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Sample records for 13th annual gmp

  1. Generics, Supergenerics and Patent Strategies--SMi's 13th Annual Meeting.

    Science.gov (United States)

    Edwards, Catherine

    2010-07-01

    SMi's 13th Annual Meeting on Generics, Supergenerics and Patent Strategies, held in London, included topics covering new trends in the generics field, the difficulties faced by companies in entering the generics market and recent developments in IP. This conference report highlights selected presentations on generics in India, protecting pharmaceutical products in China, changes in generics law and litigation in the US and Europe, challenges for market selection and entry for generics companies, the influence of changes in the healthcare market on the generics industry, supergenerics, and biosimilars.

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

    OpenAIRE

    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

    OpenAIRE

    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. Proceedings of the Annual Conference of the International Group for the Psychology of Mathematics Education (13th, Paris, France, July 9-13, 1989), Volume 2.

    Science.gov (United States)

    Vergnaud, Gerard, Ed.; Rogalski, Janine, Ed.; Artique, Michele, Ed.

    This proceedings of the annual conference of the International Group for the Psychology of Mathematics Education (PME) includes the following research papers: "Logo et Symetrie Centrale" (E. Gallou-Dumiel); "About Continuous Operator Subconstruct in Rational Numbers" (J. Gimenez); "Constructivist Epistemology and Discovery Learning in Mathematics"…

  5. Proceedings of the Annual Conference of the International Group for the Psychology of Mathematics Education (13th, Paris, France, July 9-13, 1989), Volume 3.

    Science.gov (United States)

    Vergnaud, Gerard, Ed.; Rogalski, Janine, Ed.; Artique, Michele, Ed.

    This proceedings of the annual conference of the International Group for the Psychology of Mathematics Education (PME) includes the following research papers: "A Model of Understanding Two-Digit Numeration and Computation" (H. Murray & A. Olivier); "The Computer Produces a Special Graphic Situation of Learning the Change of Coordinate System" (S.…

  6. Voyaging from the Past, to the Present, and into the Future: Knowing Your Heritage. Selected Papers from PIALA 2003, Pacific Islands Association of Libraries, Archives and Museums Annual Conference (13th, Pohnpei, Federated States of Micronesia, November 4-6, 2003)

    Science.gov (United States)

    Drake, Paul B., Ed.

    2003-01-01

    This publication follows the tradition of publishing selected papers from Pacific Islands Association of Libraries, Archives and Museums (PIALA) annual conferences. This 13th annual conference was held in Pohnpei, Federated States of Micronesia, November 4-6, 2003. The volume begins with a listing of the members of the PIALA 2003 Planning &…

  7. 13th International Symposium on Experimental Robotics

    CERN Document Server

    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...

  8. 13th International Conference on Information Technology

    CERN Document Server

    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.

  9. 13th International Conference on Chlamydomonas

    Energy Technology Data Exchange (ETDEWEB)

    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.

  10. 13th New Mexico Analysis Seminar

    CERN Document Server

    Marcantognini, Stefania; Stokolos, Alexander; Urbina, Wilfredo

    2016-01-01

    Covering a range of subjects from operator theory and classical harmonic analysis to Banach space theory, this book contains survey and expository articles by leading experts in their corresponding fields, and features fully-refereed, high-quality papers exploring new results and trends in spectral theory, mathematical physics, geometric function theory, and partial differential equations. Graduate students and researchers in analysis will find inspiration in the articles collected in this volume, which emphasize the remarkable connections between harmonic analysis and operator theory. Another shared research interest of the contributors of this volume lies in the area of applied harmonic analysis, where a new notion called chromatic derivatives has recently been introduced in communication engineering. The material for this volume is based on the 13th New Mexico Analysis Seminar held at the University of New Mexico, April 3-4, 2014 and on several special sections of the Western Spring Sectional Meeting at th...

  11. 13th International Conference Intelligent Autonomous Systems

    CERN Document Server

    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 ...

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

    OpenAIRE

    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...

  13. The 13th CAS General Assembly Convenes in Beijing

    Institute of Scientific and Technical Information of China (English)

    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.

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

    International Nuclear Information System (INIS)

    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)

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

    Directory of Open Access Journals (Sweden)

    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.

  16. The 13th international conference on scientometrics and informetrics

    DEFF Research Database (Denmark)

    Ocholla, Dennis; Ingwersen, Peter; Noyons, Ed;

    2012-01-01

    The 13th International Conference on Scientometrics and Informetrics took place in Durban, South Africa from 4 to 7 July 2011, Ocholla and Ingwersen (2011). The meeting was organised under the auspices of the International Society for Scientometrics and Informetrics (ISSI) and by the ISSI 2011...... Conference Local Organising Committee with the participation of six Universities and Research Centres in South Africa, particularly, the Durban University of Technology, the University of Cape Town, the University of KwaZulu Natal, the National Research Foundation, the University of South Africa......, the University of Pretoria and the University of Zululand. The event was hosted by the University of Zululand and Durban University of Technology....

  17. 13th International Conference on Distributed Computing and Artificial Intelligence

    CERN Document Server

    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 ...

  18. The Geobiology of Weathering: a 13th Hypothesis

    CERN Document Server

    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.

  19. 13thInternational Conference on Computer and Information Science

    CERN Document Server

    Software Engineering, Artificial Intelligence, Networking and Parallel/Distributed Computing 2012

    2013-01-01

    The purpose of the 13th  International Conference on Computer and Information Science (SNPD 2012) held on August 8-10, 2012 in Kyoto, Japan was to bring together researchers and scientists, businessmen and entrepreneurs, teachers and students to discuss the numerous fields of computer science, and to share ideas and information in a meaningful way.  Our conference officers selected the best 17 papers from those papers accepted for presentation at the conference in order to publish them in this volume.  The papers were chosen based on review scores submitted by members of the program committee, and underwent further rounds of rigorous review.   The  conference organizers selected 17 outstanding papers from SNPD 2012, all of which you will find in this volume of Springer’s Studies in Computational Intelligence.

  20. 13th Czechoslovak seminar on plasma physics and technology

    International Nuclear Information System (INIS)

    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.)

  1. 13th International Conference on X-Ray Lasers

    CERN Document Server

    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...

  2. A 13th-century kettlehat from Kodasoo, Estonia

    Directory of Open Access Journals (Sweden)

    Mäll, Jaak

    2011-12-01

    Full Text Available The oldest surviving helmet from Estonia is a 13th-century kettlehat found near village of Kodasoo in northern Estonia. Three samples from different parts of the kettlehat were investigated in a metallographic study. The raw material of the Kodasoo kettlehat was a slag-rich iron of heterogeneous carbon content. It was improved by folding and forge-welding it at least three times, resulting in layered structure with more uniformly distributed carbon content.

    El casco más antiguo encontrado en Estonia es un «chapel de fer» del siglo XIII que proviene de cerca de la aldea de Kodasoo, en el Norte de Estonia. Tres muestras de diversas partes del casco fueron investigadas mediante un estudio metalográfico. El material original del casco de Kodasoo era un hierro de contenido de carbono heterogéneo y con abundantes inclusiones de escoria. Se intentó mejorar su calidad mediante forja, doblándolo y soldándolo a la calda por lo menos tres veces, dando como resultado una estructura estratificada, con el contenido de carbono distribuido más uniformemente.

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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

  5. Series of aerial images over Monte Vista National Wildlife Refuge, acquired July 13th, 1965

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This data set includes 31 georeferenced versions of black and white aerial photographs from July 13th, 1965, taken over Monte Vista National Wildlife Refuge. A...

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

    Institute of Scientific and Technical Information of China (English)

    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

  7. 13th International Workshop on Condensed Matter Theories

    CERN Document Server

    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...

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

    Science.gov (United States)

    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.

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

    International Nuclear Information System (INIS)

    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

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

    International Nuclear Information System (INIS)

    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)

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

    Institute of Scientific and Technical Information of China (English)

    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

  12. Eight Highlights in 13th China(Humen)International Fashion Fair

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    @@ Currently,it has been learned from the Humen Fashion Fair Organizing Committee that the 13th China (Humen) International Fashion Fair (hereafter referred to the Fair for short) to be held from Nov. 21 to Nov. 25,2008 in Humen,a famous city of garments,will demonstrate eight highlights.

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

    DEFF Research Database (Denmark)

    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...

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

    International Nuclear Information System (INIS)

    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

  15. cGMP-Dependent Protein Kinases and cGMP Phosphodiesterases in Nitric Oxide and cGMP Action

    OpenAIRE

    Francis, Sharron H.; Busch, Jennifer L.; Corbin, Jackie D.

    2010-01-01

    To date, studies suggest that biological signaling by nitric oxide (NO) is primarily mediated by cGMP, which is synthesized by NO-activated guanylyl cyclases and broken down by cyclic nucleotide phosphodiesterases (PDEs). Effects of cGMP occur through three main groups of cellular targets: cGMP-dependent protein kinases (PKGs), cGMP-gated cation channels, and PDEs. cGMP binding activates PKG, which phosphorylates serines and threonines on many cellular proteins, frequently resulting in change...

  16. [Replica of a 13th century piece in the Museum of Pharmacy].

    Science.gov (United States)

    Zarranz M del, C

    1995-06-01

    Since the Museum of Pharmacy of the University of Buenos Aires has a big historical pieces collection, its Director and founddress Dr. Rosa D. de Carnevale Bonino signed an article about this subject (Revista del Museo. 1: 12/16-dic, 1986). One of such pieces, dating from the 13th century, is here depicted, as well as Alberto Magno's biography, who is intensely related with the so called "magical glass".

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

    International Nuclear Information System (INIS)

    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)

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

    Energy Technology Data Exchange (ETDEWEB)

    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)

  19. Characterisation of decorations on Iranian (10th-13th century) lustreware

    Science.gov (United States)

    Borgia, I.; Brunetti, B.; Giulivi, A.; Sgamellotti, A.; Shokouhi, F.; Oliaiy, P.; Rahighi, J.; Lamehi-Rachti, M.; Mellini, M.; Viti, C.

    It has been recently shown that lustre decoration of Medieval and Renaissance pottery consists of silver and copper nanoparticles, dispersed within the glassy matrix of the ceramic glaze. Lustre surfaces show peculiar optical effects, such as metallic reflection and iridescence. Here we report the findings of a study on lustred glazes of several shards belonging to Iranian pottery of the 10th and 13th centuries, decorated on both sides. Two different glazes, depending on the side of the sample, have been identified. Different lustre chromatic effects are characterised by the relative presence of silver- and copper-metal nanoparticles dispersed in the glassy matrix.

  20. Were Byzantine monks of the 13th-15th centuries holders of imperial grants?

    Directory of Open Access Journals (Sweden)

    Maniati-Kokkini Triantafyllitsa

    2013-01-01

    Full Text Available A small number of imperial grants to monks appear in the Byzantine sources from the late 13th to the 15th centuries, and mainly in the 14th, before the Serbian expansion in Macedonia. Especially privileged with the right of bequeathal and dedication, they were given later to the monastery to which each of the monks belonged, mostly to the Serbian monastery of Chilandar in Athos. Due to characteristic differences they represent a special category between the pronoiai of laymen and the oikonomiai of monasteries.

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

    Directory of Open Access Journals (Sweden)

    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.

  2. 13th International Symposium on Distributed Computing and Artificial Intelligence 2016

    CERN Document Server

    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).

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

    Directory of Open Access Journals (Sweden)

    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.

  4. The 13th International Workshop on Targetry and Target Chemistry Proceedings

    DEFF Research Database (Denmark)

    Haroun, Samar; Givskov, Alex David; Jensen, Mikael

    This report contains the complete proceedings of the 13th International Workshop on Targetry and Target Chemistry. The Workshop was held at Risø 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...

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

    Energy Technology Data Exchange (ETDEWEB)

    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.)

  6. Highlights and Conclusions of the Chalonge 13th Paris Cosmology Colloquium

    CERN Document Server

    de Vega, H J

    2010-01-01

    The 13th Chalonge Paris Cosmology Colloquium was held at the historic Perrault building of Observatoire de Paris on 23-25 July 2009. The Colloquium was held in the spirit of the Chalonge School, putting together real cosmological and astrophysical data and hard theory predictive approach in the framework of the Standard Model of the Universe. Peter Biermann, James Bullock, Claudio Destri, Hector de Vega, Massimo Giovannini, Sasha Kashlinsky, Eiichiro Komatsu, Anthony Lasenby and Norma G. Sanchez present here their highlights of the Colloquium. The summary and conclusions by H. J. de Vega, M. C. Falvella and N. G. Sanchez stress among other points: (i) the primordial CMB fluctuations are almost gaussian, large primordial non-gaussianity and large running index are strongly disfavored. The primordial graviton ratio r is predicted in the effective theory of inflation to be between 0.021 and 0.053, at reach of the next CMB observations. (ii) Dark energy observations are consistent with the cosmological constant. ...

  7. The 13th quality control survey for radioisotopes in vitro tests in Japan, 1991

    Energy Technology Data Exchange (ETDEWEB)

    1992-11-01

    This paper presents the results of the 13th quality control nationwide survey. Of 568 hospitals performing RI in vitro tests as of December 1991, 280 (49.3%) participated in this survey. The following 37 items were examined: ACTH, 17[alpha]-hydroxyprogesterone, [alpha]-fetoprotein, aldosterone, [beta]2-microglobulin, CA 125, CA 15-3, CA 19-9, calcitonin, CEA, cortisol, C-peptide, digoxin, esteradiol, ferritin, free triiodothyronine (T[sub 3]), free thyroxine (T[sub 4]), FSH, gastrin, GH, glucagon, IgE, insulin, LH, PAP, progesterone, prolactin, PTH, somatomedin C, T[sub 3], T[sub 4], T[sub 3] uptake, TBG, testosterone, thyroglobulin, tissue polypeptide antigen, and TSH. Reliability and safety of measurement values were evaluated based on protein components of serum samples, spontaneous reliability test, time-dependent reliability test, repeated freezing-dissolution test, and serial change test after freezing. 'Within kit variation' between facilities was great especially for ACTH, CA 15-3, CA 19-9, free T[sub 4], calcitonin, gastrin, and testosterone, and was favorable for [beta][sub 2]-microglobulin, CA 125, digoxin, cortisol, IgE, T[sub 3], T[sub 4], T[sub 3] uptake, and TBG. It was noted that there was a great difference in measurement values of free T[sub 3] and free-T[sub 4] between RIA kits and non-isotopic kits. (N.K.).

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

    International Nuclear Information System (INIS)

    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)

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

    Directory of Open Access Journals (Sweden)

    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.

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

    International Nuclear Information System (INIS)

    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.)

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

    Energy Technology Data Exchange (ETDEWEB)

    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)

  12. 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

    Institute of Scientific and Technical Information of China (English)

    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.

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

    International Nuclear Information System (INIS)

    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.)

  14. Specificity of the Cyclic GMP-Binding Activity and of a Cyclic GMP-Dependent Cyclic GMP Phosphodiesterase in Dictyostelium discoideum

    NARCIS (Netherlands)

    Haastert, Peter J.M. van; Walsum, Hans van; Meer, Rob C. van der; Bulgakov, Roman; Konijn, Theo M.

    1982-01-01

    The nucleotide specificity of the cyclic GMP-binding activity in a homogenate of Dictyostelium discoideum was determined by competition of cyclic GMP derivatives with [8-3H] cyclic GMP for the binding sites. The results indicate that cyclic GMP is bound to the binding proteins by hydrogen bonds at N

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

    Science.gov (United States)

    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

  16. PREFACE: 13th Anglo-French Physical Acoustics Conference (AFPAC2014)

    Science.gov (United States)

    Gélat, Pierre; Pinfield, Valerie; Cegla, Frederic; Saffari, Nader; Lhémery, Alain

    2015-01-01

    The 13th Anglo-French Physical Acoustics Conference (AFPAC) was held at Selsdon Park Hotel, Croydon near London, United Kingdom, on 15-17 January 2014. The venue was an excellent location to exchange ideas, regardless whether this happened in the conference room, over lunch at the drinks reception in the conservatory, in the oak panelled bar after the conference dinner or in the local pub next door. Over 45 papers were presented at the conference. There were over 60 delegates from institutions covering four countries. The invited speakers from the French side shared their knowledge about the generation of sound from supersonic jets (Prof Christophe Bailly, École Centrale de Lyon) and the application of ultrasonic microscropy in the nuclear industry (Prof Gilles Despaux, Université de Montpellier). The UK invited speakers included Prof Malcolm Povey (University of Leeds), who talked about characterisation of the nucleation of crystals using ultrasound, and Prof Bruce Drinkwater (University of Bristol), who captured the audience by speaking about "ultrasonic lassos" and ultrasonic particle manipulation. There was a strong representation of laser ultrasonics at the meeting with scientific considerations of problems and applications that range from the macro to the nanoscale. There were also numerous papers on the interaction of elastic and acoustic waves with complex materials and scattering of these waves by materials such as foams or cavitating liquids. Presentations on biomedical applications are increasingly being featured at AFPAC meetings. Talks this year covered topics such as imaging and high-intensity focused ultrasound for therapeutic applications. Finally, there were also several contributions from the field of Non-Destructive Evaluation (NDE) and Structural Health Monitoring (SHM) with talks ranging from the determination of the properties of in vivo wood to ultrasonic scattering techniques and tomographic reconstructions to recover the size and shape of

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

    OpenAIRE

    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): ...

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

    OpenAIRE

    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...

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

    Directory of Open Access Journals (Sweden)

    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.

  20. 13th FINA World Championship finals: stroke kinematical and race times according to performance, gender and event

    OpenAIRE

    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...

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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

  3. PREFACE The 13th International Conference on Rapidly Quenched and Metastable Materials

    Science.gov (United States)

    Schultz, Ludwig; Eckert, Jürgen; Battezzati, Livio; Stoica, Mihai

    2009-01-01

    The 13th International Conference on Rapidly Quenched and Metastable Materials (RQ13) took place in Dresden, Germany, 24-29 August 2008. It belongs to the triennial series of RQ meetings with a long tradition, starting in 1970 - Brela, 1975 - Boston, 1978 - Brighton, 1981 - Sendai, 1984 - Würzburg, 1987 - Montreal, 1990 - Stockholm, 1993 - Sendai, 1996 - Bratislava, 1999 - Bangalore, 2002 - Oxford, 2005 - Jeju Island. RQ13 was hosted by the Leibniz Institute of Solid State and Materials Research, IFW Dresden. Research on rapidly quenched and metastable materials is stimulated by the high demand for new materials with unique mechanical, chemical and physical properties. Topics of RQ13 conference have fallen into three parts: synthesis and processing, materials and properties, and applications of rapidly quenched and metastable materials. These topics cover exiting developments from the traditional field of rapidly quenched metals to newly emerging areas such as bulk metallic glasses and nanostructured materials. As such, the presentations reported on recent experimental and theoretical achievements in the fields of metastable materials, quasicrystals, nanometer-scale materials, magnetic materials, metallic glasses, solid state reaction, undercooling and modeling. As in the previous proceedings (RQ12), the largest number of papers is dedicated to bulk metallic glasses and magnetic materials. With respect to property characterization and applications, there are great attempts for use and application of these materials, particularly for bulk metallic glasses, as well as for further design and optimization of properties. The RQ13 conference attracted a total of 381 abstracts submitted by scientists from 38 different countries. The conference included 8 plenary talks and 25 invited keynote talks. In addition, 163 regular oral contributions were presented and more than 180 posters were presented. It was a particular highlight of the conference that Dr Ho Sou Chen was

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

    Science.gov (United States)

    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

  5. PREFACE: 13th International Conference on Muon Spin Rotation, Relaxation and Resonance

    Science.gov (United States)

    2014-12-01

    The 13th International Conference on Muon Spin Rotation, Relaxation and Resonance (μSR2014) organized by the Laboratory for Muon Spin Spectroscopy, Paul Scherrer Institute in collaboration with the University of Zurich and the University of Fribourg, was held in Grindelwald, Switzerland from 1st to 6th June 2014. The conference provided a forum for researchers from around the world with interests in the applications of μSR to study a wide range of topics including condensed matter physics, materials and molecular sciences, chemistry and biology. Polarized muons provide a unique and versatile probe of matter, enabling studies at the atomic level of electronic structure and dynamics in a wide range of systems. The conference was the thirteenth in a series, which began in Rorschach in 1978 and it took place for the third time in Switzerland. The previous conferences were held in Cancun, Mexico (2011), Tsukuba, Japan (2008), Oxford, UK (2005), Williamsburg, USA (2002), Les Diablerets, Switzerland (1999), Nikko, Japan (1996), Maui, USA (1993), Oxford, UK (1990), Uppsala, Sweden (1986), Shimoda, Japan (1983), Vancouver, Canada (1980), and Rorschach, Switzerland (1978). These conference proceedings contain 67 refereed publications from presentations covering magnetism, superconductivity, chemistry, semiconductors, biophysics and techniques. The conference logo, displayed in the front pages of these proceedings, represents both the location of μSR2014 in the Alps and the muon-spin rotation technique. The silhouette represents the famous local mountains Eiger, Mönch and Jungfrau as drawn by the Swiss painter Ferdinand Hodler and the apple with arrow is at the same time a citation of the Wilhelm Tell legend and a remembrance of the key role played by the muon spin and the asymmetric muon decay (which for the highest positron energy has an apple like shape). More than 160 participants (including 32 registered as students and 13 as accompanying persons) from 19 countries

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

    OpenAIRE

    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...

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

    CERN Document Server

    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.

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

    Data.gov (United States)

    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...

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

    Science.gov (United States)

    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

  10. 15th International Congress on Plasma Physics & 13th Latin American Workshop on Plasma Physics

    Science.gov (United States)

    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

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

    Directory of Open Access Journals (Sweden)

    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

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

    International Nuclear Information System (INIS)

    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.

  13. SOUNDS LEARNING USING TEAMS GAMES TOURNAMENT WITH FLASH CARD AS MEDIA AT THE 13th JUNIOR HIGH SCHOOL OF MAGELANG

    Directory of Open Access Journals (Sweden)

    A. Lestari

    2015-11-01

    Full Text Available The aim of this research is to know about students’ response and the effectiveness of Teams Games Tournament learning model with Flash Card as media. This research type is Quasi Experimental Nonequivalent Group Design. The subject of the study is the students of 8 E and 8 F of the 13th junior high school of Magelang. The result of cognitive assessment showed that the classical learning achievement was 87% and the gained score was 0.46 which is included in media category. Value of Sig. (2-tailed of Mann-Whitney U-Test is 0.01. It means that there is the significant difference of academic achievement between the experiment group and the control group. The result of affective and pshycomotoric assessments are included in good category at average score 75% and 71%. Correlation among cognitive, affective, and psychomotoric learning achievements was strong at R Score 0.684. Students’ response toward the learning showed average score at 74%, and it was included in good category. Based on the result, Teams Games Tournament learning model with Flash Card as media in learning about sounds at the 13th junior high school in Magelang is effective towards the learning achievement and the responses of the 8 D graders towards the learning is categorized good.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    International Nuclear Information System (INIS)

    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.

  16. 2015 Special Sessions of the 13th International Conference on Practical Applications of Agents and Multi-Agent Systems

    CERN Document Server

    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...

  17. 13th IUPAC International Congress of Pesticide Chemistry: Crop, Environment, and Public Health Protection, Technologies for a Changing World.

    Science.gov (United States)

    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

  18. Nitric oxide-cyclic GMP signaling in stem cell differentiation

    OpenAIRE

    Mujoo, Kalpana; Krumenacker, Joshua S.; Murad, Ferid

    2011-01-01

    The nitric oxide-cyclic GMP (NO-cGMP) pathway mediates important physiological functions associated with various integrative body systems including the cardiovascular and nervous systems. Furthermore, NO regulates cell growth, survival, apoptosis, proliferation and differentiation at the cellular level. To understand the significance of the NO-cGMP pathway in development and differentiation, studies have been conducted both in developing embryos and stem cells. Manipulation of the NO-cGMP pat...

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

    International Nuclear Information System (INIS)

    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

  20. Diversity of Cyclic Di-GMP-Binding Proteins and Mechanisms.

    Science.gov (United States)

    Chou, Shan-Ho; Galperin, Michael Y

    2016-01-01

    Cyclic di-GMP (c-di-GMP) synthetases and hydrolases (GGDEF, EAL, and HD-GYP domains) can be readily identified in bacterial genome sequences by using standard bioinformatic tools. In contrast, identification of c-di-GMP receptors remains a difficult task, and the current list of experimentally characterized c-di-GMP-binding proteins is likely incomplete. Several classes of c-di-GMP-binding proteins have been structurally characterized; for some others, the binding sites have been identified; and for several potential c-di-GMP receptors, the binding sites remain to be determined. We present here a comparative structural analysis of c-di-GMP-protein complexes that aims to discern the common themes in the binding mechanisms that allow c-di-GMP receptors to bind it with (sub)micromolar affinities despite the 1,000-fold excess of GTP. The available structures show that most receptors use their Arg and Asp/Glu residues to bind c-di-GMP monomers, dimers, or tetramers with stacked guanine bases. The only exception is the EAL domains that bind c-di-GMP monomers in an extended conformation. We show that in c-di-GMP-binding signature motifs, Arg residues bind to the O-6 and N-7 atoms at the Hoogsteen edge of the guanine base, while Asp/Glu residues bind the N-1 and N-2 atoms at its Watson-Crick edge. In addition, Arg residues participate in stacking interactions with the guanine bases of c-di-GMP and the aromatic rings of Tyr and Phe residues. This may account for the presence of Arg residues in the active sites of every receptor protein that binds stacked c-di-GMP. We also discuss the implications of these structural data for the improved understanding of the c-di-GMP signaling mechanisms.

  1. The Muslim mystic who fasted in his cradle – what does a 19th century MS tell of the 13th century saint?

    Directory of Open Access Journals (Sweden)

    Helena Hallenberg

    1999-01-01

    Full Text Available In this paper, the author discusses the problem of interpreting literary sources, using one specific example, namely that of a Muslim saint of the 13th century, as depicted in a 19th century manuscript. The problem is examined on three different levels: 1 the form, 2 the contents, and 3 the context. We can see that the 19th century Muslim saint does not tell so much of a 13th century saint but of the 19th century ideals — and that this statement is due to a 20th century re-reading of the sources.

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

    Science.gov (United States)

    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.

  3. 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

    NARCIS (Netherlands)

    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

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

    NARCIS (Netherlands)

    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 i

  5. Unusual guanylyl cyclases and cGMP signaling in Dictyostelium discoideum

    NARCIS (Netherlands)

    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

  6. Microbiological criteria for good manufacturing practice (GMP)

    International Nuclear Information System (INIS)

    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.)

  7. Microbiological criteria for good manufacturing practice (GMP)

    Energy Technology Data Exchange (ETDEWEB)

    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.)

  8. Solution Structure of the cGMP Binding GAF Domain from Phosphodiesterase 5: Insights into Nucleotide Specificity, Dimerization, and cGMP-Dependent Conformational Change

    Energy Technology Data Exchange (ETDEWEB)

    Heikaus, Clemens C.; Stout, Joseph R.; Sekharan, Monica R.; Eakin, Catherine M.; Rajagopal, Ponni; Brzovic, Peter S.; Beavo, Joseph A.; Klevit, Rachel E.

    2008-08-15

    Phosphodiesterase 5 (PDE5) controls intracellular levels of cGMP through its regulation of cGMP hydrolysis. Hydrolytic activity of the C-terminal catalytic domain is increased by cGMP binding to the N-terminal GAF A domain. We present the NMR solution structure of the cGMP-bound PDE5A GAF A domain. The cGMP orientation in the buried binding pocket was defined through 37 intermolecular NOEs.

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

    Directory of Open Access Journals (Sweden)

    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

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

    OpenAIRE

    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...

  11. Control of bacterial exoelectrogenesis by c-AMP-GMP.

    Science.gov (United States)

    Nelson, James W; Sudarsan, Narasimhan; Phillips, Grace E; Stav, Shira; Lünse, Christina E; McCown, Phillip J; Breaker, Ronald R

    2015-04-28

    Major changes in bacterial physiology including biofilm and spore formation involve signaling by the cyclic dinucleotides c-di-GMP and c-di-AMP. Recently, another second messenger dinucleotide, c-AMP-GMP, was found to control chemotaxis and colonization by Vibrio cholerae. We have identified a superregulon of genes controlled by c-AMP-GMP in numerous Deltaproteobacteria, including Geobacter species that use extracellular insoluble metal oxides as terminal electron acceptors. This exoelectrogenic process has been studied for its possible utility in energy production and bioremediation. Many genes involved in adhesion, pilin formation, and others that are important for exoelectrogenesis are controlled by members of a variant riboswitch class that selectively bind c-AMP-GMP. These RNAs constitute, to our knowledge, the first known specific receptors for c-AMP-GMP and reveal that this molecule is used by many bacteria to control specialized physiological processes.

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

    NARCIS (Netherlands)

    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

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

    Science.gov (United States)

    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

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

    NARCIS (Netherlands)

    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

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

    DEFF Research Database (Denmark)

    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...

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

    Science.gov (United States)

    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

  17. Precision Optics Optimization for GMp Experiment

    Science.gov (United States)

    Wang, Yang; Allada, Kalyan; Averett, Todd; Christy, Eric; Gu, Chao; Huang, Min; Wojtsekhowski, Bogdan; GMp Collaboration

    2015-04-01

    The GMp experiment aims to improve the precision on the elastic e-p cross section measurement to 2%; up to a factor of 5 better than previous measurements, with four-momentum transfer up to 14 GeV2 using the High Resolution Spectrometers (HRS) of Hall A at Jefferson Lab. These measurements will be an important benchmark for many other cross section measurements in hadron physics. To reach this goal, it is necessary to improve the precision of many instrument systems. Knowledge of the magnetic optics of HRS is critically important for precision reconstruction of the momentum and coordinates of the scattered particles at the interaction vertex. In this talk, an improved optimization method for optics will be presented in detail and the results of a study based on recent commissioning data in 2014 will be discussed.

  18. Residential Access to Bandwidth: Exploring New Paradigms. A Report of the Annual Aspen Institute Conference on Telecommunications Policy (13th, August 9-13, 1998, Aspen, Colorado).

    Science.gov (United States)

    Entman, Robert M.

    The 1998 Aspen Systems Conference on Telecommunications Policy met to consider ways of speeding the deployment of telecommunication systems that allow for robust, reliable, and innovative communications services to the home. By organizing participants into three working groups, the session was able to come up with analytical suggestions and policy…

  19. PESC '82; Annual Power Electronics Specialists Conference, 13th, Massachusetts Institute of Technology, Cambridge, MA, June 14-17, 1982, Record

    Science.gov (United States)

    Aspects of power electronics are addressed. The general topics discussed include: inverters and converters, modelling and analysis, motor drives, power conditioning appliances, power semiconductor devices, and power components and protection. Individual subjects considered include: dual-mode forward/flyback converter; a solar cell power supply system using a boost-type bidirectional DC-DC converter; complete DC analysis of the series resonant converter; variable structure control with sliding mode for DC drive speed regulation; a low-cost single-phase induction generator. Also covered are: small-signal modelling of a push-pull current-fed converter; programmable power processor for high-power space applications; high efficiency 3kW switch mode battery charger; comparison of BIMOS device types; power MOSFET temperature measurements; protection of power transistors in electric vehicle drives; general purpose variable frequency inverter using integrated power modules and LSI. For individual items see A84-18377 to A84-18408

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

    KAUST Repository

    Kim, Jeong Joo

    2016-04-09

    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. Kim et al. obtain the first crystal structure of the PKG I R domain bound with cGMP representing its activated state. It reveals a symmetric R dimer where cGMP molecules provide dimeric contacts. This R-R interaction prevents the high-affinity inhibitory interaction between R-C domain and sustains activation. © 2016 Elsevier Ltd.

  1. 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

    OpenAIRE

    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

  2. cGMP stimulation of cystic fibrosis transmembrane conductance regulator Cl- channels co-expressed with cGMP-dependent protein kinase type II but not type Ibeta

    NARCIS (Netherlands)

    A.B. Vaandrager (Arie); S.M. Lohmann (Suzanne); H.R. de Jonge (Hugo); W.C. Poller; B.C. Tilly (Bernard); A. Smolenski; S. Schneider-Rasp; A.G. Bot (Alice); M.J. Edixhoven (Marcel); B.J. Scholte (Bob); T. Jarchau; U. Walter

    1997-01-01

    textabstractIn order to investigate the involvement of cGMP-dependent protein kinase (cGK) type II in cGMP-provoked intestinal Cl- secretion, cGMP-dependent activation and phosphorylation of cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channels was ana

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

    Science.gov (United States)

    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.

  4. New GMP Models for Caucasus Region

    Science.gov (United States)

    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.

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

    International Nuclear Information System (INIS)

    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

  6. Optimization of RNA-based c-di-GMP fluorescent sensors through tuning their structural modules.

    Science.gov (United States)

    Inuzuka, Saki; Matsumura, Shigeyoshi; Ikawa, Yoshiya

    2016-08-01

    Cyclic diguanylate (c-di-GMP) is a second messenger of bacteria and its detection is an important issue in basic and applied microbiology. As c-di-GMP riboswitch ligand-binding domains (aptamer domains) capture c-di-GMP with high affinity and selectivity, they are promising platforms for the development of RNA-based c-di-GMP sensors. We analyzed two previously reported c-di-GMP sensor RNAs derived from the Vc2 riboswitch. We also designed and tested their variants, some of which showed improved properties as RNA-based c-di-GMP sensors.

