WorldWideScience

Sample records for monitoring regulations requiring

  1. Franchise Values, Regulatory Monitoring, and Capital Requirements in Optimal Bank Regulation

    DEFF Research Database (Denmark)

    Andersen, Thomas Barnebeck; Harr, Thomas

    2008-01-01

    This paper demonstrates that financial deregulation is likely to make standard prudential regulatory instruments less effective in curbing excessive risk-taking incentives among banks. This has interesting implications for optimal bank regulation. When there is an increase in competition......, the optimal capital requirement should increase, whereas regulatory auditing should decrease. In contrast, when there is an increase in gambling yields, auditing should always increase, whereas the optimal capital requirement may increase or decrease....

  2. Hydrogen Monitoring Requirements in the Global Technical Regulation on Hydrogen and Fuel Cell Vehicles: Preprint

    Energy Technology Data Exchange (ETDEWEB)

    Buttner, William; Rivkin, Carl; Burgess, Robert; Hartmann, Kevin; Bubar, Max; Post, Matthew; Boon-Brett, Lois; Weidner, Eveline; Moretto, Pietro

    2016-07-01

    The United Nations Global Technical Regulation (GTR) Number 13 (Global Technical Regulation on Hydrogen and Fuel Cell Vehicles) is the defining document regulating safety requirements in hydrogen vehicles, and in particular fuel cell electric vehicles (FCEV). GTR Number 13 has been formally implemented and will serve as the basis for the national regulatory standards for FCEV safety in North America (Canada, United States), Japan, Korea, and the European Union. The GTR defines safety requirement for these vehicles, including specifications on the allowable hydrogen levels in vehicle enclosures during in-use and post-crash conditions and on the allowable hydrogen emissions levels in vehicle exhaust during certain modes of normal operation. However, in order to be incorporated into national regulations, that is, in order to be binding, methods to verify compliance to the specific requirements must exist. In a collaborative program, the Sensor Laboratories at the National Renewable Energy Laboratory in the United States and the Joint Research Centre, Institute for Energy and Transport in the Netherlands have been evaluating and developing analytical methods that can be used to verify compliance to the hydrogen release requirement as specified in the GTR.

  3. 40 CFR 141.625 - Conditions requiring increased monitoring.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 22 2010-07-01 2010-07-01 false Conditions requiring increased monitoring. 141.625 Section 141.625 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Stage 2 Disinfection Byproducts Requirements § 141.625 Conditions...

  4. 40 CFR 433.12 - Monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... requirements. (a) In lieu of requiring monitoring for TTO, the permitting authority (or, in the case of... the permit limitation for total toxic organics (TTO), I certify that, to the best of my knowledge and... TTO standard, the industrial discharger need analyse for only those pollutants which would...

  5. Performance, requirements and testing in individual monitoring

    DEFF Research Database (Denmark)

    Julius, H.W.; Christensen, P.; Marshall, T.O.

    1990-01-01

    monitoring, and in particular of those related to the energy and angular dependences of the dosemeters. These data are important for the testing of the dosimetry systems. Finally testing programmes in individual monitoring are discussed and four categories of testing are proposed.......For implementation of the ICRP requirements in practical routine monitoring, a specification is needed with respect to specific performance criteria of the dosimetry system. This includes the dosemeters, and procedures for initial and on-going testing of the dosimetry systems to ensure...... that the specified criteria are continuously met. Methods are here specified for establishing performance criteria, and satisfying the ICRP overall accuracy requirements for individual monitoring. Furthermore, procedures are given for obtaining numerical values of the uncertainties connected with individual...

  6. 40 CFR 63.8 - Monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... standard(s) unless the Administrator— (i) Specifies or approves the use of minor changes in methodology for the specified monitoring requirements and procedures (see § 63.90(a) for definition); or (ii) Approves... (see § 63.90(a) for definition). (iii) Owners or operators with flares subject to § 63.11(b) are...

  7. 77 FR 13977 - Quality Assurance Requirements for Continuous Opacity Monitoring Systems at Stationary Sources

    Science.gov (United States)

    2012-03-08

    ... AGENCY 40 CFR Part 60 RIN 2060-AH23 Quality Assurance Requirements for Continuous Opacity Monitoring... final rule titled, ``Quality Assurance Requirements for Continuous Opacity Monitoring Systems at... electronically in www.regulations.gov or in hard copy at the Procedure 3--Quality Assurance Requirements for...

  8. 77 FR 13997 - Quality Assurance Requirements for Continuous Opacity Monitoring Systems at Stationary Sources

    Science.gov (United States)

    2012-03-08

    ... AGENCY 40 CFR Part 60 RIN 2060-AH23 Quality Assurance Requirements for Continuous Opacity Monitoring... rule titled, ``Quality Assurance Requirements for Continuous Opacity Monitoring Systems at Stationary....regulations.gov or in hard copy at the Procedure 3--Quality Assurance Requirements for Continuous Opacity...

  9. The Relationships between Monitoring, Regulation and Performance

    Science.gov (United States)

    Koriat, Asher

    2012-01-01

    The articles in this Special Issue reflect the growing interest in applying laboratory-based research to educational settings. These articles highlight the contribution of metacognitive monitoring and self-regulation to effective learning and performance. At the same time, they illustrate the methodological and theoretical challenges involved in…

  10. Review of FEWS NET biophysical monitoring requirements

    Energy Technology Data Exchange (ETDEWEB)

    Ross, K W; Underwood, L W [Science Systems and Applications, Incorporated, 411 West Michigan Street, Poplarville, MS 39470 (United States); Brown, M E [NASA-Goddard Space Flight Center, Code 614.4, Greenbelt, MD 20771 (United States); Verdin, J P [United States Geological Survey, National Integrated Drought Information System, NOAA/ESRL, 325 Broadway, Boulder, CO 80305 (United States)], E-mail: kenton_ross@ssaihq.com, E-mail: molly.brown@nasa.gov, E-mail: verdin@usgs.gov, E-mail: lauren_underwood@ssaihq.com

    2009-04-15

    The Famine Early Warning System Network (FEWS NET) provides monitoring and early warning support to decision makers responsible for responding to famine and food insecurity. FEWS NET transforms satellite remote sensing data into rainfall and vegetation information that can be used by these decision makers. The National Aeronautics and Space Administration has recently funded activities to enhance remote sensing inputs to FEWS NET. To elicit Earth observation requirements, a professional review questionnaire was disseminated to FEWS NET expert end-users; it focused upon operational requirements to determine additional useful remote sensing data and, subsequently, to assess whether such data would be beneficial as FEWS NET biophysical supplementary inputs. The review was completed by over 40 experts from around the world. Reviewers were asked to evaluate the relative importance of environmental variables and spatio-temporal requirements for Earth science data products, in particular for rainfall and vegetation products. The results showed that spatio-temporal resolution requirements are complex and need to vary according to place, time, and hazard; that high resolution remote sensing products continue to be in demand; and that rainfall and vegetation products are valued as data that provide actionable food security information.

  11. Review of FEWS NET Biophysical Monitoring Requirements

    Science.gov (United States)

    Ross, K. W.; Brown, Molly E.; Verdin, J.; Underwood, L. W.

    2009-01-01

    The Famine Early Warning System Network (FEWS NET) provides monitoring and early warning support to decision makers responsible for responding to famine and food insecurity. FEWS NET transforms satellite remote sensing data into rainfall and vegetation information that can be used by these decision makers. The National Aeronautics and Space Administration has recently funded activities to enhance remote sensing inputs to FEWS NET. To elicit Earth observation requirements, a professional review questionnaire was disseminated to FEWS NET expert end-users: it focused upon operational requirements to determine additional useful remote sensing data and; subsequently, beneficial FEWS NET biophysical supplementary inputs. The review was completed by over 40 experts from around the world, enabling a robust set of professional perspectives to be gathered and analyzed rapidly. Reviewers were asked to evaluate the relative importance of environmental variables and spatio-temporal requirements for Earth science data products, in particular for rainfall and vegetation products. The results showed that spatio-temporal resolution requirements are complex and need to vary according to place, time, and hazard: that high resolution remote sensing products continue to be in demand, and that rainfall and vegetation products were valued as data that provide actionable food security information.

  12. Requirements of CLIC Beam Loss Monitoring System

    CERN Document Server

    Sapinski, M; Holzer, EB; Jonker, M; Mallows, S; Otto, T; Welsch, C

    2010-01-01

    The Compact Linear Collider (CLIC) [1] is a proposed multi-TeV linear electron-positron collider being designed by a world-wide collaboration. It is based on a novel twobeam acceleration scheme in which two beams (drive and main beam) are placed in parallel to each other and energy is transferred from the drive beam to the main one. Beam losses on either of them can have catastrophic consequences for the machine, because of high intensity (drive beam) or high energy and small emittance (main beam). In the framework of machine protection, a Beam Loss Monitoring (BLM) system has to be put in place. This paper discusses the requirements for the beam loss system in terms of detector sensitivity, resolution, dynamic range and ability to distinguish losses originating from various sources. The two-beam module where the protection from beam losses is particularly challenging and important, is studied.

  13. 40 CFR 51.190 - Ambient air quality monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 2 2010-07-01 2010-07-01 false Ambient air quality monitoring... PROGRAMS REQUIREMENTS FOR PREPARATION, ADOPTION, AND SUBMITTAL OF IMPLEMENTATION PLANS Ambient Air Quality Surveillance § 51.190 Ambient air quality monitoring requirements. The requirements for monitoring ambient...

  14. 40 CFR 413.03 - Monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... lieu of monitoring for TTO, the control authority may allow industrial users of POTWs to make the... for total toxic organics (TTO), I certify that, to the best of my knowledge and belief, no dumping of... or leak into the wastewater. (c) If monitoring is necessary to measure compliance with the...

  15. 40 CFR 63.564 - Monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... obtained. For monitoring equipment purchased from a vendor, verification of the operational status of the... when the measured values (i.e., daily calibrations, multipoint calibrations, and performance audits... pressure device once each calendar year with a reference pressure monitor (traceable to National...

  16. 40 CFR 63.364 - Monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... obtained. For monitoring equipment purchased from a vendor, verification of the operational status of the... operated. From 15-minute or shorter period temperature values, a data acquisition system for the... year with a reference temperature monitor (traceable to National Institute of Standards and...

  17. 40 CFR 63.828 - Monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... product and packaging rotogravure or wide-web flexographic presses with intermittently-controllable work... emission monitors to measure total organic volatile matter concentration and volumetric gas flow rate...

  18. 77 FR 8209 - Quality Assurance Requirements for Continuous Opacity Monitoring Systems at Stationary Sources

    Science.gov (United States)

    2012-02-14

    ... AGENCY 40 CFR Part 60 RIN 2060-AH23 Quality Assurance Requirements for Continuous Opacity Monitoring... standards as specified in federally enforceable regulations. The quality assurance requirements will be... standards to the quality assurance requirements in Appendix F of 40 CFR Part 60 in the ``Rules and...

  19. 77 FR 18709 - Quality Assurance Requirements for Continuous Opacity Monitoring Systems at Stationary Sources

    Science.gov (United States)

    2012-03-28

    ... AGENCY 40 CFR Part 60 RIN 2060-AH23 Quality Assurance Requirements for Continuous Opacity Monitoring... final rule. SUMMARY: The EPA published a direct final rule titled ``Quality Assurance Requirements for....regulations.gov or in hard copy at the Procedure 3--Quality Assurance Requirements for Continuous Opacity...

  20. 40 CFR 63.1657 - Monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... compressed air supply for pulse-jet baghouses. (iv) An appropriate methodology for monitoring cleaning cycles... document “Fabric Filter Bag Leak Detection Guidance” (EPA-454/R-98-015). Other bag leak detection systems... baghouse for air leaks, torn or broken bags or filter media, or any other condition that may cause...

  1. 40 CFR 63.605 - Monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... rock dryer or phosphate rock calciner subject to the provisions of this subpart shall install... either the mass flow of phosphorus-bearing feed material to the dryer or calciner, or the mass flow of product from the dryer or calciner. The monitoring system shall have an accuracy of ±5 percent over...

  2. 40 CFR 52.346 - Air quality monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 3 2010-07-01 2010-07-01 false Air quality monitoring requirements. 52... (CONTINUED) APPROVAL AND PROMULGATION OF IMPLEMENTATION PLANS Colorado § 52.346 Air quality monitoring... VIII Administrator, the State submitted a revised Air Quality Monitoring State Implementation Plan....

  3. 40 CFR 146.68 - Testing and monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... to provide other site specific data; (iii) Periodic monitoring of the ground water quality in the first aquifer overlying the injection zone; (iv) Periodic monitoring of the ground water quality in the... Applicable to Class I Hazardous Waste Injection Wells § 146.68 Testing and monitoring requirements....

  4. 40 CFR 146.13 - Operating, monitoring and reporting requirements.

    Science.gov (United States)

    2010-07-01

    ...; (iii) Periodic monitoring of the ground water quality in the first aquifer overlying the injection zone; (iv) Periodic monitoring of the ground water quality in the lowermost USDW; and (v) Any additional... Applicable to Class I Wells § 146.13 Operating, monitoring and reporting requirements. (a)...

  5. 40 CFR 63.9631 - What are my monitoring requirements?

    Science.gov (United States)

    2010-07-01

    ...) through (e) and monitor the daily average secondary voltage, daily average stack outlet temperature, and... maintain a bag leak detection system to monitor the relative change in particulate matter loadings... according to the requirements in paragraphs (a)(1) through (8) of this section. (1) Monitor the...

  6. 40 CFR 429.12 - Monitoring requirements. [Reserved

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 29 2010-07-01 2010-07-01 false Monitoring requirements. 429.12 Section 429.12 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT GUIDELINES... requirements....

  7. Radiation monitoring in interventional cardiology: a requirement

    Science.gov (United States)

    Rivera, T.; Uruchurtu, E. S.

    2017-01-01

    The increasing of procedures using fluoroscopy in interventional cardiology procedures may increase medical and patients to levels of radiation that manifest in unintended outcomes. Such outcomes may include skin injury and cancer. The cardiologists and other staff members in interventional cardiology are usually working close to the area under examination and they receive the dose primarily from scattered radiation from the patient. Mexico does not have a formal policy for monitoring and recording the radiation dose delivered in hemodynamic establishments. Deterministic risk management can be improved by monitoring the radiation delivered from X-ray devices. The objective of this paper is to provide cardiologist, techniques, nurses, and all medical staff an information on DR levels, about X-ray risks and a simple a reliable method to control cumulative dose.

  8. 40 CFR 410.02 - Monitoring requirements. [Reserved

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 28 2010-07-01 2010-07-01 true Monitoring requirements. 410.02 Section 410.02 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT GUIDELINES AND STANDARDS TEXTILE MILLS POINT SOURCE CATEGORY General Provisions § 410.02 Monitoring requirements....

  9. 40 CFR 63.751 - Monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... design flow rate and organic concentration in the gas stream vented to the carbon adsorption system. The... requirements of part 60, appendix A, Method 21, sections 2, 3, 4.1, 4.2, and 4.4. The calibration gas shall either be representative of the compounds to be measured or shall be methane, and shall be at...

  10. 40 CFR 60.13 - Monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... shall be subject to the provisions of this section upon promulgation of performance specifications for... Performance Specification 1, appendix B, of this part before the performance test required under § 60.8 is... under § 60.8 or within 30 days thereafter in accordance with the applicable performance specification...

  11. Requirements to a Norwegian national automatic gamma monitoring system

    DEFF Research Database (Denmark)

    Lauritzen, B.; Jensen, Per Hedemann; Nielsen, F.

    2005-01-01

    An assessment of the overall requirements to a Norwegian gamma-monitoring network is undertaken with special emphasis on the geographical distribution of automatic gamma monitoring stations, type of detectors in such stations and the sensitivity of thesystem in terms of ambient dose equivalent rate...

  12. 50 CFR 660.312 - Vessel Monitoring System (VMS) requirements.

    Science.gov (United States)

    2010-10-01

    ... 50 Wildlife and Fisheries 9 2010-10-01 2010-10-01 false Vessel Monitoring System (VMS... West Coast Groundfish Fisheries § 660.312 Vessel Monitoring System (VMS) requirements. (a) What is a VMS? A VMS consists of a NMFS OLE type-approved mobile transceiver unit that automatically...

  13. 40 CFR 464.03 - Monitoring and reporting requirements.

    Science.gov (United States)

    2010-07-01

    ... Monitoring and reporting requirements. (a) As an alternative to monitoring for TTO (total toxic organics), an... Grease standard shall be considered equivalent to complying with the TTO standard. Alternate Oil and Grease standards are provided as substitutes for the TTO standards provided in §§ 464.15, 464.16,...

  14. 50 CFR 216.185 - Requirements for monitoring.

    Science.gov (United States)

    2010-10-01

    ... Low Frequency Active (SURTASS LFA sonar) Sonar § 216.185 Requirements for monitoring. (a) In order to... sunset); (2) Use low frequency passive SURTASS sonar to listen for vocalizing marine mammals; and (3)...

  15. Required accuracy and dose thresholds in individual monitoring

    DEFF Research Database (Denmark)

    Christensen, P.; Griffith, R.V.

    1994-01-01

    to energy and angular dependencies of the dosemeter. In type testing of personal dosimetry systems, the estimated overall standard deviation of the dosimetry system is the main parameter to be tested. An important characteristic of a personal dosimetry system is its capability of measuring low doses......The paper follows the approach given in recent revisions of CEC and IAEA recommendations on requirements in individual monitoring for external radiations. The ICRP requirements on overall accuracy for individual monitoring, as given in ICRP Publication 35 (1982), form the basis...... for the specification of detailed accuracy requirements which are needed in practical routine monitoring. The ICRP overall accuracy requirement is defined as an allowable maximum uncertainty factor at the 95% confidence level for a single measurement of the relevant dose quantity, i.e. H(p)(10) and H(p)(0.07). From...

  16. Requirements to a Norwegian National Automatic Gamma Monitoring System

    Energy Technology Data Exchange (ETDEWEB)

    Lauritzen, B.; Hedemann Jensen, P.; Nielsen, F

    2005-04-01

    An assessment of the overall requirements to a Norwegian gamma-monitoring network is undertaken with special emphasis on the geographical distribution of automatic gamma monitoring stations, type of detectors in such stations and the sensitivity of the system in terms of ambient dose equivalent rate increments above the natural background levels. The study is based upon simplified deterministic calculations of the radiological consequences of generic nuclear accident scenarios. The density of gamma monitoring stations has been estimated from an analysis of the dispersion of radioactive materials over large distances using historical weather data; the minimum density is estimated from the requirement that a radioactive plume may not slip unnoticed in between stations of the monitoring network. The sensitivity of the gamma monitoring system is obtained from the condition that events that may require protective intervention measures should be detected by the system. Action levels for possible introduction of sheltering and precautionary foodstuff restrictions are derived in terms of ambient dose equivalent rate. For emergency situations where particulates contribute with only a small fraction of the total ambient dose equivalent rate from the plume, it is concluded that measurements of dose rate are sufficient to determine the need for sheltering; simple dose rate measurements however, are inadequate to determine the need for foodstuff restrictions and spectral measurements are required. (au)

  17. Critical evaluation of factors required to terminate the postclosure monitoring period at solid waste landfills

    DEFF Research Database (Denmark)

    Barlaz, M.A.; Rooker, A.P.; Kjeldsen, Peter

    2002-01-01

    requirements and regulatory activity that might be required to prepare regulators for the large number of requests to terminate postclosure monitoring expected over the next 20 years. An approach in which the frequency and extent of postclosure monitoring is reduced as warranted by site-specific data......Regulations governing the disposal of solid waste in landfills specify that they must be monitored for 30 years after closure unless this period is extended by the governing regulatory authority. Given the wide range of conditions under which refuse is buried, technical criteria, rather than...... water or groundwater. The acceptability of gaseous releases should be evaluated against criteria for odors, the potential for subsurface migration, and greenhouse gas and ozone precursor emissions. The approach presented here must be tested on a site-specific basis to identify additional data...

