MrkH directly activates transcription of the mrkA promoter and showed that MrkH binds strongly to the mrkA regulatory region only in the presence of c-di-GMP. Furthermore, a point mutation in the putative c-di-GMP-binding domain of MrkH completely abolished its function as a transcriptional activator. In vivo analysis of the yfiN and mrkJ genes strongly indicated their c-di-GMP-specific function as diguanylate cyclase and phosphodiesterase, respectively. In addition, in vitro assays showed that purified MrkJ protein has strong c-di-GMP phosphodiesterase activity. These results demonstrate for the first time that c-di-GMP can function as an effector to stimulate the activity of a transcriptional activator, and explain how type 3 fimbriae

  5. Ab Initio QM/MM Study Shows a Highly Dissociated SN2 Hydrolysis Mechanism for the cGMP-Specific Phosphodiesterase-5.

    Li, Zhe; Wu, Yinuo; Feng, Ling-Jun; Wu, Ruibo; Luo, Hai-Bin

    2014-12-01

    Phosphodiesterases (PDEs) are the sole enzymes hydrolyzing the important second messengers cGMP and cAMP and have been identified as therapeutic targets for several diseases. The most successful examples are PDE5 inhibitors (i.e., sildenafil and tadalafil), which have been approved for the treatment of male erectile dysfunction and pulmonary hypertension. However, the side effects mostly due to nonselective inhibition toward other PDE isoforms, set back the clinical usage of PDE5 inhibitors. Until now, the exact catalytic mechanism of the substrate cGMP by PDE5 is still unclear. Herein, the first computational study on the catalytic hydrolysis mechanism of cGMP for PDE5 (catalytic domain) is performed by employing the state-of-the-art ab initio quantum mechanics/molecular mechanics (QM/MM) molecular dynamics (MD) simulations. Our simulations show a SN2 type reaction procedure via a highly dissociated transition state with a reaction barrier of 8.88 kcal/mol, which is quite different from the previously suggested hydrolysis mechanism of cAMP for PDE4. Furthermore, the subsequent ligand exchange and the release of the product GMP have also been investigated by binding energy analysis and MD simulations. It is deduced that ligand exchange would be the rate-determining step of the whole reaction, which is consistent with many previous experimental results. The obtained mechanistic insights should be valuable for not only the rational design of more specific inhibitors toward PDE5 but also understanding the general hydrolysis mechanism of cGMP-specific PDEs. PMID:26583228

  6. Evaluation of GMP-compliant culture media for in vitro expansion of human bone marrow mesenchymal stromal cells.

    Wuchter, Patrick; Vetter, Marcel; Saffrich, Rainer; Diehlmann, Anke; Bieback, Karen; Ho, Anthony D; Horn, Patrick

    2016-06-01

    Mesenchymal stromal cells (MSCs) from human bone marrow serve as a resource for cell-based therapies in regenerative medicine. Clinical applications require standardized protocols according to good manufacturing practice (GMP) guidelines. Donor variability as well as the intrinsic heterogeneity of MSC populations must be taken into consideration. The composition of the culture medium is a key factor in successful MSC expansion. The aim of this study was to comparatively assess the efficiency of xeno-free human platelet lysate (HPL)-based cell expansion with two commercially available media-StemPro MSC SFM CTS (for human ex vivo tissue and cell culture processing applications) and MSCGM (non-GMP-compliant, for research only)-in an academic setting as the first optimization step toward GMP-compliant manufacturing. We report the feasibility of MSC expansion up to the yielded cell number with all three media. MSCs exhibited the typical fibroblastoid morphology, with distinct differences in cell size depending on the medium. The differentiation capacity and characteristic immunophenotype were confirmed for all MSC populations. Proliferation was highest using StemPro MSC SFM CTS, whereas HPL medium was more cost-effective and its composition could be adjusted individually according to the respective needs. In summary, we present a comprehensive evaluation of GMP-compatible culture media for MSC expansion. Both StemPro and HPL medium proved to be suitable for clinical application and allowed sufficient cell proliferation. Specific differences were observed and should be considered according to the intended use. This study provides a detailed cost analysis and tools that may be helpful for the establishment of GMP-compliant MSC expansion. PMID:26911671

  7. Polychlorinated biphenyls PCB 153 and PCB 126 impair the glutamate-nitric oxide-cGMP pathway in cerebellar neurons in culture by different mechanisms.

    Llansola, Marta; Piedrafita, Blanca; Rodrigo, Regina; Montoliu, Carmina; Felipo, Vicente

    2009-08-01

    Polychlorinated biphenyls (PCBs) are persistent organic pollutants present in human blood and milk. Exposure to PCBs during pregnancy and lactation leads to cognitive impairment in children. Perinatal exposure to PCB 153 or PCB 126 impairs the glutamate-nitric oxide-cGMP pathway in cerebellum in vivo and learning ability in adult rats. The aims of this work were: (1) to assess whether long-term exposure of primary cultures of cerebellar neurons to PCB 153 or PCB 126 reproduces the impairment in the function of the glutamate-nitric oxide-cGMP pathway found in rat cerebellum in vivo; (2) to provide some insight on the steps of the pathway affected by these PCBs; (3) to assess whether the mechanisms of interference of the pathway are different for PCB 126 and PCB 153. Both PCB 153 and PCB 126 increase basal levels of cGMP by different mechanisms. PCB 126 increases the amount of soluble guanylate cyclase while PCB 153 does not. PCB 153 reduces the amount of calmodulin while PCB 126 does not. Also both PCBs impair the function of the glutamate-nitric oxide-cGMP pathway by different mechanisms, PCB 153 impairs nitric oxide-induced activation of soluble guanylate cyclase and increase in cGMP while PCB 126 does not. PCB 126 reduces NMDA-induced increase in calcium while PCB 153 does not. When PCB 153 and PCB 126 exhibit the same effect, PCB 126 was more potent than PCB 153, as occurs in vivo. PMID:19526286

  8. Angiotensin-converting enzyme inhibition prevents myocardial infarction-induced increase in renal cortical cGMP and cAMP phosphodiesterase activities.

    Clauss, François; Charloux, Anne; Piquard, François; Doutreleau, Stéphane; Talha, Samy; Zoll, Joffrey; Lugnier, Claire; Geny, Bernard

    2015-08-01

    We investigated whether myocardial infarction (MI) enhances renal phosphodiesterases (PDE) activities, investigating particularly the relative contribution of PDE1-5 isozymes in total PDE activity involved in both cGMP and cAMP pathways, and whether angiotensin-converting enzyme inhibition (ACEi) decreases such renal PDE hyperactivities. We also investigated whether ACEi might thereby improve atrial natriuretic peptide (ANP) efficiency. We studied renal cortical PDE1-5 isozyme activities in sham (SH)-operated, MI rats and in MI rats treated with perindopril (ACEi) 1 month after coronary artery ligation. Circulating atrial natriuretic peptide (ANP), its second intracellular messenger cyclic guanosine monophosphate (cGMP) and cGMP/ANP ratio were also determined. Cortical cGMP-PDE2 (80.3 vs. 65.1 pmol/min/mg) and cGMP-PDE1 (50.7 vs. 30.1 pmol/min/mg), and cAMP-PDE2 (161 vs. 104.1 pmol/min/mg) and cAMP-PDE4 (307.5 vs. 197.2 pmol/min/mg) activities were higher in MI than in SH rats. Despite increased ANP plasma level, ANP efficiency tended to be decreased in MI compared to SH rats. Perindopril restored PDE activities and tended to improve ANP efficiency in MI rats. One month after coronary ligation, perindopril treatment of MI rats prevents the increase in renal cortical PDE activities. This may contribute to increase renal ANP efficiency in MI rats. PMID:25939307

  9. cGMP-Manufactured Human Induced Pluripotent Stem Cells Are Available for Pre-clinical and Clinical Applications

    Behnam Ahmadian Baghbaderani

    2015-10-01

    Full Text Available The discovery of induced pluripotent stem cells (iPSCs and the concurrent development of protocols for their cell-type-specific differentiation have revolutionized our approach to cell therapy. It has now become critical to address the challenges related to the generation of iPSCs under current good manufacturing practice (cGMP compliant conditions, including tissue sourcing, manufacturing, testing, and storage. Furthermore, regarding the technical challenges, it is very important to keep the costs of manufacturing and testing reasonable and solve logistic hurdles that permit the global distribution of these products. Here we describe our efforts to develop a process for the manufacturing of iPSC master cell banks (MCBs under cGMPs and announce the availability of such banks.

  10. Use of the scanditronix MC40 cyclotron of the JRC (EC) for FDG production in compliance with the European GMP

    The Scanditronix MC40 Cyclotron of the Joint Research Centre (JRC) of the European Commission (Ispra site), installed in 1982, is used for materials research and for production of radioisotopes for medical applications (diagnosis and therapy). Regarding radioisotope production, the main ongoing projects are [18F]Fludeoxyglucose (FDG) production in collaboration with Amersham Health (Italy) and studies of methods for production of 64Cu [1], 211At [2] and 225Ac [3]. The present paper is focused on the use of MC 40 Cyclotron for routine FDG production in compliance with the European Good Manufacturing Practices (GMP) guidelines. The main components of the facility are described - the cyclotron, the target and the production and quality control laboratories. (author)

  11. Measurement of GEp/GMp in ep -> ep to Q2 = 5.6 GeV2

    Gayou, O; Jones, M K; Perdrisat, C F; Punjabi, V

    2002-01-01

    The ratio of the electric and magnetic form factors of the proton, GEp/GMp, was measured at the Thomas Jefferson National Accelerator Facility (JLab) using the recoil polarization technique. The ratio of the form factors is directly proportional to the ratio of the transverse to longitudinal components of the polarization of the recoil proton in the elastic $\\vec ep \\to e\\vec p$ reaction. The new data presented in this article span the range 3.5 < Q2 < 5.6 GeV2 and are well described by a linear Q2 fit. Also, the ratio QF2p/F1p reaches a constant value above Q2=2 GeV2.