  7. Duelund, Marts 2012, 13th edition udg. Strasbourg/Bonn : Council of Europe/ERICarts. 78 s. (Compendium of Cultural Policies and Trends in Europe)

    DEFF Research Database (Denmark)

    Duelund, Peter

    2012-01-01

    Perspective, General objectives and principles, Competence, decisionsmaking and administration, Current issues in cultural policymaking and debate,Main legal provisions in the cultural field, Financing of culture, Public institutions in the cultural infrastructure, Promoting creativity and participation......The Council of Europe/ERICarts "Compendium of Cultural Policies and Trends in Europe, 13th edition", 2012 is a web-based and permanently updated information and monitoring system of national cultural policies in Europe. It is a long term project which aims to include all 50 member states co......-operating within the context of the European Cultural Convention. Compendium cultural policy country profiles are mainly drawn up and updated by independent cultural policy experts (i.e. "the authors"), in consultation with respective ministries. The information presented in the updated country profiles is derived...

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

    DEFF Research Database (Denmark)

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

    2012-01-01

    in the development of novel antipathogenic drugs that can augment the effect of classic antibiotics by decreasing antimicrobial tolerance. The second messenger cyclic di-GMP is a positive regulator of biofilm formation, and cyclic di-GMP signaling is now regarded as a potential target for the development...... 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...

  9. 浅谈我国2010版 GMP 认证认识

    Institute of Scientific and Technical Information of China (English)

    张月秋

    2013-01-01

      毒胶囊事件引发人们深思,社会公众对制药企业的要求也日趋提高,GMP 认证势在必行,制药企业自身也加快了GMP 认证的步伐。本文回顾了我国实施 GMP 以来的发展历程。将我国新版 GMP 与美国 GMP 及欧盟 GMP 进行对比,说明我国GMP 步入世界轨道已进入重要阶段。因此,就目前我国制药企业进行 GMP 认证提出一些建议。

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

    NARCIS (Netherlands)

    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

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

    NARCIS (Netherlands)

    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

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

    International Nuclear Information System (INIS)

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

    Science.gov (United States)

    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....

  14. Compare Research of PIC/S, PIC/S GMP and Chinese GMP%PIC/S、PIC/S GMP与中国GMP的比较研究

    Institute of Scientific and Technical Information of China (English)

    张卓光

    2010-01-01

    目的 介绍药品监管公约/药品监管合作计划(PIC/S)和PIC/S GMP,比较PIC/S GMP 和中国GMP(98修订)的差异. 方法 从历史变革、机构与运行机制、宗旨以及PIC/S GMP的影响来阐述PIC/S;从人员、硬件、软件等方面来比较PIC/S GMP 与中国GMP.结果 PIC/S在药品领域倡导GMP标准、检查一致以及互认,促进了GMP发展.结论 PIC/S GMP是国际上高水平的GMP.

  15. Good Manufacturing Practice (GMP guidelines for virgin olive oil production

    Directory of Open Access Journals (Sweden)

    Petrakis, Christos

    1994-04-01

    Full Text Available This paper presents GMP guidelines for the production of virgin olive oil. Standard procedures and conditions are indicated for olive production, harvesting, transportation and storage, for oil manufacture, storage and packaging, for buildings, process logistics and the materials used throughout the production chain.

  16. cGMP signalling : different ways to create a pathway

    NARCIS (Netherlands)

    Roelofs, Jeroen; Smith, Janet L.; Haastert, Peter J.M. van

    2003-01-01

    Recently, a novel cGMP signalling cascade was uncovered in Dictyostelium, a eukaryote that diverged from the lineage leading to metazoa after plants and before yeast. In both Dictyostelium and metazoa, the ancient cAMP-binding (cNB) motif of bacterial CAP has been modified and assembled with other d

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

    Energy Technology Data Exchange (ETDEWEB)

    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)

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

    Directory of Open Access Journals (Sweden)

    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.

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

    International Nuclear Information System (INIS)

    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)

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

    Science.gov (United States)

    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.

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

    OpenAIRE

    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...

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

    Directory of Open Access Journals (Sweden)

    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.

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

    International Nuclear Information System (INIS)

    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.)

  4. 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

    CERN Document Server

    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...

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

    Science.gov (United States)

    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.

  6. 十三五时期需求结构趋势分析%Analysis of Demand Structure in the Period of 13th Five-Year Plan

    Institute of Scientific and Technical Information of China (English)

    邓娜; 陈丹; 曾思孟

    2016-01-01

    Based on the analysis of the domestic and international economic situation in the 13th five-year plan period, this paper predicts the trend of the change of demand structure. Consumption will gradually become the main driving force of economic growth. Finally, we put forward some countermeasures and suggestions to promote the steady growth and structural optimization of the three key demands of consumption, investment and export.%新世纪以来,随着经济持续快速发展和结构转型升级深入推进,我国消费、投资、出口等需求规模持续扩大,结构不断优化,“三驾马车”对经济增长的贡献发生了阶段性变化,资本形成率高、最终消费率高、净出口率低的“两高一低”特征明显。

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

    Science.gov (United States)

    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.

  8. Endo-S-c-di-GMP Analogues-Polymorphism and Binding Studies with Class I Riboswitch

    Directory of Open Access Journals (Sweden)

    Herman O. Sintim

    2012-11-01

    Full Text Available C-di-GMP, a cyclic guanine dinucleotide, has been shown to regulate biofilm formation as well as virulence gene expression in a variety of bacteria. Analogues of c-di-GMP have the potential to be used as chemical probes to study c-di-GMP signaling and could even become drug leads for the development of anti-biofilm compounds. Herein we report the synthesis and biophysical studies of a series of c-di-GMP analogues, which have both phosphate and sugar moieties simultaneously modified (called endo-S-c-di-GMP analogues. We used computational methods to predict the relative orientation of the guanine nucleobases in c-di-GMP and analogues. DOSY NMR of the endo-S-c-di-GMP series showed that the polymorphism of c-di-GMP can be tuned with conservative modifications to the phosphate and sugar moieties (conformational steering. Binding studies with Vc2 RNA (a class I c-di-GMP riboswitch revealed that conservative modifications to the phosphate and 2'-positions of c-di-GMP dramatically affected binding to class I riboswitch.

  9. Endo-S-c-di-GMP analogues-polymorphism and binding studies with class I riboswitch.

    Science.gov (United States)

    Zhou, Jie; Sayre, David A; Wang, Jingxin; Pahadi, Nirmal; Sintim, Herman O

    2012-01-01

    C-di-GMP, a cyclic guanine dinucleotide, has been shown to regulate biofilm formation as well as virulence gene expression in a variety of bacteria. Analogues of c-di-GMP have the potential to be used as chemical probes to study c-di-GMP signaling and could even become drug leads for the development of anti-biofilm compounds. Herein we report the synthesis and biophysical studies of a series of c-di-GMP analogues, which have both phosphate and sugar moieties simultaneously modified (called endo-S-c-di-GMP analogues). We used computational methods to predict the relative orientation of the guanine nucleobases in c-di-GMP and analogues. DOSY NMR of the endo-S-c-di-GMP series showed that the polymorphism of c-di-GMP can be tuned with conservative modifications to the phosphate and sugar moieties (conformational steering). Binding studies with Vc2 RNA (a class I c-di-GMP riboswitch) revealed that conservative modifications to the phosphate and 2'-positions of c-di-GMP dramatically affected binding to class I riboswitch. PMID:23143150

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

    Science.gov (United States)

    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

  11. Extracellular cGMP Modulates Learning Biphasically by Modulating Glycine Receptors, CaMKII and Glutamate-Nitric Oxide-cGMP Pathway

    Science.gov (United States)

    Cabrera-Pastor, Andrea; Malaguarnera, Michele; Taoro-Gonzalez, Lucas; Llansola, Marta; Felipo, Vicente

    2016-01-01

    It has been proposed that extracellular cGMP modulates the ability to learn a Y maze task, but the underlying mechanisms remained unknown. Here we show that extracellular cGMP, at physiological concentrations, modulates learning in the Y maze in a biphasic way by modulating the glutamate-nitric oxide-cGMP pathway in cerebellum. Extracellular cGMP reduces glycine receptors activation inducing a voltage-dependent calcium-channels-mediated increase of calcium in Purkinje neurons. This calcium increase modulates CaMKII phosphorylation in a biphasic way. When basal calcium concentration is low extracellular cGMP reduces CaMKII phosphorylation, increasing nitric oxide synthase activity, the glutamate-NO-cGMP pathway function and learning ability. When basal calcium is normal extracellular cGMP increases CaMKII phosphorylation, reducing nitric oxide synthase activity, the pathway function and learning. These data unveil new mechanisms modulating learning in the Y maze and likely other learning types which may be therapeutic targets to improve learning in pathological situations associated with altered cGMP levels. PMID:27634333

  12. Extracellular cGMP Modulates Learning Biphasically by Modulating Glycine Receptors, CaMKII and Glutamate-Nitric Oxide-cGMP Pathway.

    Science.gov (United States)

    Cabrera-Pastor, Andrea; Malaguarnera, Michele; Taoro-Gonzalez, Lucas; Llansola, Marta; Felipo, Vicente

    2016-01-01

    It has been proposed that extracellular cGMP modulates the ability to learn a Y maze task, but the underlying mechanisms remained unknown. Here we show that extracellular cGMP, at physiological concentrations, modulates learning in the Y maze in a biphasic way by modulating the glutamate-nitric oxide-cGMP pathway in cerebellum. Extracellular cGMP reduces glycine receptors activation inducing a voltage-dependent calcium-channels-mediated increase of calcium in Purkinje neurons. This calcium increase modulates CaMKII phosphorylation in a biphasic way. When basal calcium concentration is low extracellular cGMP reduces CaMKII phosphorylation, increasing nitric oxide synthase activity, the glutamate-NO-cGMP pathway function and learning ability. When basal calcium is normal extracellular cGMP increases CaMKII phosphorylation, reducing nitric oxide synthase activity, the pathway function and learning. These data unveil new mechanisms modulating learning in the Y maze and likely other learning types which may be therapeutic targets to improve learning in pathological situations associated with altered cGMP levels. PMID:27634333

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

    Science.gov (United States)

    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

  14. Annual energy review 1994

    Energy Technology Data Exchange (ETDEWEB)

    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).

  15. Design of GMP compliance radiopharmaceutical production facility in MINT

    International Nuclear Information System (INIS)

    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)

  16. cGMP in ozone and NO dependent responses

    Science.gov (United States)

    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

  17. Correlative intravital imaging of cGMP signals and vasodilation in mice

    Directory of Open Access Journals (Sweden)

    Martin eThunemann

    2014-10-01

    Full Text Available Cyclic guanosine monophosphate (cGMP is an important signaling molecule and drug target in the cardiovascular system. It is well known that stimulation of the vascular nitric oxide (NO-cGMP pathway results in vasodilation. However, the spatiotemporal dynamics of cGMP signals themselves and the cGMP concentrations within specific cardiovascular cell types in health, disease, and during pharmacotherapy with cGMP-elevating drugs are largely unknown. To facilitate the analysis of cGMP signaling in vivo, we have generated transgenic mice that express fluorescence resonance energy transfer (FRET-based cGMP sensor proteins. Here, we describe two models of intravital FRET/cGMP imaging in the vasculature of cGMP sensor mice: (1 epifluorescence-based ratio imaging in resistance-type vessels of the cremaster muscle and (2 ratio imaging by multiphoton microscopy within the walls of subcutaneous blood vessels accessed through a dorsal skinfold chamber. Both methods allow simultaneous monitoring of NO-induced cGMP transients and vasodilation in living mice. Detailed protocols of all steps necessary to perform and evaluate intravital imaging experiments of the vasculature of anesthetized mice including surgery, imaging, and data evaluation are provided. An image segmentation approach is described to estimate FRET/cGMP changes within moving structures such as the vessel wall during vasodilation. The methods presented herein should be useful to visualize cGMP or other biochemical signals that are detectable with FRET-based biosensors, such as cyclic adenosine monophosphate or Ca2+, and to correlate them with respective vascular responses. With further refinement and combination of transgenic mouse models and intravital imaging technologies, we envision an exciting future, in which we are able to ‘watch’ biochemistry, (patho physiology, and pharmacotherapy in the context of a living mammalian organism.

  18. Stimulation of innate immunity by in vivo cyclic di-GMP synthesis using adenovirus.

    Science.gov (United States)

    Koestler, Benjamin J; Seregin, Sergey S; Rastall, David P W; Aldhamen, Yasser A; Godbehere, Sarah; Amalfitano, Andrea; Waters, Christopher M

    2014-11-01

    The bacterial second messenger cyclic di-GMP (c-di-GMP) stimulates inflammation by initiating innate immune cell recruitment and triggering the release of proinflammatory cytokines and chemokines. These properties make c-di-GMP a promising candidate for use as a vaccine adjuvant, and numerous studies have demonstrated that administration of purified c-di-GMP with different antigens increases protection against infection in animal models. Here, we have developed a novel approach to produce c-di-GMP inside host cells as an adjuvant to exploit a host-pathogen interaction and initiate an innate immune response. We have demonstrated that c-di-GMP can be synthesized in vivo by transducing a diguanylate cyclase (DGC) gene into mammalian cells using an adenovirus serotype 5 (Ad5) vector. Expression of DGC led to the production of c-di-GMP in vitro and in vivo, and this was able to alter proinflammatory gene expression in murine tissues and increase the secretion of numerous cytokines and chemokines when administered to animals. Furthermore, coexpression of DGC modestly increased T-cell responses to a Clostridium difficile antigen expressed from an adenovirus vaccine, although no significant differences in antibody titers were observed. This adenovirus c-di-GMP delivery system offers a novel method to administer c-di-GMP as an adjuvant to stimulate innate immunity during vaccination.

  19. Cyclic di-GMP riboswitch-regulated type IV pili contribute to aggregation of Clostridium difficile.

    Science.gov (United States)

    Bordeleau, Eric; Purcell, Erin B; Lafontaine, Daniel A; Fortier, Louis-Charles; Tamayo, Rita; Burrus, Vincent

    2015-03-01

    Clostridium difficile is an anaerobic Gram-positive bacterium that causes intestinal infections with symptoms ranging from mild diarrhea to fulminant colitis. Cyclic diguanosine monophosphate (c-di-GMP) is a bacterial second messenger that typically regulates the switch from motile, free-living to sessile and multicellular behaviors in Gram-negative bacteria. Increased intracellular c-di-GMP concentration in C. difficile was recently shown to reduce flagellar motility and to increase cell aggregation. In this work, we investigated the role of the primary type IV pilus (T4P) locus in c-di-GMP-dependent cell aggregation. Inactivation of two T4P genes, pilA1 (CD3513) and pilB1 (CD3512), abolished pilus formation and significantly reduced cell aggregation under high c-di-GMP conditions. pilA1 is preceded by a putative c-di-GMP riboswitch, predicted to be transcriptionally active upon c-di-GMP binding. Consistent with our prediction, high intracellular c-di-GMP concentration increased transcript levels of T4P genes. In addition, single-round in vitro transcription assays confirmed that transcription downstream of the predicted transcription terminator was dose dependent and specific to c-di-GMP binding to the riboswitch aptamer. These results support a model in which T4P gene transcription is upregulated by c-di-GMP as a result of its binding to an upstream transcriptionally activating riboswitch, promoting cell aggregation in C. difficile.

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

    Science.gov (United States)

    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

  1. 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

    Directory of Open Access Journals (Sweden)

    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

  2. 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 

    OpenAIRE

    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...

  3. 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.

    Science.gov (United States)

    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.

  4. 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

    CERN Document Server

    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.

  5. Near-infrared light responsive synthetic c-di-GMP module for optogenetic applications.

    Science.gov (United States)

    Ryu, Min-Hyung; Gomelsky, Mark

    2014-11-21

    Enormous potential of cell-based therapeutics is hindered by the lack of effective means to control genetically engineered cells in mammalian tissues. Here, we describe a synthetic module for remote photocontrol of engineered cells that can be adapted for such applications. The module involves photoactivated synthesis of cyclic dimeric GMP (c-di-GMP), a stable small molecule that is not produced by higher eukaryotes and therefore is suitable for orthogonal regulation. The key component of the photocontrol module is an engineered bacteriophytochrome diguanylate cyclase, which synthesizes c-di-GMP from GTP in a light-dependent manner. Bacteriophytochromes are particularly attractive photoreceptors because they respond to light in the near-infrared window of the spectrum, where absorption by mammalian tissues is minimal, and also because their chromophore, biliverdin IXα, is naturally available in mammalian cells. The second component of the photocontrol module, a c-di-GMP phosphodiesterase, maintains near-zero background levels of c-di-GMP in the absence of light, which enhances the photodynamic range of c-di-GMP concentrations. In the E. coli model used in this study, the intracellular c-di-GMP levels could be upregulated by light by >50-fold. Various c-di-GMP-responsive proteins and riboswitches identified in bacteria can be linked downstream of the c-di-GMP-mediated photocontrol module for orthogonal regulation of biological activities in mammals as well as in other organisms lacking c-di-GMP signaling. Here, we linked the photocontrol module to a gene expression output via a c-di-GMP-responsive transcription factor and achieved a 40-fold photoactivation of gene expression.

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

    Science.gov (United States)

    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

  7. Methods and materials relating to IMPDH and GMP production

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    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

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

    OpenAIRE

    Cláudio Boghi; Dorlivete M. Shitsuka; Marcos Alexandruk; Ricardo Shitsuka; Paulo Roberto Rela

    2008-01-01

    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 stud...

  10. cGMP-binding prepares PKG for substrate binding by disclosing the C-terminal domain

    NARCIS (Netherlands)

    Alverdi, V.; Mazon, H.F.M.; Versluis, C.; Hemrika, W.; Esposito, G.; van den Heuvel, R.H.H.; Scholten, A.; Heck, A.J.R.

    2008-01-01

    Type I cyclic guanosine 3′,5′-monophosphate (cGMP)-dependent protein kinase (PKG) is involved in the nitric oxide/cGMP signaling pathway. PKG has been identified in many different species, ranging from unicelõlular organisms to mammals. The enzyme serves as one of the major receptor proteins for int

  11. Understanding the link between GMP and dough: From glutenin particles in flour towards developed dough

    NARCIS (Netherlands)

    Don, C.; Lichtendonk, W.J.; Plijter, J.J.; Hamer, R.J.

    2003-01-01

    Clear correlations exist for glutenin macropolymer (GMP) quantity and rheological properties vs. wheat quality and dough rheological properties, but real insight in understanding these links is still missing. The observation that GMP consists of glutenin particles opens up new possibilities to revea

  12. Trigger phosphodiesterases as a novel class of c-di-GMP effector proteins.

    Science.gov (United States)

    Hengge, Regine

    2016-11-01

    The bacterial second messenger c-di-GMP controls bacterial biofilm formation, motility, cell cycle progression, development and virulence. It is synthesized by diguanylate cyclases (with GGDEF domains), degraded by specific phosphodiesterases (PDEs, with EAL of HD-GYP domains) and sensed by a wide variety of c-di-GMP-binding effectors that control diverse targets. c-di-GMP-binding effectors can be riboswitches as well as proteins with highly diverse structures and functions. The latter include 'degenerate' GGDEF/EAL domain proteins that are enzymatically inactive but still able to bind c-di-GMP. Surprisingly, two enzymatically active 'trigger PDEs', the Escherichia coli proteins PdeR and PdeL, have recently been added to this list of c-di-GMP-sensing effectors. Mechanistically, trigger PDEs are multifunctional. They directly and specifically interact with a macromolecular target (e.g. with a transcription factor or directly with a promoter region), whose activity they control by their binding and degradation of c-di-GMP-their PDE activity thus represents the c-di-GMP sensor or effector function. In this process, c-di-GMP serves as a regulatory ligand, but in contrast to classical allosteric control, this ligand is also degraded. The resulting kinetics and circuitry of control are ideally suited for trigger PDEs to serve as key components in regulatory switches.This article is part of the themed issue 'The new bacteriology'.

  13. Understanding the link between GMP and dough: from glutenin particles in flour towards developed dough

    NARCIS (Netherlands)

    Don, C.; Lichtendonk, W.J.; Plijter, J.J.; Hamer, R.J.

    2003-01-01

    Clear correlations exist for glutenin macropolymer (GMP) quantity and rheological properties vs. wheat quality and dough rheological properties, but real insight in understanding these links is still missing. The observation that GMP consists of glutenin particles opens up new possibilities to revea

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

    Directory of Open Access Journals (Sweden)

    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.

  15. c-di-GMP induction of Dictyostelium cell death requires the polyketide DIF-1.

    Science.gov (United States)

    Song, Yu; Luciani, Marie-Françoise; Giusti, Corinne; Golstein, Pierre

    2015-02-15

    Cell death in the model organism Dictyostelium, as studied in monolayers in vitro, can be induced by the polyketide DIF-1 or by the cyclical dinucleotide c-di-GMP. c-di-GMP, a universal bacterial second messenger, can trigger innate immunity in bacterially infected animal cells and is involved in developmental cell death in Dictyostelium. We show here that c-di-GMP was not sufficient to induce cell death in Dictyostelium cell monolayers. Unexpectedly, it also required the DIF-1 polyketide. The latter could be exogenous, as revealed by a telling synergy between c-di-GMP and DIF-1. The required DIF-1 polyketide could also be endogenous, as shown by the inability of c-di-GMP to induce cell death in Dictyostelium HMX44A cells and DH1 cells upon pharmacological or genetic inhibition of DIF-1 biosynthesis. In these cases, c-di-GMP-induced cell death was rescued by complementation with exogenous DIF-1. Taken together, these results demonstrated that c-di-GMP could trigger cell death in Dictyostelium only in the presence of the DIF-1 polyketide or its metabolites. This identified another element of control to this cell death and perhaps also to c-di-GMP effects in other situations and organisms.

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

    Directory of Open Access Journals (Sweden)

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

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

    Science.gov (United States)

    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

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

    International Nuclear Information System (INIS)

    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

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

    Directory of Open Access Journals (Sweden)

    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.

  1. Secreted Cyclic Di-GMP Induces Stalk Cell Differentiation in the Eukaryote Dictyostelium discoideum.

    Science.gov (United States)

    Chen, Zhi-hui; Schaap, Pauline

    2016-01-01

    Cyclic di-GMP (c-di-GMP) is currently recognized as the most widely used intracellular signal molecule in prokaryotes, but roles in eukaryotes were only recently discovered. In the social amoeba Dictyostelium discoideum, c-di-GMP, produced by a prokaryote-type diguanylate cyclase, induces the differentiation of stalk cells, thereby enabling the formation of spore-bearing fruiting bodies. In this review, we summarize the currently known mechanisms that control the major life cycle transitions of Dictyostelium and focus particularly on the role of c-di-GMP in stalk formation. Stalk cell differentiation has characteristics of autophagic cell death, a process that also occurs in higher eukaryotes. We discuss the respective roles of c-di-GMP and of another signal molecule, differentiation-inducing factor 1, in autophagic cell death in vitro and in stalk formation in vivo.

  2. 75 FR 17756 - Availability of the Final General Management Plan and Environmental Impact Statement (GMP/EIS...

    Science.gov (United States)

    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...

  3. 75 FR 19422 - Availability of a Final Environmental Impact Statement for the General Management Plan (FEIS/GMP...

    Science.gov (United States)

    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...

  4. 75 FR 17761 - Termination of an Environmental Impact Statement (EIS) for the General Management Plan (GMP) for...

    Science.gov (United States)

    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...

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    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 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. PMID:24626229

  7. New insights into Legionella pneumophila biofilm regulation by c-di-GMP signaling.

    Science.gov (United States)

    Pécastaings, Sophie; Allombert, Julie; Lajoie, Barbora; Doublet, Patricia; Roques, Christine; Vianney, Anne

    2016-09-01

    The waterborne pathogen Legionella pneumophila grows as a biofilm, freely or inside amoebae. Cyclic-di-GMP (c-di-GMP), a bacterial second messenger frequently implicated in biofilm formation, is synthesized and degraded by diguanylate cyclases (DGCs) and phosphodiesterases (PDEs), respectively. To characterize the c-di-GMP-metabolizing enzymes involved in L. pneumophila biofilm regulation, the consequences on biofilm formation and the c-di-GMP concentration of each corresponding gene inactivation were assessed in the Lens strain. The results showed that one DGC and two PDEs enhance different aspects of biofilm formation, while two proteins with dual activity (DGC/PDE) inhibit biofilm growth. Surprisingly, only two mutants exhibited a change in global c-di-GMP concentration. This study highlights that specific c-di-GMP pathways control L. pneumophila biofilm formation, most likely via temporary and/or local modulation of c-di-GMP concentration. Furthermore, Lpl1054 DGC is required to enable the formation a dense biofilm in response to nitric oxide, a signal for biofilm dispersion in many other species.

  8. New insights into Legionella pneumophila biofilm regulation by c-di-GMP signaling.

    Science.gov (United States)

    Pécastaings, Sophie; Allombert, Julie; Lajoie, Barbora; Doublet, Patricia; Roques, Christine; Vianney, Anne

    2016-09-01

    The waterborne pathogen Legionella pneumophila grows as a biofilm, freely or inside amoebae. Cyclic-di-GMP (c-di-GMP), a bacterial second messenger frequently implicated in biofilm formation, is synthesized and degraded by diguanylate cyclases (DGCs) and phosphodiesterases (PDEs), respectively. To characterize the c-di-GMP-metabolizing enzymes involved in L. pneumophila biofilm regulation, the consequences on biofilm formation and the c-di-GMP concentration of each corresponding gene inactivation were assessed in the Lens strain. The results showed that one DGC and two PDEs enhance different aspects of biofilm formation, while two proteins with dual activity (DGC/PDE) inhibit biofilm growth. Surprisingly, only two mutants exhibited a change in global c-di-GMP concentration. This study highlights that specific c-di-GMP pathways control L. pneumophila biofilm formation, most likely via temporary and/or local modulation of c-di-GMP concentration. Furthermore, Lpl1054 DGC is required to enable the formation a dense biofilm in response to nitric oxide, a signal for biofilm dispersion in many other species. PMID:27494738

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

    Science.gov (United States)

    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

  10. 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

    Energy Technology Data Exchange (ETDEWEB)

    Alahari, N.; Bajeat, O.; Barue, C.; Chautard, F.; Clement, E.; Delahaye, P.; De Oliveira, F.; Dubois, M.; Fadil, M.; Fraanberg-Delahaye, H.; Jacquot, B.; Jardin, P.; Saint-Laurent, M. G.; Lecesne, N.; Leherissier, P.; Leroy, R.; Lewitowicz, M.; Lhersonneau, G.; Maunoury, L.; Mery, A. [GANIL, Bd Henri Becquerel, BP 55027, 14076 Caen Cedex 5 (France); and others

    2010-02-15

    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.

  11. 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

    International Nuclear Information System (INIS)

    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.

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

    Science.gov (United States)

    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.

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

    OpenAIRE

    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...

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

    International Nuclear Information System (INIS)

    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

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

    International Nuclear Information System (INIS)

    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

  16. Diamidinium and iminium aromatics as new aggregators of the bacterial signaling molecule, c-di-GMP.

    Science.gov (United States)

    Kelsey, Ilana; Nakayama, Shizuka; Sintim, Herman O

    2012-01-15

    C-di-GMP has emerged as an important bacterial signaling molecule that is involved in biofilm formation. Small molecules that can form biologically inactive complexes with c-di-GMP have the potential to be used as anti-biofilm agents. Herein, we report that water-soluble diamidinium/iminium aromatics (such as berenil), which are traditionally considered as minor groove binders of nucleic acids, are capable of aggregating c-di-GMP into G-quadruplexes via π-stacking interactions. PMID:22217869

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

    OpenAIRE

    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...

  18. Supercritical fluid chromatography for GMP analysis in support of pharmaceutical development and manufacturing activities.

    Science.gov (United States)

    Hicks, Michael B; Regalado, Erik L; Tan, Feng; Gong, Xiaoyi; Welch, Christopher J

    2016-01-01

    Supercritical fluid chromatography (SFC) has long been a preferred method for enantiopurity analysis in support of pharmaceutical discovery and development, but implementation of the technique in regulated GMP laboratories has been somewhat slow, owing to limitations in instrument sensitivity, reproducibility, accuracy and robustness. In recent years, commercialization of next generation analytical SFC instrumentation has addressed previous shortcomings, making the technique better suited for GMP analysis. In this study we investigate the use of modern SFC for enantiopurity analysis of several pharmaceutical intermediates and compare the results with the conventional HPLC approaches historically used for analysis in a GMP setting. The findings clearly illustrate that modern SFC now exhibits improved precision, reproducibility, accuracy and robustness; also providing superior resolution and peak capacity compared to HPLC. Based on these findings, the use of modern chiral SFC is recommended for GMP studies of stereochemistry in pharmaceutical development and manufacturing.

  19. The expanding roles of c-di-GMP in the biosynthesis of exopolysaccharides and secondary metabolites.

    Science.gov (United States)

    Liang, Zhao-Xun

    2015-05-01

    The cyclic dinucleotide c-di-GMP has emerged in the last decade as a prevalent intracellular messenger that orchestrates the transition between the motile and sessile lifestyles of many bacterial species. The motile-to-sessile transition is often associated with the formation of extracellular matrix-encased biofilm, an organized community of bacterial cells that often contributes to antibiotic resistance and host-pathogen interaction. It is increasingly clear that c-di-GMP controls motility, biofilm formation and bacterial pathogenicity partially through regulating the production of exopolysaccharides (EPS) and small-molecule secondary metabolites. This review summarizes our current understanding of the regulation of EPS biosynthesis by c-di-GMP in a diversity of bacterial species and highlights the emerging role of c-di-GMP in the biosynthesis of small-molecule secondary metabolites.

  20. Evidence for a Messenger Function of Cyclic GMP During Phosphodiesterase Induction in Dictyostelium discoideum

    NARCIS (Netherlands)

    Haastert, Peter J.M. van; Pasveer, Frank J.; Meer, Rob C. van der; Heijden, Paul R. van der; Walsum, Hans van; Konijn, Theo M.

    1982-01-01

    Chemotactic stimulation of vegetative or aggregative Dictyostelium discoideum cells induced a transient elevation of cyclic GMP levels. The addition of chemoattractants to postvegetative cells by pulsing induced phosphodiesterase activity. The following lines of evidence suggest a messenger function

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

    Science.gov (United States)

    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.

  2. The Structural Basis for the Sensing and Binding of Cyclic di-GMP by STING

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    Under a research project funded by NSFC, Prof. Su Xiaodong and his team of the National Laboratory of Protein Engineering and Plant Genetic Engineering, BIOPIC, the School of Life Sciences of Peking Uni- versity, obtained remarkable achievement and published recently a paper entitled "The Structural Basis for the Sensing and Binding of Cyclic di-GMP by STING" on online Natural Structural and Molecular Biolo- gY. STING (stimulator of interferon genes) is an essential signaling adaptor that mediates cytokine pro- duction in response to microbial invasion by directly sensing bacterial secondary messengers such as the cy- clic dinucleotide bis-(3'-5')-cyclic dimeric GMP (e-di-GMP). STING's structure and its binding mecha- nism to cyclic dinucleotides were unknown. We report here the crystal structures of the STING cytoplas- mic domain and its complex with c-di-GMP, thus providing the structural basis for understanding STING function.

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

    Science.gov (United States)

    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

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

    OpenAIRE

    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)...

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

    Science.gov (United States)

    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

  6. Targeting cyclic di-GMP signalling: a strategy to control biofilm formation?

    Science.gov (United States)

    Caly, Delphine L; Bellini, Domenico; Walsh, Martin A; Dow, J Maxwell; Ryan, Robert P

    2015-01-01

    Cyclic di-GMP is a second messenger found in almost all eubacteria that acts to regulate a wide range of functions including developmental transitions, adhesion and biofilm formation. Cyclic di-GMP is synthesised from two GTP molecules by diguanylate cyclases that have a GGDEF domain and is degraded by phosphodiesterases with either an EAL or an HD-GYP domain. Proteins with these domains often contain additional signal input domains, suggesting that their enzymatic activity may be modulated as a response to different environmental or cellular cues. Cyclic di-GMP exerts a regulatory action through binding to diverse receptors that include a small protein domain called PilZ, enzymatically inactive GGDEF, EAL or HD-GYP domains, transcription factors and riboswitches. In many bacteria, high cellular levels of cyclic di-GMP are associated with a sessile, biofilm lifestyle, whereas low levels of the nucleotide promote motility and virulence factor synthesis in pathogens. Elucidation of the roles of cyclic di-GMP signalling in biofilm formation has suggested strategies whereby modulation of the levels of the nucleotide or interference with signalling pathways may lead to inhibition of biofilm formation or promotion of biofilm dispersal. In this review we consider these approaches for the control of biofilm formation, beginning with an overview of cyclic di-GMP signalling and the different ways that it can act in regulation of biofilm dynamics.

  7. Cyclic di-GMP acts as a cell cycle oscillator to drive chromosome replication.

    Science.gov (United States)

    Lori, C; Ozaki, S; Steiner, S; Böhm, R; Abel, S; Dubey, B N; Schirmer, T; Hiller, S; Jenal, U

    2015-07-01

    Fundamental to all living organisms is the capacity to coordinate cell division and cell differentiation to generate appropriate numbers of specialized cells. Whereas eukaryotes use cyclins and cyclin-dependent kinases to balance division with cell fate decisions, equivalent regulatory systems have not been described in bacteria. Moreover, the mechanisms used by bacteria to tune division in line with developmental programs are poorly understood. Here we show that Caulobacter crescentus, a bacterium with an asymmetric division cycle, uses oscillating levels of the second messenger cyclic diguanylate (c-di-GMP) to drive its cell cycle. We demonstrate that c-di-GMP directly binds to the essential cell cycle kinase CckA to inhibit kinase activity and stimulate phosphatase activity. An upshift of c-di-GMP during the G1-S transition switches CckA from the kinase to the phosphatase mode, thereby allowing replication initiation and cell cycle progression. Finally, we show that during division, c-di-GMP imposes spatial control on CckA to install the replication asymmetry of future daughter cells. These studies reveal c-di-GMP to be a cyclin-like molecule in bacteria that coordinates chromosome replication with cell morphogenesis in Caulobacter. The observation that c-di-GMP-mediated control is conserved in the plant pathogen Agrobacterium tumefaciens suggests a general mechanism through which this global regulator of bacterial virulence and persistence coordinates behaviour and cell proliferation.