  18. Monitored Retrievable Storage System Requirements Document. Revision 1

    Energy Technology Data Exchange (ETDEWEB)

    1994-03-01

    This Monitored Retrievable Storage System Requirements Document (MRS-SRD) describes the functions to be performed and technical requirements for a Monitored Retrievable Storage (MRS) facility subelement and the On-Site Transfer and Storage (OSTS) subelement. The MRS facility subelement provides for temporary storage, at a Civilian Radioactive Waste Management System (CRWMS) operated site, of spent nuclear fuel (SNF) contained in an NRC-approved Multi-Purpose Canister (MPC) storage mode, or other NRC-approved storage modes. The OSTS subelement provides for transfer and storage, at Purchaser sites, of spent nuclear fuel (SNF) contained in MPCs. Both the MRS facility subelement and the OSTS subelement are in support of the CRWMS. The purpose of the MRS-SRD is to define the top-level requirements for the development of the MRS facility and the OSTS. These requirements include design, operation, and decommissioning requirements to the extent they impact on the physical development of the MRS facility and the OSTS. The document also presents an overall description of the MRS facility and the OSTS, their functions (derived by extending the functional analysis documented by the Physical System Requirements (PSR) Store Waste Document), their segments, and the requirements allocated to the segments. In addition, the top-level interface requirements of the MRS facility and the OSTS are included. As such, the MRS-SRD provides the technical baseline for the MRS Safety Analysis Report (SAR) design and the OSTS Safety Analysis Report design.

  19. 77 FR 51784 - Federal Acquisition Regulation; Information Collection; Qualification Requirements

    Science.gov (United States)

    2012-08-27

    .... Under the qualification requirements, an end item, or a component thereof, may be required to be... Regulation; Information Collection; Qualification Requirements AGENCY: Department of Defense (DOD), General...: Under the provisions of the Paperwork Reduction Act, the Regulatory Secretariat will be submitting...

  20. The Regulation of Health Requirements for Police and Current Problems

    National Research Council Canada - National Science Library

    Ramazan Akçan; Mahmut Şerif Yıldırım; Aykut Lale; Abdurrahman İsak; Mehmet Cavlak; Aysun Balseven Odabaşı; Ali Rıza Tümer

    2017-01-01

    Introduction: Forensic medicine specialists might prepare reports about health conditions of candidates for Police Organization or assessment of organization's staff based on The Regulation of Health Requirements for Police...

  1. IDENTIFICATION OF DOE'S POST-CLOSURE MONITORING NEEDS AND REQUIREMENTS

    Energy Technology Data Exchange (ETDEWEB)

    M.A. Ebadian, Ph.D.

    1999-01-01

    The 2006 plan sets an ambitious agenda for the U.S. Department of Energy (DOE), Office of Environmental Management (EM) and the remediation of sites contaminated by decades of nuclear weapons production activities. The plan's primary objective is to reduce overall clean up costs by first eliminating the environmental problems that are most expensive to control and safely maintain. In the context of the 2006 Plan, closure refers to the completion of area or facility specific cleanup projects. The cleanup levels are determined by the planned future use of the site or facility. Use restrictions are still undecided for most sites but are highly probable to exclude residential or agricultural activities. Most of the land will be remediated to ''industrial use'' levels with access restrictions and some areas will be closed-off through containment. Portions of the site will be reserved for waste disposal, either as a waste repository or the in-situ immobilization of contaminated soil and groundwater, and land use will be restricted to waste disposal only. The land used for waste disposal will require monitoring and maintenance activities after closure. Most of the land used for industrial use may also require such postclosure activities. The required postclosure monitoring and maintenance activities will be imposed by regulators and stakeholders. Regulators will not approve closure plans without clearly defined monitoring methods using approved technologies. Therefore, among all other more costly and labor-intensive closure-related activities, inadequate planning for monitoring and lack of appropriate monitoring technologies can prevent closure. The purpose of this project is to determine, document, and track the current and evolving postclosure monitoring requirements at DOE-EM sites. This information will aid CMST-CP in guiding its postclosure technology development and deployment efforts.

  2. 40 CFR 75.74 - Annual and ozone season monitoring and reporting requirements.

    Science.gov (United States)

    2010-07-01

    ... every five years and is also required if the flow monitor polynomial coefficients or K factor(s) are...-diluent monitoring system, each flow rate monitoring system, each moisture monitoring system and each... not apply, and, for flow rate monitoring systems, the required RATA load level(s) (or operating level...

  3. Engineering Runtime Requirements-Monitoring Systems Using MDA Technologies

    Science.gov (United States)

    Skene, James; Emmerich, Wolfgang

    The Model-Driven Architecture (MDA) technology toolset includes a language for describing the structure of meta-data, the MOF, and a language for describing consistency properties that data must exhibit, the OCL. Off-the-shelf tools can generate meta-data repositories and perform consistency checking over the data they contain. In this paper we describe how these tools can be used to implement runtime requirements monitoring of systems by modelling the required behaviour of the system, implementing a meta-data repository to collect system data, and consistency checking the repository to discover violations. We evaluate the approach by implementing a contract checker for the SLAng service-level agreement language, a language defined using a MOF meta-model, and integrating the checker into an Enterprise JavaBeans application. We discuss scalability issues resulting from immaturities in the applied technologies, leading to recommendations for their future development.

  4. 40 CFR 468.03 - Monitoring and reporting requirements.

    Science.gov (United States)

    2010-07-01

    ... monitoring procedure for TTO, indirect dischargers may monitor for oil and grease and meet the alternate... alternate monitoring oil and grease standards shall be considered to meet the TTO standard....

  5. Required accuracy and dose thresholds in individual monitoring

    DEFF Research Database (Denmark)

    Christensen, P.; Griffith, R.V.

    1994-01-01

    this uncertainty factor, a value of 21% can be evaluated for the allowable maximum overall standard deviation for dose measurements at dose levels near the annual dose limits increasing to 45% for dose levels at the lower end of the dose range required to be monitored. A method is described for evaluating...... the overall standard deviation of the dosimetry system by combining random and systematic uncertainties in quadrature, and procedures are also given for determining each individual uncertainty connected to the dose measurement. In particular, attention is paid to the evaluation of the combined uncertainty due...... to energy and angular dependencies of the dosemeter. In type testing of personal dosimetry systems, the estimated overall standard deviation of the dosimetry system is the main parameter to be tested. An important characteristic of a personal dosimetry system is its capability of measuring low doses...

  6. 47 CFR 73.53 - Requirements for authorization of antenna monitors.

    Science.gov (United States)

    2010-10-01

    ... 47 Telecommunication 4 2010-10-01 2010-10-01 false Requirements for authorization of antenna... antenna monitors. (a) Antenna monitors shall be verified for compliance with the technical requirements in...) An antenna monitor shall meet the following specifications: (1) The monitor shall be designed...

  7. GENERAL CONSIDERATIONS ON REGULATIONS AND SAFETY REQUIREMENTS FOR QUADRICYCLES

    Directory of Open Access Journals (Sweden)

    Ana Pavlovic

    2015-12-01

    Full Text Available In recent years, a new class of compact vehicles has been emerging and wide-spreading all around Europe: the quadricycle. These four-wheeled motor vehicles, originally derived from motorcycles, are a small and fuel-efficient mean of transportation used in rural or urban areas as an alternative to motorbikes or city cars. In some countries, they are also endorsed by local authorities and institutions which support small and environmentally-friendly vehicles. In this paper, several general considerations on quadricycles will be provided including the vehicle classification, evolution of regulations (as homologation, driver licence, emissions, etc, technical characteristics, safety requirements, most relevant investigations, and other additional useful information (e.g. references, links. It represents an important and actual topic of investigation for designers and manufacturers considering that the new EU regulation on the approval and market surveillance of quadricycles will soon enter in force providing conclusive requirements for functional safety environmental protection of these promising vehicles.

  8. 40 CFR 141.87 - Monitoring requirements for water quality parameters.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 22 2010-07-01 2010-07-01 false Monitoring requirements for water... § 141.87 Monitoring requirements for water quality parameters. All large water systems, and all small... representative of water quality and treatment conditions throughout the system. (d) Monitoring after State...

  9. 40 CFR 141.26 - Monitoring frequency and compliance requirements for radionuclides in community water systems.

    Science.gov (United States)

    2010-07-01

    ... for radionuclides in community water systems. (a) Monitoring and compliance requirements for gross... source of water must begin to conduct initial monitoring for the new source within the first quarter... initial monitoring requirements, a community water system having only one entry point to the distribution...

  10. 10 CFR 20.1502 - Conditions requiring individual monitoring of external and internal occupational dose.

    Science.gov (United States)

    2010-01-01

    ... 10 Energy 1 2010-01-01 2010-01-01 false Conditions requiring individual monitoring of external and internal occupational dose. 20.1502 Section 20.1502 Energy NUCLEAR REGULATORY COMMISSION STANDARDS FOR PROTECTION AGAINST RADIATION Surveys and Monitoring § 20.1502 Conditions requiring individual monitoring of external and internal...

  11. 50 CFR 216.155 - Requirements for monitoring and reporting.

    Science.gov (United States)

    2010-10-01

    ...) Systematic visual observations, by those individuals, described in paragraph (c) of this section, on pinniped... monitoring measures: (1) Visual Land-Based Monitoring. (i) Prior to each missile launch, an observer(s) will... subpart. (5) Both the 60-day and final reports will be subject to review and comment by the...

  12. Actin is required for IFT regulation in Chlamydomonas reinhardtii.

    Science.gov (United States)

    Avasthi, Prachee; Onishi, Masayuki; Karpiak, Joel; Yamamoto, Ryosuke; Mackinder, Luke; Jonikas, Martin C; Sale, Winfield S; Shoichet, Brian; Pringle, John R; Marshall, Wallace F

    2014-09-01

    Assembly of cilia and flagella requires intraflagellar transport (IFT), a highly regulated kinesin-based transport system that moves cargo from the basal body to the tip of flagella [1]. The recruitment of IFT components to basal bodies is a function of flagellar length, with increased recruitment in rapidly growing short flagella [2]. The molecular pathways regulating IFT are largely a mystery. Because actin network disruption leads to changes in ciliary length and number, actin has been proposed to have a role in ciliary assembly. However, the mechanisms involved are unknown. In Chlamydomonas reinhardtii, conventional actin is found in both the cell body and the inner dynein arm complexes within flagella [3, 4]. Previous work showed that treating Chlamydomonas cells with the actin-depolymerizing compound cytochalasin D resulted in reversible flagellar shortening [5], but how actin is related to flagellar length or assembly remains unknown. Here we utilize small-molecule inhibitors and genetic mutants to analyze the role of actin dynamics in flagellar assembly in Chlamydomonas reinhardtii. We demonstrate that actin plays a role in IFT recruitment to basal bodies during flagellar elongation and that when actin is perturbed, the normal dependence of IFT recruitment on flagellar length is lost. We also find that actin is required for sufficient entry of IFT material into flagella during assembly. These same effects are recapitulated with a myosin inhibitor, suggesting that actin may act via myosin in a pathway by which flagellar assembly is regulated by flagellar length.

  13. FTIP1 is an essential regulator required for florigen transport.

    Directory of Open Access Journals (Sweden)

    Lu Liu

    Full Text Available The capacity to respond to day length, photoperiodism, is crucial for flowering plants to adapt to seasonal change. The photoperiodic control of flowering in plants is mediated by a long-distance mobile floral stimulus called florigen that moves from leaves to the shoot apex. Although the proteins encoded by FLOWERING LOCUS T (FT in Arabidopsis and its orthologs in other plants are identified as the long-sought florigen, whether their transport is a simple diffusion process or under regulation remains elusive. Here we show that an endoplasmic reticulum (ER membrane protein, FT-INTERACTING PROTEIN 1 (FTIP1, is an essential regulator required for FT protein transport in Arabidopsis. Loss of function of FTIP1 exhibits late flowering under long days, which is partly due to the compromised FT movement to the shoot apex. FTIP1 and FT share similar mRNA expression patterns and subcellular localization, and they interact specifically in phloem companion cells. FTIP1 is required for FT export from companion cells to sieve elements, thus affecting FT transport through the phloem to the SAM. Our results provide a mechanistic understanding of florigen transport, demonstrating that FT moves in a regulated manner and that FTIP1 mediates FT transport to induce flowering.

  14. FTIP1 is an essential regulator required for florigen transport.

    Science.gov (United States)

    Liu, Lu; Liu, Chang; Hou, Xingliang; Xi, Wanyan; Shen, Lisha; Tao, Zhen; Wang, Yue; Yu, Hao

    2012-01-01

    The capacity to respond to day length, photoperiodism, is crucial for flowering plants to adapt to seasonal change. The photoperiodic control of flowering in plants is mediated by a long-distance mobile floral stimulus called florigen that moves from leaves to the shoot apex. Although the proteins encoded by FLOWERING LOCUS T (FT) in Arabidopsis and its orthologs in other plants are identified as the long-sought florigen, whether their transport is a simple diffusion process or under regulation remains elusive. Here we show that an endoplasmic reticulum (ER) membrane protein, FT-INTERACTING PROTEIN 1 (FTIP1), is an essential regulator required for FT protein transport in Arabidopsis. Loss of function of FTIP1 exhibits late flowering under long days, which is partly due to the compromised FT movement to the shoot apex. FTIP1 and FT share similar mRNA expression patterns and subcellular localization, and they interact specifically in phloem companion cells. FTIP1 is required for FT export from companion cells to sieve elements, thus affecting FT transport through the phloem to the SAM. Our results provide a mechanistic understanding of florigen transport, demonstrating that FT moves in a regulated manner and that FTIP1 mediates FT transport to induce flowering.

  15. 76 FR 50461 - Information Collection Requirement; Defense Federal Acquisition Regulation Supplement; Types of...

    Science.gov (United States)

    2011-08-15

    ... Defense Acquisition Regulations System Information Collection Requirement; Defense Federal Acquisition Regulation Supplement; Types of Contracts AGENCY: Defense Acquisition Regulations System, Department of... information collection requirement. SUMMARY: In compliance with Section 3506(c)(2)(A) of the...

  16. 78 FR 60898 - Regulation on Definition and Requirements for a Nationally Recognized Testing Laboratory...

    Science.gov (United States)

    2013-10-02

    ... Occupational Safety and Health Administration Regulation on Definition and Requirements for a Nationally..., ``definition and requirements for a nationally recognized testing laboratory'' (The Regulation). The Regulation... place all submissions, including any personal information provided, in the public docket...

  17. 40 CFR 63.1452 - What are my monitoring requirements?

    Science.gov (United States)

    2010-07-01

    ... document, “Fabric Filter Bag Leak Detection Guidance,” EPA-454/R-98-015, September 1997. You may obtain a... mechanisms. (iii) Check the compressed air supply for pulse-jet baghouses each day. (iv) Monitor...

  18. 77 FR 8160 - Quality Assurance Requirements for Continuous Opacity Monitoring Systems at Stationary Sources

    Science.gov (United States)

    2012-02-14

    ... AGENCY 40 CFR Part 60 RIN 2060-AH23 Quality Assurance Requirements for Continuous Opacity Monitoring...--Quality Assurance Requirements for Continuous Opacity Monitoring Systems at Stationary Sources Docket, EPA.... Rules establishing quality assurance requirements impose no costs independent from national emission...

  19. 40 CFR 141.88 - Monitoring requirements for lead and copper in source water.

    Science.gov (United States)

    2010-07-01

    ... § 141.88 Monitoring requirements for lead and copper in source water. (a) Sample location, collection... water samples in accordance with the following requirements regarding sample location, number of samples... 40 Protection of Environment 22 2010-07-01 2010-07-01 false Monitoring requirements for lead...

  20. 40 CFR 141.86 - Monitoring requirements for lead and copper in tap water.

    Science.gov (United States)

    2010-07-01

    ... § 141.86 Monitoring requirements for lead and copper in tap water. (a) Sample site location. (1) By the... requirements of § 141.87(e)(2), that it has re-qualified for triennial monitoring. (vii) Any water system... requirements for lead (i.e., a “lead waiver”), the water system must provide certification and...

  1. Cloning-free regulated monitoring of reporter and gene expression

    Directory of Open Access Journals (Sweden)

    Demirkaya Omer

    2009-03-01

    Full Text Available Abstract Background The majority of the promoters, their regulatory elements, and their variations in the human genome remain unknown. Reporter gene technology for transcriptional activity is a widely used tool for the study of promoter structure, gene regulation, and signaling pathways. Construction of transcriptional reporter vectors, including use of cis-acting sequences, requires cloning and time-demanding manipulations, particularly with introduced mutations. Results In this report, we describe a cloning-free strategy to generate transcriptionally-controllable linear reporter constructs. This approach was applied in common transcriptional models of inflammatory response and the interferon system. In addition, it was used to delineate minimal transcriptional activity of selected ribosomal protein promoters. The approach was tested for conversion of genes into TetO-inducible/repressible expression cassettes. Conclusion The simple introduction and tuning of any transcriptional control in the linear DNA product renders promoter activation and regulated gene studies simple and versatile.

  2. 40 CFR 63.773 - Inspection and monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... (CONTINUED) National Emission Standards for Hazardous Air Pollutants From Oil and Natural Gas Production.... Owners or operators that install and operate a flare in accordance with § 63.771(d)(1)(iii) are exempt... feasible point to the catalyst bed outlet. (C) For a flare, a heat sensing monitoring device equipped...

  3. 40 CFR 141.74 - Analytical and monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... from Ozone Science & Engineering, Pergamon Press Ltd., Fairview Park, Elmsford, New York 10523. Copies... 21, 2003, available from Industrial Test Systems, Inc., 1875 Langston St., Rock Hill, SC 29730. Free... method for use with a continuous monitoring instrument provided the chemistry, accuracy, and...

  4. 40 CFR 63.1209 - What are the monitoring requirements?

    Science.gov (United States)

    2010-07-01

    ... Administrator to use CEMS for compliance monitoring for particulate matter, mercury, semivolatile metals, low... ensure compliance with the relevant standard on a continuous basis. (B) Mercury. You may petition the... that affect adsorption and establish limits on those parameters based on the carbon used in...

  5. 40 CFR 63.11583 - What are my monitoring requirements?

    Science.gov (United States)

    2010-07-01

    ... practices at all times. (b) Operate a control device parameter (such as pressure drop or water flow, as... pressure drop. (2) Installation of the bag leak detector, parameter monitoring device, or CPMS at a... alert operators of a leak in the control device filter material. If a bag leak detection system...

  6. 40 CFR 146.23 - Operating, monitoring, and reporting requirements.

    Science.gov (United States)

    2010-07-01

    ...)); and (5) Hydrocarbon storage and enhanced recovery may be monitored on a field or project basis rather... information may be included by reference. (2) Owners or operators of hydrocarbon storage and enhanced recovery... fluid disposal operations; (ii) Monthly for enhanced recovery operations; (iii) Daily during the...

  7. 50 CFR 216.255 - Requirements for monitoring and reporting.

    Science.gov (United States)

    2010-10-01

    ... section may include retention of marine mammals without the need for a separate scientific research permit... animals sighted. (4) Conduct shipboard monitoring to reduce impacts to protected species. Trained marine..., including the time of sighting and the direction of travel, into a marine animal tracking and...