  12. Rat growth hormone-releasing factor stimulates cyclic GMP formation and phosphatidylinositol metabolism in the median eminence

    The effects of rat growth hormone releasing factor (rGRF) on somatostatin (SRIF) secretion, cyclic nucleotide production and phosphatidylinositol metabolism were investigated in the median eminence (ME), using an in vitro system. Medium was discarded and replaced by medium containing various concentrations of rGRF or rGRF plus epinephrine (E, 6x10-7M). rGRF had no effect on basal or E-stimulated release of cAMP. In the same experiments rGRF markedly stimulated SRIF release. However, GRF significantly stimulated release of both SRIF and cGMP in a dose-related manner. Maximal stimulation was observed at 10-10M GRF which also produces maximal SRIF release. 2'0-monobutyrylguanosine 3'5' cyclic phosphate fragments whereas the control, sodium butyrate in the concentration of labelled inositol phosphates ([3H]-IPs) in the ME

  13. Effects of atrial and brain natriuretic peptides upon cyclic GMP levels, potassium transport, and receptor binding in rat astrocytes

    Beaumont, K.; Tan, P.K. (Univ. of California, San Diego, La Jolla (USA))

    1990-02-01

    The ability of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) to alter cyclic GMP levels and NaKCl cotransport in rat neocortical astrocytes was determined. At concentrations of 10(-9)-10(-6) M, rat ANP99-126 (rANF), rat ANP102-126 (auriculin B), and rat ANP103-126 (atriopeptin III) stimulated 6- to 100-fold increases in cyclic GMP levels. Porcine BNP (pBNP) and rat BNP (rBNP) were 20%-90% as effective as rANF over most of this concentration range, although 10(-6) M pBNP produced a greater effect than rANF. NaKCl cotransport as measured by bumetanide-sensitive 86Rb+ influx was not altered by exposure of astrocytes to 10(-6)M rANF, pBNP, or rBNP. Both pBNP and rBNP, as well as rat ANP103-123 (atriopeptin I) and des(gl18, ser19, gly20, leu21, gly22) ANF4-23-NH2 (C-ANF4-23) strongly competed for specific 125I-rANF binding sites in astrocyte membranes with affinities ranging from 0.03 to 0.4 nM, suggesting that virtually all binding sites measured at subnanomolar concentrations of 125I-rANF were of the ANP-C (ANF-R2) receptor subtype. These receptors are thought to serve a clearance function and may be linked to a guanylate cyclase activity that is chemically and pharmacologically distinct from that coupled to ANP-A (ANF-R1) receptors. ANP receptors on astrocytes may function in limiting the access of ANP and BNP to neurons involved in body fluid and cardiovascular regulation.

  14. Automated GMP Synthesis of [18F]ICMT-11 for In Vivo Imaging of Caspase-3 Activity

    Introduction: Isatin-5-sulfonamide ([18F]ICMT-11) is a sub-nanomolar inhibitor of caspase-3 previously evaluated as an apoptosis imaging agent. Herein, an alternative radiosynthesis of [18F]ICMT-11 with increased purity and specific activity is presented. Finally, a GMP-applicable automated radiosynthesis of [18F]ICMT-11 is described. Methods: The preparation of [18F]ICMT-11 was evaluated under a variety of reaction conditions, including reaction solvent, by employing alternative phase transfer catalysts and under different deprotection conditions. Following initial investigations, the process was transferred onto a fully automated GE FASTlab synthesis platform for further development and optimisation. Results: The synthesis of [18F]ICMT-11 was successfully validated under GMP conditions, resulting in a yield of 4.6 ± 0.4 GBq with a radiochemical purity of > 98% at EOS and a specific activity of 685 ± 237 GBq/μmol within 90 min. Quality control was carried out in accordance with the European Pharmacopoeia and demonstrated that [18F]ICMT-11 can be consistently manufactured on the FASTlab to meet specifications. Conclusions: A simplified methodology for the synthesis of the apoptosis imaging agent, [18F]ICMT-11, has been achieved by the SN2 displacement of a tosylate leaving group with [18F]fluoride ion. This results in an increased purity and specific activity over the original copper catalysed “Click” synthetic stratagem reaction involving 2-[18F]fluoroethylazide with an alkyne precursor and is now suitable for routine clinical application.

  15. Strategies for selecting recombinant CHO cell lines for cGMP manufacturing: improving the efficiency of cell line generation.

    Porter, Alison J; Racher, Andrew J; Preziosi, Richard; Dickson, Alan J

    2010-01-01

    Transfectants with a wide range of cellular phenotypes are obtained during the process of cell line generation. For the successful manufacture of a therapeutic protein, a means is required to identify a cell line with desirable growth and productivity characteristics from this phenotypically wide-ranging transfectant population. This identification process is on the critical path for first-in-human studies. We have stringently examined a typical selection strategy used to isolate cell lines suitable for cGMP manufacturing. One-hundred and seventy-five transfectants were evaluated as they progressed through the different assessment stages of the selection strategy. High producing cell lines, suitable for cGMP manufacturing, were identified. However, our analyses showed that the frequency of isolation of the highest producing cell lines was low and that ranking positions were not consistent between each assessment stage, suggesting that there is potential to improve upon the strategy. Attempts to increase the frequency of isolation of the 10 highest producing cell lines, by in silico analysis of alternative selection strategies, were unsuccessful. We identified alternative strategies with similar predictive capabilities to the typical selection strategy. One alternate strategy required fewer cell lines to be progressed at the assessment stages but the stochastic nature of the models means that cell line numbers are likely to change between programs. In summary, our studies illuminate the potential for improvement to this and future selection strategies, based around use of assessments that are more informative or that reduce variance, paving the way to improved efficiency of generation of manufacturing cell lines. PMID:20623584

  16. Effects of atrial and brain natriuretic peptides upon cyclic GMP levels, potassium transport, and receptor binding in rat astrocytes

    The ability of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) to alter cyclic GMP levels and NaKCl cotransport in rat neocortical astrocytes was determined. At concentrations of 10(-9)-10(-6) M, rat ANP99-126 (rANF), rat ANP102-126 (auriculin B), and rat ANP103-126 (atriopeptin III) stimulated 6- to 100-fold increases in cyclic GMP levels. Porcine BNP (pBNP) and rat BNP (rBNP) were 20%-90% as effective as rANF over most of this concentration range, although 10(-6) M pBNP produced a greater effect than rANF. NaKCl cotransport as measured by bumetanide-sensitive 86Rb+ influx was not altered by exposure of astrocytes to 10(-6)M rANF, pBNP, or rBNP. Both pBNP and rBNP, as well as rat ANP103-123 (atriopeptin I) and des[gl18, ser19, gly20, leu21, gly22] ANF4-23-NH2 (C-ANF4-23) strongly competed for specific 125I-rANF binding sites in astrocyte membranes with affinities ranging from 0.03 to 0.4 nM, suggesting that virtually all binding sites measured at subnanomolar concentrations of 125I-rANF were of the ANP-C (ANF-R2) receptor subtype. These receptors are thought to serve a clearance function and may be linked to a guanylate cyclase activity that is chemically and pharmacologically distinct from that coupled to ANP-A (ANF-R1) receptors. ANP receptors on astrocytes may function in limiting the access of ANP and BNP to neurons involved in body fluid and cardiovascular regulation

  17. "十三五"期间中国住房与产业地产需求预测%Forecasting Demand for Housinng and Industrial Real Estate During the 13th Five-year Plan for Economic and Social Development of the People's Republic of China

    洪丽; 曾国安

    2015-01-01

    基于近十年的历史资料和新常态的经济发展速度对"十三五"期间全国保障性住房、商品住房和产业地产需求情况的预测,到"十三五"期末,住房保障覆盖率达到20%,"十三五"期间新增保障户数约612万户;商品住房需求总量约为63.2亿平方米6690万套,其中,城市化创造的住房需求为29.6亿平方米3290万套,改善型住房需求约为26亿平方米2600万套,投资和财产住房需求约为7.58亿平方米800万套,平均每年商品住宅需求量约为12.6亿平方米1338万套;"十三五"期间全国办公写字楼需求面积达到17875万平方米,年均建设3575万平方米;新增商业用房需求总面积达到43970万平方米,年均需求量约为8800万平方米;新增工业用房需求面积达到21202万平方米,年均建设4240万平方米.%Based on nearly ten years of historical data and in the case of the"new normal"of GDP growth rate being 7.5%,this paper has forecasted the demand for housing and industrial real estate during the 13th five-year plan for economic and social development of the People's Republic of China. From 2016 to 2020,affordable housing should meet the needs of 6.12 million new households and the coverage rate of affordable housing is about 20%.The commercial housing demand is about 6.32 billion square meters which can meet the needs of 66.9 million households.The commercial housing demand includes three aspects:the demand brought by the urbanization which is 2.96 billion square meters and can meet the needs of 32.9 million households, the demand for improving the level of housing which is 2.6 billion square meters and can meet the needs of 26 million households,the demand for investment which is 0.758 billion square meters.The demand for office building is 178.75 million square meters and the demand for new commercial properties is 439.7 million square meters and the demand for new industrial real estate is 212.02 million square meters during the 13th five

  18. Annual Check-up

    ... Medical Conditions Nutrition & Fitness Emotional Health Annual Check-Up Posted under Health Guides . Updated 7 January 2015. + ... I get ready for my annual medical check-up? If this is your first visit to your ...