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

    Science.gov (United States)

    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.

  9. 浅谈GMP自检%Discussion on Self-test of GMP

    Institute of Scientific and Technical Information of China (English)

    刘加宝

    2015-01-01

    GMP自检是指药品生产企业内部对药品生产实施GMP的检查,是企业执行GMP中的一项重要内容,也是日常生产质量管理工作中一项重要的质量活动,实质上就是对企业完善生产质量管理体系的自我检查。通过GMP自检,发现企业执行GMP时存在的质量风险,并通过实施纠正和预防措施,来进一步提高GMP执行的持续性、符合性、有效性,通过自检进行持续改进。%GMP pharmaceutical manufac turing self-test refers to the internal implementation of GMP for pharmaceutical production inspection, GMP is a business executive in an important part of daily production and quality management is an important quality in work activities, in essence, is for enterprises to improve production quality management system of self-examination. By GMP self-test, found that companies perform GMP quality risk exists, and through the implementation of corrective and preventive measures to further improve the sustainability of GMP execution, compliance, effectiveness, through a self-test for continuous improvement.

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

    Science.gov (United States)

    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

  11. Structural basis for c-di-GMP-mediated inside-out signaling controlling periplasmic proteolysis.

    Directory of Open Access Journals (Sweden)

    Marcos V A S Navarro

    Full Text Available The bacterial second messenger bis-(3'-5' cyclic dimeric guanosine monophosphate (c-di-GMP has emerged as a central regulator for biofilm formation. Increased cellular c-di-GMP levels lead to stable cell attachment, which in Pseudomonas fluorescens requires the transmembrane receptor LapD. LapD exhibits a conserved and widely used modular architecture containing a HAMP domain and degenerate diguanylate cyclase and phosphodiesterase domains. c-di-GMP binding to the LapD degenerate phosphodiesterase domain is communicated via the HAMP relay to the periplasmic domain, triggering sequestration of the protease LapG, thus preventing cleavage of the surface adhesin LapA. Here, we elucidate the molecular mechanism of autoinhibition and activation of LapD based on structure-function analyses and crystal structures of the entire periplasmic domain and the intracellular signaling unit in two different states. In the absence of c-di-GMP, the intracellular module assumes an inactive conformation. Binding of c-di-GMP to the phosphodiesterase domain disrupts the inactive state, permitting the formation of a trans-subunit dimer interface between adjacent phosphodiesterase domains via interactions conserved in c-di-GMP-degrading enzymes. Efficient mechanical coupling of the conformational changes across the membrane is realized through an extensively domain-swapped, unique periplasmic fold. Our structural and functional analyses identified a conserved system for the regulation of periplasmic proteases in a wide variety of bacteria, including many free-living and pathogenic species.

  12. Polyphosphate, cyclic AMP, guanosine tetraphosphate, and c-di-GMP reduce in vitro Lon activity.

    Science.gov (United States)

    Osbourne, Devon O; Soo, Valerie W C; Konieczny, Igor; Wood, Thomas K

    2014-01-01

    Lon protease is conserved from bacteria to humans and regulates cellular processes by degrading different classes of proteins including antitoxins, transcriptional activators, unfolded proteins, and free ribosomal proteins. Since we found that Lon has several putative cyclic diguanylate (c-di-GMP) binding sites and since Lon binds polyphosphate (polyP) and lipid polysaccharide, we hypothesized that Lon has an affinity for phosphate-based molecules that might regulate its activity. Hence we tested the effect of polyP, cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), guanosine tetraphosphate (ppGpp), c-di-GMP, and GMP on the ability of Lon to degrade α-casein. Inhibition of in vitro Lon activity occurred for polyP, cAMP, ppGpp, and c-di-GMP. We also demonstrated by HPLC that Lon is able to bind c-di-GMP. Therefore, four cell signals were found to regulate the activity of Lon protease.

  13. Design of GMP compliance radiopharmaceutical production facility in MINT

    International Nuclear Information System (INIS)

    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. 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)

    Science.gov (United States)

    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

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

    DEFF Research Database (Denmark)

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

    2015-01-01

    infection give rise to rugose small colony variants (RSCVs), which are hyper-biofilm-forming mutants that commonly possess mutations that increase production of the biofilm-promoting secondary messenger cyclic di-GMP (c-di-GMP). We show that RSCVs display a decreased production of acute virulence factors...... as 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......AMP), which is decreased in response to a high level of c-di-GMP. Mutations that cause reversion of the RSCV phenotype concomitantly reactivate Vfr-cAMP signaling. Attempts to uncover the mechanism underlying the observed c-di-GMP-mediated lowering of cAMP content provided evidence that it is not caused...

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

    Science.gov (United States)

    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.

  17. 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

    Institute of Scientific and Technical Information of China (English)

    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.

  18. Feasibility of 13th lymph nodes as sentinels for periampullary carcinoma%壶腹周围癌第13组淋巴结作为前哨淋巴结的可行性分析

    Institute of Scientific and Technical Information of China (English)

    白晓枫; 张业繁; 赵东兵; 王健; 赵宏

    2013-01-01

    目的 探讨第13组淋巴结是否可作为前哨淋巴结预测壶腹周围癌淋巴结转移情况.方法 选择2009年1月至2012年1月中国医学科学院肿瘤医院77例行Whipple术的壶腹周围癌.其中26例(试验组,男18例、女8例,年龄38 ~ 79岁)行Whipple术中,采用1%亚甲蓝或纳米碳混悬液术中瘤床注射,另外5l例作为对照组不注射,其中男33例、女l8例,年龄33~78岁.对染色的第13组淋巴结进行活检,之后常规行淋巴结清扫术,并对淋巴结转移情况进行研究.结果 试验组患者在瘤床注射1%亚甲蓝或纳米碳混悬液能够显示出肿瘤淋巴结的引流范围;对照组和试验组患者平均清扫淋巴结个数分别为15.8(3 ~54)个和17.6(6~ 40)个,差异无统计学意义(P=0.460).结论 第13组淋巴结可能并不适合作为前哨淋巴结用于预测胰腺及壶腹周围癌淋巴结转移情况.%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 females,age 38-79 years) of them underwent Whipple procedures during which 1% methylene blue or nanogate carbon was injected into tumor bed.The othor 51 patients as controls (33 males,18 females,age 38-78 years).The dyed 13th lymph nodes were biopsied.Then routine lymphadenectomy was performed and their pathological results were analyzed.Results In the experimental group,the lymph node drainage area of cancer was identified.However,in contrast with 51 patients in the control group,the labeling of 13th lymph nodes did not significantly increase the number of cleaned lymph nodes (15.8 (3-54) vs 17.6 (6-40),P =0.460).Conclusion 13th lymph nodes are non-suitable as sentinels for predicting general lymph nodes metastases of periampullary carcinoma.

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

    Science.gov (United States)

    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

  20. A minimalist biosensor: Quantitation of cyclic di-GMP using the conformational change of a riboswitch aptamer.

    Science.gov (United States)

    Kellenberger, Colleen A; Sales-Lee, Jade; Pan, Yuchen; Gassaway, Madalee M; Herr, Amy E; Hammond, Ming C

    2015-01-01

    Cyclic di-GMP (c-di-GMP) is a second messenger that is important in regulating bacterial physiology and behavior, including motility and virulence. Many questions remain about the role and regulation of this signaling molecule, but current methods of detection are limited by either modest sensitivity or requirements for extensive sample purification. We have taken advantage of a natural, high affinity receptor of c-di-GMP, the Vc2 riboswitch aptamer, to develop a sensitive and rapid electrophoretic mobility shift assay (EMSA) for c-di-GMP quantitation that required minimal engineering of the RNA.

  1. Liposomes loaded with a STING pathway ligand, cyclic di-GMP, enhance cancer immunotherapy against metastatic melanoma.

    Science.gov (United States)

    Nakamura, Takashi; Miyabe, Hiroko; Hyodo, Mamoru; Sato, Yusuke; Hayakawa, Yoshihiro; Harashima, Hideyoshi

    2015-10-28

    Malignant melanomas escape immunosurveillance via the loss/down-regulation of MHC-I expression. Natural killer (NK) cells have the potential to function as essential effector cells for eliminating melanomas. Cyclic di-GMP (c-di-GMP), a ligand of the stimulator of interferon genes (STING) signal pathway, can be thought of as a new class of adjuvant against cancer. However, it is yet to be tested, because technologies for delivering c-di-GMP to the cytosol are required. Herein, we report that c-di-GMP efficiently activates NK cells and induces antitumor effects against malignant melanomas when loaded in YSK05 lipid containing liposomes, by assisting in the efficient delivery of c-di-GMP to the cytosol. The intravenous administration of c-di-GMP encapsulated within YSK05-liposomes (c-di-GMP/YSK05-Lip) into mice efficiently induced the production of type I interferon (IFN) as well as the activation of NK cells, resulting in a significant antitumor effect in a lung metastasis mouse model using B16-F10. This antitumor effect was dominated by NK cells. The infiltration of NK cells was observed in the lungs with B16-F10 melanomas. These findings indicate that the c-di-GMP/YSK05-Lip induces MHC-I non-restricted antitumor immunity mediated by NK cells. Consequently, c-di-GMP/YSK05-Lip represents a potentially new adjuvant system for use in immunotherapy against malignant melanomas.

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

    Science.gov (United States)

    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.

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

    OpenAIRE

    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...

  4. 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

    Science.gov (United States)

    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

  5. Systematic Identification of Cyclic-di-GMP Binding Proteins in Vibrio cholerae Reveals a Novel Class of Cyclic-di-GMP-Binding ATPases Associated with Type II Secretion Systems.

    Science.gov (United States)

    Roelofs, Kevin G; Jones, Christopher J; Helman, Sarah R; Shang, Xiaoran; Orr, Mona W; Goodson, Jonathan R; Galperin, Michael Y; Yildiz, Fitnat H; Lee, Vincent T

    2015-10-01

    Cyclic-di-GMP (c-di-GMP) is a ubiquitous bacterial signaling molecule that regulates a variety of complex processes through a diverse set of c-di-GMP receptor proteins. We have utilized a systematic approach to identify c-di-GMP receptors from the pathogen Vibrio cholerae using the Differential Radial Capillary Action of Ligand Assay (DRaCALA). The DRaCALA screen identified a majority of known c-di-GMP binding proteins in V. cholerae and revealed a novel c-di-GMP binding protein, MshE (VC0405), an ATPase associated with the mannose sensitive hemagglutinin (MSHA) type IV pilus. The known c-di-GMP binding proteins identified by DRaCALA include diguanylate cyclases, phosphodiesterases, PilZ domain proteins and transcription factors VpsT and VpsR, indicating that the DRaCALA-based screen of open reading frame libraries is a feasible approach to uncover novel receptors of small molecule ligands. Since MshE lacks the canonical c-di-GMP-binding motifs, a truncation analysis was utilized to locate the c-di-GMP binding activity to the N-terminal T2SSE_N domain. Alignment of MshE homologs revealed candidate conserved residues responsible for c-di-GMP binding. Site-directed mutagenesis of these candidate residues revealed that the Arg9 residue is required for c-di-GMP binding. The ability of c-di-GMP binding to MshE to regulate MSHA dependent processes was evaluated. The R9A allele, in contrast to the wild type MshE, was unable to complement the ΔmshE mutant for the production of extracellular MshA to the cell surface, reduction in flagella swimming motility, attachment to surfaces and formation of biofilms. Testing homologs of MshE for binding to c-di-GMP identified the type II secretion ATPase of Pseudomonas aeruginosa (PA14_29490) as a c-di-GMP receptor, indicating that type II secretion and type IV pili are both regulated by c-di-GMP.

  6. The c-di-GMP phosphodiesterase BifA regulates biofilm development in Pseudomonas putida.

    Science.gov (United States)

    Jiménez-Fernández, Alicia; López-Sánchez, Aroa; Calero, Patricia; Govantes, Fernando

    2015-02-01

    We previously showed the isolation of biofilmpersistent Pseudomonas putida mutants that fail to undergo biofilm dispersal upon entry in stationary phase. Two such mutants were found to bear insertions in PP0914, encoding a GGDEF/EAL domain protein with high similarity to Pseudomon asaeruginosa BifA. Here we show the phenotypic characterization of a ΔbifA mutant in P. putida KT2442.This mutant displayed increased biofilm and pellicle formation, cell aggregation in liquid medium and decreased starvation-induced biofilm dispersal relative to the wild type. Unlike its P. aeruginosa counterpart, P. putida BifA did not affect swarming motility. The hyperadherent phenotype of the ΔbifA mutant correlates with a general increase in cyclic diguanylate (c-di-GMP) levels, Congo Red-binding exopolyaccharide production and transcription of the adhesin-encoding lapA gene. Integrity of the EAL motif and a modified GGDEF motif (altered to GGDQF)were crucial for BifA activity, and c-di-GMP depletion by overexpression of a heterologous c-di-GMP phosphodiesterase in the ΔbifA mutant restored wild-type biofilm dispersal and lapA expression.Our results indicate that BifA is a phosphodiesterase involved in the regulation of the c-di-GMP pool and required for the generation of the low c-di-GMP signal that triggers starvation-induced biofilm dispersal.

  7. Bacterial c-di-GMP affects hematopoietic stem/progenitors and their niches through STING.

    Science.gov (United States)

    Kobayashi, Hiroshi; Kobayashi, Chiharu I; Nakamura-Ishizu, Ayako; Karigane, Daiki; Haeno, Hiroshi; Yamamoto, Kimiyo N; Sato, Taku; Ohteki, Toshiaki; Hayakawa, Yoshihiro; Barber, Glen N; Kurokawa, Mineo; Suda, Toshio; Takubo, Keiyo

    2015-04-01

    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.

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

    Directory of Open Access Journals (Sweden)

    P.G. Carvalho-Costa

    2014-12-01

    Full Text Available 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.

  9. Temperature affects c-di-GMP signalling and biofilm formation in Vibrio cholerae.

    Science.gov (United States)

    Townsley, Loni; Yildiz, Fitnat H

    2015-11-01

    Biofilm formation is crucial to the environmental survival and transmission of Vibrio cholerae, the facultative human pathogen responsible for the disease cholera. During its infectious cycle, V. cholerae experiences fluctuations in temperature within the aquatic environment and during the transition between human host and aquatic reservoirs. In this study, we report that biofilm formation is induced at low temperatures through increased levels of the signalling molecule, cyclic diguanylate (c-di-GMP). Strains harbouring in frame deletions of all V. cholerae genes that are predicted to encode diguanylate cyclases (DGCs) or phosphodiesterases (PDEs) were screened for their involvement in low-temperature-induced biofilm formation and Vibrio polysaccharide gene expression. Of the 52 mutants tested, deletions of six DGCs and three PDEs were found to affect these phenotypes at low temperatures. Unlike wild type, a strain lacking all six DGCs did not exhibit a low-temperature-dependent increase in c-di-GMP, indicating that these DGCs are required for temperature modulation of c-di-GMP levels. We also show that temperature modulates c-di-GMP levels in a similar fashion in the Gram-negative pathogen Pseudomonas aeruginosa but not in the Gram-positive pathogen Listeria monocytogenes. This study uncovers the role of temperature in environmental regulation of biofilm formation and c-di-GMP signalling.

  10. Effects of radioprotectors on the cAMP and cGMP systems

    International Nuclear Information System (INIS)

    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.)

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  12. Cyclic di-GMP sensing via the innate immune signaling protein STING.

    Science.gov (United States)

    Yin, Qian; Tian, Yuan; Kabaleeswaran, Venkataraman; Jiang, Xiaomo; Tu, Daqi; Eck, Michael J; Chen, Zhijian J; Wu, Hao

    2012-06-29

    Detection of foreign materials is the first step of successful immune responses. Stimulator of interferon genes (STING) was shown to directly bind cyclic diguanylate monophosphate (c-di-GMP), a bacterial second messenger, and to elicit strong interferon responses. Here we elucidate the structural features of the cytosolic c-di-GMP binding domain (CBD) of STING and its complex with c-di-GMP. The CBD exhibits an α + β fold and is a dimer in the crystal and in solution. Surprisingly, one c-di-GMP molecule binds to the central crevice of a STING dimer, using a series of stacking and hydrogen bonding interactions. We show that STING is autoinhibited by an intramolecular interaction between the CBD and the C-terminal tail (CTT) and that c-di-GMP releases STING from this autoinhibition by displacing the CTT. The structures provide a remarkable example of pathogen-host interactions in which a unique microbial molecule directly engages the innate immune system.

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

    Science.gov (United States)

    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.

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

    International Nuclear Information System (INIS)

    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

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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.

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

    DEFF Research Database (Denmark)

    Kruuse, Christina Rostrup; Frandsen, E; Schifter, S;

    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 determine...... whether the pain-inducing effects of sildenafil would be reflected in plasma levels of important signalling molecules in migraine: cGMP, cyclic adenosine monophosphate (cAMP) and calcitonin gene-related peptide (CGRP). Ten healthy subjects (four women, six men) and 12 patients (12 women) suffering from...... migraine without aura were included in two separate double-blind, placebo-controlled, cross-over studies in which placebo or sildenafil 100 mg was administered orally. Plasma levels of CGRP, cAMP and cGMP were determined in blood from the antecubital vein. Despite the ability of sildenafil to induce...

  18. c-di-GMP signalling and the regulation of developmental transitions in streptomycetes.

    Science.gov (United States)

    Bush, Matthew J; Tschowri, Natalia; Schlimpert, Susan; Flärdh, Klas; Buttner, Mark J

    2015-12-01

    The complex life cycle of streptomycetes involves two distinct filamentous cell forms: the growing (or vegetative) hyphae and the reproductive (or aerial) hyphae, which differentiate into long chains of spores. Until recently, little was known about the signalling pathways that regulate the developmental transitions leading to sporulation. In this Review, we discuss important new insights into these pathways that have led to the emergence of a coherent regulatory network, focusing on the erection of aerial hyphae and the synchronous cell division event that produces dozens of unigenomic spores. In particular, we highlight the role of cyclic di-GMP (c-di-GMP) in controlling the initiation of development, and the role of the master regulator BldD in mediating c-di-GMP signalling.

  19. Cyclic-di-GMP signaling in the Gram-positive pathogen Clostridium difficile.

    Science.gov (United States)

    Bordeleau, Eric; Burrus, Vincent

    2015-11-01

    The anaerobic Gram-positive bacterium Clostridium difficile causes intestinal infections responsible for symptoms ranging from mild diarrhea to fulminant colitis. Like other bacteria, C. difficile needs to sense and integrate environmental signals in order to adapt to changes and thrive in its environment. The second messenger cyclic diguanosine monophosphate (c-di-GMP) was recently recognized as a quasi-ubiquitous phenotype coordinator in bacteria. Mostly known to be involved in the transition from motile to sessile and multicellular behaviors in Gammaproteobacteria, c-di-GMP is now known to regulate many other phenotypes from cell morphogenesis to virulence, in many Gram-negative and a few Gram-positive bacteria. Herein, we review recent advances in our understanding of c-di-GMP signaling in the lifecycle of C. difficile.

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

    Science.gov (United States)

    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

  1. Membrane targeting of cGMP-dependent protein kinase is required for cystic fibrosis transmembrane conductance regulator Cl- channel activation

    NARCIS (Netherlands)

    A.B. Vaandrager (Arie); A. Smolenski; B.C. Tilly (Bernard); A.B. Houtsmuller (Adriaan); E.M.E. Ehlert (Ehrich); A.G. Bot (Alice); M.J. Edixhoven (Marcel); W.E. Boomaars (Wendy); S.M. Lohmann (Suzanne); H.R. de Jonge (Hugo)

    1998-01-01

    textabstractA recently cloned isoform of cGMP-dependent protein kinase (cGK), designated type II, was implicated as the mediator of cGMP-provoked intestinal Cl- secretion based on its localization in the apical membrane of enterocytes and on its capacity to activate cys

  2. cGMP-dependent ABA-induced stomatal closure in the ABA-insensitive Arabidopsis mutant abi1-1.

    Science.gov (United States)

    Dubovskaya, Lyudmila V; Bakakina, Yulia S; Kolesneva, Ekaterina V; Sodel, Dmitry L; McAinsh, Martin R; Hetherington, Alistair M; Volotovski, Igor D

    2011-07-01

    • The drought hormone abscisic acid (ABA) is widely known to produce reductions in stomatal aperture in guard cells. The second messenger cyclic guanosine 3', 5'-monophosphate (cGMP) is thought to form part of the signalling pathway by which ABA induces stomatal closure. • We have examined the signalling events during cGMP-dependent ABA-induced stomatal closure in wild-type Arabidopsis plants and plants of the ABA-insensitive Arabidopsis mutant abi1-1. • We show that cGMP acts downstream of hydrogen peroxide (H(2) O(2) ) and nitric oxide (NO) in the signalling pathway by which ABA induces stomatal closure. H(2) O(2) - and NO-induced increases in the cytosolic free calcium concentration ([Ca(2+) ](cyt) ) were cGMP-dependent, positioning cGMP upstream of [Ca(2+) ](cyt) , and involved the action of the type 2C protein phosphatase ABI1. Increases in cGMP were mediated through the stimulation of guanylyl cyclase by H(2) O(2) and NO. We identify nucleoside diphosphate kinase as a new cGMP target protein in Arabidopsis. • This study positions cGMP downstream of ABA-induced changes in H(2) O(2) and NO, and upstream of increases in [Ca(2+) ](cyt) in the signalling pathway leading to stomatal closure.

  3. c-di-GMP enhances protective innate immunity in a murine model of pertussis.

    Directory of Open Access Journals (Sweden)

    Shokrollah Elahi

    Full Text Available Innate immunity represents the first line of defense against invading pathogens in the respiratory tract. Innate immune cells such as monocytes, macrophages, dendritic cells, NK cells, and granulocytes contain specific pathogen-recognition molecules which induce the production of cytokines and subsequently activate the adaptive immune response. c-di-GMP is a ubiquitous second messenger that stimulates innate immunity and regulates biofilm formation, motility and virulence in a diverse range of bacterial species with potent immunomodulatory properties. In the present study, c-di-GMP was used to enhance the innate immune response against pertussis, a respiratory infection mainly caused by Bordetella pertussis. Intranasal treatment with c-di-GMP resulted in the induction of robust innate immune responses to infection with B. pertussis characterized by enhanced recruitment of neutrophils, macrophages, natural killer cells and dendritic cells. The immune responses were associated with an earlier and more vigorous expression of Th1-type cytokines, as well as an increase in the induction of nitric oxide in the lungs of treated animals, resulting in significant reduction of bacterial numbers in the lungs of infected mice. These results demonstrate that c-di-GMP is a potent innate immune stimulatory molecule that can be used to enhance protection against bacterial respiratory infections. In addition, our data suggest that priming of the innate immune system by c-di-GMP could further skew the immune response towards a Th1 type phenotype during subsequent infection. Thus, our data suggest that c-di-GMP might be useful as an adjuvant for the next generation of acellular pertussis vaccine to mount a more protective Th1 phenotype immune response, and also in other systems where a Th1 type immune response is required.

  4. 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

    Science.gov (United States)

    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

  5. 13th CIFIT to Highlight New Features

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    On July 2, the Ministry of Commerce and the China International Fair for Investment & Trade (CIFIT) Organizing Committee held a joint press confer- ence in Beijing to discuss the upcoming CIFIT, which is slated to open on September 8, 2009. The press conference was attended by Chen Jin, Executive Vice Chairman of the CIFIT

  6. 13th Techtextil Closes with Record Figures

    Institute of Scientific and Technical Information of China (English)

    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.

  7. 13th Radiochemical Conference. Booklet of Abstracts

    International Nuclear Information System (INIS)

    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.)

  8. Mining equipment manufacturers. 13th edition

    Energy Technology Data Exchange (ETDEWEB)

    1992-01-01

    The report analyses and compares the financial ratios of mining equipment manufacturers in the United Kingdom. It includes companies who produce equipment for the mining industry such as cutting tools, drilling equipment, crushers, screens, centrifuges, tunnelling equipment, conveyors and other materials handling equipment, hydraulic equipment, filtration, pumping and mixing equipment and ventilation equipment. The report presents business ratios analysing profitability, efficiency, liquidity, employee-based ratios and a series of figures on growth rate. Accounting data for the individual companies are given, together with principal ratios, areas of activity, names of directors and company secretary, trading address and holding companies.

  9. PREFACE: 13th International Conference on Electrostatics

    Science.gov (United States)

    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

  10. 48th Annual Convention of Computer Society of India

    CERN Document Server

    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.

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

    Science.gov (United States)

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

    2015-01-01

    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 globalization status of good manufacturing practice (GMP) requirements for pharmaceutical excipients. The review outcomes demonstrate that there is a lack of accurately defined methods to evaluate and measure excipients' safety. Furthermore good manufacturing practice requirements for excipients are not effectively globalized.

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

    Directory of Open Access Journals (Sweden)

    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

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

    Directory of Open Access Journals (Sweden)

    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.

  14. 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

    Institute of Scientific and Technical Information of China (English)

    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。

  15. "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

    Institute of Scientific and Technical Information of China (English)

    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.

  16. cGMP regulates hydrogen peroxide accumulation in calcium-dependent salt resistance pathway in Arabidopsis thaliana roots.

    Science.gov (United States)

    Li, Jisheng; Wang, Xiaomin; Zhang, Yanli; Jia, Honglei; Bi, Yurong

    2011-10-01

    3',5'-cyclic guanosine monophosphate (cGMP) is an important second messenger in plants. In the present study, roles of cGMP in salt resistance in Arabidopsis roots were investigated. Arabidopsis roots were sensitive to 100 mM NaCl treatment, displaying a great increase in electrolyte leakage and Na(+)/K(+) ratio and a decrease in gene expression of the plasma membrane (PM) H(+)-ATPase. However, application of exogenous 8Br-cGMP (an analog of cGMP), H(2)O(2) or CaCl(2) alleviated the NaCl-induced injury by maintaining a lower Na(+)/K(+) ratio and increasing the PM H(+)-ATPase gene expression. In addition, the inhibition of root elongation and seed germination under salt stress was removed by 8Br-cGMP. Further study indicated that 8Br-cGMP-induced higher NADPH levels for PM NADPH oxidase to generate H(2)O(2) by regulating glucose-6-phosphate dehydrogenase (G6PDH) activity. The effect of 8Br-cGMP and H(2)O(2) on ionic homeostasis was abolished when Ca(2+) was eliminated by glycol-bis-(2-amino ethyl ether)-N,N,N',N'-tetraacetic acid (EGTA, a Ca(2+) chelator) in Arabidopsis roots under salt stress. Taken together, cGMP could regulate H(2)O(2) accumulation in salt stress, and Ca(2+) was necessary in the cGMP-mediated signaling pathway. H(2)O(2), as the downstream component of cGMP signaling pathway, stimulated PM H(+)-ATPase gene expression. Thus, ion homeostasis was modulated for salt tolerance.

  17. Bi-modal distribution of the second messenger c-di-GMP controls cell fate and asymmetry during the caulobacter cell cycle.

    Directory of Open Access Journals (Sweden)

    Sören Abel

    Full Text Available Many bacteria mediate important life-style decisions by varying levels of the second messenger c-di-GMP. Behavioral transitions result from the coordination of complex cellular processes such as motility, surface adherence or the production of virulence factors and toxins. While the regulatory mechanisms responsible for these processes have been elucidated in some cases, the global pleiotropic effects of c-di-GMP are poorly understood, primarily because c-di-GMP networks are inherently complex in most bacteria. Moreover, the quantitative relationships between cellular c-di-GMP levels and c-di-GMP dependent phenotypes are largely unknown. Here, we dissect the c-di-GMP network of Caulobacter crescentus to establish a global and quantitative view of c-di-GMP dependent processes in this organism. A genetic approach that gradually reduced the number of diguanylate cyclases identified novel c-di-GMP dependent cellular processes and unraveled c-di-GMP as an essential component of C. crescentus cell polarity and its bimodal life cycle. By varying cellular c-di-GMP concentrations, we determined dose response curves for individual c-di-GMP-dependent processes. Relating these values to c-di-GMP levels modeled for single cells progressing through the cell cycle sets a quantitative frame for the successive activation of c-di-GMP dependent processes during the C. crescentus life cycle. By reconstructing a simplified c-di-GMP network in a strain devoid of c-di-GMP we defined the minimal requirements for the oscillation of c-di-GMP levels during the C. crescentus cell cycle. Finally, we show that although all c-di-GMP dependent cellular processes were qualitatively restored by artificially adjusting c-di-GMP levels with a heterologous diguanylate cyclase, much higher levels of the second messenger are required under these conditions as compared to the contribution of homologous c-di-GMP metabolizing enzymes. These experiments suggest that a common c-di-GMP pool

  18. 经济治理现代化与“十三五”税制改革%The Mordernization of Economic Governance and the Tax System Reform During the 13th Five-Year Plan

    Institute of Scientific and Technical Information of China (English)

    王军; 汪昊; 郝胜龙

    2016-01-01

    From the perspective of modern economic governance, combined tax system structure with the specific taxes, this paper reexamines the problems of China’s current tax system from regulating income distribution gap, adjusting the economic structure, strengthening macro-control and stimulating the local economy. Based on the experiences of developed countries, this paper puts forward that China should promote an integrated national tax system which consists of optimizing tax structure, enforcing value-added tax sharing, and introducing personal income tax and real estate tax during the 13th Five-Year Plan.%从经济治理现代化的视角,从税制结构和分税制两个方面,可结合各个具体税种,重新审视我国现行税收制度在调节收入分配差距、调整经济结构、加强宏观调控和发展地方经济等方面存在的问题,在借鉴发达国家相关经验的基础上,从税制结构优化、增值税分享及个人所得税、房地产税等税种改革等方面,对我国“十三五”时期税制改革提出相关政策建议。

  19. 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.

    Science.gov (United States)

    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

  20. 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.

    Science.gov (United States)

    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.

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

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

    CERN Document Server

    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.

  3. Cyclic GMP signaling in cardiomyocytes modulates fatty acid trafficking and prevents triglyceride accumulation

    Science.gov (United States)

    While the balance between carbohydrates and fatty acids for energy production appears to be crucial for cardiac homeostasis, much remains to be learned about the molecular mechanisms underlying this relationship. Given the reported benefits of cGMP signaling on the myocardium, we investigated the im...

  4. Identification and characterization of DdPDE3, a cGMP-selective phosphodiesterase from Dictyostelium

    NARCIS (Netherlands)

    Kuwayama, H; Snippe, H; Derks, M; Roelofs, J; van Haastert, PJM

    2001-01-01

    In Dictyostelium cAMP and cGMP have important functions as first and second messengers in chemotaxis and development. Two cyclic-nucleotide phosphodiesterases (DdPDE 1 and 2) have been identified previously, an extracellular dual-specificity enzyme and an intracellular cAMP-specific enzyme (encoded

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

    DEFF Research Database (Denmark)

    Kruuse, C; Rybalkin, S D; Khurana, T S;

    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 (c...... 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.......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 (c......GMP) pathway. Immunoreactivity to phosphodiesterases 1A, 1B and 5, but not phosphodiesterase 1C was found in fractions of homogenised cerebral arteries eluted by high-pressure liquid chromatography (HPLC). Both the phosphodiesterase 1 inhibitor 8-methoxymethyl-1-methyl-3-(2methylpropyl)-xanthine (8-MM...

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

    OpenAIRE

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  8. Effect of simple shear on the physical properties of glutenin macro polymer (GMP)

    NARCIS (Netherlands)

    Peighambardoust, S.H.; Goot, van der A.J.; Hamer, R.J.; Boom, R.M.

    2005-01-01

    Gluten plays a key role in determining the end-use quality of wheat flour. The important role of the highly aggregated fraction of gluten, glutenin macropolymer (GMP), in dough properties opens up new possibilities for revealing underlying mechanisms related to dough processing. Using a new shear ce

  9. 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.

    Science.gov (United States)

    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

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    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

  12. Dimeric c-di-GMP is required for post-translational regulation of alginate production in Pseudomonas aeruginosa.

    Science.gov (United States)

    Whitney, John C; Whitfield, Gregory B; Marmont, Lindsey S; Yip, Patrick; Neculai, A Mirela; Lobsanov, Yuri D; Robinson, Howard; Ohman, Dennis E; Howell, P Lynne

    2015-05-15

    Pseudomonas aeruginosa is an opportunistic human pathogen that secretes the exopolysaccharide alginate during infection of the respiratory tract of individuals afflicted with cystic fibrosis and chronic obstructive pulmonary disease. Among the proteins required for alginate production, Alg44 has been identified as an inner membrane protein whose bis-(3',5')-cyclic dimeric guanosine monophosphate (c-di-GMP) binding activity post-translationally regulates alginate secretion. In this study, we report the 1.8 Å crystal structure of the cytoplasmic region of Alg44 in complex with dimeric self-intercalated c-di-GMP and characterize its dinucleotide-binding site using mutational analysis. The structure shows that the c-di-GMP binding region of Alg44 adopts a PilZ domain fold with a dimerization mode not previously observed for this family of proteins. Calorimetric binding analysis of residues in the c-di-GMP binding site demonstrate that mutation of Arg-17 and Arg-95 alters the binding stoichiometry between c-di-GMP and Alg44 from 2:1 to 1:1. Introduction of these mutant alleles on the P. aeruginosa chromosome show that the residues required for binding of dimeric c-di-GMP in vitro are also required for efficient alginate production in vivo. These results suggest that the dimeric form of c-di-GMP represents the biologically active signaling molecule needed for the secretion of an important virulence factor produced by P. aeruginosa.

  13. 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.

    CERN Multimedia

    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.