  8. 40 CFR Table 3 of Subpart Aaaa to... - Requirements for Validating Continuous Emission Monitoring Systems (CEMS)

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 6 2010-07-01 2010-07-01 false Requirements for Validating Continuous Emission Monitoring Systems (CEMS) 3 Table 3 of Subpart AAAA to Part 60 Protection of Environment... Continuous Emission Monitoring Systems (CEMS) For the following continuous emission monitoring systems...

  9. Rigid performance requirements assure public safety by regulations

    Energy Technology Data Exchange (ETDEWEB)

    Nickell, R.E.; Glass, R.E.

    1987-07-01

    Title 10, Part 71 of the Code of Federal Regulations (10 CFR 71) provides a set of prescriptive performance test requirements for spent nuclear fuel and high-level waste transport packaging containment systems. The hypothetical accident conditions, which involve a sequence of impact, puncture, fire, and water immersion events, are referred to as rigid because of their extremely prescriptive nature. These hypothetical accident events have now been placed within the context of real transportation accidents, at least for conventional austenitic stainless steel/ lead gamma shielded cask designs. The assurance of public safety, including the issue of safety margin for very severe accident events is discussed in this paper for both conventional and innovative cask design concepts. A particular risk assessment approach that follows from work at the Lawrence Livermore National Laboratory is suggested.

  10. Rigid performance requirements assure public safety by regulations

    Energy Technology Data Exchange (ETDEWEB)

    Nickell, R.E.; Glass, R.E.

    1987-01-01

    Title 10, Part 71 of the Code of Federal Regulations (10 CFR 71) provides a set of prescriptive performance test requirements for spent nuclear fuel and high-level waste transport packaging containment systems. The hypothetical accident conditions, which involve a sequence of impact, puncture, fire, and water immersion events, are referred to as rigid because of their extremely prescriptive nature. These hypothetical accident events have now been placed within the context of real transportation accidents, at least for conventional austenitic stainless steel/lead gamma shielded cask designs. The assurance of public safety, including the issue of safety margin for very severe accident events, is discussed in this paper for both conventional and innovative cask design concepts. A particular risk assessment approach that follows from work at the Lawrence Livermore National Laboratory is suggested. 6 refs., 2 figs.

  11. 78 FR 25445 - Federal Acquisition Regulation; Submission for OMB Review; Quality Assurance Requirements

    Science.gov (United States)

    2013-05-01

    ... Regulation; Submission for OMB Review; Quality Assurance Requirements AGENCY: Department of Defense (DOD... information collection requirement concerning quality assurance requirements. A notice was published in the..., Quality Assurance Requirements, by any of the following methods: Regulations.gov : http://www.regulations...

  12. 47 CFR 11.52 - EAS code and Attention Signal Monitoring requirements.

    Science.gov (United States)

    2010-10-01

    ... 47 Telecommunication 1 2010-10-01 2010-10-01 false EAS code and Attention Signal Monitoring... SYSTEM (EAS) Emergency Operations § 11.52 EAS code and Attention Signal Monitoring requirements. (a) EAS Participants must be capable of receiving the Attention Signal required by § 11.32(a)(9) and emergency...

  13. 40 CFR 264.97 - General ground-water monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... FACILITIES Releases From Solid Waste Management Units § 264.97 General ground-water monitoring requirements. The owner or operator must comply with the following requirements for any ground-water monitoring... 40 Protection of Environment 25 2010-07-01 2010-07-01 false General ground-water...

  14. 40 CFR 63.2269 - What are my monitoring installation, operation, and maintenance requirements?

    Science.gov (United States)

    2010-07-01

    ... Composite Wood Products Initial Compliance Requirements § 63.2269 What are my monitoring installation... moisture monitoring. For each furnish or veneer moisture meter, you must meet the requirements in... in the 25 to 35 percent (dry basis) moisture content range. For veneer redryers, use a...

  15. 40 CFR 63.1429 - Process vent monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... National Emission Standards for Hazardous Air Pollutant Emissions for Polyether Polyols Production § 63... required and should be equipped with a continuous recorder. If an acid or base absorbent is used, a...

  16. 77 FR 67366 - Federal Acquisition Regulation; Information Collection; Quality Assurance Requirements

    Science.gov (United States)

    2012-11-09

    ... Regulation; Information Collection; Quality Assurance Requirements AGENCY: Department of Defense (DOD... information collection requirement concerning quality assurance requirements. Public comments are particularly... Information Collection 9000- 0077, Quality Assurance Requirements, by any of the following methods...

  17. 77 FR 66464 - Federal Acquisition Regulation; Submission for OMB Review; Value Engineering Requirements

    Science.gov (United States)

    2012-11-05

    ... Regulation; Submission for OMB Review; Value Engineering Requirements AGENCIES: Department of Defense (DOD... approved information collection requirement concerning Value Engineering Requirements. A notice was...: Submit comments identified by Information Collection 9000- 0027, Value Engineering Requirements, by...

  18. 77 FR 43076 - Federal Acquisition Regulation; Information Collection; Value Engineering Requirements

    Science.gov (United States)

    2012-07-23

    ... Regulation; Information Collection; Value Engineering Requirements AGENCIES: Department of Defense (DOD... approved information collection requirement concerning Value Engineering Requirements. Public comments are...: Submit comments identified by Information Collection 9000- 0027, Value Engineering Requirements, by...

  19. 78 FR 16184 - Revision to Ambient Nitrogen Dioxide Monitoring Requirements

    Science.gov (United States)

    2013-03-14

    ... 98th percentile form averaged over three years and a level of 100 parts per billion (ppb), reflecting... 53 ppb. As part of the NAAQS rulemaking, the EPA promulgated revisions to requirements for minimum... groups and one citizen supporting the proposed revisions to the schedule for implementing near-road NO 2...

  20. 75 FR 81126 - Revisions to Lead Ambient Air Monitoring Requirements

    Science.gov (United States)

    2010-12-27

    ... Physicians for a Healthy Environment, Leslie and Jack Warden, WEACT for Environmental Justice and the Wasatch... requirements, the Natural Resources Defense Council (NRDC), the Missouri Coalition for the Environment... provisions of the final lead NAAQS rule. See Missouri Coalition for the Environment, et al. v. EPA, (DC...

  1. Thyroid hormone is required for hypothalamic neurons regulating cardiovascular functions.

    Science.gov (United States)

    Mittag, Jens; Lyons, David J; Sällström, Johan; Vujovic, Milica; Dudazy-Gralla, Susi; Warner, Amy; Wallis, Karin; Alkemade, Anneke; Nordström, Kristina; Monyer, Hannah; Broberger, Christian; Arner, Anders; Vennström, Björn

    2013-01-01

    Thyroid hormone is well known for its profound direct effects on cardiovascular function and metabolism. Recent evidence, however, suggests that the hormone also regulates these systems indirectly through the central nervous system. While some of the molecular mechanisms underlying the hormone's central control of metabolism have been identified, its actions in the central cardiovascular control have remained enigmatic. Here, we describe a previously unknown population of parvalbuminergic neurons in the anterior hypothalamus that requires thyroid hormone receptor signaling for proper development. Specific stereotaxic ablation of these cells in the mouse resulted in hypertension and temperature-dependent tachycardia, indicating a role in the central autonomic control of blood pressure and heart rate. Moreover, the neurons exhibited intrinsic temperature sensitivity in patch-clamping experiments, providing a new connection between cardiovascular function and core temperature. Thus, the data identify what we believe to be a novel hypothalamic cell population potentially important for understanding hypertension and indicate developmental hypothyroidism as an epigenetic risk factor for cardiovascular disorders. Furthermore, the findings may be beneficial for treatment of the recently identified patients that have a mutation in thyroid hormone receptor α1.

  2. Comparison and Cost Analysis of Drinking Water Quality Monitoring Requirements versus Practice in Seven Developing Countries

    Directory of Open Access Journals (Sweden)

    Jonny Crocker

    2014-07-01

    Full Text Available Drinking water quality monitoring programs aim to support provision of safe drinking water by informing water quality management. Little evidence or guidance exists on best monitoring practices for low resource settings. Lack of financial, human, and technological resources reduce a country’s ability to monitor water supply. Monitoring activities were characterized in Cambodia, Colombia, India (three states, Jordan, Peru, South Africa, and Uganda according to water sector responsibilities, monitoring approaches, and marginal cost. The seven study countries were selected to represent a range of low resource settings. The focus was on monitoring of microbiological parameters, such as E. coli, coliforms, and H2S-producing microorganisms. Data collection involved qualitative and quantitative methods. Across seven study countries, few distinct approaches to monitoring were observed, and in all but one country all monitoring relied on fixed laboratories for sample analysis. Compliance with monitoring requirements was highest for operational monitoring of large water supplies in urban areas. Sample transport and labor for sample collection and analysis together constitute approximately 75% of marginal costs, which exclude capital costs. There is potential for substantive optimization of monitoring programs by considering field-based testing and by fundamentally reconsidering monitoring approaches for non-piped supplies. This is the first study to look quantitatively at water quality monitoring practices in multiple developing countries.

  3. Comparison and cost analysis of drinking water quality monitoring requirements versus practice in seven developing countries.

    Science.gov (United States)

    Crocker, Jonny; Bartram, Jamie

    2014-07-18

    Drinking water quality monitoring programs aim to support provision of safe drinking water by informing water quality management. Little evidence or guidance exists on best monitoring practices for low resource settings. Lack of financial, human, and technological resources reduce a country's ability to monitor water supply. Monitoring activities were characterized in Cambodia, Colombia, India (three states), Jordan, Peru, South Africa, and Uganda according to water sector responsibilities, monitoring approaches, and marginal cost. The seven study countries were selected to represent a range of low resource settings. The focus was on monitoring of microbiological parameters, such as E. coli, coliforms, and H2S-producing microorganisms. Data collection involved qualitative and quantitative methods. Across seven study countries, few distinct approaches to monitoring were observed, and in all but one country all monitoring relied on fixed laboratories for sample analysis. Compliance with monitoring requirements was highest for operational monitoring of large water supplies in urban areas. Sample transport and labor for sample collection and analysis together constitute approximately 75% of marginal costs, which exclude capital costs. There is potential for substantive optimization of monitoring programs by considering field-based testing and by fundamentally reconsidering monitoring approaches for non-piped supplies. This is the first study to look quantitatively at water quality monitoring practices in multiple developing countries.

  4. 40 CFR Appendix P to Part 51 - Minimum Emission Monitoring Requirements

    Science.gov (United States)

    2010-07-01

    ... Requirements P Appendix P to Part 51 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS REQUIREMENTS FOR PREPARATION, ADOPTION, AND SUBMITTAL OF IMPLEMENTATION PLANS Pt. 51, App. P Appendix P to Part 51—Minimum Emission Monitoring Requirements 1.0 Purpose. This appendix P sets forth...

  5. Discovery of a Splicing Regulator Required for Cell Cycle Progression

    Energy Technology Data Exchange (ETDEWEB)

    Suvorova, Elena S.; Croken, Matthew; Kratzer, Stella; Ting, Li-Min; Conde de Felipe, Magnolia; Balu, Bharath; Markillie, Lye Meng; Weiss, Louis M.; Kim, Kami; White, Michael W.

    2013-02-01

    In the G1 phase of the cell division cycle, eukaryotic cells prepare many of the resources necessary for a new round of growth including renewal of the transcriptional and protein synthetic capacities and building the machinery for chromosome replication. The function of G1 has an early evolutionary origin and is preserved in single and multicellular organisms, although the regulatory mechanisms conducting G1 specific functions are only understood in a few model eukaryotes. Here we describe a new G1 mutant from an ancient family of apicomplexan protozoans. Toxoplasma gondii temperature-sensitive mutant 12-109C6 conditionally arrests in the G1 phase due to a single point mutation in a novel protein containing a single RNA-recognition-motif (TgRRM1). The resulting tyrosine to asparagine amino acid change in TgRRM1 causes severe temperature instability that generates an effective null phenotype for this protein when the mutant is shifted to the restrictive temperature. Orthologs of TgRRM1 are widely conserved in diverse eukaryote lineages, and the human counterpart (RBM42) can functionally replace the missing Toxoplasma factor. Transcriptome studies demonstrate that gene expression is downregulated in the mutant at the restrictive temperature due to a severe defect in splicing that affects both cell cycle and constitutively expressed mRNAs. The interaction of TgRRM1 with factors of the tri-SNP complex (U4/U6 & U5 snRNPs) indicate this factor may be required to assemble an active spliceosome. Thus, the TgRRM1 family of proteins is an unrecognized and evolutionarily conserved class of splicing regulators. This study demonstrates investigations into diverse unicellular eukaryotes, like the Apicomplexa, have the potential to yield new insights into important mechanisms conserved across modern eukaryotic kingdoms.

  6. Ground Water Monitoring Requirements for Hazardous Waste Treatment, Storage and Disposal Facilities

    Science.gov (United States)

    The groundwater monitoring requirements for hazardous waste treatment, storage and disposal facilities (TSDFs) are just one aspect of the Resource Conservation and Recovery Act (RCRA) hazardous waste management strategy for protecting human health and the

  7. 40 CFR 98.44 - Monitoring and QA/QC requirements.

    Science.gov (United States)

    2010-07-01

    ... (CONTINUED) MANDATORY GREENHOUSE GAS REPORTING Electricity Generation § 98.44 Monitoring and QA/QC requirements. Follow the applicable quality assurance procedures for CO2 emissions in appendices B, D, and G to 40 CFR part 75....

  8. Growth Conditions Regulate the Requirements for Caulobacter Chromosome Segregation

    DEFF Research Database (Denmark)

    Shebelut, Conrad W.; Jensen, Rasmus Bugge; Gitai, Zemer

    2009-01-01

    Growth environments are important metabolic and developmental regulators. Here we demonstrate a growth environment-dependent effect on Caulobacter chromosome segregation of a small-molecule inhibitor of the MreB bacterial actin cytoskeleton. Our results also implicate ParAB as important segregation...... determinants, suggesting that multiple distinct mechanisms can mediate Caulobacter chromosome segregation and that their relative contributions can be environmentally regulated....

  9. Growth Conditions Regulate the Requirements for Caulobacter Chromosome Segregation▿ †

    OpenAIRE

    Shebelut, Conrad W.; Jensen, Rasmus B.; Gitai, Zemer

    2008-01-01

    Growth environments are important metabolic and developmental regulators. Here we demonstrate a growth environment-dependent effect on Caulobacter chromosome segregation of a small-molecule inhibitor of the MreB bacterial actin cytoskeleton. Our results also implicate ParAB as important segregation determinants, suggesting that multiple distinct mechanisms can mediate Caulobacter chromosome segregation and that their relative contributions can be environmentally regulated.

  10. Growth conditions regulate the requirements for Caulobacter chromosome segregation.

    Science.gov (United States)

    Shebelut, Conrad W; Jensen, Rasmus B; Gitai, Zemer

    2009-02-01

    Growth environments are important metabolic and developmental regulators. Here we demonstrate a growth environment-dependent effect on Caulobacter chromosome segregation of a small-molecule inhibitor of the MreB bacterial actin cytoskeleton. Our results also implicate ParAB as important segregation determinants, suggesting that multiple distinct mechanisms can mediate Caulobacter chromosome segregation and that their relative contributions can be environmentally regulated.

  11. 76 FR 81408 - Contractor Legal Management Requirements; Acquisition Regulations

    Science.gov (United States)

    2011-12-28

    ... principles and contract clauses. Consequently, the Department has an ongoing obligation to monitor, supervise... covered by this part? 719.4 Are law firms that are retained by contract by the department covered by this... contract the Department awards directly to retained legal counsel exceeding $100,000. Sec. 719.4 Are law...

  12. 50 CFR 18.128 - What are the mitigation, monitoring, and reporting requirements?

    Science.gov (United States)

    2010-10-01

    ... require a monitor on the site of the activity or on board drill ships, drill rigs, aircraft, icebreakers... or require that the operator conduct activities after the female bears emerge from their dens. We... Alaska Regional Director (Attn: Marine Mammals Management Office) within 90 days after completion...

  13. 78 FR 55060 - Proposed Information Collection; Comment Request; Expanded Vessel Monitoring System Requirement...

    Science.gov (United States)

    2013-09-09

    ... commercial fishing vessels are required to install and use a vessel monitoring system (VMS) that... pilot VMS program was implemented on January 1, 2004. The pilot program required vessels registered to Pacific Coast groundfish fishery limited entry permits to carry and use VMS transceiver units...

  14. Monitoring the regulation of gene expression in a growing organ using a fluid mechanics formalism

    Directory of Open Access Journals (Sweden)

    Dreyer Erwin

    2010-03-01

    Full Text Available Abstract Background Technological advances have enabled the accurate quantification of gene expression, even within single cell types. While transcriptome analyses are routinely performed, most experimental designs only provide snapshots of gene expression. Molecular mechanisms underlying cell fate or positional signalling have been revealed through these discontinuous datasets. However, in developing multicellular structures, temporal and spatial cues, known to directly influence transcriptional networks, get entangled as the cells are displaced and expand. Access to an unbiased view of the spatiotemporal regulation of gene expression occurring during development requires a specific framework that properly quantifies the rate of change of a property in a moving and expanding element, such as a cell or an organ segment. Results We show how the rate of change in gene expression can be quantified by combining kinematics and real-time polymerase chain reaction data in a mechanistic model which considers any organ as a continuum. This framework was applied in order to assess the developmental regulation of the two reference genes Actin11 and Elongation Factor 1-β in the apex of poplar root. The growth field was determined by time-lapse photography and transcript density was obtained at high spatial resolution. The net accumulation rates of the transcripts of the two genes were found to display highly contrasted developmental profiles. Actin11 showed pulses of up and down regulation in the accelerating and decelerating parts of the growth zone while the dynamic of EF1β were much slower. This framework provides key information about gene regulation in a developing organ, such as the location, the duration and the intensity of gene induction/repression. Conclusions We demonstrated that gene expression patterns can be monitored using the continuity equation without using mutants or reporter constructions. Given the rise of imaging technologies, this

  15. An assessment of monitoring requirements and costs of 'Reduced Emissions from Deforestation and Degradation'

    Directory of Open Access Journals (Sweden)

    McCallum Ian

    2009-08-01

    Full Text Available Abstract Background Negotiations on a future climate policy framework addressing Reduced Emissions from Deforestation and Degradation (REDD are ongoing. Regardless of how such a framework will be designed, many technical solutions of estimating forest cover and forest carbon stock change exist to support policy in monitoring and accounting. These technologies typically combine remotely sensed data with ground-based inventories. In this article we assess the costs of monitoring REDD based on available technologies and requirements associated with key elements of REDD policy. Results We find that the design of a REDD policy framework (and specifically its rules can have a significant impact on monitoring costs. Costs may vary from 0.5 to 550 US$ per square kilometre depending on the required precision of carbon stock and area change detection. Moreover, they follow economies of scale, i.e. single country or project solutions will face relatively higher monitoring costs. Conclusion Although monitoring costs are relatively small compared to other cost items within a REDD system, they should be shared not only among countries but also among sectors, because an integrated monitoring system would have multiple benefits for non-REDD management. Overcoming initialization costs and unequal access to monitoring technologies is crucial for implementation of an integrated monitoring system, and demands for international cooperation.

  16. 77 FR 35925 - Fisheries of the Exclusive Economic Zone Off Alaska; Monitoring and Enforcement Requirements in...

    Science.gov (United States)

    2012-06-15

    ... conveyor belt in the factory before any sorting or processing was done. As described below, the operations... give an estimate of catch for unsampled sets. The current sampling methodologies produce accurate catch... current observer sampling station inspection and approval regulations or processes. Under both monitoring...