  19. Sildenafil citrate increases myocardial cGMP content in rat heart, decreases its hypertrophic response to isoproterenol and decreases myocardial leak of creatine kinase and troponin T

    Ketat Amal F

    2005-04-01

    Full Text Available Abstract Background Cardiac hypertrophy is a major risk factor for morbidity and mortality in a number of cardiovascular diseases. Consequently, the signaling pathways that inhibit cardiac hypertrophy are currently receiving much interest. Among them, nitric oxide (NO, signaling via cGMP and cGMP-dependent protein kinase I, has been recognized as a negative regulator of cardiac hypertrophy. The present study investigated the in-vivo effect of sildenafil as a phosphodiestrase-5A (PDE-5A inhibitor on the hypertrophic response of rat heart to isoproterenol and the relation of this effect to the level of myocardial cGMP and integrity of the constitutive nitric oxide synthase (cNOS activity. Results The results showed that daily intraperitoneal administration of sildenafil per se for 10 days was without noticeable adverse effects on survival or myocardium. Conversely, daily subcutaneous administration of isoproterenol for 10 days caused significant myocardial hypertrophy, cell injury and decline in survival. When sildenafil was injected daily, one hour before isoproterenol, survival was significantly improved and the myocardium didn't show significant hypertrophy or cell injury. Interestingly, sildenafil was accompanied by significant rise in myocardial cGMP level, a parameter which was found in the present study to possess a significant negative correlation with cardiac hypertrophy and leak of cardiac troponin T into serum. At the same time, cGMP was found to possess a positive correlation with myocardial creatine kinase activity that reflects the efficiency of the energy utilization processes in the myocardium. However, in rats given Nω-nitro-L-arginine (L-NNA as a competitive inhibitor of cNOS, sildenafil failed to show any favorable effect on survival or the myocardial injury parameters used to assess isoproterenol-induced injury. Conclusion The present study suggests that increased cardiac cGMP level by sildenafil have a cardioprotective effect

  20. The exopolysaccharide gene cluster Bcam1330-Bcam1341 is involved in Burkholderia cenocepacia biofilm formation, and its expression is regulated by c-di-GMP and Bcam1349

    Fazli, Mustafa; McCarthy, Yvonne; Givskov, Michael; Ryan, Robert P; Tolker-Nielsen, Tim

    2013-01-01

    In Burkholderia cenocepacia, the second messenger cyclic diguanosine monophosphate (c-di-GMP) has previously been shown to positively regulate biofilm formation and the expression of cellulose and type-I fimbriae genes through binding to the transcriptional regulator Bcam1349. Here, we provide...... evidence that cellulose and type-I fimbriae are not involved in B. cenocepacia biofilm formation in flow chambers, and we identify a novel Bcam1349/c-di-GMP-regulated exopolysaccharide gene cluster which is essential for B. cenocepacia biofilm formation. Overproduction of Bcam1349 in trans promotes wrinkly...... matrix exopolysaccharide and to be essential for flow-chamber biofilm formation. We demonstrate that Bcam1349 binds to the promoter region of genes in the Bcam1330-Bcam1341 cluster and that this binding is enhanced by the presence of c-di-GMP. Furthermore, we demonstrate that overproduction of both c-di-GMP...

  1. Crystallization and preliminary X-ray diffraction characterization of the XccFimXEAL–c-di-GMP and XccFimXEAL–c-di-GMP–XccPilZ complexes from Xanthomonas campestris

    Crystals of the XccFimXEAL–c-di-GMP and SeMet-XccFimXEAL–c-di-GMP–XccPilZ complexes from X. campestris diffracted to resolutions of 2.5 and 2.7 Å, respectively. c-di-GMP is a major secondary-messenger molecule in regulation of bacterial pathogenesis. Therefore, the c-di-GMP-mediated signal transduction network is of considerable interest. The PilZ domain was the first c-di-GMP receptor to be predicted and identified. However, every PilZ domain binds c-di-GMP with a different binding affinity. Intriguingly, a noncanonical PilZ domain has recently been found to serve as a mediator to link FimXEAL to the PilB or PilT ATPase to control the function of type IV pili (T4P). It is thus essential to determine the structure of the FimXEAL–PilZ complex in order to determine how the binding of c-di-GMP to the FimXEAL domain induces conformational change of the adjoining noncanonical PilZ domain, which may transmit information to PilB or PilT to control T4P function. Here, the preparation and preliminary X-ray diffraction studies of the XccFimXEAL–c-di-GMP and XccFimXEAL–c-di-GMP–XccPilZ complexes from Xcc (Xanthomonas campestris pv. campesteris) are reported. Detailed studies of these complexes may allow a more thorough understanding of how c-di-GMP transmits its effects through the degenerate EAL domain and the noncanonical PilZ domain

  2. Differential effects of chronic hyperammonemia on modulation of the glutamate-nitric oxide-cGMP pathway by metabotropic glutamate receptor 5 and low and high affinity AMPA receptors in cerebellum in vivo.

    Cabrera-Pastor, Andrea; Llansola, Marta; Reznikov, Vitaliy; Boix, Jordi; Felipo, Vicente

    2012-07-01

    Previous studies show that chronic hyperammonemia impairs learning ability of rats by impairing the glutamate-nitric oxide (NO)-cyclic guanosine mono-phosphate (cGMP) pathway in cerebellum. Three types of glutamate receptors cooperate in modulating the NO-cGMP pathway: metabotropic glutamate receptor 5 (mGluR5), (RS)-α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and N-methyl-d-aspartic acid (NMDA) receptors. The aim of this work was to assess whether hyperammonemia alters the modulation of this pathway by mGluR5 and AMPA receptors in cerebellum in vivo. The results support that in control rats: (1) low AMPA concentrations (0.1mM) activate nearly completely Ca(2+)-permeable (glutamate receptor subunit 2 (GluR2)-lacking) AMPA receptors and the NO-cGMP pathway; (2) higher AMPA concentrations (0.3 mM) also activate Ca(2+)-impermeable (GluR2-containing) AMPA receptors, leading to activation of NMDA receptors and of NO-cGMP pathway. Moreover, the data support that chronic hyperammonemia: (1) reduces glutamate release and activation of the glutamate-NO-cGMP pathway by activation of mGluR5; (2) strongly reduces the direct activation by AMPA receptors of the NO-cGMP pathway, likely due to reduced entry of Ca(2+) through GluR2-lacking, high affinity AMPA receptors; (3) strongly increases the indirect activation of the NO-cGMP pathway by high affinity AMPA receptors, likely due to increased entry of Na(+) through GluR2-lacking AMPA receptors and NMDA receptors activation; (4) reduces the indirect activation of the NO-cGMP pathway by low affinity AMPA receptors, likely due to reduced activation of NMDA receptors. PMID:22521775

  3. Compounds extracted from Phyllantus and Jatropha elliptica inhibit the binding of [3H]glutamate and [3H]GMP-PNP in rat cerebral cortex membrane.

    Martini, L H; Souza, C R; Marques, P B; Calixto, J B; Yunes, R A; Souza, D O

    2000-02-01

    Glutamate is to be considered a nociceptive neurotransmitter and glutamatergic antagonists present antinoceptive activity. In this study we investigated the effects of the naturally occurring antinociceptive compounds rutin, geraniin and quercetine extracted from Phyllanthus, as well as the diterpene jatrophone, extracted from Jatropha elliptica on the binding of [3H]glutamate and [3H]GMP-PNP [a GTP analogue which binds to extracellular site(s), modulating the glutamatergic transmission] in rat brain membrane. Jatrophone inhibited [3H]glutamate binding and geraniin inhibited [3H]GMP-PNP binding. Quercetine inhibited the binding of both ligands. These results may indicate a neurochemical parameter possibly related to the antinoceptive activity of these natural compounds. PMID:10786704

  4. Strategic Thinking on Innovation and Development of Higher Continuing Education during the 13 th Five-year Plan Period%“十三五”时期高校继续教育创新发展的战略思考

    严继昌

    2016-01-01

    The constant development of our higher education is facing new situation and challenges. The rapid development of corporate universities and the rising of vocational education groups impact the development space of higher continuing education;the development trend of online education and integration trend of continuing education are driving the transformation and upgra-ding of higher continuing education. In the Internet era, higher education should firmly grasp the opportunity of information tech-nology revolution, and be adapt to the national policy change of network education and training, to promote MOOC and Micro-lec-tures construction of degree continuing education, vigorously promoting the construction of resources, and giving impetus to estab-lishing education alliance of resources opening and online education. During the 13th Five-year Plan period, the higher continuing education should firmly establish the development ideas of innovation, coordination, green, openness and sharing, and set up the development strategy of serving the country, focusing on the cultivating mode, the content of training, the transformation of learn-ing achievements, so as to speed up the pace of reform.%当前我国高校继续教育发展面临新的形势和挑战,企业大学的快速发展、职业教育集团的兴起冲击着高校继续教育的发展空间,在线教育的发展态势、继续教育的融合趋势推动着高校继续教育的转型升级。在互联网时代,高校继续教育要紧紧抓住信息技术变革的契机,适应国家对网络教育和网络培训的政策变化,大力推进学历继续教育的慕课、微课建设,推进资源开放和在线教育联盟建设。“十三五”时期,高校继续教育要牢固树立创新、协调、绿色、开放、共享的发展理念,服务国家发展战略,重点在培养模式、培训内容、学习成果转换等方面,加快综合改革的步伐。

  5. S-nitrosothiols dilate the mesenteric artery more potently than the femoral artery by a cGMP and L-type calcium channel-dependent mechanism.

    Liu, Taiming; Schroeder, Hobe J; Zhang, Meijuan; Wilson, Sean M; Terry, Michael H; Longo, Lawrence D; Power, Gordon G; Blood, Arlin B

    2016-08-31

    S-nitrosothiols (SNOs) are metabolites of NO with potent vasodilatory activity. Our previous studies in sheep indicated that intra-arterially infused SNOs dilate the mesenteric vasculature more than the femoral vasculature. We hypothesized that the mesenteric artery is more responsive to SNO-mediated vasodilation, and investigated various steps along the NO/cGMP pathway to determine the mechanism for this difference. In anesthetized adult sheep, we monitored the conductance of mesenteric and femoral arteries during infusion of S-nitroso-l-cysteine (L-cysNO), and found mesenteric vascular conductance increased (137 ± 3%) significantly more than femoral conductance (26 ± 25%). Similar results were found in wire myography studies of isolated sheep mesenteric and femoral arteries. Vasodilation by SNOs was attenuated in both vessel types by the presence of ODQ (sGC inhibitor), and both YC-1 (sGC agonist) and 8-Br-cGMP (cGMP analog) mediated more potent relaxation in mesenteric arteries than femoral arteries. The vasodilatory difference between mesenteric and femoral arteries was eliminated by antagonists of either protein kinase G or L-type Ca(2+) channels. Western immunoblots showed a larger L-type Ca(2+)/sGC abundance ratio in mesenteric arteries than in femoral arteries. Fetal sheep mesenteric arteries were more responsive to SNOs than adult mesenteric arteries, and had a greater L-Ca(2+)/sGC ratio (p = 0.047 and r = -0.906 for correlation between Emax and L-Ca(2+)/sGC). These results suggest that mesenteric arteries, especially those in fetus, are more responsive to SNO-mediated vasodilation than femoral arteries due to a greater role of the L-type calcium channel in the NO/cGMP pathway. PMID:27235767

  6. Increased Activity of cGMP-Specific Phosphodiesterase (PDE5) Contributes to Resistance to Atrial Natriuretic Peptide Natriuresis in the Pregnant Rat

    Ni, Xi-Ping; SAFAI, MASSY; Rishi, Rahul; Baylis, Chris; Humphreys, Michael H.