  14. A direct screen for c-di-GMP modulators reveals a Salmonella Typhimurium periplasmic ʟ-arginine-sensing pathway.

    Science.gov (United States)

    Mills, Erez; Petersen, Erik; Kulasekara, Bridget R; Miller, Samuel I

    2015-06-01

    Cyclic-di-GMP (c-di-GMP) is a bacterial second messenger that transduces internal and external signals and regulates bacterial motility and biofilm formation. Some organisms encode more than 100 c-di-GMP-modulating enzymes, but only for a few has a signal been defined that modulates their activity. We developed and applied a high-throughput, real-time flow cytometry method that uses a fluorescence resonance energy transfer (FRET)-based biosensor of free c-di-GMP to screen for signals that modulate its concentration within Salmonella Typhimurium. We identified multiple compounds, including glucose, N-acetyl-d-glucosamine, salicylic acid, and ʟ-arginine, that modulated the FRET signal and therefore the free c-di-GMP concentration. By screening a library of mutants, we identified proteins required for the c-di-GMP response to each compound. Furthermore, low micromolar concentrations of ʟ-arginine induced a rapid translation-independent increase in c-di-GMP concentrations and c-di-GMP-dependent cellulose synthesis, responses that required the regulatory periplasmic domain of the diguanylate cyclase STM1987. ʟ-Arginine signaling also required the periplasmic putative ʟ-arginine-binding protein ArtI, implying that ʟ-arginine sensing occurred in the periplasm. Among the 20 commonly used amino acids, S. Typhimurium specifically responded to ʟ-arginine with an increase in c-di-GMP, suggesting that ʟ-arginine may serve as a signal during S. Typhimurium infection. Our results demonstrate that a second-messenger biosensor can be used to identify environmental signals and define pathways that alter microbial behavior.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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 (PGMP wet weight during dough processing were significant (PGMP 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.

  17. Structure of STING bound to cyclic di-GMP reveals the mechanism of cyclic dinucleotide recognition by the immune system.

    Science.gov (United States)

    Shu, Chang; Yi, Guanghui; Watts, Tylan; Kao, C Cheng; Li, Pingwei

    2012-06-24

    STING (stimulator of interferon genes) is an innate immune sensor of cyclic dinucleotides that regulates the induction of type I interferons. STING's C-terminal domain forms a V-shaped dimer and binds a cyclic diguanylate monophosphate (c-di-GMP) at the dimer interface by both direct and solvent-mediated hydrogen bonds. Guanines of c-di-GMP stack against the phenolic rings of a conserved tyrosine, and mutations at the c-di-GMP binding surface reduce nucleotide binding and affect signaling.

  18. The Path for the Development of Geological and Mineral Resources in the Guangxi Zhuang Autonomous Regionduring“the 13th Five Year Plan”%广西地矿“十三五”规划路径

    Institute of Scientific and Technical Information of China (English)

    唐善茂; 白旭宏

    2016-01-01

    During“The Twelfth FiveYear Plan”, while some achievements in geological and mineral development in Guangxi had been made, various problems had been existed. Faced with the new developing environment and target tasks, a new path for the geological development of Guangxi during “the 13th Five Year Plan” must be opened up. First, doing a good job of “addition”, it means that we should strive to raise the capacity of performing our duties. Second, “subtracting”, it means that we should remain committed to doing something but not. Third, “multiplication”, it means that we should give full play to comparative superiority. Fourth, “doing division”, it means that we should enhance our efforts to the innovation of system and mechanism.%“十二五”广西地矿工作呈现出“2个提升2个转变”主要亮点,存在“4多4少”不足。面对新的发展环境和目标任务,广西地矿“十三五”发展应开辟“加减乘除”新路径,一是做好“加法”,着力提升履职能力;二是做“减法”,坚持有所为有所不为;三是做“乘法”,发挥比较优势;四是做“除法”,加强体制机制创新。

  19. Research on Power Source Planning of Guangdong Province in the 13th Five-year Plan Period%广东"十三五"电源规划研究

    Institute of Scientific and Technical Information of China (English)

    卢洵; 林勇; 程鑫; 吴伟杰

    2016-01-01

    "十三五"是广东电力转变发展方式、优化电源结构与布局的重要机遇期,在面临经济发展方式转型、可再生能源消纳规模不断扩大、生态环境约束逐步增强的新发展时期,科学合理的电源规划工作十分重要.基于此,提出了广东电源优选方案及电源实际进度方案,研究了电源规划优化方法,并对两类规划方案技术经济进行对比.研究表明,电源优选方案将会给电网建设、调度运行、全社会资源配置等方面带来显著的社会经济效益.%The 13th five-year plan period is the important opportunity time for Guangdong electric power transferring devel-opment mode,optimizing power source structure and layout. In the new development period of transformation of economic development mode,gradual expansion of absorptive scale of renewable energy source and progressive enhancement of eco-logical environment constraint,it is very important to scientifically and reasonably implement power source planning. Therefore,this paper proposes optimization scheme for Guangdong power source and actual schedule planning,studies opti-mization methods for power source planning and compares two kinds of planning schemes in technique and economy aspects. Research indicates that the optimization scheme for power source may produce obvious social and economic benefits for pow-er grid construction,dispatching operation,social resources configuration,and so on.

  20. “十三五”时期城商行的创新发展战略%The City Commercial Bank Develop in an Innovative Way during 13th Five-Year

    Institute of Scientific and Technical Information of China (English)

    陆岷峰; 徐阳洋

    2016-01-01

    The city firm in 13th Five-Year during the planning period is facing important opportunities for development, but also by many adverse shocks. The city firm should be under the guidance of innovative thinking, and strive to develop wealth management business, asset management business, investment banking business and the establishment of these three business models to expand their own profit space. Also city commercial banks should strengthen their own management system innovation and risk mechanism innovation, improve the management system, reduce the risk, so that in the future the compli-cated and changeable market environment are better able to the sustained and healthy development.%城商行在“十三五”时期既面临着重要的发展机遇,也受到很多不利因素的冲击,城商行应该在创新思维引领下努力发展财富管理业务、资产管理业务和投资银行业务,并且建立这三者业务为一体的业务模式,扩大自己的盈利空间,同时强化自身的管理体制创新以及风险机制创新,从而在未来复杂多变的市场环境中更好地持续健康发展。

  1. EDITORIAL: `Bridging Gravitational Wave Astronomy and Observational Astrophysics', Proceedings of the 13th Gravitational Wave Data Analysis Workshop (GWDAW13) (San Juan, Puerto Rico, 19-22 January 2009), sponsored by the Center for Gravitational Wave Astronomy, The University of Texas at Brownsville and The National Astronomy and Ionosphere Center `Bridging Gravitational Wave Astronomy and Observational Astrophysics', Proceedings of the 13th Gravitational Wave Data Analysis Workshop (GWDAW13) (San Juan, Puerto Rico, 19-22 January 2009), sponsored by the Center for Gravitational Wave Astronomy, The University of Texas at Brownsville and The National Astronomy and Ionosphere Center

    Science.gov (United States)

    Díaz, Mario; Jenet, Fredrick; Mohanty, Soumya

    2009-10-01

    The 13th Gravitational Wave Data Analysis Workshop took place in San Juan, Puerto Rico on the 19-22 January 2009. This annual event has become the established venue for presenting and discussing new results and techniques in this crucial subfield of gravitational wave astronomy. A major attraction of the event is that scientists working with all possible instruments gather to discuss their projects and report on the status of their observations. The Center for Gravitational Wave Astronomy at the University of Texas at Brownsville, USA (a National Aeronautics and Space Administration University Research Center and a National Science Foundation Center for Research Excellence in Science and Technology) jointly with the National Astronomy and Ionosphere Center (which operates the Arecibo Observatory) were the proud sponsors of the gathering this time. As in previous years, GWDAW13 was well attended by more than 100 participants from over 10 countries worldwide As this issue is going to press GEO, LIGO and VIRGO are undergoing new scientific runs of their instruments with the LIGO detectors holding the promise of increasing their operational sensitivity twofold as compared with the observations finished a couple of years ago. This new cycle of observations is a major milestone compared to the previous observations which have been accomplished. Gravitational waves have not been observed yet, but the instrumental sensitivity achieved has started producing relevant astrophysical results. In particular, very recently (Nature, 20 August 2009) a letter from the LIGO Scientific Collaboration http://www.ligo.org and the VIRGO Collaboration http://www.virgo.infn.it has set the most stringent limits yet on the amount of gravitational waves that could have come from the Big Bang in the gravitational wave frequency band where current gravitational wave detectors can observe. These results have put new constraints on the physical characteristics of the early universe. The proximity

  2. cGMP-dependent protein kinase contributes to hydrogen sulfide-stimulated vasorelaxation.

    Directory of Open Access Journals (Sweden)

    Mariarosaria Bucci

    Full Text Available A growing body of evidence suggests that hydrogen sulfide (H₂S is a signaling molecule in mammalian cells. In the cardiovascular system, H₂S enhances vasodilation and angiogenesis. H₂S-induced vasodilation is hypothesized to occur through ATP-sensitive potassium channels (K(ATP; however, we recently demonstrated that it also increases cGMP levels in tissues. Herein, we studied the involvement of cGMP-dependent protein kinase-I in H₂S-induced vasorelaxation. The effect of H₂S on vessel tone was studied in phenylephrine-contracted aortic rings with or without endothelium. cGMP levels were determined in cultured cells or isolated vessel by enzyme immunoassay. Pretreatment of aortic rings with sildenafil attenuated NaHS-induced relaxation, confirming previous findings that H₂S is a phosphodiesterase inhibitor. In addition, vascular tissue levels of cGMP in cystathionine gamma lyase knockouts were lower than those in wild-type control mice. Treatment of aortic rings with NaHS, a fast releasing H₂S donor, enhanced phosphorylation of vasodilator-stimulated phosphoprotein in a time-dependent manner, suggesting that cGMP-dependent protein kinase (PKG is activated after exposure to H₂S. Incubation of aortic rings with a PKG-I inhibitor (DT-2 attenuated NaHS-stimulated relaxation. Interestingly, vasodilatory responses to a slowly releasing H₂S donor (GYY 4137 were unaffected by DT-2, suggesting that this donor dilates mouse aorta through PKG-independent pathways. Dilatory responses to NaHS and L-cysteine (a substrate for H₂S production were reduced in vessels of PKG-I knockout mice (PKG-I⁻/⁻. Moreover, glibenclamide inhibited NaHS-induced vasorelaxation in vessels from wild-type animals, but not PKG-I⁻/⁻, suggesting that there is a cross-talk between K(ATP and PKG. Our results confirm the role of cGMP in the vascular responses to NaHS and demonstrate that genetic deletion of PKG-I attenuates NaHS and L

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

    DEFF Research Database (Denmark)

    Bouzinova, Elena V.; Larsen, Per; Matchkov, Vladimir;

    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...

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

    Science.gov (United States)

    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.

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

    NARCIS (Netherlands)

    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

  6. Elevated level of the second messenger c-di-GMP in Comamonas testosteroni enhances biofilm formation and biofilm-based biodegradation of 3-chloroaniline.

    Science.gov (United States)

    Wu, Yichao; Ding, Yuanzhao; Cohen, Yehuda; Cao, Bin

    2015-02-01

    The bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP) is a ubiquitous second messenger that determines bacterial lifestyle between the planktonic and biofilm modes of life. Although the role of c-di-GMP signaling in biofilm development and dispersal has been extensively studied, how c-di-GMP signaling influences environmental bioprocess activities such as biodegradation remains unexplored. To elucidate the impacts of elevating c-di-GMP level on environmental bioprocesses, we constructed a Comamonas testosteroni strain constitutively expressing a c-di-GMP synthase YedQ from Escherichia coli and examined its capability in biofilm formation and biodegradation of 3-chloroaniline (3-CA). The high c-di-GMP strain exhibited an increased binding to Congo red dye, a decreased motility, and an enhanced biofilm formation capability. In planktonic cultures, the strain with an elevated c-di-GMP concentration and the wild type could degrade 3-CA comparably well. However, under batch growth conditions with a high surface to volume ratio, an elevated c-di-GMP concentration in C. testosteroni significantly increased the contribution of biofilms in 3-CA biodegradation. In continuous submerged biofilm reactors, C. testosteroni with an elevated c-di-GMP level exhibited an enhanced 3-CA biodegradation and a decreased cell detachment rate. Taken together, this study provides a novel strategy to enhance biofilm-based biodegradation of toxic xenobiotic compounds through manipulating bacterial c-di-GMP signaling.

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

    OpenAIRE

    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 ...

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

    Directory of Open Access Journals (Sweden)

    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.

  9. Mechanistic insights into c-di-GMP-dependent control of the biofilm regulator FleQ from Pseudomonas aeruginosa.

    Science.gov (United States)

    Matsuyama, Bruno Y; Krasteva, Petya V; Baraquet, Claudine; Harwood, Caroline S; Sondermann, Holger; Navarro, Marcos V A S

    2016-01-12

    Bacterial biofilm formation during chronic infections confers increased fitness, antibiotic tolerance, and cytotoxicity. In many pathogens, the transition from a planktonic lifestyle to collaborative, sessile biofilms represents a regulated process orchestrated by the intracellular second-messenger c-di-GMP. A main effector for c-di-GMP signaling in the opportunistic pathogen Pseudomonas aeruginosa is the transcription regulator FleQ. FleQ is a bacterial enhancer-binding protein (bEBP) with a central AAA+ ATPase σ(54)-interaction domain, flanked by a C-terminal helix-turn-helix DNA-binding motif and a divergent N-terminal receiver domain. Together with a second ATPase, FleN, FleQ regulates the expression of flagellar and exopolysaccharide biosynthesis genes in response to cellular c-di-GMP. Here we report structural and functional data that reveal an unexpected mode of c-di-GMP recognition that is associated with major conformational rearrangements in FleQ. Crystal structures of FleQ's AAA+ ATPase domain in its apo-state or bound to ADP or ATP-γ-S show conformations reminiscent of the activated ring-shaped assemblies of other bEBPs. As revealed by the structure of c-di-GMP-complexed FleQ, the second messenger interacts with the AAA+ ATPase domain at a site distinct from the ATP binding pocket. c-di-GMP interaction leads to active site obstruction, hexameric ring destabilization, and discrete quaternary structure transitions. Solution and cell-based studies confirm coupling of the ATPase active site and c-di-GMP binding, as well as the functional significance of crystallographic interprotomer interfaces. Taken together, our data offer unprecedented insight into conserved regulatory mechanisms of gene expression under direct c-di-GMP control via FleQ and FleQ-like bEBPs.

  10. Enhanced Molecular Recognition between Nucleobases and Guanine-5'-monophosphate-disodium (GMP) by Surfactant Aggregates in Aqueous Solution.

    Science.gov (United States)

    Liu, Zhang; Wang, Dong; Cao, Meiwen; Han, Yuchun; Xu, Hai; Wang, Yilin

    2015-07-15

    Only specific base pairs on DNA can bind with each other through hydrogen bonds, which is called the Watson-Crick (W/C) pairing rule. However, without the constraint of DNA chains, the nucleobases in bulk aqueous solution usually do not follow the W/C pairing rule anymore because of the strong competitive effect of water and the multi-interaction edges of nucleobases. The present work applied surfactant aggregates noncovalently functionalized by nucleotide to enhance the recognition between nucleobases without DNA chains in aqueous solution, and it revealed the effects of their self-assembling ability and morphologies on the recognition. The cationic ammonium monomeric, dimeric, and trimeric surfactants DTAB, 12-3-12, and 12-3-12-3-12 were chosen. The surfactants with guanine-5'-monophosphate-disodium (GMP) form micelles, vesicles, and fingerprint-like and plate-like aggregates bearing the hydrogen-bonding sites of GMP, respectively. The binding parameters of these aggregates with adenine (A), uracil (U), guanine (G), and cytosine(C) indicate that the surfactants can promote W/C recognitions in aqueous solution when they form vesicles (GMP/DTAB) or plate-like aggregates (GMP/12-3-12) with proper molecular packing compactness, which not only provide hydrophobic environments but also shield non-W/C recognition edges. However, the GMP/12-3-12 micelles with loose molecular packing, the GMP/12-3-12 fingerprint-like aggregates where the hydrogen bond sites of GMP are occupied by itself, and the GMP/12-3-12-3-12 vesicles with too strong self-assembling ability cannot promote W/C recognition. This work provides insight into how to design self-assemblies with the performance of enhanced molecule recognition.

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

    Science.gov (United States)

    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.

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

    OpenAIRE

    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...

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

    OpenAIRE

    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...

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

    OpenAIRE

    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...

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

    OpenAIRE

    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...

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

    Science.gov (United States)

    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

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

    OpenAIRE

    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...

  18. Glutamine inhibits ammonia-induced accumulation of cGMP in rat striatum limiting arginine supply for NO synthesis.

    Science.gov (United States)

    Hilgier, Wojciech; Freśko, Inez; Klemenska, Emilia; Beresewicz, Andrzej; Oja, Simo S; Saransaari, Pirjo; Albrecht, Jan; Zielińska, Magdalena

    2009-07-01

    Brain L-glutamine (Gln) accumulation and increased activity of the NO/cGMP pathway are immediate consequences of acute exposure to ammonia. This study tested whether excess Gln may influence NO and/or cGMP synthesis. Intrastriatal administration of the glutaminase inhibitor 6-diazo-5-oxo-L-norleucine or the system A-specific Gln uptake inhibitor methylaminoisobutyrate increased microdialysate Gln concentration and reduced basal and ammonia-induced NO and cGMP accumulation. Gln applied in vivo (via microdialysis) or in vitro (to rat brain cortical slices) reduced NO and cGMP accumulation in the presence and/or absence of ammonia, but not cGMP synthesis induced by the NO donor sodium nitroprusside. Attenuation of cGMP synthesis by Gln was prevented by administration of L-arginine (Arg). The L-arginine co-substrates of y(+)LAT2 transport system, L-leucine and cyclo-leucine, mimicked the effect of exogenous Gln, suggesting that Gln limits Arg supply for NO synthesis by interfering with y+LAT2-mediated Arg uptake across the cell membrane.

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

    DEFF Research Database (Denmark)

    Chua, Song Lin; Sivakumar, Krishnakumar; Rybtke, Morten Levin;

    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....

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

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

    2015-01-01

    Stress response plays an important role on microbial adaptation under hostile environmental conditions. It is generally unclear how the signaling transduction pathway mediates a stress response in planktonic and biofilm modes of microbial communities simultaneously. Here, we showed that metalloid 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.

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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 (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.

  4. 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

    Institute of Scientific and Technical Information of China (English)

    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。

  5. The Basic Thoughts of Targeted Poverty Alleviation during the Period of 13th Five-Year%“十三五”时期精准扶贫的基本思路

    Institute of Scientific and Technical Information of China (English)

    马燕坤; 肖金成

    2016-01-01

    “十三五”将是我国扶贫减贫,消除贫困的攻坚克难决战期。“精准扶贫”是中共中央总书记、国家主席习近平同志提出的扶贫要求。精准扶贫与粗放扶贫、“一刀切”扶贫、“漫灌”式扶贫、盲目扶贫等相对而言。精准扶贫的基本出发点是“一方水土养一方人”,但是一方水土是否能够养得了其承载的一方人口,关键要看这一方水土的经济开发潜力,而不是看已经成为现实的经济开发强度。精准扶贫的基本思路一是从以县为单元扶贫变为扶贫到户;二是从开发式扶贫转为城镇化扶贫。精准扶贫的路径有转移就业扶贫、城市建设扶贫、生态搬迁扶贫、生态工程扶贫、教育观念扶贫、保障救助扶贫等。%It will be the tackle tough period of reducing poverty reduction and eliminating poverty during the period of 13th Five-Year in China. The targeted poverty alleviation is the requirement that the General Secretary of the Communist Party of China and the Chinese President Xi Jinping put forward. The targeted poverty alleviation is mentioned relative to extensive poverty alleviation, "one-size-fits-all" poverty alleviation, "flooding" poverty alleviation and blind poverty alleviation. The basic starting point of the targeted poverty alleviation is that different water and soil raised different people. But, the key issue that whether or not one side water and soil can support the population It bears is its economic developing potentiality rather than its existing economic developing strength. The basic thoughts of the targeted poverty alleviation are from taking counties as a unit for poverty alleviation to taking households as a unit for poverty alleviation and from development-oriented poverty alleviation to urbanization-oriented poverty alleviation. The paths of the Targeted poverty alleviation mainly include transferring employment, city construction, ecological relocation

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

    KAUST Repository

    Turek, Ilona

    2016-03-05

    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. © 2016 Springer Science+Business Media Dordrecht

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

    OpenAIRE

    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...

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

    KAUST Repository

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

    International Nuclear Information System (INIS)

    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.

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

    Science.gov (United States)

    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

  11. 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?

    Directory of Open Access Journals (Sweden)

    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

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

    Directory of Open Access Journals (Sweden)

    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.

  13. Cyclic di-GMP-mediated repression of swarming motility by Pseudomonas aeruginosa PA14 requires the MotAB stator.

    Science.gov (United States)

    Kuchma, S L; Delalez, N J; Filkins, L M; Snavely, E A; Armitage, J P; O'Toole, G A

    2015-02-01

    The second messenger cyclic diguanylate (c-di-GMP) plays a critical role in the regulation of motility. In Pseudomonas aeruginosa PA14, c-di-GMP inversely controls biofilm formation and surface swarming motility, with high levels of this dinucleotide signal stimulating biofilm formation and repressing swarming. P. aeruginosa encodes two stator complexes, MotAB and MotCD, that participate in the function of its single polar flagellum. Here we show that the repression of swarming motility requires a functional MotAB stator complex. Mutating the motAB genes restores swarming motility to a strain with artificially elevated levels of c-di-GMP as well as stimulates swarming in the wild-type strain, while overexpression of MotA from a plasmid represses swarming motility. Using point mutations in MotA and the FliG rotor protein of the motor supports the conclusion that MotA-FliG interactions are critical for c-di-GMP-mediated swarming inhibition. Finally, we show that high c-di-GMP levels affect the localization of a green fluorescent protein (GFP)-MotD fusion, indicating a mechanism whereby this second messenger has an impact on MotCD function. We propose that when c-di-GMP level is high, the MotAB stator can displace MotCD from the motor, thereby affecting motor function. Our data suggest a newly identified means of c-di-GMP-mediated control of surface motility, perhaps conserved among Pseudomonas, Xanthomonas, and other organisms that encode two stator systems.

  14. Active Site Metal Occupancy and Cyclic Di-GMP Phosphodiesterase Activity of Thermotoga maritima HD-GYP.

    Science.gov (United States)

    Miner, Kyle D; Kurtz, Donald M

    2016-02-16

    HD-GYPs make up a subclass of the metal-dependent HD phosphohydrolase superfamily and catalyze conversion of cyclic di(3',5')-guanosine monophosphate (c-di-GMP) to 5'-phosphoguanylyl-(3'→5')-guanosine (pGpG) and GMP. Until now, the only reported crystal structure of an HD-GYP that also exhibits c-di-GMP phosphodiesterase activity contains a His/carboxylate ligated triiron active site. However, other structural and phylogenetic correlations indicate that some HD-GYPs contain dimetal active sites. Here we provide evidence that an HD-GYP c-di-GMP phosphodiesterase, TM0186, from Thermotoga maritima can accommodate both di- and trimetal active sites. We show that an as-isolated iron-containing TM0186 has an oxo/carboxylato-bridged diferric site, and that the reduced (diferrous) form is necessary and sufficient to catalyze conversion of c-di-GMP to pGpG, but that conversion of pGpG to GMP requires more than two metals per active site. Similar c-di-GMP phosphodiesterase activities were obtained with divalent iron or manganese. On the basis of activity correlations with several putative metal ligand residue variants and molecular dynamics simulations, we propose that TM0186 can accommodate both di- and trimetal active sites. Our results also suggest that a Glu residue conserved in a subset of HD-GYPs is required for formation of the trimetal site and can also serve as a labile ligand to the dimetal site. Given the anaerobic growth requirement of T. maritima, we suggest that this HD-GYP can function in vivo with either divalent iron or manganese occupying di- and trimetal sites.

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

    Directory of Open Access Journals (Sweden)

    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. Output targets and transcriptional regulation by a cyclic dimeric GMP-responsive circuit in the Vibrio parahaemolyticus Scr network.

    Science.gov (United States)

    Ferreira, Rosana B R; Chodur, Daniel M; Antunes, Luis Caetano M; Trimble, Michael J; McCarter, Linda L

    2012-03-01

    The Vibrio parahaemolyticus Scr system modulates decisions pertinent to surface colonization by affecting the cellular level of cyclic dimeric GMP (c-di-GMP). In this work, we explore the scope and mechanism of this regulation. Transcriptome comparison of ΔscrABC and wild-type strains revealed expression differences with respect to ∼100 genes. Elevated c-di-GMP repressed genes in the surface-sensing regulon, including those encoding the lateral flagellar and type III secretion systems and N-acetylglucosamine-binding protein GpbA while inducing genes encoding other cell surface molecules and capsular polysaccharide. The transcription of a few regulatory genes was also affected, and the role of one was characterized. Mutations in cpsQ suppressed the sticky phenotype of scr mutants. cpsQ encodes one of four V. parahaemolyticus homologs in the CsgD/VpsT family, members of which have been implicated in c-di-GMP signaling. Here, we demonstrate that CpsQ is a c-di-GMP-binding protein. By using a combination of mutant and reporter analyses, CpsQ was found to be the direct, positive regulator of cpsA transcription. This c-di-GMP-responsive regulatory circuit could be reconstituted in Escherichia coli, where a low level of this nucleotide diminished the stability of CpsQ. The molecular interplay of additional known cps regulators was defined by establishing that CpsS, another CsgD family member, repressed cpsR, and the transcription factor CpsR activated cpsQ. Thus, we are developing a connectivity map of the Scr decision-making network with respect to its wiring and output strategies for colonizing surfaces and interaction with hosts; in doing so, we have isolated and reproduced a c-di-GMP-sensitive regulatory module in the circuit.

  17. Redefining the Value of the Costumes Found in Tombs of Huangsheng of the Mid-13th Century%“福州南宋黄异墓"服饰的价值重释

    Institute of Scientific and Technical Information of China (English)

    和立勇

    2012-01-01

    From the standpoint of cultural diversity in our present society, this paper intends to redefine the multi - dimensional values of the costumes found in Tombs of Huangsheng of the m/d-13th Century in the aspects of literature study, survey of the resources of folk customs in Fujian and Taiwan, the protection and inheritance of intangible cultural heritage of clothing technology, and the construction of local knowledge. First, its value of archaeology and literature has not only enriched the proof for the research on Chinese costume studies in Southern Song Dynasty, but also filled the vacancy of archaeological literature on Fujian traditional costumes. This literature has now among the most frequently quoted in the field of clothing history study. Second, the costumes found in Tombs of Huangsheng of the mid-13~ Century show u great technology in embroidery, printing, dyeing and so on. These achievements not only reflect Fujian's advanced textile industry and the fine technology in Song Dynasty, but are also good examples for later researchers to study Fujian's local aesthetic interest in Southern Song Dynasty. Third, it also provides some valuable references for both the construction of local cultural knowledge and the promotion of local characteristics of culture.%站在当下构建文化多样性的立场,从文献索引、闽台历史民俗资源调查、服饰技艺类非物质文化遗产保护与传承以及“地方性知识”建构4个维度,对“福州南宋黄弄墓”服饰进行多维价值重释。从考古价值和文献价值上,黄弄墓服饰既丰富了中国服饰史研究中关于南宋部分的实证,又填补了闽文化传统服饰的考古文献空缺,也成为服饰史研究中最常被引用的文献;黄异墓服饰在刺绣、印染等纺织技术上的成就。既能客观反映宋代闽地纺织工业及技艺的高超,又为后人提供了研究南宋时期闽文化地方性审美意趣的真实范例;同时为

  18. 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).

    Science.gov (United States)

    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

  19. 中国GMP与欧盟GMP-ICH Q7的比较%Comparison of Chinese GMP and EU ICH Q7

    Institute of Scientific and Technical Information of China (English)

    汪敏

    2008-01-01

    了解中国GMP与国际GMP之间的差异,为法规制定和药品生产企业提供参考.从立法、指南、检查几个方面对欧盟原料药GMP-ICH Q7进行了简要介绍,并与中国GMP展开深入的对比,主要包括GMP起点、人员、厂房和设施、物料管理、验证、文件、生产管理和质量管理.中国GMP在许多重要方面与国际先进水平存在明显差距.我们共同期待我国新版GMP规范的出台.

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

    Directory of Open Access Journals (Sweden)

    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. Cyclic di-GMP mediates a histidine kinase/phosphatase switch by noncovalent domain cross-linking.

    Science.gov (United States)

    Dubey, Badri N; Lori, Christian; Ozaki, Shogo; Fucile, Geoffrey; Plaza-Menacho, Ivan; Jenal, Urs; Schirmer, Tilman

    2016-09-01

    Histidine kinases are key components of regulatory networks in bacteria. Although many of these enzymes are bifunctional, mediating both phosphorylation and dephosphorylation of downstream targets, the molecular details of this central regulatory switch are unclear. We showed recently that the universal second messenger cyclic di-guanosine monophosphate (c-di-GMP) drives Caulobacter crescentus cell cycle progression by forcing the cell cycle kinase CckA from its default kinase into phosphatase mode. We use a combination of structure determination, modeling, and functional analysis to demonstrate that c-di-GMP reciprocally regulates the two antagonistic CckA activities through noncovalent cross-linking of the catalytic domain with the dimerization histidine phosphotransfer (DHp) domain. We demonstrate that both c-di-GMP and ADP (adenosine diphosphate) promote phosphatase activity and propose that c-di-GMP stabilizes the ADP-bound quaternary structure, which allows the receiver domain to access the dimeric DHp stem for dephosphorylation. In silico analyses predict that c-di-GMP control is widespread among bacterial histidine kinases, arguing that it can replace or modulate canonical transmembrane signaling.

  2. Capture compound mass spectrometry--a powerful tool to identify novel c-di-GMP effector proteins.

    Science.gov (United States)

    Laventie, Benoît-Joseph; Nesper, Jutta; Ahrné, Erik; Glatter, Timo; Schmidt, Alexander; Jenal, Urs

    2015-01-01

    Considerable progress has been made during the last decade towards the identification and characterization of enzymes involved in the synthesis (diguanylate cyclases) and degradation (phosphodiesterases) of the second messenger c-di-GMP. In contrast, little information is available regarding the molecular mechanisms and cellular components through which this signaling molecule regulates a diverse range of cellular processes. Most of the known effector proteins belong to the PilZ family or are degenerated diguanylate cyclases or phosphodiesterases that have given up on catalysis and have adopted effector function. Thus, to better define the cellular c-di-GMP network in a wide range of bacteria experimental methods are required to identify and validate novel effectors for which reliable in silico predictions fail. We have recently developed a novel Capture Compound Mass Spectrometry (CCMS) based technology as a powerful tool to biochemically identify and characterize c-di-GMP binding proteins. This technique has previously been reported to be applicable to a wide range of organisms(1). Here we give a detailed description of the protocol that we utilize to probe such signaling components. As an example, we use Pseudomonas aeruginosa, an opportunistic pathogen in which c-di-GMP plays a critical role in virulence and biofilm control. CCMS identified 74% (38/51) of the known or predicted components of the c-di-GMP network. This study explains the CCMS procedure in detail, and establishes it as a powerful and versatile tool to identify novel components involved in small molecule signaling.

  3. ChIP-seq reveals the global regulator AlgR mediating cyclic di-GMP synthesis in Pseudomonas aeruginosa.

    Science.gov (United States)

    Kong, Weina; Zhao, Jingru; Kang, Huaping; Zhu, Miao; Zhou, Tianhong; Deng, Xin; Liang, Haihua

    2015-09-30

    AlgR is a key transcriptional regulator required for the expression of multiple virulence factors, including type IV pili and alginate in Pseudomonas aeruginosa. However, the regulon and molecular regulatory mechanism of AlgR have yet to be fully elucidated. Here, among 157 loci that were identified by a ChIP-seq assay, we characterized a gene, mucR, which encodes an enzyme that synthesizes the intracellular second messenger cyclic diguanylate (c-di-GMP). A ΔalgR strain produced lesser biofilm than did the wild-type strain, which is consistent with a phenotype controlled by c-di-GMP. AlgR positively regulates mucR via direct binding to its promoter. A ΔalgRΔmucR double mutant produced lesser biofilm than did the single ΔalgR mutant, demonstrating that c-di-GMP is a positive regulator of biofilm formation. AlgR controls the levels of c-di-GMP synthesis via direct regulation of mucR. In addition, the cognate sensor of AlgR, FimS/AlgZ, also plays an important role in P. aeruginosa virulence. Taken together, this study provides new insights into the AlgR regulon and reveals the involvement of c-di-GMP in the mechanism underlying AlgR regulation.

  4. Differential Regulation of c-di-GMP Metabolic Enzymes by Environmental Signals Modulates Biofilm Formation in Yersinia pestis.

    Science.gov (United States)

    Ren, Gai-Xian; Fan, Sai; Guo, Xiao-Peng; Chen, Shiyun; Sun, Yi-Cheng

    2016-01-01

    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 non-lethal high concentration of CaCl2, MgCl2, CuSO4, sucrose, sodium dodecyl sulfate, 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 regulate activity of DGCs and PDEs to coordinately regulate and adapt metabolism of c-di-GMP and biofilm formation to changing environments.