  17. Assessment of Unabated Facility Emission Potentials for Evaluating Airborne Radionuclide Monitoring Requirements at Pacific Northwest National Laboratory - 2007

    Energy Technology Data Exchange (ETDEWEB)

    Ballinger, Marcel Y.; Barfuss, Brad C.; Gervais, Todd L.

    2008-01-01

    Assessments were performed to evaluate compliance with the airborne radionuclide emission monitoring requirements in the National Emission Standards for Hazardous Air Pollutants (NESHAP – U.S. Code of Federal Regulations, Title 40, Part 61, Subpart H) and Washington Administrative Code (WAC) 246-247: Radiation Protection – Air Emissions. In these NESHAP assessments, potential unabated offsite doses were evaluated for emission locations at buildings that are part of the consolidated laboratory campus of the Pacific Northwest National Laboratory. This report describes the inventory-based methods and provides the results for the NESHAP assessment performed in 2007.

  18. Assessment of Unabated Facility Emission Potentials for Evaluating Airborne Radionuclide Monitoring Requirements at Pacific Northwest National Laboratory - 2010

    Energy Technology Data Exchange (ETDEWEB)

    Ballinger, Marcel Y.; Gervais, Todd L.; Barnett, J. Matthew

    2011-05-13

    Assessments were performed to evaluate compliance with the airborne radionuclide emission monitoring requirements in the National Emission Standards for Hazardous Air Pollutants ([NESHAP]; U.S. Code of Federal Regulations, Title 40, Part 61, Subpart H) and Washington Administrative Code 246-247: Radiation Protection - Air Emissions. In these NESHAP assessments, potential unabated off-site doses were evaluated for emission locations at buildings that are part of the consolidated laboratory campus of the Pacific Northwest National Laboratory. This report describes the inventory-based methods and provides the results for the NESHAP assessment performed in 2010.

  19. Board Monitoring, Regulation and Performance in the Banking Industry: Evidence from the Market for Corporate Control

    OpenAIRE

    Hagendorff, Jens; Collins, Michael; Keasey, Kevin

    2010-01-01

    Abstract Manuscript Type: Empirical Research Question/Issue: The specific monitoring effect of boards of directors versus industry regulation is unclear. In this paper, we examine how the interaction between bank-level monitoring and regulatory regimes influences the announcement period returns of acquiring banks in the US and twelve European economies. Research Findings: We study three board monitoring mechanisms (independence, CEO-chair duality and diversity) and analyze t...

  20. Thyroid hormone is required for hypothalamic neurons regulating cardiovascular functions

    NARCIS (Netherlands)

    Mittag, J.; Lyons, D.J.; Sällström, J.; Vujoviv, M.; Dudazy-Gralla, S.; Warner, A.; Wallis, K.; Alkemade, A.; Nordström, K.; Monyer, H.; Broberger, C.; Arner, A.; Vennström, B.

    2013-01-01

    Thyroid hormone is well known for its profound direct effects on cardiovascular function and metabolism. Recent evidence, however, suggests that the hormone also regulates these systems indirectly through the central nervous system. While some of the molecular mechanisms underlying the hormone’s

  1. Requirements to micro-unmanned aircraft systems in civil protection and environmental monitoring

    Energy Technology Data Exchange (ETDEWEB)

    Fischer-Stabel, Peter; Hardt, Christopher [Univ. of Applied Sciences Trier, Birkenfeld (Germany). Dept. of Environmental Planning

    2013-07-01

    Especially in application fields such as environmental monitoring or in the field of information and operations management with technical or natural disasters, increased demands on communication and sensor technology to micro unmanned aircraft systems (UAS) are given. These are currently covered by the system manufacturers, however inadequately. The use case of wildlife monitoring with micro UAS comes with some special requirements and problems, addressed in this paper. (orig.)

  2. Epigenetic Regulation in Hepatocellular Carcinoma Requires Long Noncoding RNAs

    Directory of Open Access Journals (Sweden)

    Laura Amicone

    2015-01-01

    Full Text Available Recent evidence has proven the relevance of epigenetic changes in the development of hepatocellular carcinoma (HCC, the major adult liver malignancy. Moreover, HCC onset and progression correlate with the deregulation of several long noncoding RNAs (lncRNAs, exhibiting great biological significance. As discussed in this review, many of these transcripts are able to specifically act as tumor suppressors or oncogenes by means of their role as molecular platforms. Indeed, these lncRNAs are able to bind and recruit epigenetic modifiers on specific genomic loci, ultimately resulting in regulation of the gene expression relevant in cancer development. The evidence presented in this review highlights that lncRNAs-mediated epigenetic regulation should be taken into account for potential targeted therapeutic approaches.

  3. Financial Market Regulation in Germany - Capital Requirements of Financial Institutions

    Directory of Open Access Journals (Sweden)

    Daniel Karl Detzer

    2015-03-01

    Full Text Available This paper examines capital adequacy regulation in Germany. The first part reviews capital adequacy regulation from the 1930s up to the financial crisis and identifies two main trends: a gradual softening of the eligibility criteria for equity and increasing reliance on internal risk models. While the first trend has been reversed following the financial crisis, internal risk models still play a central role. Therefore, the second part discusses the problems with the use of internal risk models and discusses the potentials of Basel 2.5 and Basel III to alleviate the identified problems. It is concluded that the relevant problems are not resolved. Therefore, in the final part some suggestions of how the problems could be addressed properly are given.

  4. 75 FR 60263 - Federal Acquisition Regulation; Offering a Construction Requirement-8(a) Program

    Science.gov (United States)

    2010-09-29

    ... Federal Acquisition Regulation; Offering a Construction Requirement--8(a) Program AGENCIES: Department of... Regulations Council (Councils) are issuing a final rule amending the Federal Acquisition Regulation (FAR) to revise FAR subpart 19.8, Contracting with the Small Business Administration (The 8(a) Program),...

  5. 13 CFR 120.1511 - Certification and other reporting and notification requirements for Other Regulated SBLCs.

    Science.gov (United States)

    2010-01-01

    ...) Certification. An SBLC seeking Other Regulated SBLC status must certify to SBA in writing that its lending... or state banking regulator that regulates the lending activities of the SBLC; (2) A statement that... may appeal this decision to the AA/CA. (g) Failure to satisfy requirements. In the event that an...

  6. 40 CFR Appendix A to Part 58 - Quality Assurance Requirements for SLAMS, SPMs and PSD Air Monitoring

    Science.gov (United States)

    2010-07-01

    ... Appendix A to Part 58—Quality Assurance Requirements for SLAMS, SPMs and PSD Air Monitoring 1. General... specifies the minimum quality system requirements applicable to SLAMS air monitoring data and PSD data for... of the air being measured. Monitoring organizations must develop quality assurance project...

  7. Awareness during anaesthesia for surgery requiring evoked potential monitoring: A pilot study

    Directory of Open Access Journals (Sweden)

    Pritish J Korula

    2017-01-01

    Full Text Available Background: Evoked potential monitoring such as somatosensory-evoked potential (SSEP or motor-evoked potential (MEP monitoring during surgical procedures in proximity to the spinal cord requires minimising the minimum alveolar concentrations (MACs below the anaesthetic concentrations normally required (1 MAC to prevent interference in amplitude and latency of evoked potentials. This could result in awareness. Our primary objective was to determine the incidence of awareness while administering low MAC inhalational anaesthetics for these unique procedures. The secondary objective was to assess the adequacy of our anaesthetic technique from neurophysiologist′s perspective. Methods: In this prospective observational pilot study, 61 American Society of Anesthesiologists 1 and 2 patients undergoing spinal surgery for whom intraoperative evoked potential monitoring was performed were included; during the maintenance phase, 0.7-0.8 MAC of isoflurane was targeted. We evaluated the intraoperative depth of anaesthesia using a bispectral (BIS index monitor as well as the patients response to surgical stimulus (PRST scoring system. Post-operatively, a modified Bruce questionnaire was used to verify awareness. The adequacy of evoked potential readings was also assessed. Results: Of the 61 patients, no patient had explicit awareness. Intraoperatively, 19 of 61 patients had a BIS value of above sixty at least once, during surgery. There was no correlation with PRST scoring and BIS during surgery. Fifty-four out of 61 patient′s evoked potential readings were deemed ′good′ or ′fair′ for the conduct of electrophysiological monitoring. Conclusions: This pilot study demonstrates that administering low MAC inhalational anaesthetics to facilitate evoked potential monitoring does not result in explicit awareness. However, larger studies are needed to verify this. The conduct of SSEP electrophysiological monitoring was satisfactory with the use of this

  8. 40 CFR 63.7188 - What are my monitoring installation, operation, and maintenance requirements?

    Science.gov (United States)

    2010-07-01

    ... Semiconductor Manufacturing Compliance Requirements § 63.7188 What are my monitoring installation, operation... emissions of your semiconductor process vent through a closed vent system to a control device, you must... evaluation procedure in § 63.7187(i) to demonstrate compliance, you must use the information from the...

  9. 40 CFR Table 6 to Subpart Jjj of... - Requirements for Validating Continuous Emission Monitoring Systems (CEMS)

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 8 2010-07-01 2010-07-01 false Requirements for Validating Continuous Emission Monitoring Systems (CEMS) 6 Table 6 to Subpart JJJ of Part 62 Protection of Environment... Combustion Units Constructed on or Before August 30, 1999 Pt. 62, Subpt. JJJ, Table 6 Table 6 to Subpart JJJ...

  10. 40 CFR Table 7 to Subpart Jjj of... - Requirements for Continuous Emission Monitoring Systems (CEMS) a

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 8 2010-07-01 2010-07-01 false Requirements for Continuous Emission Monitoring Systems (CEMS) a 7 Table 7 to Subpart JJJ of Part 62 Protection of Environment ENVIRONMENTAL... Constructed on or Before August 30, 1999 Pt. 62, Subpt. JJJ, Table 7 Table 7 to Subpart JJJ of Part 62...

  11. 10 CFR 50.65 - Requirements for monitoring the effectiveness of maintenance at nuclear power plants.

    Science.gov (United States)

    2010-01-01

    ... maintenance at nuclear power plants. 50.65 Section 50.65 Energy NUCLEAR REGULATORY COMMISSION DOMESTIC... Construction Permits § 50.65 Requirements for monitoring the effectiveness of maintenance at nuclear power..., including normal shutdown operations. (a)(1) Each holder of an operating license for a nuclear power......

  12. 40 CFR 63.996 - General monitoring requirements for control and recovery devices.

    Science.gov (United States)

    2010-07-01

    ... SOURCE CATEGORIES (CONTINUED) National Emission Standards for Closed Vent Systems, Control Devices, Recovery Devices and Routing to a Fuel Gas System or a Process § 63.996 General monitoring requirements for... cycle and the temperature of the carbon bed determined within 15 minutes of the completion of...

  13. 77 FR 59053 - Fisheries of the Exclusive Economic Zone Off Alaska; Monitoring and Enforcement Requirements in...

    Science.gov (United States)

    2012-09-26

    ... the Exclusive Economic Zone Off Alaska; Monitoring and Enforcement Requirements in the Bering Sea and... sampled, or carry one observer and use a motion-compensated scale to weigh Pacific cod before it is....S. groundfish fisheries of the exclusive economic zone off Alaska under the Fishery Management...

  14. Required sample size for monitoring stand dynamics in strict forest reserves: a case study

    Science.gov (United States)

    Diego Van Den Meersschaut; Bart De Cuyper; Kris Vandekerkhove; Noel Lust

    2000-01-01

    Stand dynamics in European strict forest reserves are commonly monitored using inventory densities of 5 to 15 percent of the total surface. The assumption that these densities guarantee a representative image of certain parameters is critically analyzed in a case study for the parameters basal area and stem number. The required sample sizes for different accuracy and...

  15. 40 CFR 141.85 - Public education and supplemental monitoring requirements.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 22 2010-07-01 2010-07-01 false Public education and supplemental monitoring requirements. 141.85 Section 141.85 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... lead more than healthy adults. Lead is stored in the bones, and it can be released later in...

  16. Functions and requirements for Hanford single-shell tank leakage detection and monitoring

    Energy Technology Data Exchange (ETDEWEB)

    Cruse, J.M.; Ohl, P.C.

    1995-04-19

    This document provides the initial functions and requirements for leakage detection and monitoring applicable to past and potential future leakage from the Hanford Site`s 149 single-shell high-level waste tanks. This mission is a part of the overall mission of the Westinghouse Hanford Company Tank Waste Remediation System division to remediate the tank waste in a safe and acceptable manner. Systems engineering principles are being applied to this effort. This document reflects the an initial step in the systems engineering approach to decompose the mission into primary functions and requirements. The document is considered approximately 30% complete relative to the effort required to produce a final version that can be used to support demonstration and/or procurement of technologies. The functions and requirements in this document apply to detection and monitoring of below ground leaks from SST containment boundaries and the resulting soil contamination. Leakage detection and monitoring is invoked in the TWRS Program in three fourth level functions: (1) Store Waste, (2) Retrieve Waste, and (3) Disposition Excess Facilities (as identified in DOE/RL-92-60 Rev. 1, Tank Waste Remediation System Functions and Requirements).

  17. 78 FR 5452 - Federal Acquisition Regulation; Submission for OMB Review; Qualification Requirements

    Science.gov (United States)

    2013-01-25

    ... award of a contract. Under the qualification requirements, an end item, or a component thereof, may be... offeror before award of a contract. Under the qualification requirements, an end item, or a component... Regulation; Submission for OMB Review; Qualification Requirements AGENCIES: Department of Defense...

  18. Metacognition in Later Adulthood: Spared Monitoring Can Benefit Older Adults' Self-regulation.

    Science.gov (United States)

    Hertzog, Christopher; Dunlosky, John

    2011-06-01

    Metacognition includes two key concepts: monitoring of internal states, and adaptive use of control strategies based on that monitoring. We review studies that indicate that aging does not materially affect the accuracy of elementary forms of monitoring encoding and retrieval states in episodic memory tasks, even though it does influence episodic memory itself. Spared monitoring accuracy can therefore serve as a basis for older adults' use of compensatory strategies to achieve learning goals, despite the influence of aging on mechanisms of learning. Metacognitive intervention studies based on this premise show greater effects on learning than traditional strategy-training approaches. Use of strategies for self-regulation, informed by monitoring, may be an important tool for older adults' effective cognitive functioning in everyday life.

  19. Condition monitoring requirements for the development of a space nuclear propulsion module

    Science.gov (United States)

    Wagner, Robert C.

    1993-01-01

    To facilitate the development of a space nuclear propulsion module for manned flights to Mars, requirements must be established early in the technology cycle. The long lead times for the acquisition of the engine system and nuclear test facilities demands that the engine system, size, performance, safety goals and condition monitoring philosophy be defined at the earliest possible time. These systems are highly complex and require a large multi-disciplinary systems engineering team to develop and track the requirements and to ensure that the as-built system reflects the intent of the mission. An effective methodology has been devised coupled with sophisticated computer tools to effectivly develop and interpret the functional requirements. These requirements can then be decomposed down to the specification level for implementation. This paper discusses the application of the methodology and the analyses to develop condition monitoring requirements under a contract with the National Aeronautics and Space Administration (NASA) Lewis Research Center (LeRC) Nuclear Propulsion Office (NPO).

  20. Monitoring Disinfection Byproduct Forming Potential with Simultaneous Absorbance Spectra and Fluorescence Excitation-Emission Mapping: Supporting Stage 2 EPA Regulation Monitoring Compliance

    Science.gov (United States)

    Gilmore, A. M.

    2011-12-01

    Water treatment plants in the United States will soon (by 2013 to 2013) be required to initialize enhanced monitoring for reducing disinfection by-products (DBPs) to meet the Stage 2 levels enforced by the EPA. The key to successfully meeting these requirements lies in the treatment plant's ability to deal with often dramatic source-water variations in natural organic matter (NOM) content. Whereas the regulated levels of NOM must be determined by measuring total organic carbon (TOC) often this parameter does not provide rapid or cost-effective qualitative or quantitative assessment of the various humic, fulvic and other aromatic NOM components. However, 2 main optical techniques namely UV absorbance and fluorescence excitation-emission mapping can be used for rapid assessment with precise identification of humic and fulvic components. This study presents data from a new type of instrument which simultaneously measures the UV-VIS absorbance spectrum and EEM. The rapid absorbance-EEM is facilitated by a single system that is more than 100 time faster than conventional scanning absorbance and fluoresence optical benches. The new system can continuously collect EEMs and absorbance spectra at a rate often greater than 1 per min with the extra capacity to monitor the UV254 absorbance and fluorescence emission spectrum excited at 254 nm in 4 ms intervals (an equivalent scan rate of 5.5 million nm/min). The EEM spectral data is corrected for all instrumental response factors including concentration dependent inner-filter effects. The accumulated EEM data sets can be modeled using conventional peak identification, PARAFAC and or PCA analysis of the fractionated samples to predict the trihalomethane forming potential (THMFP). This study compares the effectiveness of THFMP predictive models based on these three techniques and explains how these can be readily employed to facilitate the Stage 2 regulation compliance for DBP monitoring.

  1. 75 FR 14448 - Food and Drug Administration Clinical Trial Requirements, Regulations, Compliance, and Good...

    Science.gov (United States)

    2010-03-25

    ... HUMAN SERVICES Food and Drug Administration Food and Drug Administration Clinical Trial Requirements, Regulations, Compliance, and Good Clinical Practices; Public Workshop AGENCY: Food and Drug Administration, HHS. ACTION: Notice of public workshop. SUMMARY: The Food and Drug Administration (FDA) Los...

  2. Monitoring Conformance and Containment for Geological Carbon Storage: Can Technology Meet Policy and Public Requirements?

    Science.gov (United States)

    Lawton, D. C.; Osadetz, K.

    2014-12-01

    The Province of Alberta, Canada identified carbon capture and storage (CCS) as a key element of its 2008 Climate Change strategy. The target is a reduction in CO2 emissions of 139 Mt/year by 2050. To encourage uptake of CCS by industry, the province has provided partial funding to two demonstration scale projects, namely the Quest Project by Shell and partners (CCS), and the Alberta Carbon Trunk Line Project (pipeline and CO2-EOR). Important to commercial scale implementation of CCS will be the requirement to prove conformance and containment of the CO2 plume injected during the lifetime of the CCS project. This will be a challenge for monitoring programs. The Containment and Monitoring Institute (CaMI) is developing a Field Research Station (FRS) to calibrate various monitoring technologies for CO2 detection thresholds at relatively shallow depths. The objective being assessed with the FRS is sensitivity for early detection of loss of containment from a deeper CO2 storage project. In this project, two injection wells will be drilled to sandstone reservoir targets at depths of 300 m and 700 m. Up to four observation wells will be drilled with monitoring instruments installed. Time-lapse surface and borehole monitoring surveys will be undertaken to evaluate the movement and fate of the CO2 plume. These will include seismic, microseismic, cross well, electrical resistivity, electromagnetic, gravity, geodetic and geomechanical surveys. Initial baseline seismic data from the FRS will presented.

  3. Satellite data acquisition requirements for monitoring of permafrost in polar regions

    Science.gov (United States)

    Bartsch, Annett

    2015-04-01

    Requirements for space based monitoring of permafrost features had been already defined within the IGOS Cryosphere Theme Report at the start of the IPY in 2007 (IGOS, 2007). In 2012 the Polar Space Task Group (PSTG, http://www.wmo.int/pages/prog/sat/pstg_en.php) has been established as the coordinating body of space agencies, in particular the Space Task Group (STG), for space -based observations of Polar Regions after the International Polar Year (IPY) and under the auspices of the World Meteorological Organization's (WMO) Executive Council Panel of Experts on Polar Observations Research and Services (EC-PORS). The PSTG identified the need to review the requirements for permafrost monitoring and to update these requirements as necessary in 2013. Relevant surveys with focus on satellite data are already available from the ESA DUE Permafrost User requirements survey (2009), the United States National Research Council (2014) and the ESA - CliC - IPA - GTN -P workshop in February 2014. These reports have been reviewed and specific needs discussed within the community. Acquisition requirements for monitoring of especially terrain changes (incl. rock glaciers and coastal erosion) and lakes (extent, ice properties etc.) with respect to current satellite missions have been specified. Of special interest for these applications are SAR missions. Current acquisition strategies for space borne SAR data only partially cover polar permafrost regions and some of the longterm in-situ measurement sites. Many stations are located in the proximity to coastal areas and glaciers which to some extent may allow joint usage by different cryosphere applications but requirements may deviate. The results of the discussion are presented in this paper.