    2004-01-01

    Increased cGMP-specific phosphodiesterase (PDE5) activity in renal inner medullary collecting duct (IMCD) cells contributes to resistance to atrial natriuretic peptide (ANP) and the excessive sodium retention seen in experimental nephrotic syndrome and liver cirrhosis. Normal pregnancy is also accompanied by sodium retention and plasma volume expansion, and pregnant rats are resistant to the natriuretic action of ANP. The authors investigated a possible role of increased renal PDE5 activity i...

  7. Activation of a cGMP-sensitive calcium-dependent chloride channel may cause transition from calcium waves to whole-cell oscillations in smooth muscle cells

    Jacobsen, Jens Christian; Aalkjær, Christian; Nilsson, Holger;

    2007-01-01

    -frequency waves may transform into high-frequency whole-cell calcium oscillations. Simultaneously, multiple cells synchronize leading to rhythmic generation of tension. We present a mathematical model of vascular smooth muscle cells that aims at characterizing this sudden transition. Simulations show calcium...... onset of oscillations in membrane potential within the individual cell may underlie sudden intercellular synchronization and the appearance of vasomotion. Key words: Vasomotion, Chloride channel, cGMP, Mathematical model, Calcium waves....

  8. Can selective inhibitors of cyclic guanosine monophosphate (cGMP)-specific phosphadiesterase type 5 (PDE 5) offer protection against contrast induced nephropathy?

    Morcos, Sameh K.

    2014-01-01

    Parenchymal hypoxia within the renal outer medulla plays an important role in the pathogenesis of contrast induced nephropathy (CIN). Nitric oxide (NO) is crucial for medullary oxygenation by enhancing regional blood flow. Augmenting the effect of NO in the renal medulla by the use of selective inhibitors of cyclic guanosine monophosphate (cGMP)-specific phosphadiesterase type 5 (PDE 5) such as sildenafil (Viagra™), vardenafil (Levitra™) or tadalafil (Cialis™) could reduce the severity of the...

  9. Diguanylate Cyclase Null Mutant Reveals That C-Di-GMP Pathway Regulates the Motility and Adherence of the Extremophile Bacterium Acidithiobacillus caldus

    Castro, Matías; Deane, Shelly M.; Ruiz, Lina; Rawlings, Douglas E.; Guiliani, Nicolas

    2015-01-01

    An understanding of biofilm formation is relevant to the design of biological strategies to improve the efficiency of the bioleaching process and to prevent environmental damages caused by acid mine/rock drainage. For this reason, our laboratory is focused on the characterization of the molecular mechanisms involved in biofilm formation in different biomining bacteria. In many bacteria, the intracellular levels of c-di-GMP molecules regulate the transition from the motile planktonic state to ...

  10. Development of a transgenic Plasmodium berghei line (Pb pfpkg expressing the P. falciparum cGMP-dependent protein kinase, a novel antimalarial drug target.

    Rita Tewari

    Full Text Available With the inevitable selection of resistance to antimalarial drugs in treated populations, there is a need for new medicines to enter the clinic and new targets to progress through the drug discovery pipeline. In this study we set out to develop a transgenic rodent model for testing inhibitors of the Plasmodium falciparum cyclic GMP-dependent kinase in vivo. A model was needed that would allow us to investigate whether differences in amino acid sequence of this enzyme between species influences in vivo efficacy. Here we report the successful development of a transgenic P. berghei line in which the cyclic GMP-dependent protein kinase (PKG was replaced by the P. falciparum orthologue. We demonstrate that the P. falciparum orthologue was able to functionally complement the endogenous P. berghei pkg gene throughout blood stage development and early sexual development. However, subsequent development in the mosquito was severely compromised. We show that this is due to a defect in the female lineage of the transgenic by using genetic crosses with both male and female deficient P. berghei lines. This defect could be due to expression of a female-specific target in the mosquito stages of P. berghei that cannot be phosphorylated by the P. falciparum kinase. Using a previously reported anti-coccidial inhibitor of the cyclic GMP-dependent protein kinase, we show no difference in in vivo efficacy between the transgenic and control P. berghei lines. This in vivo model will be useful for screening future generations of cyclic GMP-dependent protein kinase inhibitors and allowing us to overcome any species-specific differences in the enzyme primary sequence that would influence in vivo efficacy in the rodent model. The approach will also be applicable to in vivo testing of other antimalarial compounds where the target is known.

  11. The enzymatic preparation of [α-32P]nucleoside triphosphates, cyclic [32P]AMP, and cyclic [32P]GMP

    A method has been developed for the enzymatic preparation of α-32P-labelled ribo- and deoxyribonucleoside triphosphates, cyclic [32P]AMP, and cyclic [32P]GMP of high specific radioactivity and in high yield from 32Psub(i). The method also enables the preparation of [γ-32P]ATP, [γ-32P]GTP, [γ-32P]ITP, and [γ-32P]-dATP of very high specific activity and in high yield. (Auth.)

  12. Nitric oxide synthetic pathway and cGMP levels are altered in red blood cells from end-stage renal disease patients.

    Di Pietro, Natalia; Giardinelli, Annalisa; Sirolli, Vittorio; Riganti, Chiara; Di Tomo, Pamela; Gazzano, Elena; Di Silvestre, Sara; Panknin, Christina; Cortese-Krott, Miriam M; Csonka, Csaba; Kelm, Malte; Ferdinandy, Péter; Bonomini, Mario; Pandolfi, Assunta

    2016-06-01

    Red blood cells (RBCs) enzymatically produce nitric oxide (NO) by a functional RBC-nitric oxide synthase (RBC-NOS). NO is a vascular key regulatory molecule. In RBCs its generation is complex and influenced by several factors, including insulin, acetylcholine, and calcium. NO availability is reduced in end-stage renal disease (ESRD) and associated with endothelial dysfunction. We previously demonstrated that, through increased phosphatidylserine membrane exposure, ESRD-RBCs augmented their adhesion to human cultured endothelium, in which NO bioavailability decreased. Since RBC-NOS-dependent NO production in ESRD is unknown, this study aimed to investigate RBC-NOS levels/activation, NO production/bioavailability in RBCs from healthy control subjects (C, N = 18) and ESRD patients (N = 27). Although RBC-NOS expression was lower in ESRD-RBCs, NO, cyclic guanosine monophosphate (cGMP), RBC-NOS Serine1177 phosphorylation level and eNOS/Calmodulin (CaM)/Heat Shock Protein-90 (HSP90) interaction levels were higher in ESRD-RBCs, indicating increased enzyme activation. Conversely, following RBCs stimulation with insulin or ionomycin, NO and cGMP levels were significantly lower in ESRD- than in C-RBCs, suggesting that uremia might reduce the RBC-NOS response to further stimuli. Additionally, the activity of multidrug-resistance-associated protein-4 (MRP4; cGMP-membrane transporter) was significantly lower in ESRD-RBCs, suggesting a possible compromised efflux of cGMP across the ESRD-RBCs membrane. This study for the first time showed highest basal RBC-NOS activation in ESRD-RBCs, possibly to reduce the negative impact of decreased NOS expression. It is further conceivable that high NO production only partially affects cell function of ESRD-RBCs maybe because in vivo they are unable to respond to physiologic stimuli, such as calcium and/or insulin. PMID:27206740

  13. The immunosuppressive drug azathioprine inhibits biosynthesis of the bacterial signal molecule cyclic-di-GMP by interfering with intracellular nucleotide pool availability.

    Antoniani, Davide; Rossi, Elio; Rinaldo, Serena; BOCCI, PAOLA; Lolicato, Marco; Paiardini, Alessandro; Raffaelli, Nadia; Cutruzzolà, Francesca; Landini, Paolo

    2013-01-01

    In Gram-negative bacteria, production of the signal molecule c-di-GMP by diguanylate cyclases (DGCs) is a key trigger for biofilm formation, which, in turn, is often required for the development of chronic bacterial infections. Thus, DGCs represent interesting targets for new chemotherapeutic drugs with anti-biofilm activity. We searched for inhibitors of the WspR protein, a Pseudomonas aeruginosa DGC involved in biofilm formation and production of virulence factors, using a set of microbiolo...

  14. The cyclic-di-GMP diguanylate cyclase CdgA has a role in biofilm formation and exopolysaccharide production in Azospirillum brasilense.