  5. Cyclic di-GMP mediates a histidine kinase/phosphatase switch by noncovalent domain cross-linking

    Science.gov (United States)

    Dubey, Badri N.; Lori, Christian; Ozaki, Shogo; Fucile, Geoffrey; Plaza-Menacho, Ivan; Jenal, Urs; Schirmer, Tilman

    2016-01-01

    Histidine kinases are key components of regulatory networks in bacteria. Although many of these enzymes are bifunctional, mediating both phosphorylation and dephosphorylation of downstream targets, the molecular details of this central regulatory switch are unclear. We showed recently that the universal second messenger cyclic di–guanosine monophosphate (c-di-GMP) drives Caulobacter crescentus cell cycle progression by forcing the cell cycle kinase CckA from its default kinase into phosphatase mode. We use a combination of structure determination, modeling, and functional analysis to demonstrate that c-di-GMP reciprocally regulates the two antagonistic CckA activities through noncovalent cross-linking of the catalytic domain with the dimerization histidine phosphotransfer (DHp) domain. We demonstrate that both c-di-GMP and ADP (adenosine diphosphate) promote phosphatase activity and propose that c-di-GMP stabilizes the ADP-bound quaternary structure, which allows the receiver domain to access the dimeric DHp stem for dephosphorylation. In silico analyses predict that c-di-GMP control is widespread among bacterial histidine kinases, arguing that it can replace or modulate canonical transmembrane signaling. PMID:27652341

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

    Directory of Open Access Journals (Sweden)

    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.

  7. Structural analysis of the STING adaptor protein reveals a hydrophobic dimer interface and mode of cyclic di-GMP binding.

    Science.gov (United States)

    Ouyang, Songying; Song, Xianqiang; Wang, Yaya; Ru, Heng; Shaw, Neil; Jiang, Yan; Niu, Fengfeng; Zhu, Yanping; Qiu, Weicheng; Parvatiyar, Kislay; Li, Yang; Zhang, Rongguang; Cheng, Genhong; Liu, Zhi-Jie

    2012-06-29

    STING is an essential signaling molecule for DNA and cyclic di-GMP (c-di-GMP)-mediated type I interferon (IFN) production via TANK-binding kinase 1 (TBK1) and interferon regulatory factor 3 (IRF3) pathway. It contains an N-terminal transmembrane region and a cytosolic C-terminal domain (CTD). Here, we describe crystal structures of STING CTD alone and complexed with c-di-GMP in a unique binding mode. The strictly conserved aa 153-173 region was shown to be cytosolic and participated in dimerization via hydrophobic interactions. The STING CTD functions as a dimer and the dimerization was independent of posttranslational modifications. Binding of c-di-GMP enhanced interaction of a shorter construct of STING CTD (residues 139-344) with TBK1. This suggests an extra TBK1 binding site, other than serine 358. This study provides a glimpse into the unique architecture of STING and sheds light on the mechanism of c-di-GMP-mediated TBK1 signaling.

  8. Cyclic di-GMP mediates a histidine kinase/phosphatase switch by noncovalent domain cross-linking.

    Science.gov (United States)

    Dubey, Badri N; Lori, Christian; Ozaki, Shogo; Fucile, Geoffrey; Plaza-Menacho, Ivan; Jenal, Urs; Schirmer, Tilman

    2016-09-01

    Histidine kinases are key components of regulatory networks in bacteria. Although many of these enzymes are bifunctional, mediating both phosphorylation and dephosphorylation of downstream targets, the molecular details of this central regulatory switch are unclear. We showed recently that the universal second messenger cyclic di-guanosine monophosphate (c-di-GMP) drives Caulobacter crescentus cell cycle progression by forcing the cell cycle kinase CckA from its default kinase into phosphatase mode. We use a combination of structure determination, modeling, and functional analysis to demonstrate that c-di-GMP reciprocally regulates the two antagonistic CckA activities through noncovalent cross-linking of the catalytic domain with the dimerization histidine phosphotransfer (DHp) domain. We demonstrate that both c-di-GMP and ADP (adenosine diphosphate) promote phosphatase activity and propose that c-di-GMP stabilizes the ADP-bound quaternary structure, which allows the receiver domain to access the dimeric DHp stem for dephosphorylation. In silico analyses predict that c-di-GMP control is widespread among bacterial histidine kinases, arguing that it can replace or modulate canonical transmembrane signaling. PMID:27652341

  9. 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

    Institute of Scientific and Technical Information of China (English)

    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。

  10. 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

    Institute of Scientific and Technical Information of China (English)

    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.

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

    Directory of Open Access Journals (Sweden)

    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

  12. Defining specificity determinants of cGMP mediated gustatory sensory transduction in Caenorhabditis elegans.

    Science.gov (United States)

    Smith, Heidi K; Luo, Linjiao; O'Halloran, Damien; Guo, Dagang; Huang, Xin-Yun; Samuel, Aravinthan D T; Hobert, Oliver

    2013-08-01

    Cyclic guanosine monophosphate (cGMP) is a key secondary messenger used in signal transduction in various types of sensory neurons. The importance of cGMP in the ASE gustatory receptor neurons of the nematode Caenorhabditis elegans was deduced by the observation that multiple receptor-type guanylyl cyclases (rGCs), encoded by the gcy genes, and two presently known cyclic nucleotide-gated ion channel subunits, encoded by the tax-2 and tax-4 genes, are essential for ASE-mediated gustatory behavior. We describe here specific mechanistic features of cGMP-mediated signal transduction in the ASE neurons. First, we assess the specificity of the sensory functions of individual rGC proteins. We have previously shown that multiple rGC proteins are expressed in a left/right asymmetric manner in the functionally lateralized ASE neurons and are required to sense distinct salt cues. Through domain swap experiments among three different rGC proteins, we show here that the specificity of individual rGC proteins lies in their extracellular domains and not in their intracellular, signal-transducing domains. Furthermore, we find that rGC proteins are also sufficient to confer salt sensory responses to other neurons. Both findings support the hypothesis that rGC proteins are salt receptor proteins. Second, we identify a novel, likely downstream effector of the rGC proteins in gustatory signal transduction, a previously uncharacterized cyclic nucleotide-gated (CNG) ion channel, encoded by the che-6 locus. che-6 mutants show defects in gustatory sensory transduction that are similar to defects observed in animals lacking the tax-2 and tax-4 CNG channels. In contrast, thermosensory signal transduction, which also requires tax-2 and tax-4, does not require che-6, but requires another CNG, cng-3. We propose that CHE-6 may form together with two other CNG subunits, TAX-2 and TAX-4, a gustatory neuron-specific heteromeric CNG channel complex. PMID:23695300

  13. GMP cryopreservation of large volumes of cells for regenerative medicine: active control of the freezing process.

    Science.gov (United States)

    Massie, Isobel; Selden, Clare; Hodgson, Humphrey; Fuller, Barry; Gibbons, Stephanie; Morris, G John

    2014-09-01

    Cryopreservation protocols are increasingly required in regenerative medicine applications but must deliver functional products at clinical scale and comply with Good Manufacturing Process (GMP). While GMP cryopreservation is achievable on a small scale using a Stirling cryocooler-based controlled rate freezer (CRF) (EF600), successful large-scale GMP cryopreservation is more challenging due to heat transfer issues and control of ice nucleation, both complex events that impact success. We have developed a large-scale cryocooler-based CRF (VIA Freeze) that can process larger volumes and have evaluated it using alginate-encapsulated liver cell (HepG2) spheroids (ELS). It is anticipated that ELS will comprise the cellular component of a bioartificial liver and will be required in volumes of ∼2 L for clinical use. Sample temperatures and Stirling cryocooler power consumption was recorded throughout cooling runs for both small (500 μL) and large (200 mL) volume samples. ELS recoveries were assessed using viability (FDA/PI staining with image analysis), cell number (nuclei count), and function (protein secretion), along with cryoscanning electron microscopy and freeze substitution techniques to identify possible injury mechanisms. Slow cooling profiles were successfully applied to samples in both the EF600 and the VIA Freeze, and a number of cooling and warming profiles were evaluated. An optimized cooling protocol with a nonlinear cooling profile from ice nucleation to -60°C was implemented in both the EF600 and VIA Freeze. In the VIA Freeze the nucleation of ice is detected by the control software, allowing both noninvasive detection of the nucleation event for quality control purposes and the potential to modify the cooling profile following ice nucleation in an active manner. When processing 200 mL of ELS in the VIA Freeze-viabilities at 93.4% ± 7.4%, viable cell numbers at 14.3 ± 1.7 million nuclei/mL alginate, and protein secretion at 10.5 ± 1.7

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

    OpenAIRE

    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...

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  16. Mutational analysis of structural elements in a class-I cyclic di-GMP riboswitch to elucidate its regulatory mechanism.

    Science.gov (United States)

    Inuzuka, Saki; Nishimura, Kei-Ichiro; Kakizawa, Hitoshi; Fujita, Yuki; Furuta, Hiroyuki; Matsumura, Shigeyoshi; Ikawa, Yoshiya

    2016-09-01

    The Vc2 riboswitch possesses an aptamer domain belonging to the class-I c-di-GMP riboswitch family. This domain has been analysed and the molecular mechanism by which it recognizes the c-di-GMP ligand has been elucidated. On the other hand, the regulatory mechanism of the full-length Vc2 riboswitch to control its downstream open reading frame (ORF) remains largely unknown. In this study, we performed in vivo reporter assays and in vitro biochemical analyses of the full-length riboswitch and its aptamer domain. We evaluated the results of in vivo and in vitro analyses to elucidate the regulatory mechanism of the Vc2 riboswitch. The present results suggest that recognition of c-di-GMP ligand by the Vc2 riboswitch aptamer domain downregulates expression of its downstream ORF primarily at the translational level.

  17. Differential Contribution of the Guanylyl Cyclase-Cyclic GMP-Protein Kinase G Pathway to the Proliferation of Neural Stem Cells Stimulated by Nitric Oxide

    Directory of Open Access Journals (Sweden)

    Bruno P. Carreira

    2012-02-01

    Full Text Available Nitric oxide (NO is an important inflammatory mediator involved in the initial boost in the proliferation of neural stem cells following brain injury. However, the mechanisms underlying the proliferative effect of NO are still unclear. The aim of this work was to investigate whether cyclic GMP (cGMP and the cGMP-dependent kinase (PKG are involved in the proliferative effect triggered by NO in neural stem cells. For this purpose, cultures of neural stem cells isolated from the mouse subventricular zone (SVZ were used. We observed that long-term exposure to the NO donor (24 h, NOC-18, increased the proliferation of SVZ cells in a cGMP-dependent manner, since the guanylate cyclase inhibitor, ODQ, prevented cell proliferation. Similarly to NOC-18, the cGMP analogue, 8-Br-cGMP, also increased cell proliferation. Interestingly, shorter exposures to NO (6 h increased cell proliferation in a cGMP-independent manner via the ERK/MAP kinase pathway. The selective inhibitor of PKG, KT5823, prevented the proliferative effect induced by NO at 24 h but not at 6 h. In conclusion, the proliferative effect of NO is initially mediated by the ERK/MAPK pathway, and at later stages by the GC/cGMP/PKG pathway. Thus, our work shows that NO induces neural stem cell proliferation by targeting these two pathways in a biphasic manner.

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

    DEFF Research Database (Denmark)

    Christensen, Louise D.; van Gennip, Maria; Rybtke, Morten Theil;

    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...

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

    Science.gov (United States)

    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. 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

    Directory of Open Access Journals (Sweden)

    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

  1. 新版GMP与Simatic WinCC%The New Version of GMP and Simatic WinCC

    Institute of Scientific and Technical Information of China (English)

    孙鸿祥

    2011-01-01

    介绍新版GMP发布的背景及对制药行业自动化系统提出的新要求。GMP认证规范以及Simatic WinCC对于FDA 21 CFR Part 11法规的遵从情况。%This essay maily describe the background of new released GMP and fresh demand on automation system of pharmaceutical industry.The certification specifications of GMP and compliance response FDA 21 CFR Part 11 for Simatic WinCC are introduced in detail.

  2. Formation and dimerization of the phosphodiesterase active site of the Pseudomonas aeruginosa MorA, a bi-functional c-di-GMP regulator.

    Science.gov (United States)

    Phippen, Curtis William; Mikolajek, Halina; Schlaefli, Henry George; Keevil, Charles William; Webb, Jeremy Stephen; Tews, Ivo

    2014-12-20

    Diguanylate cyclases (DGC) and phosphodiesterases (PDE), respectively synthesise and hydrolyse the secondary messenger cyclic dimeric GMP (c-di-GMP), and both activities are often found in a single protein. Intracellular c-di-GMP levels in turn regulate bacterial motility, virulence and biofilm formation. We report the first structure of a tandem DGC-PDE fragment, in which the catalytic domains are shown to be active. Two phosphodiesterase states are distinguished by active site formation. The structures, in the presence or absence of c-di-GMP, suggest that dimerisation and binding pocket formation are linked, with dimerisation being required for catalytic activity. An understanding of PDE activation is important, as biofilm dispersal via c-di-GMP hydrolysis has therapeutic effects on chronic infections.

  3. 医药企业的GMP也要随着时代的步伐而前进%GMP Certification of Pharmaceutical Enterprises Need to Advance with Times

    Institute of Scientific and Technical Information of China (English)

    张公绪; 孙静

    2003-01-01

    @@ 由于特大药难的影响,1962年美国首创GMP,1969年世界卫生组织(WHO)公布了WHO的GMP.1975年WHO向各成员国推荐GMP,1977年WHO再次推荐GMP并将GMP制定为法规.在以后的年代里,许多国家都建立了本国的GMP.我国在1988年建立了GMP(药品生产质量管理规范).迄今我国GMP认证通过的医药企业已有1440家.

  4. Bestrophin-3 (vitelliform macular dystrophy 2-like 3 protein) is essential for the cGMP-dependent calcium-activated chloride conductance in vascular smooth muscle cells

    DEFF Research Database (Denmark)

    Matchkov, Vladimir; Larsen, Per; Bouzinova, Elena V.;

    2008-01-01

    have recently characterized a cGMP-dependent Ca(2+)-activated Cl(-) current with unique characteristics in smooth muscle cells. This novel current has been shown to coexist with a "classic" (cGMP-independent) Ca(2+)-activated Cl(-) current and to have characteristics distinct from those previously...... known for Ca(2+)-activated Cl(-) currents. Here, we suggest that a bestrophin, a product of the Best gene family, is responsible for the cGMP-dependent Ca(2+)-activated Cl(-) current based on similarities between the membrane currents produced by heterologous expressions of bestrophins and the cGMP......-dependent Ca(2+)-activated Cl(-) current. This is supported by similarities in the distribution pattern of the cGMP-dependent Ca(2+)-activated Cl(-) current and bestrophin-3 (the product of Best-3 gene) expression in different smooth muscle. Furthermore, downregulation of Best-3 gene expression with small...

  5. Hybrid promiscuous (Hypr) GGDEF enzymes produce cyclic AMP-GMP (3', 3'-cGAMP).

    Science.gov (United States)

    Hallberg, Zachary F; Wang, Xin C; Wright, Todd A; Nan, Beiyan; Ad, Omer; Yeo, Jongchan; Hammond, Ming C

    2016-02-16

    Over 30 years ago, GGDEF domain-containing enzymes were shown to be diguanylate cyclases that produce cyclic di-GMP (cdiG), a second messenger that modulates the key bacterial lifestyle transition from a motile to sessile biofilm-forming state. Since then, the ubiquity of genes encoding GGDEF proteins in bacterial genomes has established the dominance of cdiG signaling in bacteria. However, the observation that proteobacteria encode a large number of GGDEF proteins, nearing 1% of coding sequences in some cases, raises the question of why bacteria need so many GGDEF enzymes. In this study, we reveal that a subfamily of GGDEF enzymes synthesizes the asymmetric signaling molecule cyclic AMP-GMP (cAG or 3', 3'-cGAMP). This discovery is unexpected because GGDEF enzymes function as symmetric homodimers, with each monomer binding to one substrate NTP. Detailed analysis of the enzyme from Geobacter sulfurreducens showed it is a dinucleotide cyclase capable of switching the major cyclic dinucleotide (CDN) produced based on ATP-to-GTP ratios. We then establish through bioinformatics and activity assays that hybrid CDN-producing and promiscuous substrate-binding (Hypr) GGDEF enzymes are found in other deltaproteobacteria. Finally, we validated the predictive power of our analysis by showing that cAG is present in surface-grown Myxococcus xanthus. This study reveals that GGDEF enzymes make alternative cyclic dinucleotides to cdiG and expands the role of this widely distributed enzyme family to include regulation of cAG signaling.

  6. Extracellular dGMP enhances Deinococcus radiodurans tolerance to oxidative stress.

    Directory of Open Access Journals (Sweden)

    Mingfeng Li

    Full Text Available Free extracellular DNA provides nutrition to bacteria and promotes bacterial evolution by inducing excessive mutagenesis of the genome. To understand the influence of extracellular DNA fragments on D. radiodurans, we investigated cell growth and survival after extracellular DNA or dNMPs treatment. The results showed that the extracellular DNA fragments inhibited the growth of D. radiodurans. Interestingly, dGMP, a DNA component, enhanced D. radiodurans tolerance to H(2O(2 and gamma-radiation significantly. Further experiments indicated that extracellular dGMP stimulated the activity of one catalase (KatA, DR1998, and induced gene transcription including the extracellular nuclease (drb0067. When this only extracellular nuclease gene (drb0067 in D. radiodurans was deleted, the mutant strain showed more sensitive to H(2O(2 and gamma-radiation than the wild type strain. These results suggest that DRB0067 plays an important role in oxidative stress resistance. Taken together, we proposed a new anti-oxidation mechanism in D. radiodurans. This mechanism acts to increase expression levels of DRB0067 which then secretes active nuclease to degrade extracellular DNA fragments. The extracellular nuclease has a two-fold benefit, creating more free dNTPs for further cell protection and the removal of extracellular DNA fragments.

  7. C-di-GMP Synthesis: Structural Aspects of Evolution, Catalysis and Regulation.

    Science.gov (United States)

    Schirmer, Tilman

    2016-09-25

    Cellular levels of the second messenger cyclic di-guanosine monophosphate (c-di-GMP) are determined by the antagonistic activities of diguanylate cyclases and specific phosphodiesterases. In a given bacterial organism, there are often multiple variants of the two enzymes, which are tightly regulated by a variety of external and internal cues due to the presence of specialized sensory or regulatory domains. Dependent on the second messenger level, specific c-di-GMP receptors then control fundamental cellular processes, such as bacterial life style, biofilm formation, and cell cycle control. Here, I review the large body of data on structure-function relationships in diguanylate cyclases. Although the catalytic GGDEF domain is related to the respective domain of adenylate cyclases, the catalyzed intermolecular condensation reaction of two GTP molecules requires the formation of a competent GGDEF dimer with the two substrate molecules juxtaposed. This prerequisite appears to constitute the basis for GGDEF regulation with signal-induced changes within the homotypic dimer of the input domain (PAS, GAF, HAMP, etc.), which are structurally coupled with the arrangement of the GGDEF domains via a rigid coiled-coil linker. Alternatively, phosphorylation of a Rec input domain can drive GGDEF dimerization. Both mechanisms allow modular combination of input and output function that appears advantageous for evolution and rationalizes the striking similarities in domain architecture found in diguanylate cyclases and histidine kinases. PMID:27498163

  8. Systems Pharmacology and Rational Polypharmacy: Nitric Oxide−Cyclic GMP Signaling Pathway as an Illustrative Example and Derivation of the General Case

    Science.gov (United States)

    Garmaroudi, Farshid S.; Handy, Diane E.; Liu, Yang-Yu; Loscalzo, Joseph

    2016-01-01

    Impaired nitric oxide (NO˙)-cyclic guanosine 3', 5'-monophosphate (cGMP) signaling has been observed in many cardiovascular disorders, including heart failure and pulmonary arterial hypertension. There are several enzymatic determinants of cGMP levels in this pathway, including soluble guanylyl cyclase (sGC) itself, the NO˙-activated form of sGC, and phosphodiesterase(s) (PDE). Therapies for some of these disorders with PDE inhibitors have been successful at increasing cGMP levels in both cardiac and vascular tissues. However, at the systems level, it is not clear whether perturbation of PDE alone, under oxidative stress, is the best approach for increasing cGMP levels as compared with perturbation of other potential pathway targets, either alone or in combination. Here, we develop a model-based approach to perturbing this pathway, focusing on single reactions, pairs of reactions, or trios of reactions as targets, then monitoring the theoretical effects of these interventions on cGMP levels. Single perturbations of all reaction steps within this pathway demonstrated that three reaction steps, including the oxidation of sGC, NO˙ dissociation from sGC, and cGMP degradation by PDE, exerted a dominant influence on cGMP accumulation relative to other reaction steps. Furthermore, among all possible single, paired, and triple perturbations of this pathway, the combined perturbations of these three reaction steps had the greatest impact on cGMP accumulation. These computational findings were confirmed in cell-based experiments. We conclude that a combined perturbation of the oxidatively-impaired NO˙-cGMP signaling pathway is a better approach to the restoration of cGMP levels as compared with corresponding individual perturbations. This approach may also yield improved therapeutic responses in other complex pharmacologically amenable pathways. PMID:26985825

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

    Science.gov (United States)

    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.

  10. Oligoribonuclease is the primary degradative enzyme for pGpG in Pseudomonas aeruginosa that is required for cyclic-di-GMP turnover.

    Science.gov (United States)

    Orr, Mona W; Donaldson, Gregory P; Severin, Geoffrey B; Wang, Jingxin; Sintim, Herman O; Waters, Christopher M; Lee, Vincent T

    2015-09-01

    The bacterial second messenger cyclic di-GMP (c-di-GMP) controls biofilm formation and other phenotypes relevant to pathogenesis. Cyclic-di-GMP is synthesized by diguanylate cyclases (DGCs). Phosphodiesterases (PDE-As) end signaling by linearizing c-di-GMP to 5'-phosphoguanylyl-(3',5')-guanosine (pGpG), which is then hydrolyzed to two GMP molecules by yet unidentified enzymes termed PDE-Bs. We show that pGpG inhibits a PDE-A from Pseudomonas aeruginosa. In a dual DGC and PDE-A reaction, excess pGpG extends the half-life of c-di-GMP, indicating that removal of pGpG is critical for c-di-GMP homeostasis. Thus, we sought to identify the PDE-B enzyme(s) responsible for pGpG degradation. A differential radial capillary action of ligand assay-based screen for pGpG binding proteins identified oligoribonuclease (Orn), an exoribonuclease that hydrolyzes two- to five-nucleotide-long RNAs. Purified Orn rapidly converts pGpG into GMP. To determine whether Orn is the primary enzyme responsible for degrading pGpG, we assayed cell lysates of WT and ∆orn strains of P. aeruginosa PA14 for pGpG stability. The lysates from ∆orn showed 25-fold decrease in pGpG hydrolysis. Complementation with WT, but not active site mutants, restored hydrolysis. Accumulation of pGpG in the ∆orn strain could inhibit PDE-As, increasing c-di-GMP concentration. In support, we observed increased transcription from the c-di-GMP-regulated pel promoter. Additionally, the c-di-GMP-governed auto-aggregation and biofilm phenotypes were elevated in the ∆orn strain in a pel-dependent manner. Finally, we directly detect elevated pGpG and c-di-GMP in the ∆orn strain. Thus, we identified that Orn serves as the primary PDE-B enzyme that removes pGpG, which is necessary to complete the final step in the c-di-GMP degradation pathway.

  11. Relationship between Adaptation of the Folic Acid and the cAMP Mediated cGMP Response in Dictyostelium

    NARCIS (Netherlands)

    Haastert, Peter J.M. van

    1983-01-01

    Chemotactic stimulation of post-vegetative Dictyostelium cells with folic acid or aggregative cells with cAMP results in a fast transient cGMP response which peaks at 10 s; basal levels are recovered in about 30-40 s. Stimulation with folic acid or cAMP rapidly desensitizes the cells for equal or lo

  12. Systematic Nomenclature for GGDEF and EAL Domain-Containing Cyclic Di-GMP Turnover Proteins of Escherichia coli.

    Science.gov (United States)

    Hengge, Regine; Galperin, Michael Y; Ghigo, Jean-Marc; Gomelsky, Mark; Green, Jeffrey; Hughes, Kelly T; Jenal, Urs; Landini, Paolo

    2016-01-01

    In recent years, Escherichia coli has served as one of a few model bacterial species for studying cyclic di-GMP (c-di-GMP) signaling. The widely used E. coli K-12 laboratory strains possess 29 genes encoding proteins with GGDEF and/or EAL domains, which include 12 diguanylate cyclases (DGC), 13 c-di-GMP-specific phosphodiesterases (PDE), and 4 "degenerate" enzymatically inactive proteins. In addition, six new GGDEF and EAL (GGDEF/EAL) domain-encoding genes, which encode two DGCs and four PDEs, have recently been found in genomic analyses of commensal and pathogenic E. coli strains. As a group of researchers who have been studying the molecular mechanisms and the genomic basis of c-di-GMP signaling in E. coli, we now propose a general and systematic dgc and pde nomenclature for the enzymatically active GGDEF/EAL domain-encoding genes of this model species. This nomenclature is intuitive and easy to memorize, and it can also be applied to additional genes and proteins that might be discovered in various strains of E. coli in future studies.

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

    Science.gov (United States)

    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.

  14. The cyclic GMP/protein kinase G pathway as a therapeutic target in head and neck squamous cell carcinoma.

    Science.gov (United States)

    Tuttle, Traci R; Mierzwa, Michelle L; Wells, Susanne I; Fox, Sejal R; Ben-Jonathan, Nira

    2016-01-28

    Head and neck squamous cell carcinoma (HNSCC) is an aggressive disease with high mortality. Treatments, which can result in significant morbidity, have not substantially changed in three decades. The second messenger cyclic GMP (cGMP), which targets protein kinase G (PKG), is generated by guanylate cyclases (GCs), and is rapidly hydrolyzed by phosphodiesterases (PDEs). Activation of the cGMP/PKG pathway is antineoplastic in several cancer types, but its impact on HNSCC has not been fully exploited. We found differential expression of critical components of this pathway in four HNSCC cell lines. Several activators of soluble GC (sGC), as well as inhibitors of PDE5, increased intracellular cGMP, reduced cell viability, and induced apoptosis in HNSCC cells. The apoptotic effects of the sGC activator BAY 41-2272 and the PDE5 inhibitor Tadalafil (Cialis) were mediated by PKG. Furthermore, Tadalafil substantially reduced the growth of CAL27-derived tumors in athymic mice. Several drugs which either activate sGC or inhibit PDE5 are approved for treatment of nonmalignant conditions. These drugs could be repurposed as novel and effective therapeutics in patients with head and neck cancer.

  15. Synthesis of Triazole-Linked Analogues of c-di-GMP and Their Interactions with Diguanylate Cyclase.

    Science.gov (United States)

    Fernicola, Silvia; Torquati, Ilaria; Paiardini, Alessandro; Giardina, Giorgio; Rampioni, Giordano; Messina, Marco; Leoni, Livia; Del Bello, Fabio; Petrelli, Riccardo; Rinaldo, Serena; Cappellacci, Loredana; Cutruzzolà, Francesca

    2015-10-22

    Cyclic di-GMP (c-di-GMP) is a widespread second messenger that plays a key role in bacterial biofilm formation. The compound's ability to assume multiple conformations allows it to interact with a diverse set of target macromolecules. Here, we analyzed the binding mode of c-di-GMP to the allosteric inhibitory site (I-site) of diguanylate cyclases (DGCs) and compared it to the conformation adopted in the catalytic site of the EAL phosphodiesterases (PDEs). An array of novel molecules has been designed and synthesized by simplifying the native c-di-GMP structure and replacing the charged phosphodiester backbone with an isosteric nonhydrolyzable 1,2,3-triazole moiety. We developed the first neutral small molecule able to selectively target DGCs discriminating between the I-site of DGCs and the active site of PDEs; this molecule represents a novel tool for mechanistic studies, particularly on those proteins bearing both DGC and PDE modules, and for future optimization studies to target DGCs in vivo.

  16. Mechanistic insight into the conserved allosteric regulation of periplasmic proteolysis by the signaling molecule cyclic-di-GMP.

    Science.gov (United States)

    Chatterjee, Debashree; Cooley, Richard B; Boyd, Chelsea D; Mehl, Ryan A; O'Toole, George A; Sondermann, Holger

    2014-01-01

    Stable surface adhesion of cells is one of the early pivotal steps in bacterial biofilm formation, a prevalent adaptation strategy in response to changing environments. In Pseudomonas fluorescens, this process is regulated by the Lap system and the second messenger cyclic-di-GMP. High cytoplasmic levels of cyclic-di-GMP activate the transmembrane receptor LapD that in turn recruits the periplasmic protease LapG, preventing it from cleaving a cell surface-bound adhesin, thereby promoting cell adhesion. In this study, we elucidate the molecular basis of LapG regulation by LapD and reveal a remarkably sensitive switching mechanism that is controlled by LapD's HAMP domain. LapD appears to act as a coincidence detector, whereby a weak interaction of LapG with LapD transmits a transient outside-in signal that is reinforced only when cyclic-di-GMP levels increase. Given the conservation of key elements of this receptor system in many bacterial species, the results are broadly relevant for cyclic-di-GMP- and HAMP domain-regulated transmembrane signaling.

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

    DEFF Research Database (Denmark)

    Matchkov, Vladimir; Aalkjær, Christian; Nilsson, Holger

    2004-01-01

    differed from those of the calcium-activated chloride current in pulmonary myocytes, which was cGMP-independent, exhibited a high sensitivity to inhibition by niflumic acid, was unaffected by zinc ions, and showed outward current rectification as has previously been reported for this current. Under...

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

    Science.gov (United States)

    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

  19. Improved GMP-compliant multi-dose production and quality control of 6-[18F]fluoro-L-DOPA

    NARCIS (Netherlands)

    Luurtsema, Geert; Boersma, Hendrikus; Schepers, Marianne; de Vries, Michel; Maas, Bram; Zijlma, Rolf; de Vries, Erik; Elsinga, Philippus

    2016-01-01

    Background: 6-[18F]Fluoro-L-3,4-dihydroxyphenylalanine (FDOPA) is a frequently used radiopharmaceutical for detecting neuroendocrine and brain tumors and for the differential diagnosis of Parkinson’s disease. To meet the demand for FDOPA, a high-yield GMP-compliant production method is required. The

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

    NARCIS (Netherlands)

    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

  1. Ozone and nitric oxide induce cGMP-dependent and -independent transcription of defence genes in tobacco.

    Science.gov (United States)

    Pasqualini, Stefania; Meier, Stuart; Gehring, Chris; Madeo, Laura; Fornaciari, Marco; Romano, Bruno; Ederli, Luisa

    2009-03-01

    Here, we analyse the temporal signatures of ozone (O3)-induced hydrogen peroxide(H2O2) and nitric oxide (NO) and the role of the second messenger guanosine3′,5′-cyclic monophosphate (cGMP) in transcriptional changes of genes diagnostic for biotic and abiotic stress responses. Within 90 min O3 induced H2O2 and NO peaks and we demonstrate that NO donors cause rapid H2O2 accumulation in tobacco (Nicotiana tabacum) leaf. Ozone also causes highly significant, late (> 2 h) and sustained cGMP increases, suggesting that the second messenger may not be required in all early (< 2 h) responses to O3,but is essential and sufficient for the induction of some O3-dependent pathways.This hypothesis was tested resolving the time course of O3-induced transcript accumulation of alternative oxidase (AOX1a), glutathione peroxidase (GPX),aminocyclopropancarboxylic acid synthase (ACS2) that is critical for the synthesis of ethylene, phenylalanine ammonia lyase (PALa) and the pathogenesis-related protein PR1a.The data show that early O3 and NO caused transcriptional activation of the scavenger encoding proteins AOX1a, GPX and the induction of ethylene production through ACS2 are cGMP independent. By contrast, the early response of PALa and the late response of PR1a show critical dependence on cGMP.

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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…

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

    NARCIS (Netherlands)

    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

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

    Science.gov (United States)

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

    2016-07-06

    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.

  6. Relationship of plasma levels of LP-PLA2, GMP, LPA and platelet activation markers with transient ischemic attack

    Institute of Scientific and Technical Information of China (English)

    Hang Li

    2015-01-01

    Objective:To explore the relationship of plasma levels of LP-PLA2, GMP, LPA and platelet activation markers with transient ischemic attack(TIA).Methods:A total of 87 cases with TIA were collected, and all patients were treated with ozagrel (120mg/times/day for 2 weeks). Fasting blood were collected in 24 h after attack and 7d, 14 days after treatment respectively, then flow cytometry was used to detect indicators of platelet activation, such as CD63, PAC-1, GPⅡb /Ⅲa, and the plasma levels of LP-PLA2, GMP, LPA. The dynamic changes of the such indexes and the differences between different populations were analyzed.Results:The plasma levels of LP-PLA2, GMP, LPA and the platelet activation indexes including CD63, PAC-1, GPⅡb/Ⅲa of patients with TIA were all significantly higher than control group (allP<0.05). One day after treatment of sodium Ozagrel, the plasma levels of LP-PLA2, GMP, LPA and the platelet activation indexes such as CD63, PAC-1, GPⅡb/Ⅲa of patients with TIA began to decline, and returned to normal in the treatment of 7 d or 14 d.Conclusions:Platelet activation plays an important role in the occurrence of TIA, and antiplatelet therapy can effectively inhibit platelet activation, which has positive significance in the treatment of TIA.

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

    Directory of Open Access Journals (Sweden)

    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.

  8. Brain Region-Specific Effects of cGMP-Dependent Kinase II Knockout on AMPA Receptor Trafficking and Animal Behavior

    Science.gov (United States)

    Kim, Seonil; Pick, Joseph E.; Abera, Sinedu; Khatri, Latika; Ferreira, Danielle D. P.; Sathler, Matheus F.; Morison, Sage L.; Hofmann, Franz; Ziff, Edward B.