  4. A Next Generation Air Monitor: Combining Orion and ISS Requirements for a Common Major Constituent Analyzer

    Science.gov (United States)

    Burchfield, David E.; Tissandier, Michael; Niu, William Hsein-Chi; Lewis, John F.

    2013-01-01

    The Major Constituent Analyzer (MCA) is a mass spectrometer-based instrument designed to provide critical monitoring of six major atmospheric constituents; nitrogen, oxygen, hydrogen, carbon dioxide, methane, and water vapor on-board the International Space Station. The analyzer has been an integral part of the Environmental Control and Life Support System (ECLSS) since the station went on-line. The Orion Air Monitor (OAM) was derived from the MCA and heavily optimized for reduced mass, lower power, faster water vapor response, and maintenance-free operation. The resulting OAM is approximately the size of the analyzer portion of the MCA, orbital-replacement unit 02 (ORU 02), while incorporating the functions of three other modules: Data Processing and Communication (ORU 01), Verification Gas Assembly (ORU 08), and Low Voltage Power Supply (ORU 04). The overlap in MCA and OAM requirements makes it possible to derive a common Air Monitor design that spans both applications while minimally impacting the weight and power limits imposed by the Multipurpose Crew Vehicle (MPCV). Benefits to ISS include the retirement of ORUs 01, 04, and 08, reducing up-mass and eliminating EEE parts obsolescence issues through the extended ISS mission phases. Benefits to MPCV and future deployed habitats under the Constellation program include greater interchangeability across ECLSS subsystems. This paper discusses the results of the requirements development study, where a superset of ISS and Orion air monitoring requirements were distilled; evaluated against increases in OAM functionality, mass, and power; and traded-off where possible using simple operating mode modifications. A system architecture and preliminary design addressing the common requirements will be presented.

  5. 77 FR 43523 - Revisions to the Unregulated Contaminant Monitoring Regulation (UCMR 3) for Public Water Systems

    Science.gov (United States)

    2012-07-25

    ... AGENCY 40 CFR Parts 141 and 142 RIN 2040-AF10 Revisions to the Unregulated Contaminant Monitoring Regulation (UCMR 3) for Public Water Systems Correction In rule document 2012-9978 appearing on pages 26072... table entitled ``EXHIBIT 1--APPLICABILITY OF UCMR 3 TO WATER UTILITIES BY SYSTEM TYPE AND SIZE'' should...

  6. 77 FR 39182 - Revisions to the Unregulated Contaminant Monitoring Regulation (UCMR 3) for Public Water Systems

    Science.gov (United States)

    2012-07-02

    ... AGENCY 40 CFR Parts 141 and 142 RIN 2040-AF10 Revisions to the Unregulated Contaminant Monitoring Regulation (UCMR 3) for Public Water Systems Correction In rule document 2012-9978 appearing on pages 26072... table entitled ``EXHIBIT 1--APPLICABILITY OF UCMR 3 TO WATER UTILITIES BY SYSTEM TYPE AND SIZE'' should...

  7. Improving Self-Monitoring and Self-Regulation: From Cognitive Psychology to the Classroom

    Science.gov (United States)

    de Bruin, Anique B. H.; van Gog, Tamara

    2012-01-01

    Although there is abundant experimental metamemory research on the relation between students' monitoring, regulation of learning, and learning outcomes, relatively little of this work has influenced educational research and practice. Metamemory research, traditionally based on experimental paradigms from cognitive psychology, can potentially…

  8. Improving Self-Monitoring and Self-Regulation: From Cognitive Psychology to the Classroom

    Science.gov (United States)

    de Bruin, Anique B. H.; van Gog, Tamara

    2012-01-01

    Although there is abundant experimental metamemory research on the relation between students' monitoring, regulation of learning, and learning outcomes, relatively little of this work has influenced educational research and practice. Metamemory research, traditionally based on experimental paradigms from cognitive psychology, can potentially…

  9. Monitoring accuracy and self-regulation when learning to play a chess endgame

    NARCIS (Netherlands)

    De Bruin, Anique BH; Rikers, Remy MJP; Schmidt, Henk G

    2005-01-01

    In order to examine the effect of monitoring and self-regulation on skill acquisition, the present study asked novice chess players to provide judgments of learning (JOLs) and to select moves for restudy after studying an endgame of chess. In four groups, we varied the JOL instruction (present versu

  10. Endogenous monitoring: a new challenge for the regulation of energy externalities

    Energy Technology Data Exchange (ETDEWEB)

    Millock, K

    1999-11-01

    The present article studies the problem of regulating externalities from energy use, with particular emphasis on the problem of information. A standard economics. The article shows how the optimal choice of instruments used to regulate the externalities from energy use will change with the more widespread adoption of modern monitoring technologies. We will illustrate how the application of incentive policies conditioned on monitoring can contribute to the implementation of flexible mechanisms for greenhouse gas abatement. Section 2 briefly summarises the economic framework for the regulation of externalities resulting from energy use. In Section 3, an illustrative model is presented in which the introduction of monitoring of emissions is endogenous and conditions the best type of instrument used by the regulator to internalize the externalities from energy use. Section 4 then examines the implications of this framework for the application of the flexible incentives introduced in the Kyoto Protocol. It shows how the success of project-based approaches, such as Joint Implementation and the Clean Development Mechanism, depends upon the implementation of appropriate incentives for monitoring. Section 5 concludes. (author)

  11. 26 CFR 55.6001-1 - Notice or regulations requiring records, statements, and special returns.

    Science.gov (United States)

    2010-04-01

    ... INVESTMENT TRUSTS AND REGULATED INVESTMENT COMPANIES Procedure and Administration § 55.6001-1 Notice or regulations requiring records, statements, and special returns. (a) In general. Any person subject to tax... retained so long as the contents thereof may become material in the administration of any internal...

  12. 76 FR 14562 - Federal Acquisition Regulation; Additional Requirements for Market Research

    Science.gov (United States)

    2011-03-16

    ... 52 RIN 9000-AL50 Federal Acquisition Regulation; Additional Requirements for Market Research AGENCY... interim rule amending the Federal Acquisition Regulation (FAR) to implement section 826, Market Research... items engages in market research as necessary before making purchases. DATES: Effective Date: April...

  13. 40 CFR Table 6 to Subpart Bbbb of... - Model Rule-Requirements for Validating Continuous Emission Monitoring Systems (CEMS)

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 6 2010-07-01 2010-07-01 false Model Rule-Requirements for Validating Continuous Emission Monitoring Systems (CEMS) 6 Table 6 to Subpart BBBB of Part 60 Protection of Environment...—Requirements for Validating Continuous Emission Monitoring Systems (CEMS) For the following continuous...

  14. Impact of shock requiring norepinephrine on the accuracy and reliability of subcutaneous continuous glucose monitoring.

    Science.gov (United States)

    Holzinger, Ulrike; Warszawska, Joanna; Kitzberger, Reinhard; Herkner, Harald; Metnitz, Philipp G H; Madl, Christian

    2009-08-01

    To evaluate the impact of circulatory shock requiring norepinephrine therapy on the accuracy and reliability of a subcutaneous continuous glucose monitoring system (CGMS) in critically ill patients. A prospective, validation study of a medical intensive care unit at a university hospital was carried out. Continuous glucose monitoring was performed subcutaneously in 50 consecutive patients on intensive insulin therapy (IIT), who were assessed according to the a priori strata of circulatory shock requiring norepinephrine therapy or not. A total of 736 pairs of sensor glucose (SG)/blood glucose (BG) values were analysed (502 without and 234 with norepinephrine therapy). For all values, repeated measures Bland-Altman analysis showed a mean difference of 0.08 mmol/l (limits of agreement: -1.26 and 1.43 mmol/l). Circulatory shock requiring norepinephrine therapy did not influence the relation of arterial BG with SG in a multivariable random effects linear regression analysis. The covariates norepinephrine dose, body mass index (BMI), glucose level and severity of illness also had no influence. Insulin titration grid analysis showed that 98.6% of the data points were in the acceptable treatment zone. No data were in the life-threatening zone. Circulatory shock requiring norepinephrine therapy, as well as other covariates, had no influence on the accuracy and reliability of the CGMS in critically ill patients.

  15. Resolution requirements for monitor viewing of digital flat-panel detector radiographs: a contrast detail analysis

    Energy Technology Data Exchange (ETDEWEB)

    Peer, Siegfried; Giacomuzzi, Salvatore M.; Peer, Regina; Gassner, Eva; Steingruber, Iris; Jaschke, Werner [Department of Radiology, University Hospital, Anichstrasse 35, 6020 Innsbruck (Austria)

    2003-02-01

    With the introduction of digital flat-panel detector systems into clinical practice, the still unresolved question of resolution requirements for picture archiving communication system (PACS) workstation monitors has gained new momentum. This contrast detail analysis was thus performed to define the differences in observer performance in the detection of small low-contrast objects on clinical 1K and 2K monitor workstations. Images of the CDRAD 2.0 phantom were acquired at varying exposures on an indirect-type digital flat-panel detector. Three observers evaluated a total of 15 images each with respect to the threshold contrast for each detail size. The numbers of correctly identified objects were determined for all image subsets. No significant difference in the correct detection ratio was detected among the observers; however, the difference between the two types of workstations (1K vs 2K monitors) despite less than 3% was significant at a 95% confidence level. Slight but statistically significant differences exist in the detection of low-contrast nodular details visualized on 1K- and 2K-monitor workstations. Further work is needed to see if this result holds true also for comparison of clinical flat-panel detector images and may, for example, exert an influence on the diagnostic accuracy of chest X-ray readings. (orig.)

  16. To What Extent is Drinking Water Tested in Sub-Saharan Africa? A Comparative Analysis of Regulated Water Quality Monitoring.

    Science.gov (United States)

    Peletz, Rachel; Kumpel, Emily; Bonham, Mateyo; Rahman, Zarah; Khush, Ranjiv

    2016-03-02

    Water quality information is important for guiding water safety management and preventing water-related diseases. To assess the current status of regulated water quality monitoring in sub-Saharan Africa, we evaluated testing programs for fecal contamination in 72 institutions (water suppliers and public health agencies) across 10 countries. Data were collected through written surveys, in-person interviews, and analysis of microbial water quality testing levels. Though most institutions did not achieve the testing levels specified by applicable standards or World Health Organization (WHO) Guidelines, 85% of institutions had conducted some microbial water testing in the previous year. Institutions were more likely to meet testing targets if they were suppliers (as compared to surveillance agencies), served larger populations, operated in urban settings, and had higher water quality budgets (all p water providers and rural public health offices will require greater attention and additional resources to achieve regulatory compliance for water quality monitoring in sub-Saharan Africa. The cost-effectiveness of water quality monitoring should be improved by the application of risk-based water management approaches. Efforts to strengthen monitoring capacity should pay greater attention to program sustainability and institutional commitment to water safety.

  17. To What Extent is Drinking Water Tested in Sub-Saharan Africa? A Comparative Analysis of Regulated Water Quality Monitoring

    Directory of Open Access Journals (Sweden)

    Rachel Peletz

    2016-03-01

    Full Text Available Water quality information is important for guiding water safety management and preventing water-related diseases. To assess the current status of regulated water quality monitoring in sub-Saharan Africa, we evaluated testing programs for fecal contamination in 72 institutions (water suppliers and public health agencies across 10 countries. Data were collected through written surveys, in-person interviews, and analysis of microbial water quality testing levels. Though most institutions did not achieve the testing levels specified by applicable standards or World Health Organization (WHO Guidelines, 85% of institutions had conducted some microbial water testing in the previous year. Institutions were more likely to meet testing targets if they were suppliers (as compared to surveillance agencies, served larger populations, operated in urban settings, and had higher water quality budgets (all p < 0.05. Our results indicate that smaller water providers and rural public health offices will require greater attention and additional resources to achieve regulatory compliance for water quality monitoring in sub-Saharan Africa. The cost-effectiveness of water quality monitoring should be improved by the application of risk-based water management approaches. Efforts to strengthen monitoring capacity should pay greater attention to program sustainability and institutional commitment to water safety.

  18. 31 CFR 103.120 - Anti-money laundering program requirements for financial institutions regulated by a Federal...

    Science.gov (United States)

    2010-07-01

    ... requirements for financial institutions regulated by a Federal functional regulator or a self-regulatory... financial institutions regulated by a Federal functional regulator or a self-regulatory organization, and... futures commission merchants. A financial institution regulated by a self-regulatory organization shall...

  19. Low Cost Environmental Sensors for Spaceflight: NMP Space Environmental Monitor (SEM) Requirements

    Science.gov (United States)

    Garrett, Henry B.; Buehler, Martin G.; Brinza, D.; Patel, J. U.

    2005-01-01

    An outstanding problem in spaceflight is the lack of adequate sensors for monitoring the space environment and its effects on engineering systems. By adequate, we mean low cost in terms of mission impact (e.g., low price, low mass/size, low power, low data rate, and low design impact). The New Millennium Program (NMP) is investigating the development of such a low-cost Space Environmental Monitor (SEM) package for inclusion on its technology validation flights. This effort follows from the need by NMP to characterize the space environment during testing so that potential users can extrapolate the test results to end-use conditions. The immediate objective of this effort is to develop a small diagnostic sensor package that could be obtained from commercial sources. Environments being considered are: contamination, atomic oxygen, ionizing radiation, cosmic radiation, EMI, and temperature. This talk describes the requirements and rational for selecting these environments and reviews a preliminary design that includes a micro-controller data logger with data storage and interfaces to the sensors and spacecraft. If successful, such a sensor package could be the basis of a unique, long term program for monitoring the effects of the space environment on spacecraft systems.

  20. 40 CFR Table 7 to Subpart Bbbb of... - Model Rule-Requirements for Continuous Emission Monitoring Systems (CEMS)

    Science.gov (United States)

    2010-07-01

    ... Emission Monitoring Systems (CEMS) 7 Table 7 to Subpart BBBB of Part 60 Protection of Environment...—Requirements for Continuous Emission Monitoring Systems (CEMS) For the following pollutants Use the following span values for CEMS Use the following performance specifications in appendix B of this part for...

  1. 40 CFR Table 4 to Subpart Ffff of... - Model Rule-Requirements for Continuous Emission Monitoring Systems (CEMS)

    Science.gov (United States)

    2010-07-01

    ... Emission Monitoring Systems (CEMS) 4 Table 4 to Subpart FFFF of Part 60 Protection of Environment...—Model Rule—Requirements for Continuous Emission Monitoring Systems (CEMS) As stated in § 60.3039, you... CEMS Use the following performance specifications (P.S.) in appendix B of this part for your CEMS...

  2. 40 CFR Table 6 to Subpart Ppp of... - Process Vents From Continuous Unit Operations-Monitoring, Recordkeeping, and Reporting Requirements

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 11 2010-07-01 2010-07-01 true Process Vents From Continuous Unit Operations-Monitoring, Recordkeeping, and Reporting Requirements 6 Table 6 to Subpart PPP of Part 63... Subpart PPP of Part 63—Process Vents From Continuous Unit Operations—Monitoring, Recordkeeping, and...

  3. 40 CFR Table 7 to Subpart Ggg of... - Wastewater-Inspection and Monitoring Requirements for Waste Management Units

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 11 2010-07-01 2010-07-01 true Wastewater-Inspection and Monitoring..., Subpt. GGG, Table 7 Table 7 to Subpart GGG of Part 63—Wastewater—Inspection and Monitoring Requirements...(b)(7)63.1256(b)(8) Inspect wastewater tank for control equipment failures and improper...

  4. 77 FR 39959 - Draft Guidance To Implement Requirements for the Treatment of Air Quality Monitoring Data...

    Science.gov (United States)

    2012-07-06

    ... Air Quality Monitoring Data Influenced by Exceptional Events AGENCY: Environmental Protection Agency... for the Treatment of Air Quality Monitoring Data Influenced by Exceptional Events and associated... Treatment of Air Quality Monitoring Data Influenced by Exceptional Events and associated attachments and...

  5. PR72, a novel regulator of Wnt signaling required for Naked cuticle function

    NARCIS (Netherlands)

    Creyghton, M.P.; Roël, G.; Eichhorn, P.J.A.; Hijmans, E.M.; Maurer, I.; Destrée, O.; Bernards, R.A.

    2005-01-01

    The Wnt signaling cascade is a central regulator of cell fate determination during embryonic development, whose deregulation contributes to oncogenesis. Naked cuticle is the first Wnt-induced antagonist found in this pathway, establishing a negative-feedback loop that limits the Wnt signal required

  6. Capital Regulation, Liquidity Requirements and Taxation in a Dynamic Model of Banking

    NARCIS (Netherlands)

    Di Nicolo, G.; Gamba, A.; Lucchetta, M.

    2011-01-01

    This paper formulates a dynamic model of a bank exposed to both credit and liquidity risk, which can resolve financial distress in three costly forms: fire sales, bond issuance ad equity issuance. We use the model to analyze the impact of capital regulation, liquidity requirements and taxation on ba

  7. Capital Regulation, Liquidity Requirements and Taxation in a Dynamic Model of Banking

    NARCIS (Netherlands)

    Di Nicolo, G.; Gamba, A.; Lucchetta, M.

    2011-01-01

    This paper formulates a dynamic model of a bank exposed to both credit and liquidity risk, which can resolve financial distress in three costly forms: fire sales, bond issuance and equity issuance. We use the model to analyze the impact of capital regulation, liquidity requirements and taxation on b

  8. 77 FR 34315 - National Pollutant Discharge Elimination System-Proposed Regulations to Establish Requirements...

    Science.gov (United States)

    2012-06-11

    .... In other words, the cooling tower was in place but cooling water was used in a single pass mode, with...--Proposed Regulations to Establish Requirements for Cooling Water Intake Structures at Existing Facilities... proposed standards for cooling water intake structures at all existing power generating facilities...

  9. 78 FR 73104 - Federal Travel Regulation (FTR); Agency Requirements for Payment of Expenses Connected With the...

    Science.gov (United States)

    2013-12-05

    ...] RIN 3090-AJ37 Federal Travel Regulation (FTR); Agency Requirements for Payment of Expenses Connected... family member. The employee may elect an alternate destination, but it must be approved by your agency...) The new CONUS location if in transit; or (c) An alternate destination as approved by the agency....

  10. 50 CFR 25.23 - What are the general regulations and information collection requirements?

    Science.gov (United States)

    2010-10-01

    ... collect information to assist us in administering our programs in accordance with statutory authorities... information collection requirements? 25.23 Section 25.23 Wildlife and Fisheries UNITED STATES FISH AND... PROVISIONS Administrative Provisions § 25.23 What are the general regulations and information...

  11. PR72, a novel regulator of Wnt signaling required for Naked cuticle function

    NARCIS (Netherlands)

    Creyghton, M.P.; Roël, G.; Eichhorn, P.J.A.; Hijmans, E.M.; Maurer, I.; Destrée, O.; Bernards, R.A.