    Ramírez-Mata, Alberto; López-Lara, Lilia I; Xiqui-Vázquez, Ma Luisa; Jijón-Moreno, Saúl; Romero-Osorio, Angelica; Baca, Beatriz E

    2016-04-01

    In bacteria, proteins containing GGDEF domains are involved in production of the second messenger c-di-GMP. Here we report that the cdgA gene encoding diguanylate cyclase A (CdgA) is involved in biofilm formation and exopolysaccharide (EPS) production in Azospirillum brasilense Sp7. Biofilm quantification using crystal violet staining revealed that inactivation of cdgA decreased biofilm formation. In addition, confocal laser scanning microscopy analysis of green-fluorescent protein-labeled bacteria showed that, during static growth, the biofilms had differential levels of development: bacteria harboring a cdgA mutation exhibited biofilms with considerably reduced thickness compared with those of the wild-type Sp7 strain. Moreover, DNA-specific staining and treatment with DNase I, and epifluorescence studies demonstrated that extracellular DNA and EPS are components of the biofilm matrix in Azospirillum. After expression and purification of the CdgA protein, diguanylate cyclase activity was detected. The enzymatic activity of CdgA-producing cyclic c-di-GMP was determined using GTP as a substrate and flavin adenine dinucleotide (FAD(+)) and Mg(2)(+) as cofactors. Together, our results revealed that A. brasilense possesses a functional c-di-GMP biosynthesis pathway. PMID:26708984

  15. Biosynthetic Enzyme GMP Synthetase Cooperates with Ubiquitin-Specific Protease 7 in Transcriptional Regulation of Ecdysteroid Target Genes▿

    van der Knaap, Jan A.; Kozhevnikova, Elena; Langenberg, Karin; Moshkin, Yuri M.; Verrijzer, C. Peter

    2010-01-01

    Drosophila GMP synthetase binds ubiquitin-specific protease 7 (USP7) and is required for its ability to deubiquitylate histone H2B. Previously, we showed that the GMPS/USP7 complex cooperates with the Polycomb silencing system through removal of the active ubiquitin mark from histone H2B (H2Bub). Here, we explored the interplay between GMPS and USP7 further and assessed their role in hormone-regulated gene expression. Genetic analysis established a strong cooperation between GMPS and USP7, which is counteracted by the histone H2B ubiquitin ligase BRE1. Loss of either GMPS or USP7 led to increased levels of histone H2Bub in mutant animals. These in vivo analyses complement our earlier biochemical results, establishing that GMPS/USP7 mediates histone H2B deubiquitylation. We found that GMPS/USP7 binds ecdysone-regulated loci and that mutants display severe misregulation of ecdysone target genes. Ecdysone receptor (EcR) interacts biochemically and genetically with GMPS/USP7. Genetic and gene expression analyses suggested that GMPS/USP7 acts as a transcriptional corepressor. These results revealed the cooperation between a biosynthetic enzyme and a ubiquitin protease in developmental gene control by hormone receptors. PMID:19995917

  16. Aversive Behavior in the Nematode C. elegans Is Modulated by cGMP and a Neuronal Gap Junction Network.

    Krzyzanowski, Michelle C; Woldemariam, Sarah; Wood, Jordan F; Chaubey, Aditi H; Brueggemann, Chantal; Bowitch, Alexander; Bethke, Mary; L'Etoile, Noelle D; Ferkey, Denise M

    2016-07-01

    All animals rely on their ability to sense and respond to their environment to survive. However, the suitability of a behavioral response is context-dependent, and must reflect both an animal's life history and its present internal state. Based on the integration of these variables, an animal's needs can be prioritized to optimize survival strategies. Nociceptive sensory systems detect harmful stimuli and allow for the initiation of protective behavioral responses. The polymodal ASH sensory neurons are the primary nociceptors in C. elegans. We show here that the guanylyl cyclase ODR-1 functions non-cell-autonomously to downregulate ASH-mediated aversive behaviors and that ectopic cGMP generation in ASH is sufficient to dampen ASH sensitivity. We define a gap junction neural network that regulates nociception and propose that decentralized regulation of ASH signaling can allow for rapid correlation between an animal's internal state and its behavioral output, lending modulatory flexibility to this hard-wired nociceptive neural circuit. PMID:27459302

  17. Nitric oxide regulates BDNF release from nodose ganglion neurons in a pattern-dependent and cGMP-independent manner.

    Hsieh, Hui-ya; Robertson, Carolyn L; Vermehren-Schmaedick, Anke; Balkowiec, Agnieszka

    2010-05-01

    Activity of arterial baroreceptors is modulated by neurohumoral factors, including nitric oxide (NO), released from endothelial cells. Baroreceptor reflex responses can also be modulated by NO signaling in the brainstem nucleus tractus solitarius (NTS), the primary central target of cardiovascular afferents. Our recent studies indicate that brain-derived neurotrophic factor (BDNF) is abundantly expressed by developing and adult baroreceptor afferents in vivo, and released from cultured nodose ganglion (NG) neurons by patterns of baroreceptor activity. Using electrical field stimulation and ELISA in situ, we show that exogenous NO nearly abolishes BDNF release from newborn rat NG neurons in vitro stimulated with single pulses delivered at 6 Hz, but not 2-pulse bursts delivered at the same 6-Hz frequency, that corresponds to a rat heart rate. Application of L-NAME, a specific inhibitor of endogenous NO synthases, does not have any significant effect on activity-dependent BDNF release, but leads to upregulation of BDNF expression in an activity-dependent manner. The latter effect suggests a novel mechanism of homeostatic regulation of activity-dependent BDNF expression with endogenous NO as a key player. The exogenous NO-mediated effect does not involve the cGMP-protein kinase G (PKG) pathway, but is largely inhibited by N-ethylmaleimide and TEMPOL that are known to prevent S-nitrosylation. Together, our current data identify previously unknown mechanisms regulating BDNF availability, and point to NO as a likely regulator of BDNF at baroafferent synapses in the NTS. PMID:19937808

  18. Involvement of NO/cGMP pathway in the antidepressant-like effect of gabapentin in mouse forced swimming test.

    Ostadhadi, Sattar; Kordjazy, Nastaran; Haj-Mirzaian, Arya; Ameli, Sanaz; Akhlaghipour, Golnoosh; Dehpour, AhmadReza

    2016-04-01

    Based on clinical studies regarding the beneficial effect of gabapentin in depression, we aimed to evaluate the antidepressant-like properties of gabapentin in mice and also the participation of nitric oxide (NO)/cyclic guanosine monophosphate pathway in this effect. The following drugs were used in this study: gabapentin; N(G)-nitro-L-arginine methyl ester (L-NAME), a non-specific NO synthase (NOS) inhibitor; 7-nitroindazole, a specific neuronal NOS inhibitor; aminoguanidine, a specific inducible NOS inhibitor; L-arginine, a NO precursor; and sildenafil, a phosphodiestrase inhibitor. Finally, we studied the behavioral effects through the forced swimming test (FST) and the changes of the hippocampus NO level through nitrite assay. The immobility time was significantly reduced after gabapentin administration. Co-administration of non-effective doses of gabapentin and L-NAME or 7-nitroindazole (7-NI) resulted in antidepressant-like effect in FST, while aminoguanidine did not affect the immobility time of gabapentin-treated mice. Furthermore, the antidepressant-like property of gabapentin was prevented by L-arginine or sildenafil. Also, the hippocampal nitrite level was significantly lower in gabapentin-treated mice relative to saline-injected mice, and co-administration of 7-NI with sub-effective gabapentin caused a significant decrease in hippocampal nitrite levels. Our results indicate that the antidepressant-like effect of gabapentin in the mice FST model is mediated at least in part through nitric oxide/cyclic guanosine monophosphate (cGMP) pathway. PMID:26753696

  19. Nitric oxide inhibits neuroendocrine CaV1 L-channel gating via cGMP-dependent protein kinase in cell-attached patches of bovine chromaffin cells

    Carabelli, Valentina; D'Ascenzo, Marcello; Carbone, Emilio; Grassi, Claudio

    2002-01-01

    Nitric oxide (NO) regulates the release of catecholamines from the adrenal medulla but the molecular targets of its action are not yet well identified. Here we show that the NO donor sodium nitroprusside (SNP, 200 μM) causes a marked depression of the single CaV1 L-channel activity in cell-attached patches of bovine chromaffin cells. SNP action was complete within 3-5 min of cell superfusion. In multichannel patches the open probability (NPo) decreased by ∼60 % between 0 and +20 mV. Averaged currents over a number of traces were proportionally reduced and showed no drastic changes to their time course. In single-channel patches the open probability (Po) at +10 mV decreased by the same amount as that of multichannel patches (∼61 %). Such a reduction was mainly associated with an increased probability of null sweeps and a prolongation of mean shut times, while first latency, mean open time and single-channel conductance were not significantly affected. Addition of the NO scavenger carboxy-PTIO or cell treatment with the guanylate cyclase inhibitor ODQ prevented the SNP-induced inhibition. 8-Bromo-cyclicGMP (8-Br-cGMP; 400 μM) mimicked the action of the NO donor and the protein kinase G blocker KT-5823 prevented this effect. The depressive action of SNP was preserved after blocking the cAMP-dependent up-regulatory pathway with the protein kinase A inhibitor H89. Similarly, the inhibitory action of 8-Br-cGMP proceeded regardless of the elevation of cAMP levels, suggesting that cGMP/PKG and cAMP/PKA act independently on L-channel gating. The inhibitory action of 8-Br-cGMP was also independent of the G protein-induced inhibition of L-channels mediated by purinergic and opiodergic autoreceptors. Since Ca2+ channels contribute critically to both the local production of NO and catecholamine release, the NO/PKG-mediated inhibition of neuroendocrine L-channels described here may represent an important autocrine signalling mechanism for controlling the rate of

  20. Nitric oxide inhibits neuroendocrine Ca(V)1 L-channel gating via cGMP-dependent protein kinase in cell-attached patches of bovine chromaffin cells.