    2016-01-01

    Phosphorylation of GluA1, a subunit of AMPA receptors (AMPARs), is critical for AMPAR synaptic trafficking and control of synaptic transmission. cGMP-dependent protein kinase II (cGKII) mediates this phosphorylation, and cGKII knockout (KO) affects GluA1 phosphorylation and alters animal behavior. Notably, GluA1 phosphorylation in the KO…

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

    Science.gov (United States)

    Dawis, S M; Graeff, R M; Heyman, R A; Walseth, T F; Goldberg, N D

    1988-06-25

    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 (protein-bound and unbound) concentrations of the guanine nucleotide metabolic intermediates; and changes in the concentration of metabolic (unbound) GDP calculated from the fraction of the alpha-GDP that undergoes labeling with 18O. The latter is interpreted to reflect the state of the equilibrium between GDP- and GTP-complexed forms of G-protein. 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 (0.6-3 log photons.micron-2.s-1) 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.(ABSTRACT TRUNCATED AT 250 WORDS)

  10. Genetic Dissection of the Regulatory Network Associated with High C-di-GMP Levels in Pseudomonas putida KT2440

    Directory of Open Access Journals (Sweden)

    María Isabel Ramos-González

    2016-07-01

    Full Text Available Most bacteria grow in nature forming multicellular structures named biofilms. The bacterial second messenger cyclic diguanosine monophosphate (c-di-GMP is a key player in the regulation of the transition from planktonic to sessile lifestyles and this regulation is crucial in the development of biofilms. In Pseudomonas putida KT2440, Rup4959, a multidomain response regulator with diguanylate cyclase activity, when overexpressed causes an increment in the intracellular levels of c-di-GMP that gives rise to a pleiotropic phenotype consisting of increased biofilm formation and crinkly colony morphology. In a broad genomic screen we have isolated mutant derivatives that lose the crinkly morphology, designed as cfc (crinkle free colony. A total of nineteen different genes have been identified as being related with the emergence of the cfc phenotype either because the expression or functionality of Rup4959 is compromised, or due to a lack of transduction of the c-di-GMP signal to downstream elements involved in the acquisition of the phenotype. Discernment between these possibilities was investigated by using a c-di-GMP biosensor and by HPLC-MS quantification of the second messenger. Interestingly five of the identified genes encode proteins with AAA+ ATPase domain. Among the bacterial determinants found in this screen are the global transcriptional regulators GacA, AlgU and FleQ and two enzymes involved in the arginine biosynthesis pathway. We present evidences that this pathway seems to be an important element to both the availability of the free pool of the second messenger c-di-GMP and to its further transduction as a signal for biosynthesis of biopolimers. In addition we have identified an uncharacterized hybrid sensor histidine kinase whose phosphoaceptor conserved histidine residue has been shown in this work to be required for in vivo activation of the orphan response regulator Rup4959, which suggests these two elements constitute a two

  11. Genetic Dissection of the Regulatory Network Associated with High c-di-GMP Levels in Pseudomonas putida KT2440

    Science.gov (United States)

    Ramos-González, María Isabel; Travieso, María L.; Soriano, María I.; Matilla, Miguel A.; Huertas-Rosales, Óscar; Barrientos-Moreno, Laura; Tagua, Víctor G.; Espinosa-Urgel, Manuel

    2016-01-01

    Most bacteria grow in nature forming multicellular structures named biofilms. The bacterial second messenger cyclic diguanosine monophosphate (c-di-GMP) is a key player in the regulation of the transition from planktonic to sessile lifestyles and this regulation is crucial in the development of biofilms. In Pseudomonas putida KT2440, Rup4959, a multidomain response regulator with diguanylate cyclase activity, when overexpressed causes an increment in the intracellular levels of c-di-GMP that gives rise to a pleiotropic phenotype consisting of increased biofilm formation and crinkly colony morphology. In a broad genomic screen we have isolated mutant derivatives that lose the crinkly morphology, designed as cfc (crinkle free colony). A total of 19 different genes have been identified as being related with the emergence of the cfc phenotype either because the expression or functionality of Rup4959 is compromised, or due to a lack of transduction of the c-di-GMP signal to downstream elements involved in the acquisition of the phenotype. Discernment between these possibilities was investigated by using a c-di-GMP biosensor and by HPLC-MS quantification of the second messenger. Interestingly five of the identified genes encode proteins with AAA+ ATPase domain. Among the bacterial determinants found in this screen are the global transcriptional regulators GacA, AlgU and FleQ and two enzymes involved in the arginine biosynthesis pathway. We present evidences that this pathway seems to be an important element to both the availability of the free pool of the second messenger c-di-GMP and to its further transduction as a signal for biosynthesis of biopolimers. In addition we have identified an uncharacterized hybrid sensor histidine kinase whose phosphoaceptor conserved histidine residue has been shown in this work to be required for in vivo activation of the orphan response regulator Rup4959, which suggests these two elements constitute a two-component phosphorelay system

  12. Genetic Dissection of the Regulatory Network Associated with High c-di-GMP Levels in Pseudomonas putida KT2440.

    Science.gov (United States)

    Ramos-González, María Isabel; Travieso, María L; Soriano, María I; Matilla, Miguel A; Huertas-Rosales, Óscar; Barrientos-Moreno, Laura; Tagua, Víctor G; Espinosa-Urgel, Manuel

    2016-01-01

    Most bacteria grow in nature forming multicellular structures named biofilms. The bacterial second messenger cyclic diguanosine monophosphate (c-di-GMP) is a key player in the regulation of the transition from planktonic to sessile lifestyles and this regulation is crucial in the development of biofilms. In Pseudomonas putida KT2440, Rup4959, a multidomain response regulator with diguanylate cyclase activity, when overexpressed causes an increment in the intracellular levels of c-di-GMP that gives rise to a pleiotropic phenotype consisting of increased biofilm formation and crinkly colony morphology. In a broad genomic screen we have isolated mutant derivatives that lose the crinkly morphology, designed as cfc (crinkle free colony). A total of 19 different genes have been identified as being related with the emergence of the cfc phenotype either because the expression or functionality of Rup4959 is compromised, or due to a lack of transduction of the c-di-GMP signal to downstream elements involved in the acquisition of the phenotype. Discernment between these possibilities was investigated by using a c-di-GMP biosensor and by HPLC-MS quantification of the second messenger. Interestingly five of the identified genes encode proteins with AAA+ ATPase domain. Among the bacterial determinants found in this screen are the global transcriptional regulators GacA, AlgU and FleQ and two enzymes involved in the arginine biosynthesis pathway. We present evidences that this pathway seems to be an important element to both the availability of the free pool of the second messenger c-di-GMP and to its further transduction as a signal for biosynthesis of biopolimers. In addition we have identified an uncharacterized hybrid sensor histidine kinase whose phosphoaceptor conserved histidine residue has been shown in this work to be required for in vivo activation of the orphan response regulator Rup4959, which suggests these two elements constitute a two-component phosphorelay system.

  13. cGMP activates a pH-sensitive leak K+ current in the presumed cholinergic neuron of basal forebrain.

    Science.gov (United States)

    Toyoda, Hiroki; Saito, Mitsuru; Sato, Hajime; Dempo, Yoshie; Ohashi, Atsuko; Hirai, Toshihiro; Maeda, Yoshinobu; Kaneko, Takeshi; Kang, Youngnam

    2008-05-01

    In an earlier study, we demonstrated that nitric oxide (NO) causes the long-lasting membrane hyperpolarization in the presumed basal forebrain cholinergic (BFC) neurons by cGMP-PKG-dependent activation of leak K+ currents in slice preparations. In the present study, we investigated the ionic mechanisms underlying the long-lasting membrane hyperpolarization with special interest in the pH sensitivity because 8-Br-cGMP-induced K+ current displayed Goldman-Hodgkin-Katz rectification characteristic of TWIK-related acid-sensitive K+ (TASK) channels. When examined with the ramp command pulse depolarizing from -130 to -40 mV, the presumed BFC neurons displayed a pH-sensitive leak K+ current that was larger in response to pH decrease from 8.3 to 7.3 than in response to pH decrease from 7.3 to 6.3. This K+ current was similar to TASK1 current in its pH sensitivity, whereas it was highly sensitive to Ba(2+), unlike TASK1 current. The 8-Br-cGMP-induced K+ currents in the presumed BFC neurons were almost completely inhibited by lowering external pH to 6.3 as well as by bath application of 100 microM Ba(2+), consistent with the nature of the leak K+ current expressed in the presumed BFC neurons. After 8-Br-cGMP application, the K+ current obtained by pH decrease from 7.3 to 6.3 was larger than that obtained by pH decrease from pH 8.3 to 7.3, contrary to the case seen in the control condition. These observations strongly suggest that 8-Br-cGMP activates a pH- and Ba(2+)-sensitive leak K+ current expressed in the presumed BFC neurons by modulating its pH sensitivity.

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

    KAUST Repository

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  17. The Importance of cGMP Signaling in Sensory Cilia for Body Size Regulation in Caenorhabditis elegans.

    Science.gov (United States)

    Fujiwara, Manabi; Hino, Takahiro; Miyamoto, Ryuta; Inada, Hitoshi; Mori, Ikue; Koga, Makoto; Miyahara, Koji; Ohshima, Yasumi; Ishihara, Takeshi

    2015-12-01

    The body size of Caenorhabditis elegans is thought to be controlled by sensory inputs because many mutants with sensory cilium structure defects exhibit small body size. The EGL-4 cGMP-dependent protein kinase acts in sensory neurons to reduce body size when animals fail to perceive sensory signals. In addition to body size control, EGL-4 regulates various other behavioral and developmental pathways, including those involved in the regulation of egg laying and chemotaxis behavior. Here we have identified gcy-12, which encodes a receptor-type guanylyl cyclase, as a gene involved in the sensory regulation of body size. Analyses with GFP fusion constructs showed that gcy-12 is expressed in several sensory neurons and localizes to sensory cilia. Genetic analyses indicated that GCY-12 acts upstream of EGL-4 in body size control but does not affect other EGL-4 functions. Our studies indicate that the function of the GCY-12 guanylyl cyclase is to provide cGMP to the EGL-4 cGMP-dependent kinase only for limited tasks including body size regulation. We also found that the PDE-2 cyclic nucleotide phosphodiesterase negatively regulates EGL-4 in controlling body size. Thus, the cGMP level is precisely controlled by GCY-12 and PDE-2 to determine body size through EGL-4, and the defects in the sensory cilium structure may disturb the balanced control of the cGMP level. The large number of guanylyl cyclases encoded in the C. elegans genome suggests that EGL-4 exerts pleiotropic effects by partnering with different guanylyl cyclases for different downstream functions.

  18. Structures of the activator of K. pneumonia biofilm formation, MrkH, indicates PilZ domains involved in c-di-GMP and DNA binding.

    Science.gov (United States)

    Schumacher, Maria A; Zeng, Wenjie

    2016-09-01

    The pathogenesis of Klebsiella pneumonia is linked to the bacteria's ability to form biofilms. Mannose-resistant Klebsiella-like (Mrk) hemagglutinins are critical for K pneumonia biofilm development, and the expression of the genes encoding these proteins is activated by a 3',5'-cyclic diguanylic acid (c-di-GMP)-regulated transcription factor, MrkH. To gain insight into MrkH function, we performed structural and biochemical analyses. Data revealed MrkH to be a monomer with a two-domain architecture consisting of a PilZ C-domain connected to an N domain that unexpectedly also harbors a PilZ-like fold. Comparison of apo- and c-di-GMP-bound MrkH structures reveals a large 138° interdomain rotation that is induced by binding an intercalated c-di-GMP dimer. c-di-GMP interacts with PilZ C-domain motifs 1 and 2 (RxxxR and D/NxSxxG) and a newly described c-di-GMP-binding motif in the MrkH N domain. Strikingly, these c-di-GMP-binding motifs also stabilize an open state conformation in apo MrkH via contacts from the PilZ motif 1 to residues in the C-domain motif 2 and the c-di-GMP-binding N-domain motif. Use of the same regions in apo structure stabilization and c-di-GMP interaction allows distinction between the states. Indeed, domain reorientation by c-di-GMP complexation with MrkH, which leads to a highly compacted structure, suggests a mechanism by which the protein is activated to bind DNA. To our knowledge, MrkH represents the first instance of specific DNA binding mediated by PilZ domains. The MrkH structures also pave the way for the rational design of inhibitors that target K pneumonia biofilm formation.

  19. The Xanthomonas oryzae pv. oryzae PilZ Domain Proteins Function Differentially in Cyclic di-GMP Binding and Regulation of Virulence and Motility.

    Science.gov (United States)

    Yang, Fenghuan; Tian, Fang; Chen, Huamin; Hutchins, William; Yang, Ching-Hong; He, Chenyang

    2015-07-01

    The PilZ domain proteins have been demonstrated to be one of the major types of receptors mediating cyclic di-GMP (c-di-GMP) signaling pathways in several pathogenic bacteria. However, little is known about the function of PilZ domain proteins in c-di-GMP regulation of virulence in the bacterial blight pathogen of rice Xanthomonas oryzae pv. oryzae. Here, the roles of PilZ domain proteins PXO_00049 and PXO_02374 in c-di-GMP binding, regulation of virulence and motility, and subcellular localization were characterized in comparison with PXO_02715, identified previously as an interactor with the c-di-GMP receptor Filp to regulate virulence. The c-di-GMP binding motifs in the PilZ domains were conserved in PXO_00049 and PXO_02374 but were less well conserved in PXO_02715. PXO_00049 and PXO_02374 but not PXO_02715 proteins bound to c-di-GMP with high affinity in vitro, and the R(141) and R(10) residues in the PilZ domains of PXO_00049 and PXO_02374, respectively, were crucial for c-di-GMP binding. Gene deletion of PXO_00049 and PXO_02374 resulted in significant increases in virulence and hrp gene transcription, indicating their negative regulation of virulence via type III secretion system expression. All mutants showed significant changes in sliding motility but not exopolysaccharide production and biofilm formation. In trans expression of the full-length open reading frame (ORF) of each gene in the relevant mutants led to restoration of the phenotype to wild-type levels. Moreover, PXO_00049 and PXO_02374 displayed mainly multisite subcellular localizations, whereas PXO_02715 showed nonpolar distributions in the X. oryzae pv. oryzae cells. Therefore, this study demonstrated the different functions of the PilZ domain proteins in mediation of c-di-GMP regulation of virulence and motility in X. oryzae pv. oryzae.

  20. Time-dependent inhibitory effects of cGMP-analogues on thrombin-induced platelet-derived microparticles formation, platelet aggregation, and P-selectin expression.

    Science.gov (United States)

    Nygaard, Gyrid; Herfindal, Lars; Kopperud, Reidun; Aragay, Anna M; Holmsen, Holm; Døskeland, Stein Ove; Kleppe, Rune; Selheim, Frode

    2014-07-01

    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.

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

    Science.gov (United States)

    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.

  2. Cleanrooms and tissue banking how happy I could be with either GMP or GTP?

    Science.gov (United States)

    Klykens, J; Pirnay, J-P; Verbeken, G; Giet, O; Baudoux, E; Jashari, R; Vanderkelen, A; Ectors, N

    2013-12-01

    The regulatory framework of tissue banking introduces a number of requirements for monitoring cleanrooms for processing tissue or cell grafts. Although a number of requirements were clearly defined, some requirements are open for interpretation. This study aims to contribute to the interpretation of GMP or GTP guidelines for tissue banking. Based on the experience of the participating centers, the results of the monitoring program were evaluated to determine the feasibility of a cleanroom in tissue banking and the monitoring program. Also the microbial efficacy of a laminar airflow cabinet and an incubator in a cleanroom environment was evaluated. This study indicated that a monitoring program of a cleanroom at rest in combination with (final) product testing is a feasible approach. Although no statistical significance (0.90 contamination of an incubator in a cleanroom is limited but requires closed containers for tissue and cell products.

  3. cGMP-dependent protein kinase: linking foraging to energy homeostasis.

    Science.gov (United States)

    Kaun, Karla R; Sokolowski, Marla B

    2009-01-01

    Successful foraging is necessary for procurement of nutritional resources essential for an animal's survival. Maintenance of foraging and food acquisition is dependent on the ability to balance food intake and energy expenditure. This review examines the role of cGMP-dependent protein kinase (PKG) as a regulator of foraging behaviour, food acquisition, and energy balance. The role of PKG in food-related behaviours is highly conserved among worms, flies, bees, ants, and mammals. A growing body of literature suggests that PKG plays an integral role in the component behaviours and physiologies underlying foraging behaviour. These include energy acquisition, nutrient absorption, nutrient allocation, nutrient storage, and energy use. New evidence suggests that PKG mediates both neural and physiological mechanisms underlying these processes. This review illustrates how investigating the role of PKG in energy homeostasis in a diversity of organisms can offer a broad perspective on the mechanisms mediating energy balance.

  4. Cyclic GMP signaling in cardiomyocytes modulates fatty acid trafficking and prevents triglyceride accumulation.

    Science.gov (United States)

    Khairallah, Ramzi J; Khairallah, Maya; Gélinas, Roselle; Bouchard, Bertrand; Young, Martin E; Allen, Bruce G; Lopaschuk, Gary D; Deschepper, Christian F; Des Rosiers, Christine

    2008-08-01

    While the balance between carbohydrates and fatty acids for energy production appears to be crucial for cardiac homeostasis, much remains to be learned about the molecular mechanisms underlying this relationship. Given the reported benefits of cGMP signaling on the myocardium, we investigated the impact of its chronic activation on cardiac energy metabolism using mice overexpressing a constitutively active cytoplasmic guanylate cyclase (GC(+/0)) in cardiomyocytes. Ex vivo working GC(+/0) heart perfusions with (13)C-labeled substrates revealed an altered pattern of exogenous substrate fuel selection compared to controls, namely a 38+/-9% lower contribution of exogenous fatty acids to acetyl-CoA formation, while that of carbohydrates remains unchanged despite a two-fold increase in glycolysis. The lower contribution of exogenous fatty acids to energy production is not associated with changes in energy demand or supply (contractile function, oxygen consumption, tissue acetyl-CoA or CoA levels, citric acid cycle flux rate) or in the regulation of beta-oxidation (acetyl-CoA carboxylase activity, tissue malonyl-CoA levels). However, GC(+/0) hearts show a two-fold increase in the incorporation of exogenous oleate into triglycerides. Furthermore, the following molecular data are consistent with a concomitant increase in triglyceride hydrolysis: (i) increased abundance of hormone sensitive lipase (HSL) protein (24+/-11%) and mRNA (22+/-4%) as well as (ii) several phosphorylation events related to HSL inhibitory (AMPK) and activation (ERK 1/2) sites, which should contribute to enhance its activity. These changes in exogenous fatty acid trafficking in GC(+/0) hearts appear to be functionally relevant, as demonstrated by their resistance to fasting-induced triglyceride accumulation. While the documented metabolic profile of GC(+/0) mouse hearts is partly reminiscent of hypertrophied hearts, the observed changes in lipid trafficking have not been previously documented, and may

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

    KAUST Repository

    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.

  6. Hybrid promiscuous (Hypr) GGDEF enzymes produce cyclic AMP-GMP (3', 3'-cGAMP).

    Science.gov (United States)

    Hallberg, Zachary F; Wang, Xin C; Wright, Todd A; Nan, Beiyan; Ad, Omer; Yeo, Jongchan; Hammond, Ming C

    2016-02-16

    Over 30 years ago, GGDEF domain-containing enzymes were shown to be diguanylate cyclases that produce cyclic di-GMP (cdiG), a second messenger that modulates the key bacterial lifestyle transition from a motile to sessile biofilm-forming state. Since then, the ubiquity of genes encoding GGDEF proteins in bacterial genomes has established the dominance of cdiG signaling in bacteria. However, the observation that proteobacteria encode a large number of GGDEF proteins, nearing 1% of coding sequences in some cases, raises the question of why bacteria need so many GGDEF enzymes. In this study, we reveal that a subfamily of GGDEF enzymes synthesizes the asymmetric signaling molecule cyclic AMP-GMP (cAG or 3', 3'-cGAMP). This discovery is unexpected because GGDEF enzymes function as symmetric homodimers, with each monomer binding to one substrate NTP. Detailed analysis of the enzyme from Geobacter sulfurreducens showed it is a dinucleotide cyclase capable of switching the major cyclic dinucleotide (CDN) produced based on ATP-to-GTP ratios. We then establish through bioinformatics and activity assays that hybrid CDN-producing and promiscuous substrate-binding (Hypr) GGDEF enzymes are found in other deltaproteobacteria. Finally, we validated the predictive power of our analysis by showing that cAG is present in surface-grown Myxococcus xanthus. This study reveals that GGDEF enzymes make alternative cyclic dinucleotides to cdiG and expands the role of this widely distributed enzyme family to include regulation of cAG signaling. PMID:26839412

  7. A label-free and self-assembled electrochemical biosensor for highly sensitive detection of cyclic diguanylate monophosphate (c-di-GMP) based on RNA riboswitch.

    Science.gov (United States)

    Xie, Qingyun; Zhao, Fulin; Liu, Hongrui; Shan, Yanke; Liu, Fei

    2015-07-01

    Cyclic diguanylate monophosphate (c-di-GMP) is an important second messenger that regulates a variety of complex physiological processes involved in motility, virulence, biofilm formation and cell cycle progression in several bacteria. Herein we report a simple label-free and self-assembled RNA riboswitch-based biosensor for sensitive and selective detection of c-di-GMP. The detectable concentration range of c-di-GMP is from 50 nM to 1 μM with a detection limit of 50 nM.

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

    OpenAIRE

    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...

  9. CRP-Cyclic AMP Regulates the Expression of Type 3 Fimbriae via Cyclic di-GMP in Klebsiella pneumoniae.

    Science.gov (United States)

    Lin, Ching-Ting; Lin, Tien-Huang; Wu, Chien-Chen; Wan, Lei; Huang, Chun-Fa; Peng, Hwei-Ling

    2016-01-01

    Klebsiella pneumoniae is the predominant pathogen isolated from liver abscesses of diabetic patients in Asian countries. However, the effects of elevated blood glucose levels on the virulence of this pathogen remain largely unknown. Type 3 fimbriae, encoded by the mrkABCDF genes, are important virulence factors in K. pneumoniae pathogenesis. In this study, the effects of exogenous glucose and the intracellular cyclic AMP (cAMP) signaling pathway on type 3 fimbriae expression regulation were investigated. The production of MrkA, the major subunit of type 3 fimbriae, was increased in glucose-rich medium, whereas cAMP supplementation reversed the effect. MrkA production was markedly increased by cyaA or crp deletion, but slightly decreased by cpdA deletion. In addition, the mRNA levels of mrkABCDF genes and the activity of PmrkA were increased in Δcrp strain, as well as the mRNA levels of mrkHIJ genes that encode cyclic di-GMP (c-di-GMP)-related regulatory proteins that influence type 3 fimbriae expression. Moreover, the activities of PmrkHI and PmrkJ were decreased in ΔlacZΔcrp strain. These results indicate that CRP-cAMP down-regulates mrkABCDF and mrkHIJ at the transcriptional level. Further deletion of mrkH or mrkI in Δcrp strain diminished the production of MrkA, indicating that MrkH and MrkI are required for the CRP regulation of type 3 fimbriae expression. Furthermore, the high activity of PmrkHI in the ΔlacZΔcrp strain was diminished in ΔlacZΔcrpΔmrkHI, but increased in the ΔlacZΔcrpΔmrkJ strain. Deletion of crp increased the intracellular c-di-GMP concentration and reduced the phosphodiesterase activity. Moreover, we found that the mRNA levels of multiple genes related to c-di-GMP metabolism were altered in Δcrp strain. These indicate that CRP regulates type 3 fimbriae expression indirectly via the c-di-GMP signaling pathway. In conclusion, we found evidence of a coordinated regulation of type 3 fimbriae expression by the CRP-cAMP and c-di-GMP

  10. CRP-Cyclic AMP Regulates the Expression of Type 3 Fimbriae via Cyclic di-GMP in Klebsiella pneumoniae.

    Science.gov (United States)

    Lin, Ching-Ting; Lin, Tien-Huang; Wu, Chien-Chen; Wan, Lei; Huang, Chun-Fa; Peng, Hwei-Ling

    2016-01-01

    Klebsiella pneumoniae is the predominant pathogen isolated from liver abscesses of diabetic patients in Asian countries. However, the effects of elevated blood glucose levels on the virulence of this pathogen remain largely unknown. Type 3 fimbriae, encoded by the mrkABCDF genes, are important virulence factors in K. pneumoniae pathogenesis. In this study, the effects of exogenous glucose and the intracellular cyclic AMP (cAMP) signaling pathway on type 3 fimbriae expression regulation were investigated. The production of MrkA, the major subunit of type 3 fimbriae, was increased in glucose-rich medium, whereas cAMP supplementation reversed the effect. MrkA production was markedly increased by cyaA or crp deletion, but slightly decreased by cpdA deletion. In addition, the mRNA levels of mrkABCDF genes and the activity of PmrkA were increased in Δcrp strain, as well as the mRNA levels of mrkHIJ genes that encode cyclic di-GMP (c-di-GMP)-related regulatory proteins that influence type 3 fimbriae expression. Moreover, the activities of PmrkHI and PmrkJ were decreased in ΔlacZΔcrp strain. These results indicate that CRP-cAMP down-regulates mrkABCDF and mrkHIJ at the transcriptional level. Further deletion of mrkH or mrkI in Δcrp strain diminished the production of MrkA, indicating that MrkH and MrkI are required for the CRP regulation of type 3 fimbriae expression. Furthermore, the high activity of PmrkHI in the ΔlacZΔcrp strain was diminished in ΔlacZΔcrpΔmrkHI, but increased in the ΔlacZΔcrpΔmrkJ strain. Deletion of crp increased the intracellular c-di-GMP concentration and reduced the phosphodiesterase activity. Moreover, we found that the mRNA levels of multiple genes related to c-di-GMP metabolism were altered in Δcrp strain. These indicate that CRP regulates type 3 fimbriae expression indirectly via the c-di-GMP signaling pathway. In conclusion, we found evidence of a coordinated regulation of type 3 fimbriae expression by the CRP-cAMP and c-di-GMP

  11. Potentiation of cGMP signaling increases oxygen delivery and oxidative metabolism in contracting skeletal muscle of older but not young humans

    DEFF Research Database (Denmark)

    Nyberg, Michael Permin; Piil, Peter Bergmann; Egelund, Jon;

    2015-01-01

    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...... in the older subjects correlated with the increase in leg O2 uptake (r (2) = 0.843). These findings suggest an insufficient O2 delivery to the contracting skeletal muscle of aged individuals and that reduced cGMP availability is a novel mechanism underlying impaired skeletal muscle perfusion with advancing age....

  12. The exopolysaccharide gene cluster Bcam1330-Bcam1341 is involved in Burkholderia cenocepacia biofilm formation, and its expression is regulated by c-di-GMP and Bcam1349

    DEFF Research Database (Denmark)

    Fazli, Mustafa; McCarthy, Yvonne; Givskov, Michael;

    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...

  13. “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

    Institute of Scientific and Technical Information of China (English)

    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.

  14. 浅谈制药企业实施GMP的三要素%Three Elements for Pharmaceutical Companies to Implement GMP

    Institute of Scientific and Technical Information of China (English)

    葛兰英

    2014-01-01

    This paper reviews the development since China implement GMP, elaborates the three elements of pharmaceutical companies to implement GMP and the quality of personnel is a key element.%本文回顾了我国实施GMP以来的发展历程,阐述了制药企业实施GMP的三要素及人员素质是关键要素。

  15. Identification of PDE9 as a cGMP-specific phosphodiesterase in germinal vesicle oocytes: A proposed role in the resumption of meiosis

    Science.gov (United States)

    Hanna, Carol B.; Yao, Shan; Wu, Xuemei; Jensen, Jeffrey T.

    2012-01-01

    Objective To identify a cGMP-specific phosphodiesterase (PDE) in non-human primate germinal vesicle (GV) oocytes and establish a proposed effect on oocyte maturation through preliminary experiments in mouse GV oocytes. Design Controlled non-human primate and rodent experiments. Setting Academic research institution. Animals Rhesus macaques and B6/129F1 mice. Interventions Rhesus macaques were stimulated with FSH to collect GV oocytes and cumulus for gene expression analysis. Female mice were stimulated with PMSG to collect GV oocytes. Main Outcome Measures PDE transcript expression in primate GV oocytes and cumulus cells. Fluorescence polarization (FP) measurements of PDE3A activity. Spontaneous resumption of meiosis in mouse GV oocytes. Results Five PDE transcripts were detected in Rhesus GV oocytes, only PDE9A was cGMP-specific. FP assays indicated cGMP has an inhibitory effect on PDE3A while the PDE9 inhibitor, BAY73-6691, did not. Similarly, BAY73-6691, had little effect on preventing spontaneous maturation in oocytes, but did augment the inhibitory effects of cGMP. Inclusion of 0µM (control), 10µM, 100µM, and 1 mM BAY73-6691 significantly increased the proportion of mouse oocytes maintaining GV arrest in the presence of the cGMP analog 8-Br-cGMP at: 100µM (8.8%, 11.4%, 18.8%, and 28%), 500µM (21.1%, 38.1%, 74.5%,and 66.5%), and 1 mM (57.8%, 74.5%, 93.9%, and 94.0%) respectively, when P<0.05. Conclusions PDE9 is a cGMP-specific hydrolyzing enzyme present in primate oocytes, and PDE9 antagonists augment the inhibitory effect of cGMP during spontaneous in vitro maturation of GV mouse oocytes. PMID:22704629

  16. 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

    Institute of Scientific and Technical Information of China (English)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  19. Identification of a negative feedback loop between cyclic di-GMP-induced levels of IFI16 and p202 cytosolic DNA sensors and STING.

    Science.gov (United States)

    Panchanathan, Ravichandran; Liu, Hongzhu; Xin, Duan; Choubey, Divaker

    2014-10-01

    A host type I IFN response is induced by cytosolic sensing of the bacterial second messenger cyclic-di-GMP (c-di-GMP) by STING (stimulator of IFN genes). Because the STING, an adaptor protein, links the cytosolic detection of DNA by the cytosolic DNA sensors such as the IFN-inducible human IFI16 and murine p202 proteins to the TBK1/IRF3 axis, we investigated whether c-di-GMP-induced signaling could regulate expression of IFI16 and p202 proteins. Here, we report that activation of c-di-GMP-induced signaling in human and murine cells increased steady-state levels of IFI16 and p202 proteins. The increase was c-di-GMP concentration- and time-dependent. Unexpectedly, treatment of cells with type I IFN decreased levels of the adaptor protein STING. Therefore, we investigated whether the IFI16 or p202 protein could regulate the expression of STING and activation of the TBK1/IRF3 axis. We found that constitutive knockdown of IFI16 or p202 expression in cells increased steady-state levels of STING. Additionally, the knockdown of IFI16 resulted in activation of the TBK1/IRF3 axis. Accordingly, increased levels of the IFI16 or p202 protein in cells decreased STING levels. Together, our observations identify a novel negative feedback loop between c-di-GMP-induced levels of IFI16 and p202 cytosolic DNA sensors and the adaptor protein STING.

  20. Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa.

    Science.gov (United States)

    Rugjee, Kushal N; An, Shi-Qi; Ryan, Robert P

    2016-01-01

    Bacterial resistance to traditional antibiotics has driven research attempts to identify new drug targets in recently discovered regulatory pathways. Regulatory systems that utilize intracellular cyclic di-GMP (c-di-GMP) as a second messenger are one such class of target. c-di-GMP is a signaling molecule found in almost all bacteria that acts to regulate an extensive range of processes including antibiotic resistance, biofilm formation and virulence. The understanding of how c-di-GMP signaling controls aspects of antibiotic resistant biofilm development has suggested approaches whereby alteration of the cellular concentrations of the nucleotide or disruption of these signaling pathways may lead to reduced biofilm formation or increased susceptibility of the biofilms to antibiotics. We describe a simple high-throughput bioreporter protocol, based on green fluorescent protein (GFP), whose expression is under the control of the c-di-GMP responsive promoter cdrA, to rapidly screen for small molecules with the potential to modulate c-di-GMP cellular levels in Pseudomonas aeruginosa (P. aeruginosa). This simple protocol can screen upwards of 3,500 compounds within 48 hours and has the ability to be adapted to multiple microorganisms.

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

    Science.gov (United States)

    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

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

    Directory of Open Access Journals (Sweden)

    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.

  3. Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa

    Science.gov (United States)

    Rugjee, Kushal N.; An, Shi-qi; Ryan, Robert P.

    2016-01-01

    Bacterial resistance to traditional antibiotics has driven research attempts to identify new drug targets in recently discovered regulatory pathways. Regulatory systems that utilize intracellular cyclic di-GMP (c-di-GMP) as a second messenger are one such class of target. c-di-GMP is a signaling molecule found in almost all bacteria that acts to regulate an extensive range of processes including antibiotic resistance, biofilm formation and virulence. The understanding of how c-di-GMP signaling controls aspects of antibiotic resistant biofilm development has suggested approaches whereby alteration of the cellular concentrations of the nucleotide or disruption of these signaling pathways may lead to reduced biofilm formation or increased susceptibility of the biofilms to antibiotics. We describe a simple high-throughput bioreporter protocol, based on green fluorescent protein (GFP), whose expression is under the control of the c-di-GMP responsive promoter cdrA, to rapidly screen for small molecules with the potential to modulate c-di-GMP cellular levels in Pseudomonas aeruginosa (P. aeruginosa). This simple protocol can screen upwards of 3,500 compounds within 48 hours and has the ability to be adapted to multiple microorganisms. PMID:27404278

  4. Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa.