    2005-01-01

    The Wnt signaling cascade is a central regulator of cell fate determination during embryonic development, whose deregulation contributes to oncogenesis. Naked cuticle is the first Wnt-induced antagonist found in this pathway, establishing a negative-feedback loop that limits the Wnt signal required

  12. 75 FR 51824 - Food and Drug Administration Clinical Trial Requirements, Regulations, Compliance, and Good...

    Science.gov (United States)

    2010-08-23

    ... HUMAN SERVICES Food and Drug Administration Food and Drug Administration Clinical Trial Requirements, Regulations, Compliance, and Good Clinical Practice; Public Workshop AGENCY: Food and Drug Administration, HHS. ACTION: Notice of public workshop. ] SUMMARY: The Food and Drug Administration (FDA) New Jersey...

  13. Sterol homeostasis requires regulated degradation of squalene monooxygenase by the ubiquitin ligase Doa10/Teb4

    DEFF Research Database (Denmark)

    Foresti, Ombretta; Ruggiano, Annamaria; Hannibal-Bach, Hans K

    2013-01-01

    ligase implicated in a branch of the endoplasmic reticulum (ER)-associated protein degradation (ERAD) pathway. Since the other branch of ERAD is required for HMGR regulation, our results reveal a fundamental role for ERAD in sterol homeostasis, with the two branches of this pathway acting together...

  14. Oogenesis requires germ cell-specific transcriptional regulators Sohlh1 and Lhx8

    Science.gov (United States)

    Pangas, Stephanie A.; Choi, Youngsok; Ballow, Daniel J.; Zhao, Yangu; Westphal, Heiner; Matzuk, Martin M.; Rajkovic, Aleksandar

    2006-01-01

    Mammalian oogenesis requires oocyte-specific transcriptional regulators. The full complement of oocyte-specific transcription factors is unknown. Here, we describe the finding that Sohlh1, a spermatogenesis and oogenesis basic helix–loop–helix transcription factor in females, is preferentially expressed in oocytes and required for oogenesis. Sohlh1 disruption perturbs follicular formation in part by causing down-regulation of two genes that are known to disrupt folliculogenesis: newborn ovary homeobox gene (Nobox) and factor in the germ-line alpha (Figla). In addition, we show that Lhx8 is downstream of Sohlh1 and critical in fertility. Thus, Sohlh1 and Lhx8 are two germ cell-specific, critical regulators of oogenesis. PMID:16690745

  15. Additional requirements for charitable hospitals; community health needs assessments for charitable hospitals; requirement of a section 4959 excise tax return and time for filing the return. Final regulations and removal of temporary regulations.

    Science.gov (United States)

    2014-12-31

    This document contains final regulations that provide guidance regarding the requirements for charitable hospital organizations added by the Patient Protection and Affordable Care Act of 2010. The regulations will affect charitable hospital organizations

  16. Stochastic modeling to identify requirements for centralized monitoring of distributed wastewater treatment.

    Science.gov (United States)

    Hug, T; Maurer, M

    2012-01-01

    Distributed (decentralized) wastewater treatment can, in many situations, be a valuable alternative to a centralized sewer network and wastewater treatment plant. However, it is critical for its acceptance whether the same overall treatment performance can be achieved without on-site staff, and whether its performance can be measured. In this paper we argue and illustrate that the system performance depends not only on the design performance and reliability of the individual treatment units, but also significantly on the monitoring scheme, i.e. on the reliability of the process information. For this purpose, we present a simple model of a fleet of identical treatment units. Thereby, their performance depends on four stochastic variables: the reliability of the treatment unit, the respond time for the repair of failed units, the reliability of on-line sensors, and the frequency of routine inspections. The simulated scenarios show a significant difference between the true performance and the observations by the sensors and inspections. The results also illustrate the trade-off between investing in reactor and sensor technology and in human interventions in order to achieve a certain target performance. Modeling can quantify such effects and thereby support the identification of requirements for the centralized monitoring of distributed treatment units. The model approach is generic and can be extended and applied to various distributed wastewater treatment technologies and contexts.

  17. Fluorescence monitoring of riboswitch transcription regulation using a dual molecular beacon assay.

    Science.gov (United States)

    Chinnappan, Raja; Dubé, Audrey; Lemay, Jean-François; Lafontaine, Daniel A

    2013-05-01

    Riboswitches are mRNA elements that specifically bind cellular metabolites and control gene expression by modifying their structure. As riboswitches often control essential genes in pathogenic bacteria, riboswitches have been proposed as new targets for antibiotics. High-throughput screening provides a powerful approach to identify riboswitch ligand analogs that could act as powerful antibacterial drugs. Biochemical assays have already been used to find riboswitch-binding analogs, but those methods do take into account the transcriptional context for riboswitch regulation. As the importance of co-transcriptional ligand binding has been shown for several riboswitches, it is vital to develop an assay that screens riboswitch-binding analogs during the transcriptional process. Here, we describe the development of a dual molecular beacon system monitoring the transcriptional regulation activity of the Bacillus subtilis pbuE adenine riboswitch. This system relies on two molecular beacons that enable the monitoring of transcription efficiency, as well as the regulatory activity of the riboswitch. Different analogs were tested using our system, and a good correlation was observed between riboswitch activity and reported metabolite affinities. This method is specific, reliable and could be applied at the high-throughput level for the identification of new potential antibiotics targeting any riboswitch-regulating gene expression at the mRNA level.

  18. [Better AVK treatment with self monitoring. Dosage can be regulated in time].

    Science.gov (United States)

    Stigendal, L; André, U; Christenson, B

    1999-05-19

    As long-term anticoagulant treatment, with warfarin for instance, is associated with a risk of both thrombotic and thrombolytic complications, blood testing for dose regulation is necessary at 3-8-week intervals, which is expensive and inconvenient for patients who must take time off work and travel to and fro. A new technique, using small portable monitors designed for home use by patients, makes self-management of anticoagulant treatment possible. In Germany, over 25,000 patients had their own monitor by the end of 1998. After appropriate instruction, the German patients are able to monitor their prothrombin time and adjust their anticoagulant treatment accordingly. In case of problems they contact their GP. In a two-year pilot study conducted at the Anticoagulation Clinic of Sahlgrenska University Hospital, Gothenburg, in 1996-98, where 51 patients on long-term anticoagulant treatment were trained in self-management, the results of over 1,000 patient-hours of treatment showed self-management to be at least as safe as management by the clinic. The level of patient satisfaction is high, in terms of safety and freedom from regular hospital attendance during working hours, and the convenience of self-monitoring on holiday or business trips. As the patients do their testing once a week, the risk of complications is also reduced.

  19. Flow cytometry quality requirements for monitoring of minimal disease in plasma cell myeloma.

    Science.gov (United States)

    Oldaker, Teri A; Wallace, Paul K; Barnett, David

    2016-01-01

    Current therapeutic approaches for plasma cell myeloma (PCM) attain an overall survival of more than 6 years for the majority of newly diagnosed patients. However, PFS and OS are the only accepted FDA clinical endpoints for demonstrating drug efficacy before they can be become frontline therapeutic options. There is, however, recognition that the increasing gap between drug development and approval for mainstream therapeutic use needs to be shortened. As such regulatory bodies such as the FDA are now considering whether biomarker response evaluation, as in measurement of minimal residual disease (MRD) as assessed by flow cytometry (FC), can provide an early, robust prediction of survival and therefore improve the drug approval process. Recently, FC MRD using a standardized eight-color antibody methodology has been shown to have a minimum sensitivity of 0.01% and an upper sensitivity of 0.001%. To ensure that all laboratories using this approach achieve the same levels of sensitivity it is crucially important to have standardized quality management procedures in place. This manuscript accompanies those published in this special issue and describes the minimum that is required for validating and quality monitoring of this highly specific test to ensure any laboratory, irrespective of location, will achieve the expected quality standards required. © 2015 International Clinical Cytometry Society.

  20. Electronic Nose Testing Procedure for the Definition of Minimum Performance Requirements for Environmental Odor Monitoring.

    Science.gov (United States)

    Eusebio, Lidia; Capelli, Laura; Sironi, Selena

    2016-09-21

    Despite initial enthusiasm towards electronic noses and their possible application in different fields, and quite a lot of promising results, several criticalities emerge from most published research studies, and, as a matter of fact, the diffusion of electronic noses in real-life applications is still very limited. In general, a first step towards large-scale-diffusion of an analysis method, is standardization. The aim of this paper is describing the experimental procedure adopted in order to evaluate electronic nose performances, with the final purpose of establishing minimum performance requirements, which is considered to be a first crucial step towards standardization of the specific case of electronic nose application for environmental odor monitoring at receptors. Based on the experimental results of the performance testing of a commercialized electronic nose type with respect to three criteria (i.e., response invariability to variable atmospheric conditions, instrumental detection limit, and odor classification accuracy), it was possible to hypothesize a logic that could be adopted for the definition of minimum performance requirements, according to the idea that these are technologically achievable.

  1. Electronic Nose Testing Procedure for the Definition of Minimum Performance Requirements for Environmental Odor Monitoring

    Directory of Open Access Journals (Sweden)

    Lidia Eusebio

    2016-09-01

    Full Text Available Despite initial enthusiasm towards electronic noses and their possible application in different fields, and quite a lot of promising results, several criticalities emerge from most published research studies, and, as a matter of fact, the diffusion of electronic noses in real-life applications is still very limited. In general, a first step towards large-scale-diffusion of an analysis method, is standardization. The aim of this paper is describing the experimental procedure adopted in order to evaluate electronic nose performances, with the final purpose of establishing minimum performance requirements, which is considered to be a first crucial step towards standardization of the specific case of electronic nose application for environmental odor monitoring at receptors. Based on the experimental results of the performance testing of a commercialized electronic nose type with respect to three criteria (i.e., response invariability to variable atmospheric conditions, instrumental detection limit, and odor classification accuracy, it was possible to hypothesize a logic that could be adopted for the definition of minimum performance requirements, according to the idea that these are technologically achievable.

  2. Electronic Nose Testing Procedure for the Definition of Minimum Performance Requirements for Environmental Odor Monitoring

    Science.gov (United States)

    Eusebio, Lidia; Capelli, Laura; Sironi, Selena

    2016-01-01

    Despite initial enthusiasm towards electronic noses and their possible application in different fields, and quite a lot of promising results, several criticalities emerge from most published research studies, and, as a matter of fact, the diffusion of electronic noses in real-life applications is still very limited. In general, a first step towards large-scale-diffusion of an analysis method, is standardization. The aim of this paper is describing the experimental procedure adopted in order to evaluate electronic nose performances, with the final purpose of establishing minimum performance requirements, which is considered to be a first crucial step towards standardization of the specific case of electronic nose application for environmental odor monitoring at receptors. Based on the experimental results of the performance testing of a commercialized electronic nose type with respect to three criteria (i.e., response invariability to variable atmospheric conditions, instrumental detection limit, and odor classification accuracy), it was possible to hypothesize a logic that could be adopted for the definition of minimum performance requirements, according to the idea that these are technologically achievable. PMID:27657086

  3. Endocytosis-dependent coordination of multiple actin regulators is required for wound healing.

    Science.gov (United States)

    Matsubayashi, Yutaka; Coulson-Gilmer, Camilla; Millard, Tom H

    2015-08-01

    The ability to heal wounds efficiently is essential for life. After wounding of an epithelium, the cells bordering the wound form dynamic actin protrusions and/or a contractile actomyosin cable, and these actin structures drive wound closure. Despite their importance in wound healing, the molecular mechanisms that regulate the assembly of these actin structures at wound edges are not well understood. In this paper, using Drosophila melanogaster embryos, we demonstrate that Diaphanous, SCAR, and WASp play distinct but overlapping roles in regulating actin assembly during wound healing. Moreover, we show that endocytosis is essential for wound edge actin assembly and wound closure. We identify adherens junctions (AJs) as a key target of endocytosis during wound healing and propose that endocytic remodeling of AJs is required to form "signaling centers" along the wound edge that control actin assembly. We conclude that coordination of actin assembly, AJ remodeling, and membrane traffic is required for the construction of a motile leading edge during wound healing.

  4. Monitoring of transcriptional regulation in Pichia pastoris under protein production conditions

    Directory of Open Access Journals (Sweden)

    Bhattacharyya Anamitra

    2007-06-01

    Full Text Available Abstract Background It has become evident that host cells react to recombinant protein production with a variety of metabolic and intrinsic stresses such as the unfolded protein response (UPR pathway. Additionally, environmental conditions such as growth temperature may have a strong impact on cell physiology and specific productivity. However, there is little information about the molecular reactions of the host cells on a genomic level, especially in context to recombinant protein secretion. For the first time, we monitored transcriptional regulation of a subset of marker genes in the common production host Pichia pastoris to gain insights into the general physiological status of the cells under protein production conditions, with the main focus on secretion stress related genes. Results Overexpression of the UPR activating transcription factor Hac1p was employed to identify UPR target genes in P. pastoris and the responses were compared to those known for Saccharomyces cerevisiae. Most of the folding/secretion related genes showed similar regulation patterns in both yeasts, whereas genes associated with the general stress response were differentially regulated. Secretion of an antibody Fab fragment led to induction of UPR target genes in P. pastoris, however not to the same magnitude as Hac1p overproduction. Overexpression of S. cerevisiae protein disulfide isomerase (PDI1 enhances Fab secretion rates 1.9 fold, but did not relief UPR stress. Reduction of cultivation temperature from 25°C to 20°C led to a 1.4-fold increase of specific product secretion rate in chemostat cultivations, although the transcriptional levels of the product genes (Fab light and heavy chain were significantly reduced at the lower temperature. A subset of folding related genes appeared to be down-regulated at the reduced temperature, whereas transcription of components of the ER associated degradation and the secretory transport was enhanced. Conclusion Monitoring of

  5. Reliability of a rating procedure to monitor industry self-regulation codes governing alcohol advertising content.

    Science.gov (United States)

    Babor, Thomas F; Xuan, Ziming; Proctor, Dwayne

    2008-03-01

    The purposes of this study were to develop reliable procedures to monitor the content of alcohol advertisements broadcast on television and in other media, and to detect violations of the content guidelines of the alcohol industry's self-regulation codes. A set of rating-scale items was developed to measure the content guidelines of the 1997 version of the U.S. Beer Institute Code. Six focus groups were conducted with 60 college students to evaluate the face validity of the items and the feasibility of the procedure. A test-retest reliability study was then conducted with 74 participants, who rated five alcohol advertisements on two occasions separated by 1 week. Average correlations across all advertisements using three reliability statistics (r, rho, and kappa) were almost all statistically significant and the kappas were good for most items, which indicated high test-retest agreement. We also found high interrater reliabilities (intraclass correlations) among raters for item-level and guideline-level violations, indicating that regardless of the specific item, raters were consistent in their general evaluations of the advertisements. Naïve (untrained) raters can provide consistent (reliable) ratings of the main content guidelines proposed in the U.S. Beer Institute Code. The rating procedure may have future applications for monitoring compliance with industry self-regulation codes and for conducting research on the ways in which alcohol advertisements are perceived by young adults and other vulnerable populations.

  6. RpoS is required for natural transformation of Vibrio cholerae through regulation of chitinases.

    Science.gov (United States)

    Dalia, Ankur B

    2016-11-01

    Vibrio species naturally reside in the aquatic environment and a major metabolite in this habitat is the chitinous exoskeletons of crustacean zooplankton. In addition to serving as a nutrient, chitin-derived oligosaccharides also induce natural genetic competence in many Vibrio spp., a physiological state in which bacteria take up DNA from the extracellular environment and can integrate it into their chromosome by homologous recombination. Another inducing cue required for competence are quorum-sensing autoinducers. The alternative sigma factor RpoS is critical for natural transformation in Vibrio cholerae, and it was previously presumed to exert this effect through regulation of quorum sensing. Here, we show that RpoS does not affect quorum sensing-dependent regulation of competence. Instead, we show that an rpoS mutant has reduced chitinase activity, which is required to liberate the soluble chitin oligosaccharides that serve as an inducing cue for competence. Consistent with this, we demonstrate that RpoS is required for growth of V. cholerae on insoluble chitin. RpoS also regulates the mucosal escape response in pathogenic strains of V. cholerae. Thus, in addition to promoting egress from its human host, RpoS may also prime this pathogen for successful reentry into the aquatic environment. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

  7. REQUIREMENTS ECO-DESIGN VACUUMS BY COMMISSION REGULATION (EU NO 666/2013

    Directory of Open Access Journals (Sweden)

    Przemysław Kurczewski

    2014-10-01

    Full Text Available For several years, the executive acts related to the Directive of the European Parliament and of the Council 2009/125/EC of 21 October 2009 establishing a framework for the setting of ecodesign requirements for energy-related products are systematically adopted as a part of European Union legislation. Especially the Regulation 666/2013 concerning the eco-design requirements for vacuum cleaners introduced in July 2013 is strongly discussed. This article presents the background to the Directive 2009/125/EC and explains the concept that underlies the formulation of ecodesign requirements for different powered energy products, with special focus on the role of environmental life cycle assessment (LCA.

  8. Cost-optimal levels of minimum energy performance requirements in the Danish Building Regulations

    Energy Technology Data Exchange (ETDEWEB)

    Aggerholm, S.

    2013-09-15

    The purpose of the report is to analyse the cost optimality of the energy requirements in the Danish Building Regulations 2010, BR10 to new building and to existing buildings undergoing major renovation. The energy requirements in the Danish Building Regulations have by tradition always been based on the cost and benefits related to the private economical or financial perspective. Macro economical calculations have in the past only been made in addition. The cost optimum used in this report is thus based on the financial perspective. Due to the high energy taxes in Denmark there is a significant difference between the consumer price and the macro economical for energy. Energy taxes are also paid by commercial consumers when the energy is used for building operation e.g. heating, lighting, ventilation etc. In relation to the new housing examples the present minimum energy requirements in BR 10 all shows gaps that are negative with a deviation of up till 16 % from the point of cost optimality. With the planned tightening of the requirements to new houses in 2015 and in 2020, the energy requirements can be expected to be tighter than the cost optimal point, if the costs for the needed improvements don't decrease correspondingly. In relation to the new office building there is a gap of 31 % to the point of cost optimality in relation to the 2010 requirement. In relation to the 2015 and 2020 requirements there are negative gaps to the point of cost optimality based on today's prices. If the gaps for all the new buildings are weighted to an average based on mix of building types and heat supply for new buildings in Denmark there is a gap of 3 % in average for the new building. The excessive tightness with today's prices is 34 % in relation to the 2015 requirement and 49 % in relation to the 2020 requirement. The component requirement to elements in the building envelope and to installations in existing buildings adds up to significant energy efficiency

  9. Exploring the structural requirements for jasmonates and related compounds as novel plant growth regulators

    Science.gov (United States)

    Chen, Ke-Xian

    2009-01-01

    Jasmonates and related compounds have been highlighted recently in the field of plant physiology and plant molecular biology due to their significant regulatory roles in the signaling pathway for the diverse aspects of plant development and survival. Though a considerable amount of studies concerning their biological effects in different plants have been widely reported, the molecular details of the signaling mechanism are still poorly understood. This review sheds new light on the structural requirements for the bioactivity/property of jasmonic acid derivatives in current computational perspective, which differs from previous research that mainly focus on their biological evaluation, gene and metabolic regulation and the enzymes in their biosynthesis. The computational results may contribute to further understanding the mechanism of drug-receptor interactions in their signaling pathway and designing novel plant growth regulators as high effective ecological pesticides. PMID:20009552

  10. Engineered hybrid cardiac patches with multifunctional electronics for online monitoring and regulation of tissue function

    Science.gov (United States)

    Feiner, Ron; Engel, Leeya; Fleischer, Sharon; Malki, Maayan; Gal, Idan; Shapira, Assaf; Shacham-Diamand, Yosi; Dvir, Tal

    2016-01-01

    In cardiac tissue engineering approaches to treat myocardial infarction, cardiac cells are seeded within three-dimensional porous scaffolds to create functional cardiac patches. However, current cardiac patches do not allow for online monitoring and reporting of engineered-tissue performance, and do not interfere to deliver signals for patch activation or to enable its integration with the host. Here, we report an engineered cardiac patch that integrates cardiac cells with flexible, free-standing electronics and a 3D nanocomposite scaffold. The patch exhibited robust electronic properties, enabling the recording of cellular electrical activities and the on-demand provision of electrical stimulation for synchronizing cell contraction. We also show that electroactive polymers containing biological factors can be deposited on designated electrodes to release drugs in the patch microenvironment on-demand. We expect that the integration of complex electronics within cardiac patches will eventually provide therapeutic control and regulation of cardiac function. PMID:26974408

  11. Radioactivity in drinking water: regulations, monitoring results and radiation protection issues

    Directory of Open Access Journals (Sweden)

    Cristina Nuccetelli

    2012-12-01

    Full Text Available INTRODUCTION: Drinking waters usually contain several natural radionuclides: tritium, radon, radium, uranium isotopes, etc. Their concentrations vary widely since they depend on the nature of the aquifer, namely, the prevailing lithology and whether there is air in it or not. AIMS: In this work a broad overview of the radioactivity in drinking water is presented: national and international regulations, for limiting the presence of radioactivity in waters intended for human consumption; results of extensive campaigns for monitoring radioactivity in drinking waters, including mineral bottled waters, carried out throughout the world in recent years; a draft of guidelines for the planning of campaigns to measure radioactivity in drinking water proposed by the Environmental Protection Agency (ARPA of Lombardia.