    Carabelli, Valentina; D'Ascenzo, Marcello; Carbone, Emilio; Grassi, Claudio

    2002-06-01

    Nitric oxide (NO) regulates the release of catecholamines from the adrenal medulla but the molecular targets of its action are not yet well identified. Here we show that the NO donor sodium nitroprusside (SNP, 200 microM) causes a marked depression of the single Ca(V)1 L-channel activity in cell-attached patches of bovine chromaffin cells. SNP action was complete within 3-5 min of cell superfusion. In multichannel patches the open probability (NP(o)) decreased by approximately 60 % between 0 and +20 mV. Averaged currents over a number of traces were proportionally reduced and showed no drastic changes to their time course. In single-channel patches the open probability (P(o)) at +10 mV decreased by the same amount as that of multichannel patches (approximately 61 %). Such a reduction was mainly associated with an increased probability of null sweeps and a prolongation of mean shut times, while first latency, mean open time and single-channel conductance were not significantly affected. Addition of the NO scavenger carboxy-PTIO or cell treatment with the guanylate cyclase inhibitor ODQ prevented the SNP-induced inhibition. 8-Bromo-cyclicGMP (8-Br-cGMP; 400 microM) mimicked the action of the NO donor and the protein kinase G blocker KT-5823 prevented this effect. The depressive action of SNP was preserved after blocking the cAMP-dependent up-regulatory pathway with the protein kinase A inhibitor H89. Similarly, the inhibitory action of 8-Br-cGMP proceeded regardless of the elevation of cAMP levels, suggesting that cGMP/PKG and cAMP/PKA act independently on L-channel gating. The inhibitory action of 8-Br-cGMP was also independent of the G protein-induced inhibition of L-channels mediated by purinergic and opiodergic autoreceptors. Since Ca(2+) channels contribute critically to both the local production of NO and catecholamine release, the NO/PKG-mediated inhibition of neuroendocrine L-channels described here may represent an important autocrine signalling mechanism

  1. Annual report 1973

    The GKSS scientific annual report summarizes the problems and results of the research and development projects of 1973. In contrast to earlier annual reports, a comprehensive description of the research facilities is not included. The annual report was extended by the paragraph 'Financial Report 1973' in the chapter 'Development of Geesthacht Research Centre'. The financial report gives a survey of the financial transactions and the major operations of the year under review. (orig./AK)

  2. Annual Statistical Supplement, 2015

    Social Security Administration — The Annual Statistical Supplement, 2015 includes the most comprehensive data available on the Social Security and Supplemental Security Income programs. More than...

  3. Annual Statistical Supplement, 2011

    Social Security Administration — The Annual Statistical Supplement, 2011 includes the most comprehensive data available on the Social Security and Supplemental Security Income programs. More than...

  4. Annual Statistical Supplement, 2003

    Social Security Administration — The Annual Statistical Supplement, 2003 includes the most comprehensive data available on the Social Security and Supplemental Security Income programs. More than...

  5. Annual Statistical Supplement, 2004

    Social Security Administration — The Annual Statistical Supplement, 2004 includes the most comprehensive data available on the Social Security and Supplemental Security Income programs. More than...

  6. Annual Statistical Supplement, 2008

    Social Security Administration — The Annual Statistical Supplement, 2008 includes the most comprehensive data available on the Social Security and Supplemental Security Income programs. More than...

  7. Annual Statistical Supplement, 2002

    Social Security Administration — The Annual Statistical Supplement, 2002 includes the most comprehensive data available on the Social Security and Supplemental Security Income programs. More than...

  8. Annual Statistical Supplement, 2001

    Social Security Administration — The Annual Statistical Supplement, 2001 includes the most comprehensive data available on the Social Security and Supplemental Security Income programs. More than...

  9. Annual Statistical Supplement, 2006

    Social Security Administration — The Annual Statistical Supplement, 2006 includes the most comprehensive data available on the Social Security and Supplemental Security Income programs. More than...

  10. Annual Statistical Supplement, 2000

    Social Security Administration — The Annual Statistical Supplement, 2000 includes the most comprehensive data available on the Social Security and Supplemental Security Income programs. More than...

  11. Annual Statistical Supplement, 2009

    Social Security Administration — The Annual Statistical Supplement, 2009 includes the most comprehensive data available on the Social Security and Supplemental Security Income programs. More than...

  12. Annual Statistical Supplement, 2010

    Social Security Administration — The Annual Statistical Supplement, 2010 includes the most comprehensive data available on the Social Security and Supplemental Security Income programs. More than...

  13. Annual Statistical Supplement, 2007

    Social Security Administration — The Annual Statistical Supplement, 2007 includes the most comprehensive data available on the Social Security and Supplemental Security Income programs. More than...

  14. Annual Statistical Supplement, 2014

    Social Security Administration — The Annual Statistical Supplement, 2014 includes the most comprehensive data available on the Social Security and Supplemental Security Income programs. More than...

  15. Annual Statistical Supplement, 2005

    Social Security Administration — The Annual Statistical Supplement, 2005 includes the most comprehensive data available on the Social Security and Supplemental Security Income programs. More than...

  16. Le jugement divin, variations byzantines. Duel judiciaire et épreuve du fer rouge dans la pratique judiciaire à Byzance (XIIIe-XIVe siècles Byzantine Variations of the Divine Judgment: The Trial by Duel and the Ordeal of the Red-Hot Iron in Byzantine Judicial Practice (13th-14th Centuries

    Georgios Katsenis

    2009-03-01

    Full Text Available Le contact du monde byzantin avec les Croisades et l’Orient latin eut des répercussions dans le domaine de la justice byzantine. Au lieu des méthodes de preuve établies par le droit romain, l’application du duel judiciaire et de l’épreuve du fer rouge apparaît sous une forme presque identique à l’occidentale. L’absence d’un pouvoir central efficace et la crise conséquente des règles du droit romain permettaient leur diffusion à travers le monde byzantin. Malgré les réactions des savants ecclésiastiques, comme Démétrios Chomatianos, l’épreuve du fer rouge était largement appliquée en Epire pendant le XIIIe siècle. En Nicée, dans l’affaire de lèse-majesté (1252 où Michel Paléologue, le futur empereur, fut accusé – et durant l’instruction de laquelle un duel judiciaire eut lieu – des pressions furent exercées sur l’accusé pour qu’il subisse l’ordalie. Bien que plus tard Michel ait aboli les ordalies et le duel judiciaire, leur existence ne disparaîtra pas : pendant le XIVe siècle, l’application du fer rouge est attestée par l’empereur Jean Cantacuzène et la pratique du « chaudron » figure dans le Code d’Etienne Dušan, roi des Serbes.The contact of the Byzantine world with the Crusades and the Latin East provoked repercussions in the domain of Byzantine justice. Instead of the methods of proof established by the Roman law, the application of judicial combat and that of trial by hot iron appear in form almost similar to the Western. The lack of a strong central power and the subsequent crisis of the rules of Roman law permitted their diffusion throughout the Byzantine world. Despite the reaction of ecclesiastical scholars, like Demetrios Chomatianos, trial by hot iron was widely applied in Epirus during the 13th century. In Nicea, in the case of high treason (1252, where Michael Palaiologos, the future emperor, was accused – and where a judicial combat took place during the

  17. Annual Report 2001

    This Annual Report of the Institute of Atomic Energy describes the results of the research works carried out at the Institute at 2001. As in the preceding years the authors of the individual scientific reports published in this Annual Report are fully responsible for their content and layout. The Report contains the information on other activities of the Institute as well. (author)

  18. Annual Report 1997

    This Annual Report of the Institute of Atomic Energy describes the results of the research works carried out at the Institute at 1997. As in the preceding years the authors of the individual scientific reports published in this Annual Report are fully responsible for their content and layout. The Report contains the information on other activities of the Institute as well. (author)

  19. Natural gas annual 1991

    The Natural Gas Annual 1991 provides information on the supply and disposition of natural gas to a wide audience including industry, consumers Federal and State agencies, and education institutions. This report, the Natural Gas Annual 1991 Supplement: Company Profiles, presents a detailed profile of selected companies

  20. Annual Report 1997

    Golnik, N.; Mika, J.R.; Wieteska, K. [eds.

    1998-12-31

    This Annual Report of the Institute of Atomic Energy describes the results of the research works carried out at the Institute at 1997. As in the preceding years the authors of the individual scientific reports published in this Annual Report are fully responsible for their content and layout. The Report contains the information on other activities of the Institute as well. (author)

  1. Annual Energy Review, 2008

    None

    2009-06-01

    The Annual Energy Review (AER) is the Energy Information Administration's (EIA) primary report of annual historical energy statistics. For many series, data begin with the year 1949. Included are statistics on total energy production, consumption, trade, and energy prices; overviews of petroleum, natural gas, coal, electricity, nuclear energy, renewable energy, and international energy; financial and environment indicators; and data unit conversions.

  2. Systemic Induction of NO-, Redox-, and cGMP Signaling in the Pumpkin Extrafascicular Phloem upon Local Leaf Wounding.

    Gaupels, Frank; Furch, Alexandra C U; Zimmermann, Matthias R; Chen, Faxing; Kaever, Volkhard; Buhtz, Anja; Kehr, Julia; Sarioglu, Hakan; Kogel, Karl-Heinz; Durner, Jörg

    2016-01-01

    Cucurbits developed the unique extrafascicular phloem (EFP) as a defensive structure against herbivorous animals. Mechanical leaf injury was previously shown to induce a systemic wound response in the EFP of pumpkin (Cucurbita maxima). Here, we demonstrate that the phloem antioxidant system and protein modifications by NO are strongly regulated during this process. Activities of the central antioxidant enzymes dehydroascorbate reductase, glutathione reductase and ascorbate reductase were rapidly down-regulated at 30 min with a second minimum at 24 h after wounding. As a consequence levels of total ascorbate and glutathione also decreased with similar bi-phasic kinetics. These results hint toward a wound-induced shift in the redox status of the EFP. Nitric oxide (NO) is another important player in stress-induced redox signaling in plants. Therefore, we analyzed NO-dependent protein modifications in the EFP. Six to forty eight hours after leaf damage total S-nitrosothiol content and protein S-nitrosylation were clearly reduced, which was contrasted by a pronounced increase in protein tyrosine nitration. Collectively, these findings suggest that NO-dependent S-nitrosylation turned into peroxynitrite-mediated protein nitration upon a stress-induced redox shift probably involving the accumulation of reactive oxygen species within the EFP. Using the biotin switch assay and anti-nitrotyrosine antibodies we identified 9 candidate S-nitrosylated and 6 candidate tyrosine-nitrated phloem proteins. The wound-responsive Phloem Protein 16-1 (PP16-1) and Cyclophilin 18 (CYP18) as well as the 26.5 kD isoform of Phloem Protein 2 (PP2) were amenable to both NO modifications and could represent important redox-sensors within the cucurbit EFP. We also found that leaf injury triggered the systemic accumulation of cyclic guanosine monophosphate (cGMP) in the EFP and discuss the possible function of this second messenger in systemic NO and redox signaling within the EFP. PMID:26904092

  3. Annually laminated speleotherms : a review

    Baker, Andy; Smith, Claire; Jex, Catherine; Fairchild, I. J.; Genty, Dominique; Fuller, Lisa

    2008-01-01

    This review of annually laminated speleothems firstly considers the four types of annual laminae found within speleothems: fluorescent laminae formed by annual variations in organic matter flux; visible or petrographic laminae, formed by annual variations in calcite texture or fabric; calcite-aragonite couplets; and finally trace element laminae. The methods available to confirm the annual nature, or otherwise, of lamina deposition are reviewed. We consider the use of annual laminae ...