    Science.gov (United States)

    Rugjee, Kushal N; An, Shi-Qi; Ryan, Robert P

    2016-01-01

    Bacterial resistance to traditional antibiotics has driven research attempts to identify new drug targets in recently discovered regulatory pathways. Regulatory systems that utilize intracellular cyclic di-GMP (c-di-GMP) as a second messenger are one such class of target. c-di-GMP is a signaling molecule found in almost all bacteria that acts to regulate an extensive range of processes including antibiotic resistance, biofilm formation and virulence. The understanding of how c-di-GMP signaling controls aspects of antibiotic resistant biofilm development has suggested approaches whereby alteration of the cellular concentrations of the nucleotide or disruption of these signaling pathways may lead to reduced biofilm formation or increased susceptibility of the biofilms to antibiotics. We describe a simple high-throughput bioreporter protocol, based on green fluorescent protein (GFP), whose expression is under the control of the c-di-GMP responsive promoter cdrA, to rapidly screen for small molecules with the potential to modulate c-di-GMP cellular levels in Pseudomonas aeruginosa (P. aeruginosa). This simple protocol can screen upwards of 3,500 compounds within 48 hours and has the ability to be adapted to multiple microorganisms. PMID:27404278

  5. Comparative Study of GMP Used in China and Japan%我国GMP与日本GMP的比较研究

    Institute of Scientific and Technical Information of China (English)

    梁毅; 郦旻; 付倩

    2012-01-01

    目的:对日本GMP的发展、框架、内容和特色进行研究与探讨.方法:就日本GMP与我国新修订的GMP有关内容进行对比研究结合.结论:借鉴日本GMP,对我国GMP实施有很大的帮助.

  6. In Vivo Synthesis of Cyclic-di-GMP Using a Recombinant Adenovirus Preferentially Improves Adaptive Immune Responses against Extracellular Antigens.

    Science.gov (United States)

    Alyaqoub, Fadel S; Aldhamen, Yasser A; Koestler, Benjamin J; Bruger, Eric L; Seregin, Sergey S; Pereira-Hicks, Cristiane; Godbehere, Sarah; Waters, Christopher M; Amalfitano, Andrea

    2016-02-15

    There is a compelling need for more effective vaccine adjuvants to augment induction of Ag-specific adaptive immune responses. Recent reports suggested the bacterial second messenger bis-(3'-5')-cyclic-dimeric-guanosine monophosphate (c-di-GMP) acts as an innate immune system modulator. We recently incorporated a Vibrio cholerae diguanylate cyclase into an adenovirus vaccine, fostering production of c-di-GMP as well as proinflammatory responses in mice. In this study, we recombined a more potent diguanylate cyclase gene, VCA0848, into a nonreplicating adenovirus serotype 5 (AdVCA0848) that produces elevated amounts of c-di-GMP when expressed in mammalian cells in vivo. This novel platform further improved induction of type I IFN-β and activation of innate and adaptive immune cells early after administration into mice as compared with control vectors. Coadministration of the extracellular protein OVA and the AdVCA0848 adjuvant significantly improved OVA-specific T cell responses as detected by IFN-γ and IL-2 ELISPOT, while also improving OVA-specific humoral B cell adaptive responses. In addition, we found that coadministration of AdVCA0848 with another adenovirus serotype 5 vector expressing the HIV-1-derived Gag Ag or the Clostridium difficile-derived toxin B resulted in significant inhibitory effects on the induction of Gag and toxin B-specific adaptive immune responses. As a proof of principle, these data confirm that in vivo synthesis of c-di-GMP stimulates strong innate immune responses that correlate with enhanced adaptive immune responses to concomitantly administered extracellular Ag, which can be used as an adjuvant to heighten effective immune responses for protein-based vaccine platforms against microbial infections and cancers.

  7. Cardioprotection by H2S engages a cGMP-dependent protein kinase G/phospholamban pathway

    Science.gov (United States)

    Bibli, Sofia-Iris; Andreadou, Ioanna; Chatzianastasiou, Athanasia; Tzimas, Christos; Sanoudou, Despina; Kranias, Evangelia; Brouckaert, Peter; Coletta, Ciro; Szabo, Csaba; Kremastinos, Dimitrios Th.; Iliodromitis, Efstathios K.; Papapetropoulos, Andreas

    2015-01-01

    Aims H2S is known to confer cardioprotection; however, the pathways mediating its effects in vivo remain incompletely understood. The purpose of the present study is to evaluate the contribution of cGMP-regulated pathways in the infarct-limiting effect of H2S in vivo. Methods and results Anaesthetized rabbits were subjected to myocardial ischaemia (I)/reperfusion (R), and infarct size was determined in control or H2S-exposed groups. The H2S donor sodium hydrosulfide (NaHS, an agent that generates H2S) increased cardiac cGMP and reduced the infarct size. The cGMP-dependent protein kinase (PKG)-I inhibitor DT2 abrogated the protective effect of NaHS, whereas the control peptide TAT or l-nitroarginine methyl ester (l-NAME) did not alter the effect of NaHS. Moreover, the KATP channel inhibitor, glibenclamide, partially reversed the effects of NaHS, whereas inhibition of mitochondrial KATP did not modify the NaHS response. NaHS enhanced phosphorylation of phospholamban (PLN), in a PKG-dependent manner. To further investigate the role of PLN in H2S-mediated cardioprotection, wild-type and PLN KO mice underwent I/R. NaHS did not exert cardioprotection in PLN KO mice. Unlike what was observed in rabbits, genetic or pharmacological inhibition of eNOS abolished the infarct-limiting effect of NaHS in mice. Conclusions Our findings demonstrate (i) that administration of NaHS induces cardioprotection via a cGMP/PKG/PLN pathway and (ii) contribution of nitric oxide to the H2S response is species-specific. PMID:25870184

  8. Both cyclic-AMP and cyclic-GMP can act as regulators of the phenylpropanoid pathway in Arabidopsis thaliana seedlings.

    Science.gov (United States)

    Pietrowska-Borek, Małgorzata; Nuc, Katarzyna

    2013-09-01

    Cyclic nucleotides (cAMP and cGMP) are important signaling molecules that control a range of cellular functions and modulate different reactions. It is known that under abiotic or biotic stress plant cells synthesize these nucleotides and that they also enhance the activity of the phenylpropanoid pathway. Wondering what is the relation between these two facts, we investigated how the exogenously applied membrane-permeable derivatives, 8-Br-cAMP or 8-Br-cGMP, which are believed to act as the original cyclic nucleotides, affect the expression of the genes for and the specific activity of three enzymes of the phenylpropanoid pathway in Arabidopsis thaliana seedlings. We found that the expression of the genes of phenylalanine ammonia-lyase (PAL2), 4-coumarate:coenzyme A ligase (4CL1) and chalcone synthase (CHS), and the specific activities of PAL (EC 4.3.1.5), 4CL (EC 6.2.1.12) and CHS (EC 2.3.1.74) were induced in the same way by either of these cyclic nucleotides used at 5 μM concentration. None of the possible cAMP and cGMP degradation products (AMP, GMP, adenosine or guanosine) evoked such effects. Expression of PAL1, 4CL2 and 4CL3 were practically not affected. Although the investigated nucleotides induced rapid expression of the aforementioned enzymes, they did not affect the level of anthocyanins within the same period. We discuss the effects exerted by the exogenously administered cyclic nucleotides, their relation with stress and the role which the phenylpropanoid pathways the cyclic nucleotides may play in plants.

  9. Nitric oxide increases mitochondrial respiration in a cGMP-dependent manner in the callus from Arabidopsis thaliana.

    Science.gov (United States)

    Wang, Xiaomin; Li, Jisheng; Liu, Jie; He, Wenliang; Bi, Yurong

    2010-11-01

    Nitric oxide (NO) acts as a key molecule in many physiological processes in plants. In this study, the roles of NO in mitochondrial respiration were investigated in the calli from wild-type Arabidopsis and NO associated 1 mutant (Atnoa1) which has a reduced endogenous NO level. Long-term exposure of wild-type Arabidopsis callus to sodium nitroprusside (SNP) increased mitochondrial respiration in both cytochrome and alternative pathways. In Atnoa1 callus, the capacity of both the cytochrome pathway and the alternative pathway was lower than that in wild-type callus. Further study indicated that NO enhanced the transcript abundance of genes encoding mitochondrial respiration-chain proteins as well as the protein expression of the NADH-ubiquinone reductase 75 kDa subunit and the alternative oxidase 1/2 in wild-type and Atnoa1 calli. 2-Phenyl-4,4,5,5-tetremethy-limidazolinone-1-oxyl-3-oxide (PTIO), a NO scavenger, inhibited the effects of NO in both calli. Co-incubation of callus with 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), a guanylate cyclase inhibitor, also abolished NO effects. The membrane-permeable cGMP analog 8Br-cGMP mimicked NO effects. Moreover, the alternative pathway showed a higher sensitivity to the cellular cGMP changes than the cytochrome pathway did in gene transcription, protein expression and O(2) consumption. Taken together, NO could enhance mitochondrial respiration in both cytochrome and alternative pathways in a cGMP-dependent manner in Arabidopsis.

  10. NZ-GMP Approved Serum Improve hDPSC Osteogenic Commitment and Increase Angiogenic Factor Expression

    Science.gov (United States)

    Spina, Anna; Montella, Roberta; Liccardo, Davide; De Rosa, Alfredo; Laino, Luigi; Mitsiadis, Thimios A.; La Noce, Marcella

    2016-01-01

    Human dental pulp stem cells (hDPSCs), selected from the stromal-vascular fraction of dental pulp, are ecto-mesenchymal stem cells deriving from neural crests, successfully used in human bone tissue engineering. For their use in human therapy GMP procedures are required. For instance, the use of fetal bovine serum (FBS) is strongly discouraged in clinical practice due to its high risk of prions and other infections for human health. Alternatively, clinical grade sera have been suggested, including the New Zealand FBS (NZ-FBS). Therefore, the aim of this study was to evaluate the behavior of hDPSCs expanded in culture medium containing NZ-FBS. Since it was widely demonstrated hDPSCs display relevant capabilities to differentiate into osteogenic and angiogenic lineages, we performed a comparative study to assess if these features are also retained by cultivating the cells with a safer serum never tested on this cell line. hDPSCs were grown using NZ-FBS and conventional (C-FBS) for 7, 14, and 21 days, in both 2D and 3D cultures. Growth curves, expression of bone-related markers, calcification and angiogenesis were evaluated. NZ-FBS induced significant cell growth with respect to C-FBS and promoted an earlier increase expression of osteogenic markers, in particular of those involved in the formation of mineralized matrix (BSP and OPN) within 14 days. In addition, hDPSCs cultured in presence of NZ-FBS were found to produce higher mRNA levels of the angiogenic factors, such as VEGF and PDGFA. Taken together, our results highlight that hDPSCs proliferate, enhance their osteogenic commitment and increase angiogenic factors in NZ-FBS containing medium. These features have also been found when hDPSC were seeded on the clinical-grade collagen I scaffold (Bio-Gide®), leading to the conclusion that for human therapy some procedures and above all the use of GMP-approved materials have no negative impact. PMID:27594842

  11. cGMP production of patient-specific iPSCs and photoreceptor precursor cells to treat retinal degenerative blindness.

    Science.gov (United States)

    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

  12. GMP synthase is essential for viability and infectivity of Trypanosoma brucei despite a redundant purine salvage pathway.

    Science.gov (United States)

    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. Islet isolation and GMP, ISO 9001:2000: what do we need--a 3-year experience.

    Science.gov (United States)

    Hengster, P; Hermann, M; Pirkebner, D; Draxl, A; Margreiter, R

    2005-10-01

    Pancreatic islet cell isolation and transplantation has been performed for many years at several institutions. Although all institutions aim to produce high-quality islets, applied standards widely deviate from standards in the pharmaceutical industry. The legal situation within the European Union has changed requirements for setting up and running such a laboratory. The process is now clearly defined as a production of a pharmaceutical and therefore must be licensed by federal authorities. Analysis of workload for establishing an islet isolation program that fulfil GMP and ISO 9001 criteria including an estimation of costs and the impact of such a system on the isolation process. The definition of quality parameters and documentation is a central issue of all islet isolation laboratories. Therefore, GMP and ISO 9001:2000 do not add additional work per se. On the other hand, clear guidelines, a clear policy, working place descriptions, forms, checklists, and, particularly standard operating procedures, are instrumental for smooth functioning within the department. Collection of data such as errors, improvement measures, and preventive measures reduces subsequent costs. A clear definition of responsibilities minimizes organizational problems. Steering of inspection devices prevents bias errors and validating the processes clearly points out incorrect assumptions. Documentation helps to prove the correctness of the production at any time and is of use also for scientific evaluations. We strongly feel that GMP criteria are mandatory and together with an ISO 9001:2000 quality management system offers significant advantages for the process of islet isolation and a continuous improvement process.

  14. ADOPTION OF GOOD MANAGEMENT PRACTICE (GMP IN SMALL AND MEDIUM SCALE VANNAMEI SHRIMP FARMS ON THE NORTHERN SHORE EAST JAVA

    Directory of Open Access Journals (Sweden)

    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

  15. The structure of substrate-free microbial ribonuclease binase and of its complexes with 3'GMP and sulfate ions.

    Science.gov (United States)

    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

  16. Homologs of the LapD-LapG c-di-GMP Effector System Control Biofilm Formation by Bordetella bronchiseptica.

    Directory of Open Access Journals (Sweden)

    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

    Science.gov (United States)

    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. Next-generation RNA-based fluorescent biosensors enable anaerobic detection of cyclic di-GMP.

    Science.gov (United States)

    Wang, Xin C; Wilson, Stephen C; Hammond, Ming C

    2016-09-30

    Bacteria occupy a diverse set of environmental niches with differing oxygen availability. Anaerobic environments such as mammalian digestive tracts and industrial reactors harbor an abundance of both obligate and facultative anaerobes, many of which play significant roles in human health and biomanufacturing. Studying bacterial function under partial or fully anaerobic conditions, however, is challenging given the paucity of suitable live-cell imaging tools. Here, we introduce a series of RNA-based fluorescent biosensors that respond selectively to cyclic di-GMP, an intracellular bacterial second messenger that controls cellular motility and biofilm formation. We demonstrate the utility of these biosensors in vivo under both aerobic and anaerobic conditions, and we show that biosensor expression does not interfere with the native motility phenotype. Together, our results attest to the effectiveness and versatility of RNA-based fluorescent biosensors, priming further development and application of these and other analogous sensors to study host-microbial and microbial-microbial interactions through small molecule signals. PMID:27382070

  19. Diurnal cycles in serotonin acetyltransferase activity and cyclic GMP content of cultured chick pineal glands.

    Science.gov (United States)

    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

  20. GMP-grade platelet lysate enhances proliferation and migration of tenon fibroblasts.

    Science.gov (United States)

    Carducci, Augusto; Scafetta, Gaia; Siciliano, Camilla; Carnevale, Roberto; Rosa, Paolo; Coccia, Andrea; Mangino, Giorgio; Bordin, Antonella; Vingolo, Enzo Maria; Pierelli, Luca; Lendaro, Eugenio; Ragona, Giuseppe; Frati, Giacomo; De Falco, Elena

    2016-01-01

    Tenon's fibroblasts (TFs), widely employed as in vitro model for many ophthalmological studies, are routinely cultured with FBS. Platelet Lysate (PL), a hemoderivate enriched with growth factors and cytokines has been largely tested in several clinical applications and as substitute of FBS in culture. Here, we investigate whether PL can exert biological effects on TF populations similarly to other cell types. Results show that PL significantly enhances cell proliferation and migration vs. FBS, without influencing cell size/granularity. Upregulation of EGF, VEGF, KDR, MMP2-9, FAK mRNA levels also occurs and phosphorylation of AKT but not of ERK1/2 is significantly enhanced. The inhibition of the PI3kinase/AKT pathway with the specific inhibitor wortmannin, decreases PL-induced cell migration but not proliferation. Condition supernatants containing PL show increased bioavailability of Nitric Oxide and reduced levels of 8-Iso-PGF2-alpha, correlating with cell proliferation and migration. Pro-angiogenic/inflammatory soluble factors (GRO, Angiogenin, EGF, I-309, PARC) are exclusively or greater expressed in media containing PL than FBS. GMP-grade PL preparations positively influence in vitro biological effects of TFs representing a suitable and safer alternative to FBS. PMID:26709648

  1. Inherent regulation of EAL domain-catalyzed hydrolysis of second messenger cyclic di-GMP.

    Science.gov (United States)

    Sundriyal, Amit; Massa, Claudia; Samoray, Dietrich; Zehender, Fabian; Sharpe, Timothy; Jenal, Urs; Schirmer, Tilman

    2014-03-01

    The universal second messenger cyclic di-GMP (cdG) is involved in the regulation of a diverse range of cellular processes in bacteria. The intracellular concentration of the dinucleotide is determined by the opposing actions of diguanylate cyclases and cdG-specific phosphodiesterases (PDEs). Whereas most PDEs have accessory domains that are involved in the regulation of their activity, the regulatory mechanism of this class of enzymes has remained unclear. Here, we use biophysical and functional analyses to show that the isolated EAL domain of a PDE from Escherichia coli (YahA) is in a fast thermodynamic monomer-dimer equilibrium, and that the domain is active only in its dimeric state. Furthermore, our data indicate thermodynamic coupling between substrate binding and EAL dimerization with the dimerization affinity being increased about 100-fold upon substrate binding. Crystal structures of the YahA-EAL domain determined under various conditions (apo, Mg(2+), cdG·Ca(2+) complex) confirm structural coupling between the dimer interface and the catalytic center. The built-in regulatory properties of the EAL domain probably facilitate its modular, functional combination with the diverse repertoire of accessory domains.

  2. Diguanylate cyclase null mutant reveals that C-Di-GMP pathway regulates the motility and adherence of the extremophile bacterium Acidithiobacillus caldus.

    Science.gov (United States)

    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.

  3. C-di-GMP Regulates Motile to Sessile Transition by Modulating MshA Pili Biogenesis and Near-Surface Motility Behavior in Vibrio cholerae.

    Science.gov (United States)

    Jones, Christopher J; Utada, Andrew; Davis, Kimberly R; Thongsomboon, Wiriya; Zamorano Sanchez, David; Banakar, Vinita; Cegelski, Lynette; Wong, Gerard C L; Yildiz, Fitnat H

    2015-10-01

    In many bacteria, including Vibrio cholerae, cyclic dimeric guanosine monophosphate (c-di-GMP) controls the motile to biofilm life style switch. Yet, little is known about how this occurs. In this study, we report that changes in c-di-GMP concentration impact the biosynthesis of the MshA pili, resulting in altered motility and biofilm phenotypes in V. cholerae. Previously, we reported that cdgJ encodes a c-di-GMP phosphodiesterase and a ΔcdgJ mutant has reduced motility and enhanced biofilm formation. Here we show that loss of the genes required for the mannose-sensitive hemagglutinin (MshA) pilus biogenesis restores motility in the ΔcdgJ mutant. Mutations of the predicted ATPase proteins mshE or pilT, responsible for polymerizing and depolymerizing MshA pili, impair near surface motility behavior and initial surface attachment dynamics. A ΔcdgJ mutant has enhanced surface attachment, while the ΔcdgJmshA mutant phenocopies the high motility and low attachment phenotypes observed in a ΔmshA strain. Elevated concentrations of c-di-GMP enhance surface MshA pilus production. MshE, but not PilT binds c-di-GMP directly, establishing a mechanism for c-di-GMP signaling input in MshA pilus production. Collectively, our results suggest that the dynamic nature of the MshA pilus established by the assembly and disassembly of pilin subunits is essential for transition from the motile to sessile lifestyle and that c-di-GMP affects MshA pilus assembly and function through direct interactions with the MshE ATPase.

  4. Diguanylate cyclase null mutant reveals that C-Di-GMP pathway regulates the motility and adherence of the extremophile bacterium Acidithiobacillus caldus.

    Directory of Open Access Journals (Sweden)

    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.

  5. Annual report

    International Nuclear Information System (INIS)

    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

  6. Raising the standard: changes to the Australian Code of Good Manufacturing Practice (cGMP) for human blood and blood components, human tissues and human cellular therapy products.

    Science.gov (United States)

    Wright, Craig; Velickovic, Zlatibor; Brown, Ross; Larsen, Stephen; Macpherson, Janet L; Gibson, John; Rasko, John E J

    2014-04-01

    In Australia, manufacture of blood, tissues and biologicals must comply with the federal laws and meet the requirements of the Therapeutic Goods Administration (TGA) Manufacturing Principles as outlined in the current Code of Good Manufacturing Practice (cGMP). The Therapeutic Goods Order (TGO) No. 88 was announced concurrently with the new cGMP, as a new standard for therapeutic goods. This order constitutes a minimum standard for human blood, tissues and cellular therapeutic goods aimed at minimising the risk of infectious disease transmission. The order sets out specific requirements relating to donor selection, donor testing and minimisation of infectious disease transmission from collection and manufacture of these products. The Therapeutic Goods Manufacturing Principles Determination No. 1 of 2013 references the human blood and blood components, human tissues and human cellular therapy products 2013 (2013 cGMP). The name change for the 2013 cGMP has allowed a broadening of the scope of products to include human cellular therapy products. It is difficult to directly compare versions of the code as deletion of some clauses has not changed the requirements to be met, as they are found elsewhere amongst the various guidelines provided. Many sections that were specific for blood and blood components are now less prescriptive and apply to a wider range of cellular therapies, but the general overall intent remains the same. Use of 'should' throughout the document instead of 'must' allows flexibility for alternative processes, but these systems will still require justification by relevant logical argument and validation data to be acceptable to TGA. The cGMP has seemingly evolved so that specific issues identified at audit over the last decade have now been formalised in the new version. There is a notable risk management approach applied to most areas that refer to process justification and decision making. These requirements commenced on 31 May 2013 and a 12 month

  7. Time-dependent inhibitory effects of cGMP-analogues on thrombin-induced platelet-derived microparticles formation, platelet aggregation, and P-selectin expression

    Energy Technology Data Exchange (ETDEWEB)

    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.

  8. Characterization of the cGMP-dependent protein kinase SmcGK1 of Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    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

  9. The impact of cGMP compliance on consumer confidence in dietary supplement products.

    Science.gov (United States)

    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

  10. The impact of cGMP compliance on consumer confidence in dietary supplement products

    International Nuclear Information System (INIS)

    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

  11. Bacterial rotary export ATPases are allosterically regulated by the nucleotide second messenger cyclic-di-GMP.

    Science.gov (United States)

    Trampari, Eleftheria; Stevenson, Clare E M; Little, Richard H; Wilhelm, Thomas; Lawson, David M; Malone, Jacob G

    2015-10-01

    The widespread second messenger molecule cyclic di-GMP (cdG) regulates the transition from motile and virulent lifestyles to sessile, biofilm-forming ones in a wide range of bacteria. Many pathogenic and commensal bacterial-host interactions are known to be controlled by cdG signaling. Although the biochemistry of cyclic dinucleotide metabolism is well understood, much remains to be discovered about the downstream signaling pathways that induce bacterial responses upon cdG binding. As part of our ongoing research into the role of cdG signaling in plant-associated Pseudomonas species, we carried out an affinity capture screen for cdG binding proteins in the model organism Pseudomonas fluorescens SBW25. The flagella export AAA+ ATPase FliI was identified as a result of this screen and subsequently shown to bind specifically to the cdG molecule, with a KD in the low micromolar range. The interaction between FliI and cdG appears to be very widespread. In addition to FliI homologs from diverse bacterial species, high affinity binding was also observed for the type III secretion system homolog HrcN and the type VI ATPase ClpB2. The addition of cdG was shown to inhibit FliI and HrcN ATPase activity in vitro. Finally, a combination of site-specific mutagenesis, mass spectrometry, and in silico analysis was used to predict that cdG binds to FliI in a pocket of highly conserved residues at the interface between two FliI subunits. Our results suggest a novel, fundamental role for cdG in controlling the function of multiple important bacterial export pathways, through direct allosteric control of export ATPase proteins.

  12. The cGMP signaling pathway affects feeding behavior in the necromenic nematode Pristionchus pacificus.

    Directory of Open Access Journals (Sweden)

    Silvina M Kroetz

    Full Text Available BACKGROUND: The genetic tractability and the species-specific association with beetles make the nematode Pristionchus pacificus an exciting emerging model organism for comparative studies in development and behavior. P. pacificus differs from Caenorhabditis elegans (a bacterial feeder by its buccal teeth and the lack of pharyngeal grinders, but almost nothing is known about which genes coordinate P. pacificus feeding behaviors, such as pharyngeal pumping rate, locomotion, and fat storage. METHODOLOGY/PRINCIPAL FINDINGS: We analyzed P. pacificus pharyngeal pumping rate and locomotion behavior on and off food, as well as on different species of bacteria (Escherichia coli, Bacillus subtilis, and Caulobacter crescentus. We found that the cGMP-dependent protein kinase G (PKG Ppa-EGL-4 in P. pacificus plays an important role in regulating the pumping rate, mouth form dimorphism, the duration of forward locomotion, and the amount of fat stored in intestine. In addition, Ppa-EGL-4 interacts with Ppa-OBI-1, a recently identified protein involved in chemosensation, to influence feeding and locomotion behavior. We also found that C. crescentus NA1000 increased pharyngeal pumping as well as fat storage in P. pacificus. CONCLUSIONS: The PKG EGL-4 has conserved functions in regulating feeding behavior in both C. elegans and P. pacificus nematodes. The Ppa-EGL-4 also has been co-opted during evolution to regulate P. pacificus mouth form dimorphism that indirectly affect pharyngeal pumping rate. Specifically, the lack of Ppa-EGL-4 function increases pharyngeal pumping, time spent in forward locomotion, and fat storage, in part as a result of higher food intake. Ppa-OBI-1 functions upstream or parallel to Ppa-EGL-4. The beetle-associated omnivorous P. pacificus respond differently to changes in food state and food quality compared to the exclusively bacteriovorous C. elegans.

  13. GMP-compliant isolation and large-scale expansion of bone marrow-derived MSC.

    Directory of Open Access Journals (Sweden)

    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

  14. Regulation of Motility and Phenazine Pigment Production by FliA Is Cyclic-di-GMP Dependent in Pseudomonas aeruginosa PAO1

    Science.gov (United States)

    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

  15. Regulation of Motility and Phenazine Pigment Production by FliA Is Cyclic-di-GMP Dependent in Pseudomonas aeruginosa PAO1.

    Science.gov (United States)

    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.

  16. 制药企业实施新版GMP的思考%Thoughts on Application of New Version of GMP to Pharmaceutical Enterprises

    Institute of Scientific and Technical Information of China (English)

    黄海燕; 马丽芳

    2012-01-01

    Relevant suggestion on the application of new version of GMP was presented according to the rules of new version of GMP and the problems met during the process of application of GMP to pharmaceutical Enterprises.In order to successfully propel the application of new version of GMP,it needed to realize rules of GMP deeply,and combine it to the practice during manufacturing.Besides,it was also useful to construct quality management system,which emphasized on control of process and self inspection.%根据新版GMP的相关规定及当前制药企业实施GMP过程中存在的一些问题,本文对新版药品生产质量管理规范的实施提出了相关的建议。要顺利推进新版GMP实施,必须以深刻认识GMP规范;与生产实际相结合;建设质量管理体系;强调过程控制、加强GMP自检为基础。

  17. Derivation of vascular endothelial cells from human embryonic stem cells under GMP-compliant conditions: towards clinical studies in ischaemic disease.

    Science.gov (United States)

    Kaupisch, A; Kennedy, L; Stelmanis, V; Tye, B; Kane, N M; Mountford, J C; Courtney, A; Baker, A H

    2012-10-01

    Revascularisation of ischaemic tissue remains an area of substantial unmet clinical need in cardiovascular disease. Strategies to induce therapeutic angiogenesis are therefore attractive. Our recent focus has been on human embryonic stem cell (hESC) strategies since hESC can be maintained in a pluripotent state or differentiated into any desired cell type, including endothelial cells (EC), under defined differentiation culture conditions. We recently published a protocol for non-good manufacturing practice (GMP) feeder- and serum-free hESC-EC-directed monolayer differentiation to vascular EC demonstrating the potential to generate hESC-derived EC in a GMP-compliant manner suitable for use in clinical trials. In this study we modified that laboratory protocol to GMP compliance. EC production was confirmed by flow cytometry, qRT-PCR and production of vascular structures in Matrigel®, yielding approximately 30 % mature VE-cadherin(+)/PECAM-1(+) cells using the GMP-compliant hESC line RC13. In conclusion, we have successfully demonstrated the production of vascular EC under GMP-compliant conditions suitable for clinical evaluation.

  18. 13th AINSE radiation biology conference: conference handbook

    International Nuclear Information System (INIS)

    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

  19. Prague Overview Week, September 13th-20th, 2003

    CERN Multimedia

    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...

  20. 13th workshop on what comes beyond the standard models

    CERN Document Server

    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...

  1. MANAGEMENT BOARD MEETING OF 13th MARCH 2003

    CERN Multimedia

    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...

  2. 13th Conference of the Canadian Number Theory Association

    CERN Document Server

    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...

  3. Proceedings of the 13th International Stellarator Workshop

    International Nuclear Information System (INIS)

    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

  4. The 13th Tihany Symposium on Radiation Chemistry

    Science.gov (United States)

    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.

  5. 13th European Conference on Mathematics for Industry

    CERN Document Server

    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.

  6. 13th Forum for Specification and Design Languages (FDL) conference

    CERN Document Server

    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...

  7. 13th International Mass Spectrometry Conference. Book of Abstracts

    International Nuclear Information System (INIS)

    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.)

  8. 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

    Directory of Open Access Journals (Sweden)

    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

  9. Participation of the NO/cGMP/K+ATP pathway in the antinociception induced by Walker tumor bearing in rats

    International Nuclear Information System (INIS)

    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

  10. Lentivirus-induced 'Smart' dendritic cells: Pharmacodynamics and GMP-compliant production for immunotherapy against TRP2-positive melanoma.

    Science.gov (United States)

    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

  11. The Role of Aquaporin 1 Activated by cGMP in Myocardial Edema Caused by Cardiopulmonary Bypass in Sheep

    Directory of Open Access Journals (Sweden)

    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.

  12. Food GMP Certification System in Taiwan%我国台湾的食品GMP认证制度

    Institute of Scientific and Technical Information of China (English)

    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.

  13. Resveratrol enhances on the proliferation and osteoblastic differentiation of BMSCs through MAPK and ER/NO/cGMP pathways

    Institute of Scientific and Technical Information of China (English)

    Ya-linLI; Li-huaSONG; Zhi-jieDAI; Zhou-shengXIAO

    2005-01-01

    AIM The stimulatory effect of resveratrol ( Resv, a natural phytoestrogen) has been reported on osteoblasts in vitro, whereas cyclosporine A (CsA, an immunosuppressant) was associated with bone loss in vivo. This project is intend to investigate the in vitro effect of Resv and CsA on osteoblastic differentiation in bone marrow-derived mesenchymal stem cells (BMSCs), and tries to identify whether the MAPK or ER/NO/cGMP pathway is involved in the stimulatory effect of Resv on BMSCs.

  14. Receptor-mediated rapid action of 1 alpha,25-dihydroxycholecalciferol: increase of intracellular cGMP in human skin fibroblasts.

    OpenAIRE

    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...

  15. GMP-compliant automated synthesis of [{sup 18}F]AV-45 (Florbetapir F 18) for imaging {beta}-amyloid plaques in human brain

    Energy Technology Data Exchange (ETDEWEB)

    Yao, C.-H. [Department of Nuclear Medicine and Molecular Imaging Center, Chang Gung Memorial Hospital, Taiwan (China); Lin, K.-J. [Department of Nuclear Medicine and Molecular Imaging Center, Chang Gung Memorial Hospital, Taiwan (China); Department of Medical Imaging and Radiological Sciences, Chang Gung University, 259 Wen-Hua 1st Road, Kweishan, Taoyuan 333, Taiwan (China); Weng, C.-C. [Department of Medical Imaging and Radiological Sciences, Chang Gung University, 259 Wen-Hua 1st Road, Kweishan, Taoyuan 333, Taiwan (China); Hsiao, I.-T. [Department of Nuclear Medicine and Molecular Imaging Center, Chang Gung Memorial Hospital, Taiwan (China); Department of Medical Imaging and Radiological Sciences, Chang Gung University, 259 Wen-Hua 1st Road, Kweishan, Taoyuan 333, Taiwan (China); Ting, Y.-S. [Department of Nuclear Medicine and Molecular Imaging Center, Chang Gung Memorial Hospital, Taiwan (China); Yen, T.-C. [Department of Nuclear Medicine and Molecular Imaging Center, Chang Gung Memorial Hospital, Taiwan (China); Department of Medical Imaging and Radiological Sciences, Chang Gung University, 259 Wen-Hua 1st Road, Kweishan, Taoyuan 333, Taiwan (China); Jan, T.-R. [Department and Graduate Institute of Veterinary Medicine, National Taiwan University, Taipei, Taiwan (China); Skovronsky, Daniel [Avid Radiopharmaceuticals, Inc., Philadelphia, PA 19104 (United States); Kung, M.-P. [Department of Nuclear Medicine and Molecular Imaging Center, Chang Gung Memorial Hospital, Taiwan (China); Department of Radiology, University of Pennsylvania, Philadelphia, PA 19104 (United States); Wey, S.-P., E-mail: spwey@mail.cgu.edu.t [Department of Nuclear Medicine and Molecular Imaging Center, Chang Gung Memorial Hospital, Taiwan (China); Department of Medical Imaging and Radiological Sciences, Chang Gung University, 259 Wen-Hua 1st Road, Kweishan, Taoyuan 333, Taiwan (China)

    2010-12-15

    We report herein the Good Manufacturing Practice (GMP)-compliant automated synthesis of {sup 18}F-labeled styrylpyridine, AV-45 (Florbetapir), a novel tracer for positron emission tomography (PET) imaging of {beta}-amyloid (A{beta}) plaques in the brain of Alzheimer's disease patients. [{sup 18}F]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 [{sup 18}F]AV-45 reached as high as 470{+-}135 TBq/mmol (n=19). The present studies show that [{sup 18}F]AV-45 can be manufactured under GMP-compliant conditions and could be widely available for routine clinical use.

  16. Activation of a cGMP-sensitive calcium-dependent chloride channel may cause transition from calcium waves to whole-cell oscillations in smooth muscle cells

    DEFF Research Database (Denmark)

    Jacobsen, Jens Christian; Aalkjær, Christian; Nilsson, Holger;

    2007-01-01

    waves sweeping through the cytoplasm when the SR is stimulated to release calcium. A rise in cyclic guanosine monophosphate (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 uniform opening of L-type 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....