  12. Global infant formula: monitoring and regulating the impacts to protect human health.

    Science.gov (United States)

    Kent, George

    2015-01-01

    Worldwide promotion of infant formula and other commercial baby foods is leading to increased use of these products, raising concerns about their impact on the health of infants. These products are made and marketed through a global system that extends beyond the control of separate nations. As the industry is increasingly globalized, there is a growing need for guidance, monitoring, and regulation. This study suggests a path toward achieving better control of infant formula and other baby foods to ensure that infants and young children everywhere are well nourished. The negotiation of a new Optional Protocol on Children's Nutrition, to be linked to the most relevant human rights treaty, the Convention on the Rights of the Child, would bring the major issues relating to infant formula and other baby foods to the attention of the global community and all national governments.

  13. Wireless sensor network for helicopter rotor blade vibration monitoring: Requirements definition and technological aspects

    NARCIS (Netherlands)

    Sanchez Ramirez, Andrea; Das, Kallol; Loendersloot, Richard; Tinga, Tiedo; Havinga, Paul; Basu, Biswajit

    2013-01-01

    The main rotor accounts for the largest vibration source for a helicopter fuselage and its components. However, accurate blade monitoring has been limited due to the practical restrictions on instrumenting rotating blades. The use of Wireless Sensor Networks (WSNs) for real time vibration monitoring

  14. 40 CFR 63.11527 - What are the monitoring requirements for new and existing sources?

    Science.gov (United States)

    2010-07-01

    ... paragraph (b)(1) of this section. (4) When operating a CPMS, if the 3-hour average pressure drop or scrubber.... Manufacturer's specifications for pressure drop and liquid flow rate will be used to determine normal... filters—(1) Visual monitoring. You must conduct visual monitoring of the monovent or fabric filter...

  15. 40 CFR Table 4 of Subpart Aaaa to... - Requirements for Continuous Emission Monitoring Systems (CEMS)

    Science.gov (United States)

    2010-07-01

    ... Monitoring Systems (CEMS) 4 Table 4 of Subpart AAAA to Part 60 Protection of Environment ENVIRONMENTAL... Monitoring Systems (CEMS) For the following pollutants Use the following span values for your CEMS Use the following performance specifications in appendix B of this part for your CEMS If needed to meet...

  16. Lung surfactant levels are regulated by Ig-Hepta/GPR116 by monitoring surfactant protein D.

    Directory of Open Access Journals (Sweden)

    Taku Fukuzawa

    Full Text Available Lung surfactant is a complex mixture of lipids and proteins, which is secreted from the alveolar type II epithelial cell and coats the surface of alveoli as a thin layer. It plays a crucial role in the prevention of alveolar collapse through its ability to reduce surface tension. Under normal conditions, surfactant homeostasis is maintained by balancing its release and the uptake by the type II cell for recycling and the internalization by alveolar macrophages for degradation. Little is known about how the surfactant pool is monitored and regulated. Here we show, by an analysis of gene-targeted mice exhibiting massive accumulation of surfactant, that Ig-Hepta/GPR116, an orphan receptor, is expressed on the type II cell and sensing the amount of surfactant by monitoring one of its protein components, surfactant protein D, and its deletion results in a pulmonary alveolar proteinosis and emphysema-like pathology. By a coexpression experiment with Sp-D and the extracellular region of Ig-Hepta/GPR116 followed by immunoprecipitation, we identified Sp-D as the ligand of Ig-Hepta/GPR116. Analyses of surfactant metabolism in Ig-Hepta(+/+ and Ig-Hepta(-/- mice by using radioactive tracers indicated that the Ig-Hepta/GPR116 signaling system exerts attenuating effects on (i balanced synthesis of surfactant lipids and proteins and (ii surfactant secretion, and (iii a stimulating effect on recycling (uptake in response to elevated levels of Sp-D in alveolar space.

  17. Lung surfactant levels are regulated by Ig-Hepta/GPR116 by monitoring surfactant protein D.

    Science.gov (United States)

    Fukuzawa, Taku; Ishida, Junji; Kato, Akira; Ichinose, Taro; Ariestanti, Donna Maretta; Takahashi, Tomoya; Ito, Kunitoshi; Abe, Jumpei; Suzuki, Tomohiro; Wakana, Shigeharu; Fukamizu, Akiyoshi; Nakamura, Nobuhiro; Hirose, Shigehisa

    2013-01-01

    Lung surfactant is a complex mixture of lipids and proteins, which is secreted from the alveolar type II epithelial cell and coats the surface of alveoli as a thin layer. It plays a crucial role in the prevention of alveolar collapse through its ability to reduce surface tension. Under normal conditions, surfactant homeostasis is maintained by balancing its release and the uptake by the type II cell for recycling and the internalization by alveolar macrophages for degradation. Little is known about how the surfactant pool is monitored and regulated. Here we show, by an analysis of gene-targeted mice exhibiting massive accumulation of surfactant, that Ig-Hepta/GPR116, an orphan receptor, is expressed on the type II cell and sensing the amount of surfactant by monitoring one of its protein components, surfactant protein D, and its deletion results in a pulmonary alveolar proteinosis and emphysema-like pathology. By a coexpression experiment with Sp-D and the extracellular region of Ig-Hepta/GPR116 followed by immunoprecipitation, we identified Sp-D as the ligand of Ig-Hepta/GPR116. Analyses of surfactant metabolism in Ig-Hepta(+/+) and Ig-Hepta(-/-) mice by using radioactive tracers indicated that the Ig-Hepta/GPR116 signaling system exerts attenuating effects on (i) balanced synthesis of surfactant lipids and proteins and (ii) surfactant secretion, and (iii) a stimulating effect on recycling (uptake) in response to elevated levels of Sp-D in alveolar space.

  18. Interpretation of Section 52.21(i) (3) of the Regulations for the PSD Requirement

    Science.gov (United States)

    This document may be of assistance in applying the New Source Review (NSR) air permitting regulations including the Prevention of Significant Deterioration (PSD) requirements. This document is part of the NSR Policy and Guidance Database. Some documents in the database are a scanned or retyped version of a paper photocopy of the original. Although we have taken considerable effort to quality assure the documents, some may contain typographical errors. Contact the office that issued the document if you need a copy of the original.

  19. Agreement that the PSD Regulations Require a Source to Commence Construction

    Science.gov (United States)

    This document may be of assistance in applying the New Source Review (NSR) air permitting regulations including the Prevention of Significant Deterioration (PSD) requirements. This document is part of the NSR Policy and Guidance Database. Some documents in the database are a scanned or retyped version of a paper photocopy of the original. Although we have taken considerable effort to quality assure the documents, some may contain typographical errors. Contact the office that issued the document if you need a copy of the original.

  20. Iron-regulated biofilm formation in Staphylococcus aureus Newman requires ica and the secreted protein Emp.

    Science.gov (United States)

    Johnson, Miranda; Cockayne, Alan; Morrissey, Julie A

    2008-04-01

    Staphylococcus aureus biofilm formation is induced in iron-restricted growth conditions in vitro. In this study, we showed that Emp and Eap play important roles in low-iron-induced biofilm formation of S. aureus Newman. Eap and Emp are secreted proteins which are non-covalently attached to the S. aureus cell surface and have previously been implicated in a number of aspects of S. aureus pathogenesis. We showed here that the transcription of these important virulence factors is induced by growth in low-iron medium, reflective of the in vivo environment. Our results show that iron regulation of Eap and Emp is Fur independent. However, Fur is required for full induction of eap and emp expression in low-iron conditions. In this study, we demonstrated that in addition to Fur, low-iron-induced biofilm formation requires Sae, Agr, and SarA. In iron-restricted growth conditions, Sae and Agr are essential for Emp and Eap expression and hence for biofilm formation, whereas SarA appears to have a less-significant role. We also showed that expression of the ica operon is required for biofilm formation in iron-restricted growth conditions. We demonstrated that in fact, ica is required for the expression of the important multifunctional virulence determinants eap and emp.

  1. Fis is required for proper regulation of ssaG expression in Salmonella enterica serovar Typhimurium.

    Science.gov (United States)

    Lim, Sangyong; Kim, Byeongkwan; Choi, Hyun-Sook; Lee, Younghoon; Ryu, Sangryeol

    2006-07-01

    Salmonella pathogenicity island 2 (SPI2) encodes a type III secretion system (TTSS) necessary for bacterial survival and replication in intracellular environment of host cells. SPI2 genes are transcribed preferentially after Salmonella enters the host cells. Transcriptional regulation of ssaG encoding the component of SPI2-TTSS apparatus was studied in vivo and in vitro. Fis, one of the major components of bacterial nucleoid, activated the stationary phase-specific expression of ssaG when Salmonella was grown in LB media. Gel-shift and footprinting analysis showed Fis bound to four distinct sites of the ssaG promoter region with different affinities. All four Fis-binding sites were required for timely transcription activation of ssaG after Salmonella entered macrophage cells. Gentamicin protection experiments using bacteria grown to stationary phase prior to infection showed that the ability of the fis mutant strain to replicate within the RAW264.7 macrophage cells was lower than the wild type. These observations confirm that Fis plays an important role in regulations of SPI2 as well as SPI1 for an efficient regulation of the virulence genes.

  2. Medical regulation and health outcomes: the effect of the physician examination requirement.

    Science.gov (United States)

    Cotet, Anca M; Benjamin, Daniel K

    2013-04-01

    This article investigates the effect on health outcomes of the regulation prohibiting physicians from prescribing drugs without a prior physical examination. This requirement could improve health by reducing illegal access to prescription drugs. However, it reduces access to health care by making it more difficult for patients and physicians to use many forms of telemedicine. Thus, this regulation generates a trade-off between access and safety. Using matching techniques, we find that the physician examination requirement leads to an increase of 1% in mortality rates from disease, the equivalent of 8.5 more deaths per 100,000 people, and a decrease of 6.7% in injury mortality, the equivalent of 2.5 deaths per 100,000 people. The magnitude of these effects is larger in rural areas and in areas with low physician density and is accompanied by an 18% increase in the number of days lost each month to illness. Copyright © 2012 John Wiley & Sons, Ltd.

  3. Extracellular Matrix-Regulated Gene Expression RequiresCooperation of SWI/SNF and Transcription Factors

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Ren; Spencer, Virginia A.; Bissell, Mina J.

    2006-05-25

    Extracellular cues play crucial roles in the transcriptional regulation of tissue-specific genes, but whether and how these signals lead to chromatin remodeling is not understood and subject to debate. Using chromatin immunoprecipitation (ChIP) assays and mammary-specific genes as models, we show here that extracellular matrix (ECM) molecules and prolactin cooperate to induce histone acetylation and binding of transcription factors and the SWI/SNF complex to the {beta}- and ?-casein promoters. Introduction of a dominant negative Brg1, an ATPase subunit of SWI/SNF complex, significantly reduced both {beta}- and ?-casein expression, suggesting that SWI/SNF-dependent chromatin remodeling is required for transcription of mammary-specific genes. ChIP analyses demonstrated that the ATPase activity of SWI/SNF is necessary for recruitment of RNA transcriptional machinery, but not for binding of transcription factors or for histone acetylation. Coimmunoprecipitation analyses showed that the SWI/SNF complex is associated with STAT5, C/EBP{beta}, and glucocorticoid receptor (GR). Thus, ECM- and prolactin-regulated transcription of the mammary-specific casein genes requires the concerted action of chromatin remodeling enzymes and transcription factors.

  4. 40 CFR 98.344 - Monitoring and QA/QC requirements.

    Science.gov (United States)

    2010-07-01

    ...) of this section, you may use total gaseous organic concentration analyzers and calculate the methane... composition monitor determines CH4 concentration on a dry basis, moisture content. (1) ASME...

  5. 50 CFR 18.118 - What are the mitigation, monitoring, and reporting requirements?

    Science.gov (United States)

    2010-10-01

    ... active seismic-source vessels and/or exploratory drilling operations. No more than four simultaneous... area. Monitors will act as an early detection system in regard to proximate bear activity to...

  6. UHRF1 regulation of Dnmt1 is required for pre-gastrula zebrafish development.

    Science.gov (United States)

    Kent, Brandon; Magnani, Elena; Walsh, Martin J; Sadler, Kirsten C

    2016-04-01

    Landmark epigenetic events underlie early embryonic development, yet how epigenetic modifiers are regulated to achieve rapid epigenome re-patterning is not known. Uhrf1 and DNA methyltransferase 1 (Dnmt1) are known to largely mediate maintenance DNA methylation and Uhrf1 is also required for both Dnmt1 localization and stability. Here, we investigate how these two key epigenetic modifiers regulate early zebrafish development and characterize the developmental consequences of disrupting their homeostatic relationship. Unlike Uhrf1 knockdown, which causes developmental arrest and death prior to gastrulation, overexpression of human UHRF1 (WT-UHRF1) caused asymmetric epiboly, inefficient gastrulation and multi-systemic defects. UHRF1 phosphorylation was previously demonstrated as essential for zebrafish embryogenesis, and we found that penetrance of the asymmetric epiboly phenotype was significantly increased in embryos injected with mRNA encoding non-phosphorylatable UHRF1 (UHRF1(S661A)). Surprisingly, both WT-UHRF1 and UHRF1(S661A) overexpression caused DNA hypomethylation. However, since other approaches that caused an equivalent degree of DNA hypomethylation did not cause the asymmetric epiboly phenotype, we conclude that bulk DNA methylation is not the primary mechanism. Instead, UHRF1(S661A) overexpression resulted in accumulation of Dnmt1 protein and the overexpression of both WT and a catalytically inactive Dnmt1 phenocopied the assymetric epiboly phenotype. Dnmt1 knockdown suppressed the phenotype caused by UHRF1(S661A) overexpression, and Uhrf1 knockdown suppressed the effect of Dnmt1 overexpression. Therefore, we conclude that the interaction between these two proteins is the mechanism underlying the gastrulation defects. This indicates that Dnmt1 stability requires UHRF1 phosphorylation and that crosstalk between the proteins is essential for the function of these two important epigenetic regulators during gastrulation.

  7. Leptin is required for hypothalamic regulation of miRNAs targeting POMC 3’UTR

    Directory of Open Access Journals (Sweden)

    Adel eDerghal

    2015-05-01

    Full Text Available The central nervous system monitors modifications in metabolic parameters or hormone levels and elicits adaptive responses such as food intake regulation. Particularly, within the hypothalamus, leptin modulates the activity of pro-opiomelanocortin (POMC neurons which are critical regulators of energy balance. Consistent with a pivotal role of the melanocortin system in the control of energy homeostasis, disruption of the POMC gene causes hyperphagia and obesity. MicroRNAs (miRNAs are short noncoding RNA molecules that post-transcriptionally repress the expression of genes by binding to 3'-untranslated regions (3'UTR of the target mRNAs. However, little is known regarding the role of miRNAs that target POMC 3’UTR in the central control energy homeostasis. Particularly, their interaction with the leptin signaling pathway remain unclear. First, we used common prediction programs to search for potential miRNAs target sites on 3'UTR of POMC mRNA. This screening identified a set of conserved miRNAs seed sequences for mir-383, mir-384-3p and mir-488. We observed that mir-383, mir-384-3p and mir-488 are up-regulated in the hypothalamus of leptin deficient ob/ob mice. In accordance with these observations, we also showed that mir-383, mir-384-3p and mir-488 were increased in db/db mice that exhibit a non-functional leptin receptor. The intraperitoneal injection of leptin down-regulated the expression of these miRNAs of interest in the hypothalamus of ob/ob mice showing the involvement of leptin in the expression of mir-383, mir-384-3p and mir-488. Finally, the evaluation of responsivity to intracerebroventricular administration of leptin exhibited that a chronic treatment with leptin decreased mir-488 expression in hypothalamus of C57BL/6 mice. In summary, these results suggest that leptin modulates the expression of miRNAs that target POMC mRNA in hypothalamus.

  8. Leptin is required for hypothalamic regulation of miRNAs targeting POMC 3′UTR

    Science.gov (United States)

    Derghal, Adel; Djelloul, Mehdi; Airault, Coraline; Pierre, Clément; Dallaporta, Michel; Troadec, Jean-Denis; Tillement, Vanessa; Tardivel, Catherine; Bariohay, Bruno; Trouslard, Jérôme; Mounien, Lourdes

    2015-01-01

    The central nervous system (CNS) monitors modifications in metabolic parameters or hormone levels and elicits adaptive responses such as food intake regulation. Particularly, within the hypothalamus, leptin modulates the activity of pro-opiomelanocortin (POMC) neurons which are critical regulators of energy balance. Consistent with a pivotal role of the melanocortin system in the control of energy homeostasis, disruption of the POMC gene causes hyperphagia and obesity. MicroRNAs (miRNAs) are short noncoding RNA molecules that post-transcriptionally repress the expression of genes by binding to 3′-untranslated regions (3′UTR) of the target mRNAs. However, little is known regarding the role of miRNAs that target POMC 3′UTR in the central control energy homeostasis. Particularly, their interaction with the leptin signaling pathway remain unclear. First, we used common prediction programs to search for potential miRNAs target sites on 3′UTR of POMC mRNA. This screening identified a set of conserved miRNAs seed sequences for mir-383, mir-384-3p, and mir-488. We observed that mir-383, mir-384-3p, and mir-488 are up-regulated in the hypothalamus of leptin deficient ob/ob mice. In accordance with these observations, we also showed that mir-383, mir-384-3p, and mir-488 were increased in db/db mice that exhibit a non-functional leptin receptor. The intraperitoneal injection of leptin down-regulated the expression of these miRNAs of interest in the hypothalamus of ob/ob mice showing the involvement of leptin in the expression of mir-383, mir-384-3p, and mir-488. Finally, the evaluation of responsivity to intracerebroventricular administration of leptin exhibited that a chronic treatment with leptin decreased mir-488 expression in hypothalamus of C57BL/6 mice. In summary, these results suggest that leptin modulates the expression of miRNAs that target POMC mRNA in hypothalamus. PMID:25999818

  9. Radioactivity monitoring and import regulation of the contaminated foodstuffs in Japan following the Chernobyl nuclear power plant accident

    Energy Technology Data Exchange (ETDEWEB)

    Izumo, Yoshiro [Institute of Public Health, Tokyo (Japan)

    1997-03-01

    Radioactivity monitoring and import regulation of the contaminated foodstuffs executed by Minstry of Health and Welfare following the Chernobyl nuclear plant accident were reviewed as follows; (1) background of socio-psychological effects and environmental radioactivity leading to the regulation (to may 3, 1986); (2) intial intervention for imported foodstuffs in Japan (may 8, `86), and (3) in european countries (to may 31, `86), immediately after the Accident, respectively; (4) determination of the interim driven intervention level for radionuclides in imported foodstuffs (({sup 134}Cs + {sup 137}Cs): 370 Bq/Kg) and activation of the monitoring, (5) outline of the monitoring with elapsed time, number of foodstuffs monitored, number of foodstuffs exceeded radioactivity of the intervention level and re-exported; (6) guideline in international trade of radioactive contaminated foodstuffs adopted by CODEX Alimentarius Commission (FAO/WHO) and the intervention level recommended by ICRP following the Accident; (7) discussion for problems and scopes in future based on the results of monitoring. As the results, a number of imported foodstuffs (about 75,000 samples at present) has been monitored, 55 samples exceeding the interim intervention level were re-exported to each export`s country, and socio-psychological doubts for radioactive contamination of imported foodstuffs have been dispersed. In addition, problems for several factors based on calculation of the interim intervention level, radioactivity level of foodstuffs exceeding about 50 Bq/Kg as radiocesiums and necessity of monitoring for the other radionuclides in foods except radiocesiums were also discussed. (author)

  10. Resolution requirements for monitor viewing of digital flat-panel detector radiographs: a contrast detail analysis

    Science.gov (United States)

    Peer, Siegfried; Steingruber, Iris; Gassner, Eva; Peer, Regina; Giacomuzzi, Salvatore M.