  4. The regulatory role of the NO/cGMP signal transduction cascade during larval attachment and metamorphosis of the barnacle Balanus (=Amphibalanus) amphitrite

    Zhang, Y.

    2012-08-01

    The barnacle Balanus amphitrite is among the most dominant fouling species on intertidal rocky shores in tropical and subtropical areas and is thus a target organism in antifouling research. After being released from adults, the swimming nauplius undertakes six molting cycles and then transforms into a cyprid. Using paired antennules, a competent cyprid actively explores and selects a suitable substratum for attachment and metamorphosis (collectively known as settlement). This selection process involves the reception of exogenous signals and subsequent endogenous signal transduction. To investigate the involvement of nitric oxide (NO) and cyclic GMP (cGMP) during larval settlement of B. amphitrite, we examined the effects of an NO donor and an NO scavenger, two nitric oxide synthase (NOS) inhibitors and a soluble guanylyl cyclase (sGC) inhibitor on settling cyprids. We found that the NO donor sodium nitroprusside (SNP) inhibited larval settlement in a dose-dependent manner. In contrast, both the NO scavenger carboxy-PTIO and the NOS inhibitors aminoguanidine hemisulfate (AGH) and S-methylisothiourea sulfate (SMIS) significantly accelerated larval settlement. Suppression of the downstream guanylyl cyclase (GC) activity using a GC-selective inhibitor ODQ could also significantly accelerate larval settlement. Interestingly, the settlement inhibition effects of SNP could be attenuated by ODQ at all concentrations tested. In the developmental expression profiling of NOS and sGC, the lowest expression of both genes was detected in the cyprid stage, a crucial stage for the larval decision to attach and metamorphose. In summary, we concluded that NO regulates larval settlement via mediating downstream cGMP signaling.

  5. Identification of A cGMP-dependent protein kinase substrate in nerve terminals as a new member of the septin family

    Full text: The cGMP signalling pathway plays a role in neurotransmitter release and learning and memory. The major mechanism by which cGMP achieves its effects is through activation of cGMP-dependent protein kinase (PKG) and phosphorylation of its substrates. More than 40 relatively PKG specific substrates have recently been found in brain in our laboratory. Our overall aim is to identify which of these proteins are found in nerve terminals and to purify and identify them. In this study we focussed on a 40 kDa PKG substrate in nerve terminals. This P40 was purified from an extract of rat brain peripheral membrane proteins by ion-exchange chromatography. The column fractions which contain P40 were phosphorylated in the presence of Mg2+-[γ-32P]ATP and purified protein kinases PKG, PKA or PKC. P40 was only phosphorylated by PKG in vitro, and not by PKA or PKC. We excised P40 from an SDS-PAGE gel and digested it with trypsin and subjected it to amino acid sequencing. This revealed P40 to be a new member of the septin family of proteins. Other septins, including Nedd5 in brain, play an essential role in cell division at the cytokinesis stage. We immunoprecipitated [γ-32P-ATP-labelled P40, which was phosphorylated by PKG in vitro and also a 40 kDa protein from intact synaptosomes, which were labelled by 32Pi by using two antibodies against Nedd5 (which crossreact with the other septins). The peptide maps under all these conditions were similar, suggesting P40 is phosphorylated in intact nerve terminals. These results show that P40 is a new member of the septin family and a PKG substrate in nerve terminals. Copyright (1998) Australian Neuroscience Society

  6. Participation of the NO/cGMP/K{sup +}ATP pathway in the antinociception induced by Walker tumor bearing in rats

    Barbosa, A.L.R.; Pinheiro, C.A.; Oliveira, G.J.; Torres, J.N.L.; Moraes, M.O.; Ribeiro, R.A.; Vale, M.L.; Souza, M.H.L.P. [Departamento de Fisiologia e Farmacologia, Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, CE (Brazil)

    2012-03-30

    Implantation of Walker 256 tumor decreases acute systemic inflammation in rats. Inflammatory hyperalgesia is one of the most important events of acute inflammation. The L-arginine/NO/cGMP/K{sup +}ATP pathway has been proposed as the mechanism of peripheral antinociception mediated by several drugs and physical exercise. The objective of this study was to investigate a possible involvement of the NO/cGMP/K{sup +}ATP pathway in antinociception induced in Walker 256 tumor-bearing male Wistar rats (180-220 g). The groups consisted of 5-6 animals. Mechanical inflammatory hypernociception was evaluated using an electronic version of the von Frey test. Walker tumor (4th and 7th day post-implantation) reduced prostaglandin E{sub 2}- (PGE{sub 2}, 400 ng/paw; 50 µL; intraplantar injection) and carrageenan-induced hypernociception (500 µg/paw; 100 µL; intraplantar injection). Walker tumor-induced analgesia was reversed (99.3% for carrageenan and 77.2% for PGE{sub 2}) by a selective inhibitor of nitric oxide synthase (L-NAME; 90 mg/kg, ip) and L-arginine (200 mg/kg, ip), which prevented (80% for carrageenan and 65% for PGE{sub 2}) the effect of L-NAME. Treatment with the soluble guanylyl cyclase inhibitor ODQ (100% for carrageenan and 95% for PGE{sub 2}; 8 µg/paw) and the ATP-sensitive K{sup +} channel (KATP) blocker glibenclamide (87.5% for carrageenan and 100% for PGE{sub 2}; 160 µg/paw) reversed the antinociceptive effect of tumor bearing in a statistically significant manner (P < 0.05). The present study confirmed an intrinsic peripheral antinociceptive effect of Walker tumor bearing in rats. This antinociceptive effect seemed to be mediated by activation of the NO/cGMP pathway followed by the opening of KATP channels.

  7. Rats with minimal hepatic encephalopathy show reduced cGMP-dependent protein kinase activity in hypothalamus correlating with circadian rhythms alterations.

    Felipo, Vicente; Piedrafita, Blanca; Barios, Juan A; Agustí, Ana; Ahabrach, Hanan; Romero-Vives, María; Barrio, Luis C; Rey, Beatriz; Gaztelu, Jose M; Llansola, Marta

    2015-01-01

    Patients with liver cirrhosis show disturbances in sleep and in its circadian rhythms which are an early sign of minimal hepatic encephalopathy (MHE). The mechanisms of these disturbances are poorly understood. Rats with porta-caval shunt (PCS), a model of MHE, show sleep disturbances reproducing those of cirrhotic patients. The aims of this work were to characterize the alterations in circadian rhythms in PCS rats and analyze the underlying mechanisms. To reach these aims, we analyzed in control and PCS rats: (a) daily rhythms of spontaneous and rewarding activity and of temperature, (b) timing of the onset of activity following turning-off the light, (c) synchronization to light after a phase advance and (d) the molecular mechanisms contributing to these alterations in circadian rhythms. PCS rats show altered circadian rhythms of spontaneous and rewarding activities (wheel running). PCS rats show more rest bouts during the active phase, more errors in the onset of motor activity and need less time to re-synchronize after a phase advance than control rats. Circadian rhythm of body temperature is also slightly altered in PCS rats. The internal period length (tau) of circadian rhythm of motor activity is longer in PCS rats. We analyzed some mechanisms by which hypothalamus modulate circadian rhythms. PCS rats show increased content of cGMP in hypothalamus while the activity of cGMP-dependent protein kinase was reduced by 41% compared to control rats. Altered cGMP-PKG pathway in hypothalamus would contribute to altered circadian rhythms and synchronization to light. PMID:26203935

  8. Time-lapse imaging as a tool to investigate contractility of the epididymal duct--effects of cGMP signaling.

    Andrea Mietens

    Full Text Available The well orchestrated function of epididymal smooth muscle cells ensures transit of spermatozoa through the epididymal duct during which spermatozoa acquire motility and fertilizing capacity. Relaxation of smooth muscle cells is mediated by cGMP signaling and components of this pathway are found within the male reproductive tract. Whereas contractile function of caudal parts of the rat epididymal duct can be examined in organ bath studies, caput and corpus regions are fragile and make it difficult to mount them in an organ bath. We developed an ex vivo time-lapse imaging-based approach to investigate the contractile pattern in these parts of the epididymal duct. Collagen-embedding allowed immobilization without impeding contractility or diffusion of drugs towards the duct and therefore facilitated subsequent movie analyses. The contractile pattern was made visible by placing virtual sections through the acquired image stack to track wall movements over time. By this, simultaneous evaluation of contractile activity at different positions of the observed duct segment was possible. With each contraction translating into a spike, drug-induced alterations in contraction frequency could be assessed easily. Peristaltic contractions were also detectable and throughout all regions in the proximal epididymis we found regular spontaneous contractile activity that elicited movement of intraluminal contents. Stimulating cGMP production by natriuretic peptide ANP or inhibiting degradation of cGMP by the phosphodiesterase 5 inhibitor sildenafil significantly reduced contractile frequency in isolated duct segments from caput and corpus. RT-PCR analysis after laser-capture microdissection localized the corresponding molecules to the smooth muscle layer of the duct. Our time-lapse imaging approach proved to be feasible to assess contractile function in all regions of the epididymal duct under near physiological conditions and provides a tool to evaluate acute

  9. Levosimendan Relaxes Pulmonary Arteries and Veins in Precision-Cut Lung Slices - The Role of KATP-Channels, cAMP and cGMP.