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

    Science.gov (United States)

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

    2015-08-01

    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. However, freshly dispersed biofilm cells exhibit a physiology distinct from biofilm and planktonic cells, and they might have a clinically relevant role in infections. Here we present in vitro and in vivo protocols for the generation and characterization of dispersed cells from Pseudomonas aeruginosa 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 identified and quantified, serving as potential targets for treating the dispersed cells. The in vitro protocol can be completed within 4 d, whereas the in vivo protocol requires 7 d.

  18. Development and Application of GMP Management Information System%兽药GMP信息管理系统的开发与应用

    Institute of Scientific and Technical Information of China (English)

    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。

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

    DEFF Research Database (Denmark)

    Chua, Song Lin; Hultqvist, Louise D; Yuan, Mingjun;

    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...

  20. Cyclic diGMP regulates production of sortase substrates of Clostridium difficile and their surface exposure through ZmpI protease-mediated cleavage.

    Science.gov (United States)

    Peltier, Johann; Shaw, Helen A; Couchman, Edward C; Dawson, Lisa F; Yu, Lu; Choudhary, Jyoti S; Kaever, Volkhard; Wren, Brendan W; Fairweather, Neil F

    2015-10-01

    In Gram-positive pathogens, surface proteins may be covalently anchored to the bacterial peptidoglycan by sortase, a cysteine transpeptidase enzyme. In contrast to other Gram-positive bacteria, only one single sortase enzyme, SrtB, is conserved between strains of Clostridium difficile. Sortase-mediated peptidase activity has been reported in vitro, and seven potential substrates have been identified. Here, we demonstrate the functionality of sortase in C. difficile. We identify two sortase-anchored proteins, the putative adhesins CD2831 and CD3246, and determine the cell wall anchor structure of CD2831. The C-terminal PPKTG sorting motif of CD2831 is cleaved between the threonine and glycine residues, and the carboxyl group of threonine is amide-linked to the side chain amino group of diaminopimelic acid within the peptidoglycan peptide stem. We show that CD2831 protein levels are elevated in the presence of high intracellular cyclic diGMP (c-diGMP) concentrations, in agreement with the control of CD2831 expression by a c-diGMP-dependent type II riboswitch. Low c-diGMP levels induce the release of CD2831 and presumably CD3246 from the surface of cells. This regulation is mediated by proteolytic cleavage of CD2831 and CD3246 by the zinc metalloprotease ZmpI, whose expression is controlled by a type I c-diGMP riboswitch. These data reveal a novel regulatory mechanism for expression of two sortase substrates by the secondary messenger c-diGMP, on which surface anchoring is dependent. PMID:26283789

  1. The Research on the Training Method and Control of China Sanshou Team Preparing for the 13th Asian Games%中国散打集训队备战第13届亚运会训练监控方法与手段的研究

    Institute of Scientific and Technical Information of China (English)

    赵光圣; 邱丕相; 陈文鹤; 袁海萍; 刘建奋; 邱俊; 郭玉成; 曾民

    2001-01-01

    为了更好地备战第13届亚运会散打比赛,首次对训练监控的方法与手段进行了研究。结果表明,教练员、科研人员、队医三位一体的监控模式是有效的;从身体素质、身体形态、心理、生理、生化、技术、膳食营养等方面监控训练过程是科学、合理的。%In order to prepare for sanshou match in the 13th Asian Games, wushu sports management center organizes a research group and makes the study on training method and control for the first time. The result shows that the pattern of coach, researchers and doctor participating in training is effective. It is reasonable to supervise the training from physical fitness, body physique, psychology, physiology, biochemistry, technique, diet and nourishment.

  2. 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

    Directory of Open Access Journals (Sweden)

    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.

  3. Activation of a cGMP-sensitive calcium-dependent chloride channel may cause transition from calcium waves to whole cell oscillations in smooth muscle cells

    DEFF Research Database (Denmark)

    Jacobsen, Jens Christian Brings; Aalkjær, Christian; Nilsson, Holger;

    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...

  4. The absence of the Pseudomonas aeruginosa OprF protein leads to increased biofilm formation through variation in c-di-GMP level.

    Science.gov (United States)

    Bouffartigues, Emeline; Moscoso, Joana A; Duchesne, Rachel; Rosay, Thibaut; Fito-Boncompte, Laurène; Gicquel, Gwendoline; Maillot, Olivier; Bénard, Magalie; Bazire, Alexis; Brenner-Weiss, Gerald; Lesouhaitier, Olivier; Lerouge, Patrice; Dufour, Alain; Orange, Nicole; Feuilloley, Marc G J; Overhage, Joerg; Filloux, Alain; Chevalier, Sylvie

    2015-01-01

    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.

  5. The absence of the Pseudomonas aeruginosa OprF protein leads to increased biofilm formation through variation in c-di-GMP level

    Directory of Open Access Journals (Sweden)

    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.

  6. Heme oxygenase-1 is involved in nitric oxide- and cGMP-induced α-Amy2/54 gene expression in GA-treated wheat aleurone layers.

    Science.gov (United States)

    Wu, Mingzhu; Wang, Fangquan; Zhang, Chen; Xie, Yanjie; Han, Bin; Huang, Jingjing; Shen, Wenbiao

    2013-01-01

    Here, α-Amy2/54 gene expression was used as a molecular probe to investigate the interrelationship among nitric oxide (NO), cyclic GMP (cGMP), and heme oxygenase-1 (HO-1) in GA-treated wheat aleurone layers. The inducible expressions of α-Amy2/54 and α-amylase activity were respectively amplified by two NO-releasing compounds, sodium nitroprusside (SNP) and spermine NONOate, in a GA-dependent fashion. Similar responses were observed when an inducer of HO-1, hemin-or one of its catalytic products, carbon monoxide (CO) in aqueous solution-was respectively added. The SNP-induced responses, mimicked by 8-bromoguanosine 3',5'-cyclic monophosphate (8-Br-cGMP), a cGMP derivative, were NO-dependent. This conclusion was supported by the fact that endogenous NO overproduction was rapidly induced by SNP, and thereafter induction of α-Amy2/54 gene expression and increased α-amylase activity were sensitive to the NO scavenger. We further observed that the above induction triggered by SNP and 8-Br-cGMP was partially prevented by zinc protoporphyrin IX (ZnPPIX), an inhibitor of HO-1. These blocking effects were clearly reversed by CO, confirming that the above response was HO-1-specific. Further analyses showed that both SNP and 8-Br-cGMP rapidly up-regulated HO-1 gene expression and increased HO activity, and SNP responses were sensitive to cPTIO and the guanylate cyclase inhibitor 6-anilino-5,8-quinolinedione (LY83583). Molecular evidence confirmed that GA-induced GAMYB and ABA-triggered PKABA1 transcripts were up-regulated or down-regulated by SNP, 8-Br-cGMP or CO cotreated with GA. Contrasting changes were observed when cPTIO, LY83583, or ZnPPIX was added. Together, our results suggested that HO-1 is involved in NO- and cGMP-induced α-Amy2/54 gene expression in GA-treated aleurone layers.

  7. STING activator c-di-GMP enhances the anti-tumor effects of peptide vaccines in melanoma-bearing mice.

    Science.gov (United States)

    Wang, Zili; Celis, Esteban

    2015-08-01

    Therapeutic vaccines to induce anti-tumor CD8 T cells have been used in clinical trials for advanced melanoma patients, but the clinical response rate and overall survival time have not improved much. We believe that these dismal outcomes are caused by inadequate number of antigen-specific CD8 T cells generated by most vaccines. In contrast, huge CD8 T cell responses readily occur during acute viral infections. High levels of type-I interferon (IFN-I) are produced during these infections, and this cytokine not only exhibits anti-viral activity but also promotes CD8 T cell responses. The studies described here were performed to determine whether promoting the production of IFN-I could enhance the potency of a peptide vaccine. We report that cyclic diguanylate monophosphate (c-di-GMP), which activates the stimulator of interferon genes, potentiated the immunogenicity and anti-tumor effects of a peptide vaccine against mouse B16 melanoma. The synergistic effects of c-di-GMP required co-administration of costimulatory anti-CD40 antibody, the adjuvant poly-IC, and were mediated in part by IFN-I. These findings demonstrate that peptides representing CD8 T cell epitopes can be effective inducers of large CD8 T cell responses in vaccination strategies that mimic acute viral infections.

  8. Rapid and efficient conversion of integration-free human induced pluripotent stem cells to GMP-grade culture conditions.

    Directory of Open Access Journals (Sweden)

    Jens Durruthy-Durruthy

    Full Text Available Data suggest that clinical applications of human induced pluripotent stem cells (hiPSCs will be realized. Nonetheless, clinical applications will require hiPSCs that are free of exogenous DNA and that can be manufactured through Good Manufacturing Practice (GMP. Optimally, derivation of hiPSCs should be rapid and efficient in order to minimize manipulations, reduce potential for accumulation of mutations and minimize financial costs. Previous studies reported the use of modified synthetic mRNAs to reprogram fibroblasts to a pluripotent state. Here, we provide an optimized, fully chemically defined and feeder-free protocol for the derivation of hiPSCs using synthetic mRNAs. The protocol results in derivation of fully reprogrammed hiPSC lines from adult dermal fibroblasts in less than two weeks. The hiPSC lines were successfully tested for their identity, purity, stability and safety at a GMP facility and cryopreserved. To our knowledge, as a proof of principle, these are the first integration-free iPSCs lines that were reproducibly generated through synthetic mRNA reprogramming that could be putatively used for clinical purposes.

  9. Design and package of a {sup 14}CO{sub 2} field analyzer The Global Monitor Platform (GMP)

    Energy Technology Data Exchange (ETDEWEB)

    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.

  10. The phytosulfokine (PSK) receptor is capable of guanylate cyclase activity and enabling cyclic GMP-dependent signaling in plants

    KAUST Repository

    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.

  11. Evaluation of Tissue Homogenization to Support the Generation of GMP-Compliant Mesenchymal Stromal Cells from the Umbilical Cord

    Directory of Open Access Journals (Sweden)

    Ryan J. Emnett

    2016-01-01

    Full Text Available Recent studies have demonstrated that the umbilical cord (UC is an excellent source of mesenchymal stromal cells (MSCs. However, current protocols for extracting and culturing UC-MSCs do not meet current good manufacturing practice (cGMP standards, in part due to the use of xenogeneic reagents. To support the development of a cGMP-compliant method, we have examined an enzyme-free isolation method utilizing tissue homogenization (t-H followed by culture in human platelet lysate (PL supplemented media. The yield and viability of cells after t-H were comparable to those obtained after collagenase digestion (Col-D. Importantly, kinetic analysis of cultured cells showed logarithmic growth over 10 tested passages, although the rate of cell division was lower for t-H as compared to Col-D. This slower growth of t-H-derived cells was also reflected in their longer population doubling time. Interestingly, there was no difference in the expression of mesenchymal markers and trilineage differentiation potential of cells generated using either method. Finally, t-H-derived cells had greater clonogenic potential compared to Col-D/FBS but not Col-D/PL and were able to maintain CFU-F capacity through P7. This bench scale study demonstrates the possibility of generating therapeutic doses of good quality UC-MSCs within a reasonable length of time using t-H and PL.

  12. Derivation of xeno-free and GMP-grade human embryonic stem cells--platforms for future clinical applications.

    Directory of Open Access Journals (Sweden)

    Shelly E Tannenbaum

    Full Text Available 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 lines in adherence to regulations for embryo procurement, and good tissue, manufacturing and laboratory practices. To minimize freezing and thawing, we continuously expanded the lines from initial outgrowths and samples were cryopreserved as early stocks and banks. Batch release criteria included DNA-fingerprinting and HLA-typing for identity, characterization of pluripotency-associated marker expression, proliferation, karyotyping and differentiation in-vitro and in-vivo. These hESCs may be valuable for regenerative therapy. The ethical, scientific and regulatory methodology presented here may serve for development of additional clinical-grade hESCs.

  13. DNA-Mediated Cyclic GMP-AMP Synthase-Dependent and -Independent Regulation of Innate Immune Responses.

    Science.gov (United States)

    Motani, Kou; Ito, Shinji; Nagata, Shigekazu

    2015-05-15

    Cytoplasmic DNA activates cyclic GMP-AMP synthase (cGAS) to produce cyclic 2'-5'3'-5'GMP-AMP dinucleotide (2'5 'cGAMP). The binding of 2'5'cGAMP to an adaptor protein, stimulator of IFN genes (STING), activates a transcription factor, IFN regulatory factor 3, leading to the induction of IFN and chemokine gene expression. In this study, we found that the 2'5'cGAMP-dependent STING activation induced highly upregulated CXCL10 gene expression. Formation of a distinct STING dimer, which was detected by native PAGE, was induced by 2'5'cGAMP, but not 3'-5'3'-5'cGAMP. Analysis of DNase II(-/-) mice, which constitutively produce IFN-β and CXCL10, showed the accumulation of 2'5'cGAMP in their fetal livers and spleens, suggesting that the undigested DNA accumulating in DNase II(-/-) cells may have leaked from the lysosomes into the cytoplasm. The DNase II(-/-) mouse embryonic fibroblasts produced 2'5'cGAMP in a cGAS-dependent manner during apoptotic cell engulfment. However, cGAS deficiency did not impair the STING-dependent upregulation of CXCL10 in DNase II(-/-) mouse embryonic fibroblasts that was induced by apoptotic cell engulfment or DNA lipofection. These results suggest the involvement of a cGAS-independent additional DNA sensor(s) that induces the STING-dependent activation of innate immunity.

  14. "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

    Institute of Scientific and Technical Information of China (English)

    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

  15. Effects of Kaempferia parviflora Wall. Ex. Baker and sildenafil citrate on cGMP level, cardiac function, and intracellular Ca2+ regulation in rat hearts.

    Science.gov (United States)

    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

  16. Crystallization and preliminary X-ray analysis of the flagellar motor ‘brake’ molecule YcgR with c-di-GMP from Escherichia coli

    International Nuclear Information System (INIS)

    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

  17. Isotype-specific activation of cystic fibrosis transmembrane conductance regulator-chloride channels by cGMP-dependent protein kinase II

    NARCIS (Netherlands)

    P.J. French (Pim); J. Bijman (Jan); M.J. Edixhoven (Marcel); A.B. Vaandrager (Arie); B.J. Scholte (Bob); S.M. Lohmann (Suzanne); A.C. Nairn; H.R. de Jonge (Hugo)

    1995-01-01

    textabstractType II cGMP-dependent protein kinase (cGKII) isolated from pig intestinal brush borders and type I alpha cGK (cGKI) purified from bovine lung were compared for their ability to activate the cystic fibrosis transmembrane conductance regulator (CFTR)-Cl- channel in excis

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

    DEFF Research Database (Denmark)

    Fazli, Mustafa; O'Connell, Aileen; Nilsson, Martin;

    2011-01-01

    of a number of components, including cellulose and fimbriae. It was demonstrated that the Bcam1349 protein binds to the promoter region of the cellulose synthase genes, and that this binding is enhanced by the presence of c-di-GMP. The bcam1349 mutant showed reduced virulence in a Galleria mellonella wax moth...

  19. The NO-cGMP-PKG Signaling Pathway Regulates Synaptic Plasticity and Fear Memory Consolidation in the Lateral Amygdala via Activation of ERK/MAP Kinase

    Science.gov (United States)

    Ota, Kristie T.; Pierre, Vicki J.; Ploski, Jonathan E.; Queen, Kaila; Schafe, Glenn E.

    2008-01-01

    Recent studies have shown that nitric oxide (NO) signaling plays a crucial role in memory consolidation of Pavlovian fear conditioning and in synaptic plasticity in the lateral amygdala (LA). In the present experiments, we examined the role of the cGMP-dependent protein kinase (PKG), a downstream effector of NO, in fear memory consolidation and…

  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

    Science.gov (United States)

    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…

  1. Endogenous type II cGMP-dependent protein kinase exists as a dimer in membranes and can Be functionally distinguished from the type I isoforms

    NARCIS (Netherlands)

    A.B. Vaandrager (Arie); M.J. Edixhoven (Marcel); A.G. Bot (Alice); M.A. Kroos (Marian); T. Jarchau; S. Lohmann; H.G. Genieser; H.R. de Jonge (Hugo)

    1997-01-01

    textabstractIn mammalian tissues two types of cGMP-dependent protein kinase (cGK) have been identified. In contrast to the dimeric cGK I, cGK II purified from pig intestine was shown previously to behave as a monomer. However, recombinant rat cGK II was found to have hy

  2. The Selection of the Candidates Leaving for Britain to Study during the Ruling Period of the 13th Dalai Lama and their Departure%十三世达赖喇嘛时期西藏赴英留学生的选派考

    Institute of Scientific and Technical Information of China (English)

    朗杰紫丹

    2015-01-01

    十三世达赖喇嘛在什么样的背景下,产生了派遣留学生到西方学习的想法;他推行的新政措施的重要内容之一,派遣留学生一事,又是怎么实施的;最终怎么选定或产生这四位赴英留学生的;他们又是什么时候离开西藏启程去英国等问题,说法各异,与史实有一定的出入.文章以藏文、汉文和英文资料为依据,结合人物访谈资料,对十三世达赖喇嘛时期西藏赴英留学生的选定和启程等方面,提出了自己的观点.%The questions like: what was the social background when the 13th Dalai Lama came up the idea of sending Tibetan students studying in the West? Was it one of the important components of his new deal? How was it undertaken? How eventually were the four candidates selected studying in Britain? When did they leave Tibet for Britain? and so on and so forth, have different answers, which have some discrepancies with the histori-cal facts. This article provides the author's own view of the historical event by referring to the Tibetan and Chi-nese, and English documents and literatures combined with some interview data.

  3. MrkH, a novel c-di-GMP-dependent transcriptional activator, controls Klebsiella pneumoniae biofilm formation by regulating type 3 fimbriae expression.

    Directory of Open Access Journals (Sweden)

    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

  4. YfiBNR mediates cyclic di-GMP dependent small colony variant formation and persistence in Pseudomonas aeruginosa.

    Directory of Open Access Journals (Sweden)

    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

  5. Evaluation of GMP-compliant culture media for in vitro expansion of human bone marrow mesenchymal stromal cells.

    Science.gov (United States)

    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.

  6. Three cyanobacteriochromes work together to form a light color-sensitive input system for c-di-GMP signaling of cell aggregation.

    Science.gov (United States)

    Enomoto, Gen; Ni-Ni-Win; Narikawa, Rei; Ikeuchi, Masahiko

    2015-06-30

    Cyanobacteriochromes (CBCRs) are cyanobacterial photoreceptors that have diverse spectral properties and domain compositions. Although large numbers of CBCR genes exist in cyanobacterial genomes, no studies have assessed whether multiple CBCRs work together. We recently showed that the diguanylate cyclase (DGC) activity of the CBCR SesA from Thermosynechococcus elongatus is activated by blue-light irradiation and that, when irradiated, SesA, via its product cyclic dimeric GMP (c-di-GMP), induces aggregation of Thermosynechococcus vulcanus cells at a temperature that is suboptimum for single-cell viability. For this report, we first characterize the photobiochemical properties of two additional CBCRs, SesB and SesC. Blue/teal light-responsive SesB has only c-di-GMP phosphodiesterase (PDE) activity, which is up-regulated by teal light and GTP. Blue/green light-responsive SesC has DGC and PDE activities. Its DGC activity is enhanced by blue light, whereas its PDE activity is enhanced by green light. A ΔsesB mutant cannot suppress cell aggregation under teal-green light. A ΔsesC mutant shows a less sensitive cell-aggregation response to ambient light. ΔsesA/ΔsesB/ΔsesC shows partial cell aggregation, which is accompanied by the loss of color dependency, implying that a nonphotoresponsive DGC(s) producing c-di-GMP can also induce the aggregation. The results suggest that SesB enhances the light color dependency of cell aggregation by degrading c-di-GMP, is particularly effective under teal light, and, therefore, seems to counteract the induction of cell aggregation by SesA. In addition, SesC seems to improve signaling specificity as an auxiliary backup to SesA/SesB activities. The coordinated action of these three CBCRs highlights why so many different CBCRs exist.

  7. Effects of papaverine and vasointestinal polypeptide on penile and vascular cAMP and cGMP in control and diabetic animals: an in vitro study.

    Science.gov (United States)

    Miller, M A; Morgan, R J; Thompson, C S; Mikhailidis, D P; Jeremy, J Y

    1995-06-01

    Adenosine 3'5'-cyclic monophosphate (cAMP) and guanosine 3'5'-cyclic monophosphate (cGMP) mediate penile erection. We have previously established that adenylate and guanylate cyclase activity is elevated in the diabetic rat penis and aorta. This study investigates the action of papaverine and vasoactive intestinal polypeptide (VIP) on these cyclases. The aortae and penes of Sprague Dawley rats (n = 7) were stimulated with VIP and papaverine. Diabetes mellitus (DM) was induced in Sprague Dawley rats (n = 7) with streptozotocin and the penile and aortic tissues were treated with VIP. The penes, aortae and carotid arteries of New Zealand White rabbits were similarly processed. cAMP and cGMP generation was measured by radioimmunoassay. In all tissues: VIP stimulated cAMP synthesis; VIP did not increase cGMP levels; papaverine was without effect on either cAMP or cGMP synthesis. VIP-stimulated cAMP was significantly enhanced in the diabetic rat penis and aorta; there was also a significant elevation in the basal levels of cGMP in these tissues. These data: (1) consolidate that cAMP is a mediator of penile erection, (2) indicate that papaverine and VIP elicit erection by different mechanisms, (3) suggest that an enhanced penile capacity to generate cAMP in DM may constitute an adaptive response to counteract the previously reported reduction in VIP content and VIP receptors, and (4) indicate that the penile and vascular tissues of the rabbit respond in a similar manner to VIP and papaverine. PMID:7496446

  8. Structures of the activator of K. pneumonia biofilm formation, MrkH, indicates PilZ domains involved in c-di-GMP and DNA binding

    Science.gov (United States)

    Schumacher, Maria A.; Zeng, Wenjie

    2016-01-01

    The pathogenesis of Klebsiella pneumonia is linked to the bacteria’s ability to form biofilms. Mannose-resistant Klebsiella-like (Mrk) hemagglutinins are critical for K. pneumonia biofilm development, and the expression of the genes encoding these proteins is activated by a 3′,5′-cyclic diguanylic acid (c-di-GMP)–regulated transcription factor, MrkH. To gain insight into MrkH function, we performed structural and biochemical analyses. Data revealed MrkH to be a monomer with a two-domain architecture consisting of a PilZ C-domain connected to an N domain that unexpectedly also harbors a PilZ-like fold. Comparison of apo- and c-di-GMP–bound MrkH structures reveals a large 138° interdomain rotation that is induced by binding an intercalated c-di-GMP dimer. c-di-GMP interacts with PilZ C-domain motifs 1 and 2 (RxxxR and D/NxSxxG) and a newly described c-di-GMP–binding motif in the MrkH N domain. Strikingly, these c-di-GMP–binding motifs also stabilize an open state conformation in apo MrkH via contacts from the PilZ motif 1 to residues in the C-domain motif 2 and the c-di-GMP–binding N-domain motif. Use of the same regions in apo structure stabilization and c-di-GMP interaction allows distinction between the states. Indeed, domain reorientation by c-di-GMP complexation with MrkH, which leads to a highly compacted structure, suggests a mechanism by which the protein is activated to bind DNA. To our knowledge, MrkH represents the first instance of specific DNA binding mediated by PilZ domains. The MrkH structures also pave the way for the rational design of inhibitors that target K. pneumonia biofilm formation. PMID:27551088

  9. Ab Initio QM/MM Study Shows a Highly Dissociated SN2 Hydrolysis Mechanism for the cGMP-Specific Phosphodiesterase-5.

    Science.gov (United States)

    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

  10. Angiotensin-converting enzyme inhibition prevents myocardial infarction-induced increase in renal cortical cGMP and cAMP phosphodiesterase activities.

    Science.gov (United States)

    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

  11. Angiotensin-converting enzyme inhibition prevents myocardial infarction-induced increase in renal cortical cGMP and cAMP phosphodiesterase activities.

    Science.gov (United States)

    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.

  12. Annual Check-up

    Science.gov (United States)

    ... 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 ...

  13. GMP facilities for manufacturing of advanced therapy medicinal products for clinical trials: an overview for clinical researchers.

    Science.gov (United States)

    Alici, Evren; Blomberg, Pontus

    2010-12-01

    To be able to produce advanced therapy medicinal products, compliance with regulatory standards while maintaining flexibility is mandatory. For this purpose, careful planning is vital in the design or upgrade of a facility. Similarly, extensive foresight is elemental to anticipate upcoming needs and requirements. Failing this may lead to the facility's in-ability to meet the demands. In this chapter we aimed to outline the current issues with regards to the European Union Directives (EUD) and the proposal for Advanced Therapies, which are of importance to cellular and gene therapy facilities in Europe. This chapter is an attempt to elucidate what the minimum requirements for GMP facilities for cell and gene therapy products are and what is considered necessary to comply with the regulations in Europe.

  14. Measurement of GEp/GMp in ep -> ep to Q2 = 5.6 GeV2

    CERN Document Server

    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.

  15. cGMP-Manufactured Human Induced Pluripotent Stem Cells Are Available for Pre-clinical and Clinical Applications

    Directory of Open Access Journals (Sweden)

    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.

  16. Molecular basis for the specific recognition of the metazoan cyclic GMP-AMP by the innate immune adaptor protein STING.

    Science.gov (United States)

    Shi, Heping; Wu, Jiaxi; Chen, Zhijian J; Chen, Chuo

    2015-07-21

    Cyclic GMP-AMP containing a unique combination of mixed phosphodiester linkages (2'3'-cGAMP) is an endogenous second messenger molecule that activates the type-I IFN pathway upon binding to the homodimer of the adaptor protein STING on the surface of endoplasmic reticulum membrane. However, the preferential binding of the asymmetric ligand 2'3'-cGAMP to the symmetric dimer of STING represents a physicochemical enigma. Here we show that 2'3'-cGAMP, but not its linkage isomers, adopts an organized free-ligand conformation that resembles the STING-bound conformation and pays low entropy and enthalpy costs in converting into the active conformation. Our results demonstrate that analyses of free-ligand conformations can be as important as analyses of protein conformations in understanding protein-ligand interactions.

  17. Effects of atrial and brain natriuretic peptides upon cyclic GMP levels, potassium transport, and receptor binding in rat astrocytes

    International Nuclear Information System (INIS)

    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

  18. Strategies for selecting recombinant CHO cell lines for cGMP manufacturing: improving the efficiency of cell line generation.

    Science.gov (United States)

    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

  19. 带状发展:“十三五:中国文化产业发展新趋势%New trend in the ribbon development of China’s culture industry in the period of the 13 th Five-Year Plan

    Institute of Scientific and Technical Information of China (English)

    范建华

    2015-01-01

    “三五”是中国文化产业发展的重要拐点,带状发展成为中国现阶段文化产业发展的新趋势。在带状发展的空间布局下,我国的文化产业必将走向国际市场,丰富国内市场,提升产品升级换代,出现历史性跨越发展和几何级数倍增的新特点;而大数据时代的到来势必要求我们做好顶层设计和规划,实现大数据分析、决策、定位和拓展。在这个过程中,必须突破现行行政区划限制、突破产业门类分割、突破市场壁垒束缚、突破管理体制制约,最终使中国的文化产业发展形成统一市场、统一标准、统一国民待遇的公平竞争大格局。%The period of the 13 th Five-Year Plan is an important turning point in the development of China's culture industry,when ribbon development has become its new trend.Under this new spatial layout,the culture industry will present the following new features:quickened steps for entering the international market,a more diversified domestic market,the quickened upgrading of products,a his-toric leap in development and multiplied increase in productivity.Meanwhile,the arrival of the era of big data requires us to be a good top-level designer and planner to achieve big data analytics,decision-making,positioning and expansion.To attain this goal,we must break the restrictions of the existing administrative divisions,the segmentation of industrial categories,the shackles of market barriers as well as the constraints of the management system.Thus,China's culture industry will develop into a strong one while entering the large international market with fair competition and established market criteria as well as uniform regulations and treatment for all.

  20. 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

    Institute of Scientific and Technical Information of China (English)

    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。

  1. Stimulus Response of Au-NPs@GMP-Tb Core-Shell Nanoparticles: Toward Colorimetric and Fluorescent Dual-Mode Sensing of Alkaline Phosphatase Activity in Algal Blooms of a Freshwater Lake.

    Science.gov (United States)

    Zhang, Xiaolei; Deng, Jingjing; Xue, Yumeng; Shi, Guoyue; Zhou, Tianshu

    2016-01-19

    In this study, we demonstrate a colorimetric and fluorescent dual-mode method for alkaline phosphatase activity (APA) sensing in freshwater lake with stimuli-responsive gold nanoparticles@terbium-guanosine monophosphate (Au-NPs@GMP-Tb) core-shell nanoparticles. Initially, the core-shell nanoparticles were fabricated based on Au-NPs decorated with a fluorescent GMP-Tb shell. Upon being excited at 290 nm, the as-formed Au-NPs@GMP-Tb core-shell nanoparticles emit green fluorescence, and the decorated GMP-Tb shell causes the aggregation of Au-NPs. However, the addition of ALP destroys GMP-Tb shell, resulting in the release of Au-NPs from the shell into the solvent. As a consequence, the aggregated Au-NPs solubilizes with the changes in the UV-vis spectrum of the dispersion, and in the meantime, the fluorescence of GMP-Tb shell turns off, which constitutes a new mechanism for colorimetric and fluorescent dual-mode sensing of APA. With the method developed here, we could monitor the dynamic change of APA during an algal bloom of a freshwater lake, both by the naked eye and further confirmed by fluorometric determination. This study not only offers a new method for on-site visible detection of APA but also provides a strategy for dual-mode sensing mechanisms by the rational design of the excellent optical properties of Au-NPs and the adaptive inclusion properties of the luminescent infinite coordination polymers.

  2. Analysis of the overseas GMP inspection and fee systems%国外药品GMP检查及收费标准探讨

    Institute of Scientific and Technical Information of China (English)

    翁新愚; 陈玉文; 毕开顺

    2012-01-01

    通过分析欧洲药品质量管理局、澳大利亚治疗产品管理局、新加坡卫生科学局以及美国食品药品管理局的国外GMP检查要求及收费制度,为我国开展国外药品GMP检查提供参考依据.%We analyzed the overseas GMP inspection and fee systems of the European Directorate for the Quality of Medicines & HealthCare, Australia Therapeutic Goods Administration, Singapore Health Science Authority and US Food and Drug Administration. This analysis will provide reference for Chinese overseas GMP inspection.

  3. Annual Statistical Supplement, 2014

    Data.gov (United States)

    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...

  4. Annual Statistical Supplement, 2015

    Data.gov (United States)

    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...

  5. Annual Statistical Supplement, 2010

    Data.gov (United States)

    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...

  6. Annual Statistical Supplement, 2000

    Data.gov (United States)

    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...

  7. Annual Statistical Supplement, 2004

    Data.gov (United States)

    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...

  8. Annual Statistical Supplement, 2001

    Data.gov (United States)

    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, 2003

    Data.gov (United States)

    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...

  10. Annual Statistical Supplement, 2005

    Data.gov (United States)

    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...

  11. Annual Statistical Supplement, 2007

    Data.gov (United States)

    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...

  12. Annual Statistical Supplement, 2008

    Data.gov (United States)

    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...

  13. Annual Statistical Supplement, 2009

    Data.gov (United States)

    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...

  14. Annual Statistical Supplement, 2002

    Data.gov (United States)

    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...

  15. Annual Statistical Supplement, 2006

    Data.gov (United States)

    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...

  16. Annual Statistical Supplement, 2011

    Data.gov (United States)

    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...

  17. S-nitrosothiols dilate the mesenteric artery more potently than the femoral artery by a cGMP and L-type calcium channel-dependent mechanism.

    Science.gov (United States)

    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.

  18. Can selective inhibitors of cyclic guanosine monophosphate (cGMP)-specific phosphadiesterase type 5 (PDE 5) offer protection against contrast induced nephropathy?

    OpenAIRE

    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...

  19. Host cell contact induces expression of virulence factors and VieA, a cyclic di-GMP phosphodiesterase, in Vibrio cholerae.

    Science.gov (United States)

    Dey, Amit K; Bhagat, Abha; Chowdhury, Rukhsana

    2013-05-01

    Vibrio cholerae, a noninvasive bacterium, colonizes the intestinal epithelium and secretes cholera toxin (CT), a potent enterotoxin that causes the severe fluid loss characteristic of the disease cholera. In this study, we demonstrate that adherence of V. cholerae to the intestinal epithelial cell line INT 407 strongly induces the expression of the major virulence genes ctxAB and tcpA and the virulence regulatory gene toxT. No induction of toxR and tcpP, which encode transcriptional activators of toxT, was observed in adhered bacteria, and the adherence-dependent upregulation of toxT expression was independent of ToxR and TcpP. A sharp increase in the expression of the vieA gene, which encodes a cyclic di-GMP (c-di-GMP) phosphodiesterase, was observed in INT 407-adhered V. cholerae immediately after infection. Induction of toxT, ctxAB, and tcpA in INT 407-adhered vieA mutant strain O395 ΔvieA was consistently lower than in the parent strain, although no effect was observed in unadhered bacteria, suggesting that VieA has a role in the upregulation of toxT expression specifically in host cell-adhered V. cholerae. Furthermore, though VieA has both a DNA binding helix-turn-helix domain and an EAL domain conferring c-di-GMP phosphodiesterase activity, the c-di-GMP phosphodiesterase activity of VieA is necessary and sufficient for the upregulation of toxT expression.

  20. S-nitrosothiols dilate the mesenteric artery more potently than the femoral artery by a cGMP and L-type calcium channel-dependent mechanism.

    Science.gov (United States)

    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