    Annette D Rieg

    Full Text Available Levosimendan is approved for left heart failure and is also used in right heart failure to reduce right ventricular afterload. Despite the fact that pulmonary arteries (PAs and pulmonary veins (PVs contribute to cardiac load, their responses to levosimendan are largely unknown.Levosimendan-induced vasorelaxation of PAs and PVs was studied in precision-cut lung slices from guinea pigs by videomicroscopy; baseline luminal area was defined as 100%. Intracellular cAMP- and cGMP-levels were measured by ELISA and NO end products were determined by the Griess reaction.Levosimendan relaxed control PVs (116% and those pre-constricted with an endothelinA-receptor agonist (119%. PAs were only relaxed if pre-constricted (115%. Inhibition of KATP-channels (glibenclamide, adenyl cyclase (SQ 22536 and protein kinase G (KT 5823 largely attenuated the levosimendan-induced relaxation in control PVs, as well as in pre-constricted PAs and PVs. Inhibition of BKCa (2+-channels (iberiotoxin and Kv-channels (4-aminopyridine only contributed to the relaxant effect of levosimendan in pre-constricted PAs. In both PAs and PVs, levosimendan increased intracellular cAMP- and cGMP-levels, whereas NO end products remained unchanged. Notably, basal NO-levels were higher in PVs. The KATP-channel activator levcromakalim relaxed PAs dependent on cAMP/PKA/PKG and increased cAMP-levels in PAs.Levosimendan initiates complex and divergent signaling pathways in PAs and PVs. Levosimendan relaxes PAs and PVs primarily via KATP-channels and cAMP/cGMP; in PAs, BKCa (2+- and Kv-channels are also involved. Our findings with levcromakalim do further suggest that in PAs the activation of KATP-channels leads to the production of cAMP/PKA/PKG. In conclusion, these results suggest that levosimendan might reduce right ventricular afterload by relaxation of PAs as well as pulmonary hydrostatic pressure and pulmonary edema by relaxation of PVs.

  10. cGMP-dependent protein kinase Iβ regulates breast cancer cell migration and invasion via interaction with the actin/myosin-associated protein caldesmon

    Schwappacher, Raphaela; Rangaswami, Hema; Su-Yuo, Jacqueline; Hassad, Aaron; Spitler, Ryan; Casteel, Darren E.

    2013-01-01

    The two isoforms of type I cGMP-dependent protein kinase (PKGIα and PKGIβ) differ in their first ∼100 amino acids, giving each isoform unique dimerization and autoinhibitory domains. The dimerization domains form coiled-coil structures and serve as platforms for isoform-specific protein–protein interactions. Using the PKGIβ dimerization domain as an affinity probe in a proteomic screen, we identified the actin/myosin-associated protein caldesmon (CaD) as a PKGIβ-specific binding protein. PKGI...

  11. Activation of the nitric oxide-cGMP pathway reduces phasic contractions in neonatal rat bladder strips via protein kinase G

    Artim, Debra E.; Kullmann, F. Aura; Daugherty, Stephanie L.; Wu, Hsi-Yang; de Groat, William C.

    2009-01-01

    Nitric oxide (NO), a neurotransmitter in the lower urinary tract, stimulates soluble guanylyl cyclase (sGC) and in turn cGMP-dependent protein kinase G (PKG) to modulate a number of downstream targets. NO donors reduce bladder hyperactivity in some pathological models but do not affect normal bladder activity in the adult rat. In this study, the NO donor S-nitroso-N-acetyl-dl-penicillamine (SNAP; 100 μM) decreased the amplitude and frequency of spontaneous and carbachol-enhanced contractions ...

  12. 兽药GMP验收应注意的问题%Problems which should be paid attention in qualifying veterinary drug GMP

    王增梁

    2004-01-01

    兽药GMP(GMP为Good Manufacturing Practice for Drugs的缩写)是《兽药生产质量管理规范》的简称,可译为优良兽药的生产实践。兽药GMP是生产出优质兽药的标准,实施兽药GMP的宗旨是为了保证兽药生产质量的均一性、稳定性和安全性,

  13. Characterization of the binding of cAMP and cGMP to the CRP*598 mutant of the E. coli cAMP receptor protein.

    Ren, Y. L.; Garges, S; Adhya, S; Krakow, J. S.

    1990-01-01

    Wild type cAMP receptor protein (CRP) activates in vitro lac transcription only in the presence of cAMP. In contrast the mutant CRP*598 (Arg-142 to His, Ala-144 to Thr) can activate lac transcription in the absence of cyclic nucleotide or at concentrations of cAMP below that required by CRP. To further characterize the properties of CRP*598, the binding of cAMP and cGMP to CRP and CRP*598 has been determined. The intrinsic binding constant (K) values obtained for cAMP binding are: CRP, 1.9 x ...

  14. Evaluation of a Cyclic GMP-Dependent Protein Kinase Inhibitor in Treatment of Murine Toxoplasmosis: Gamma Interferon Is Required for Efficacy

    Nare, Bakela; Allocco, John J.; Liberator, Paul A.; Donald, Robert G. K.

    2002-01-01

    The trisubstituted pyrrole 4-[2-(4-fluorophenyl)-5-(1-methylpiperidine-4-yl)-1H-pyrrol-3-yl]pyridine (compound 1) is a potent inhibitor of cyclic GMP-dependent protein kinases from Apicomplexan protozoa and displays cytostatic activity against Toxoplasma gondii in vitro. Compound 1 has now been evaluated against T. gondii infections in the mouse and appeared to protect the animals when given intraperitoneally at 50 mg/kg twice daily for 10 days. However, samples from brain, spleen, and lung t...

  15. Measurement of GEp/GMp in e vector p → ep vector to Q2 = 5.6 GeV2

    The ratio of the electric and magnetic form factors of the proton, GEp/GMp, was measured at the Thomas Jefferson National Accelerator Facility (JLab) using the recoil polarization technique. The ratio of the form factors is directly proportional to the ratio of the transverse to longitudinal components of the polarization of the recoil proton in the elastic e vector p → ep vector reaction. The new data presented in this article span the range 3.5 2 2 and are well described by a linear Q2 fit. Also, the ratio QF2p/F1p reaches a constant value above Q2 = 2 GeV2. (authors)

  16. Comparison of phosphorylation of ribosomal proteins from HeLa and Krebs II ascites-tumour cells by cyclic AMP-dependent and cyclic GMP-dependent protein kinases

    Issinger, O G; Beier, H; Speichermann, N; Flokerzi, V; Hofmann, F

    1980-01-01

    Phosphorylation of eukaryotic ribosomal proteins in vitro by essentially homogeneous preparations of cyclic AMP-dependent protein kinase catalytic subunit and cyclic GMP-dependent protein kinase was compared. Each protein kinase was added at a concentration of 30nM. Ribosomal proteins were...... cyclic AMP-dependent enzyme. Between 0.1 and 0.2 mol of phosphate was incorporated/mol of these phosphorylated proteins. With the exception of protein S7, the same proteins were also major substrates for the cyclic GMP-dependent protein kinase. Time courses of the phosphorylation of individual proteins...... from the small and large ribosomal subunits in the presence of either protein kinase suggested four types of phosphorylation reactions: (1) proteins S2, S10 and L5 were preferably phosphorylated by the cyclic GMP-dependent protein kinase; (2) proteins S3 and L6 were phosphorylated at very similar rates...

  17. 就业发展“十三五”基本思路与目标--构建更高质量的充分就业型社会%Goals and Outlines of Employment Development in the 13th Five-Year Plan:Constructing a higher-quality labor market with full employment

    胡鞍钢; 杨竺松; 鄢一龙

    2015-01-01

    基于“十二五”时期我国就业发展相关规划指标和统计数据,对“十二五”时期就业目标完成情况进行中期评估,结果显示:“十二五”规划城镇新增就业的目标提前实现,就业结构进一步优化,我国城镇新增就业岗位更加多元。“十三五”时期,我国就业形势总体将趋于缓和,同时也面临多方面机遇和挑战。“十三五”期间我国应以实现更加充分的、更高质量的就业作为就业发展的总体目标,坚持就业优先,以创业带动就业,继续实施积极就业政策,着力做好重点群体就业工作,健全促进就业创业体制机制,使就业发展与经济增长、结构升级协调推进的基本思路,着力完成六项重点任务:以创业为新的就业增长极,打造创业型社会;继续实施就业优先战略和更加积极的就业政策;继续做好高校毕业生、农村转移劳动力、城镇困难人员、退役军人等重点群体的就业工作,构建青年友好型就业创业体系;加强职业技能培训,建设现代职业教育体系;健全人力资源市场,完善就业服务体系;构建和谐劳资关系,完善就业援助机制。%Based on the related planning targets and statistics ,this article conducts the midterm appraisal of the employment in the 12th Five-Year Plan .The results show that the goal of increasing employment in ur-ban area has been accomplished in advance and that the employment structure has been ameliorated with di-versified employment positions provided .In prospect of the 13th Five-Year ,the overall trend of employ-ment will be steady while opportunities and challenges go hand in hand .To cope with the circumstances , the goal of developing fuller and higher-quality employment should be set .Furthermore ,to match the de-velopment of employment with economic grow th and structural improvement ,there are six major focuses in employment promotion .First

  18. KLE annual report 1985

    Report on legal and economic fundamentals, licensing procedures, the state of construction, waste management schemes, financing, staff requirements, public relations, annual balances, cost-benefit calculations for 1985. (HP)

  19. Annual General Canvass Statistics

    National Oceanic and Atmospheric Administration, Department of Commerce — This data set contains annual quantities and value for all seafood products that are landed and sold by established seafood dealers and brokers in the Southeast...

  20. Annual report 1980

    This annual report contains a description of the named institute, the research programm, reports from the scientific establishments, a description of different cooperations, and a list of scientific publications. (HSI)