    2002-05-01

    Since the introduction of digital flat panel detectors into clinical routine the discussion on monitor specifications for primary soft copy reading has gained new impetus. Major concerns exist for viewing of tiny opacities such as pulmonary nodules. In this study CDRAD phantom images were acquired on a caesium iodid/amorphous silicon detector at varying exposure levels. Images were read three times by three observers on a clinical 1K and 2K monitor workstation. All typical workstation functions such as magnification and window/level setting were applied during image reading. Correct detection ratios were calculated according to the CDRAD evaluation manual. Observer ratings were highest for high dose exposure and 2K monitor reading. No significant difference was detected in the correct detection ratio of observers. However, the difference between the two types of workstations (1K versus 2K monitors) despite less than 3% was significant at a 95% confidence level. This is in good accordance with recently published clinical studies. However, further clinical work will be needed to strengthen this laboratory based impression. Given these subtle differences in low contrast detail detection on 1K and 2K clinical PACS workstation we should probably rethink the recommendations of various national boards for the use of 2K monitors.

  11. Spindle assembly checkpoint robustness requires Tpr-mediated regulation of Mad1/Mad2 proteostasis.

    Science.gov (United States)

    Schweizer, Nina; Ferrás, Cristina; Kern, David M; Logarinho, Elsa; Cheeseman, Iain M; Maiato, Helder

    2013-12-23

    Tpr is a conserved nuclear pore complex (NPC) protein implicated in the spindle assembly checkpoint (SAC) by an unknown mechanism. Here, we show that Tpr is required for normal SAC response by stabilizing Mad1 and Mad2 before mitosis. Tpr coimmunoprecipitated with Mad1 and Mad2 (hereafter designated as Tpr/Mad1/Mad2 or TM2 complex) during interphase and mitosis, and is required for Mad1–c-Mad2 recruitment to NPCs. Interestingly, Tpr was normally undetectable at kinetochores and dispensable for Mad1, but not for Mad2, kinetochore localization, which suggests that SAC robustness depends on Mad2 levels at kinetochores. Protein half-life measurements demonstrate that Tpr stabilizes Mad1 and Mad2, ensuring normal Mad1–c-Mad2 production in an mRNA- and kinetochore-independent manner. Overexpression of GFP-Mad2 restored normal SAC response and Mad2 kinetochore levels in Tpr-depleted cells. Mechanistically, we provide evidence that Tpr might spatially regulate SAC proteostasis through the SUMO-isopeptidases SENP1 and SENP2 at NPCs. Thus, Tpr is a kinetochore-independent, rate-limiting factor required to mount and sustain a robust SAC response.

  12. Regulating Effluents From India’s Textile Sector: New Commands and Compliance Monitoring for Zero Liquid Discharge

    Directory of Open Access Journals (Sweden)

    Jenny Grönwall

    2017-06-01

    Full Text Available In 2016 large parts of India had experienced failing monsoons for two consecutive years. Textiles production was a natural target in the quest for solutions when re-distribution of water between sectors became imperative, and it seemed likely that the zero liquid discharge (ZLD approach would be mainstreamed nationwide. However, when amended standards for discharge of effluents were enacted it was without strict requirements for reuse of water in-house; the legislator chose to make ZLD applicable only to large units and refrained from making it mandatory. The carrying capacity of the environment in sensitive or otherwise critical areas may henceforth be taken into account by the executive, but the textile sector’s wastewater is not yet regarded the resource that a circular economy calls for. This paper examines the command-side of regulation by shedding light on the applicable law, the reform steps taken in 2014–16 and how judicial interventions influenced these. It also seeks to contribute to the understanding of enforcement control by discussing what role court-established committees are playing in implementation and monitoring of compliance, based on an in-depth case study of Tirupur, India’s ‘knit city’.

  13. 40 CFR 281.40 - Requirements for compliance monitoring program and authority.

    Science.gov (United States)

    2010-07-01

    ... inspection procedures to determine, independent of information supplied by regulated persons, compliance with..., these procedures must be conducted in a manner (for example, using proper “chain of custody”...

  14. Development of LC/MS/MS Methods for Implementation in US EPA’s Drinking Water Unregulated Contaminant Monitoring Regulations

    Science.gov (United States)

    Well-characterized and standardized methods are the foundation upon which monitoring of regulated and unregulated contaminants in drinking water are based. To obtain reliable, high quality data for trace analysis of contaminants, these methods must be rugged, selective and sensit...

  15. Meeting the International Health Regulations (2005) surveillance core capacity requirements at the subnational level in Europe

    DEFF Research Database (Denmark)

    Ziemann, Alexandra; Rosenkötter, Nicole; Riesgo, Luis Garcia-Castrillo;

    2015-01-01

    public health emergencies of international concern: (i) can syndromic surveillance support countries, especially the subnational level, to meet the International Health Regulations (2005) core surveillance capacity requirements, (ii) are European syndromic surveillance systems comparable to enable cross......-border surveillance, and (iii) at which administrative level should syndromic surveillance best be applied? DISCUSSION: Despite the ongoing criticism on the usefulness of syndromic surveillance which is related to its clinically nonspecific output, we demonstrate that it was a suitable supplement for timely...... assessment of the impact of three different public health emergencies affecting Europe. Subnational syndromic surveillance analysis in some cases proved to be of advantage for detecting an event earlier compared to national level analysis. However, in many cases, syndromic surveillance did not detect local...

  16. Spindly/CCDC99 is required for efficient chromosome congression and mitotic checkpoint regulation.

    Science.gov (United States)

    Barisic, Marin; Sohm, Bénédicte; Mikolcevic, Petra; Wandke, Cornelia; Rauch, Veronika; Ringer, Thomas; Hess, Michael; Bonn, Günther; Geley, Stephan

    2010-06-15

    Spindly recruits a fraction of cytoplasmic dynein to kinetochores for poleward movement of chromosomes and control of mitotic checkpoint signaling. Here we show that human Spindly is a cell cycle-regulated mitotic phosphoprotein that interacts with the Rod/ZW10/Zwilch (RZZ) complex. The kinetochore levels of Spindly are regulated by microtubule attachment and biorientation induced tension. Deletion mutants lacking the N-terminal half of the protein (NDelta253), or the conserved Spindly box (DeltaSB), strongly localized to kinetochores and failed to respond to attachment or tension. In addition, these mutants prevented the removal of the RZZ complex and that of MAD2 from bioriented chromosomes and caused cells to arrest at metaphase, showing that RZZ-Spindly has to be removed from kinetochores to terminate mitotic checkpoint signaling. Depletion of Spindly by RNAi, however, caused cells to arrest in prometaphase because of a delay in microtubule attachment. Surprisingly, this defect was alleviated by codepletion of ZW10. Thus, Spindly is not only required for kinetochore localization of dynein but is a functional component of a mechanism that couples dynein-dependent poleward movement of chromosomes to their efficient attachment to microtubules.

  17. Interdomain allosteric regulation of Polo kinase by Aurora B and Map205 is required for cytokinesis.

    Science.gov (United States)

    Kachaner, David; Pinson, Xavier; El Kadhi, Khaled Ben; Normandin, Karine; Talje, Lama; Lavoie, Hugo; Lépine, Guillaume; Carréno, Sébastien; Kwok, Benjamin H; Hickson, Gilles R; Archambault, Vincent

    2014-10-27

    Drosophila melanogaster Polo and its human orthologue Polo-like kinase 1 fulfill essential roles during cell division. Members of the Polo-like kinase (Plk) family contain an N-terminal kinase domain (KD) and a C-terminal Polo-Box domain (PBD), which mediates protein interactions. How Plks are regulated in cytokinesis is poorly understood. Here we show that phosphorylation of Polo by Aurora B is required for cytokinesis. This phosphorylation in the activation loop of the KD promotes the dissociation of Polo from the PBD-bound microtubule-associated protein Map205, which acts as an allosteric inhibitor of Polo kinase activity. This mechanism allows the release of active Polo from microtubules of the central spindle and its recruitment to the site of cytokinesis. Failure in Polo phosphorylation results in both early and late cytokinesis defects. Importantly, the antagonistic regulation of Polo by Aurora B and Map205 in cytokinesis reveals that interdomain allosteric mechanisms can play important roles in controlling the cellular functions of Plks.

  18. Learned stressor resistance requires extracellular signal-regulated kinase in the prefrontal cortex

    Directory of Open Access Journals (Sweden)

    John Paul Christianson

    2014-10-01

    Full Text Available Behaviorally controllable stressors confer protection from the neurochemical and behavioral consequences of future uncontrollable stressors, a phenomenon termed behavioral immunization. Recent data implicate neuroplasticity within the ventromedial prefrontal cortex (mPFC as critical to behavioral immunization. Adult, male Sprague-Dawley rats were exposed to a series of controllable tailshocks and one week later to uncontrollable tailshocks, followed 24h later by social exploration and shuttlebox escape tests. To test the involvement of N-methyl-D-aspartate receptors (NMDAR and the extracellular signal-regulated kinase (ERK cascade in behavioral immunization, either D-AP5 or the MEK inhibitor U0126 was injected to the prelimbic (PL or infralimbic (IL mPFC prior to controllable stress exposure. Phosphorylated ERK and P70S6K, regulators of transcription and translation, were quantified by Western blot or immunohistochemistry after controllable or uncontrollable tailshocks. Prior controllable stress prevented the social exploration and shuttlebox performance deficits caused by the later uncontrollable stressor, and this effect was blocked by injections of D-AP5 into mPFC. A significant increase in phosphorylated ERK1 and ERK2, but not P70S6K, occurred within the PL and IL in rats exposed to controllable stress, but not to uncontrollable stress. However, U0126 only prevented behavioral immunization when injected to the PL. We provide evidence that NMDAR and ERK dependent plasticity within the PL region is required for behavioral immunization, a learned form of stressor resistance.

  19. Use of models to support the monitoring requirements in the water framework directive

    NARCIS (Netherlands)

    Højberg, A.L.; Refsgaard, J.C.; Geer, F. van; Jørgensen, L.F.; Zsuffa, I.

    2007-01-01

    Implementation of the EU Water Framework Directive (WFD) poses many new challenges to European water managers. Monitoring programmes play a key role to assess the status and identify possible trends in the environmental conditions of river basins; to gain new knowledge on water processes and to

  20. Linking requirements with capabilities for deforestation monitoring in the context of the UNFCCC-REDD process

    Energy Technology Data Exchange (ETDEWEB)

    Herold, Martin [GOFC-GOLD Land Cover Project Office, Department of Earth Observation, Friedrich-Schiller University, Jena 07743 (Germany); Johns, Tracy [Joanneum Research, Institute for Energy Research, Graz 8010 (Austria)

    2007-10-15

    As the United Nations Framework Convention on Climate Change nears the end of a two-year period of evaluation of the issue of reducing emissions from deforestation in developing countries, participating countries have agreed on the need to address this globally important source of greenhouse gas emissions. Negotiations on policy frameworks to monitor and reduce deforestation rely on an understanding of the scientific and technical capacity to support these efforts. Current UNFCCC programs to improve observation of land and forest cover change can provide valuable input to a future policy mechanism focusing on deforestation emissions from developing countries. Countries participating in the current debate have officially referenced the value of remote sensing tools and methods for deforestation monitoring, and have identified specific needs and goals related to their implementation. Based on these identified needs, this paper outlines recommendations for a monitoring framework that can be globally applied with sufficient levels of accuracy and certainty. This framework can serve as a starting point for monitoring programs, and can be modified in response to expected progress in establishing an international policy framework for reducing emissions from deforestation.

  1. 40 CFR 63.9804 - What are my monitoring system installation, operation, and maintenance requirements?

    Science.gov (United States)

    2010-07-01

    ..., oxidation, and galvanic corrosion. (c) For each pressure CPMS that is used to monitor the pressure drop... where it is easily recognized by plant operating personnel. (6) For positive pressure fabric filter... pressure or induced air fabric filters, the bag leak detector must be installed downstream of the...

  2. 75 FR 43487 - Proposed Information Collection; Comment Request; Vessel Monitoring System Requirements in...

    Science.gov (United States)

    2010-07-26

    ... vessel monitoring system (VMS) units on their vessels when directed to do so by NOAA enforcement personnel. VMS units automatically send periodic reports on the position of the vessel. NOAA uses the..., VMS installation and annual maintenance are considered public burden. This request combines three...

  3. 77 FR 36488 - Proposed Information Collection; Comment Request; Vessel Monitoring System Requirements Under the...

    Science.gov (United States)

    2012-06-19

    ... near real-time satellite-based position-fixing transmitters (``Vessel Monitoring System-VMS-units'') at... must transmit: (1) ``On/off reports'' to NMFS whenever the VMS unit is turned off while the vessel is in port, (2) ``activation reports'' to NMFS prior to the first use of a VMS unit, and (3)...

  4. New Approach to Purging Monitoring Wells: Lower Flow Rates Reduce Required Purging Volumes and Sample Turbidity

    Science.gov (United States)

    It is generally accepted that monitoring wells must be purged to access formation water to obtain “representative” ground water quality samples. Historically anywhere from 3 to 5 well casing volumes have been removed prior to sample collection to evacuate the standing well water...

  5. 40 CFR Table 3 to Subpart Eeee of... - Requirements for Continuous Emission Monitoring Systems (CEMS)

    Science.gov (United States)

    2010-07-01

    ... Monitoring Systems (CEMS) 3 Table 3 to Subpart EEEE of Part 60 Protection of Environment ENVIRONMENTAL... Systems (CEMS) As stated in § 60.2940, you must comply with the following: For the followingpollutants Use the following span values for your CEMS Use the following performance specifications (P.S.)...

  6. Use of models to support the monitoring requirements in the water framework directive

    NARCIS (Netherlands)

    Højberg, A.L.; Refsgaard, J.C.; Geer, F. van; Jørgensen, L.F.; Zsuffa, I.

    2007-01-01

    Implementation of the EU Water Framework Directive (WFD) poses many new challenges to European water managers. Monitoring programmes play a key role to assess the status and identify possible trends in the environmental conditions of river basins; to gain new knowledge on water processes and to asse

  7. 40 CFR Table 4 to Subpart Ggg of... - Monitoring Requirements for Control Devices a

    Science.gov (United States)

    2010-07-01

    ... a diversion was detected at any time during each hour. 2. Valves sealed closed with car-seal or lock... heater fuel Temperature monitoring device... cycle(s) 1. For each regeneration cycle, record the total regeneration stream mass or volumetric...

  8. 40 CFR Table 3 to Subpart Mmm of... - Monitoring Requirements for Control Devices a

    Science.gov (United States)

    2010-07-01

    .... 2. Valves sealed closed with car-seal or lock-and-key configuration 2. Monthly inspections of sealed... the primary fuel Temperature monitoring device installed in firebox b Combustion temperature Every 15... volumetric flow during carbon bed regeneration cycle(s) 1. For each regeneration cycle, record the...

  9. SMART MONITORING AND DECISION MAKING FOR REGULATING ANNULUS BOTTOM HOLE PRESSURE WHILE DRILLING OIL WELLS

    Directory of Open Access Journals (Sweden)

    M. P. Vega

    Full Text Available Abstract Real time measurements and development of sensor technology are research issues associated with robustness and safety during oil well drilling operations, making feasible the diagnosis of problems and the development of a regulatory strategy. The major objective of this paper is to use an experimental plant and also field data, collected from a basin operation, offshore Brazil, for implementing smart monitoring and decision making, in order to assure drilling inside operational window, despite the commonly observed disturbances that produce fluctuations in the well annulus bottom hole pressure. Using real time measurements, the performance of a continuous automated drilling unit is analyzed under a scenario of varying levels of rate of penetration; aiming pressure set point tracking (inside the operational drilling window and also rejecting kick, a phenomenon that occurs when the annulus bottom hole pressure is inferior to the porous pressure, producing the migration of reservoir fluids into the annulus region. Finally, an empirical model was built, using real experimental data from offshore Brazil basins, enabling diagnosing and regulating a real drilling site by employing classic and advanced control strategies.

  10. 40 CFR 141.563 - What follow-up action is my system required to take based on continuous turbidity monitoring?

    Science.gov (United States)

    2010-07-01

    ... combined filter effluent (CFE) for systems with 2 filters that monitor CFE in lieu of individual filters... consecutive recordings 15 minutes apart at the same filter (or CFE for systems with 2 filters that monitor CFE... as specified in paragraph (c) of this section was required. Systems with 2 filters that monitor CFE...

  11. Th9 cell development requires a BATF-regulated transcriptional network.

    Science.gov (United States)

    Jabeen, Rukhsana; Goswami, Ritobrata; Awe, Olufolakemi; Kulkarni, Aishwarya; Nguyen, Evelyn T; Attenasio, Andrea; Walsh, Daniel; Olson, Matthew R; Kim, Myung H; Tepper, Robert S; Sun, Jie; Kim, Chang H; Taparowsky, Elizabeth J; Zhou, Baohua; Kaplan, Mark H

    2013-11-01

    T helper 9 (Th9) cells are specialized for the production of IL-9, promote allergic inflammation in mice, and are associated with allergic disease in humans. It has not been determined whether Th9 cells express a characteristic transcriptional signature. In this study, we performed microarray analysis to identify genes enriched in Th9 cells compared with other Th subsets. This analysis defined a transcriptional regulatory network required for the expression of a subset of Th9-enriched genes. The activator protein 1 (AP1) family transcription factor BATF (B cell, activating transcription factor–like) was among the genes enriched in Th9 cells and was required for the expression of IL-9 and other Th9-associated genes in both human and mouse T cells. The expression of BATF was increased in Th9 cultures derived from atopic infants compared with Th9 cultures from control infants. T cells deficient in BATF expression had a diminished capacity to promote allergic inflammation compared with wild-type controls. Moreover, mouse Th9 cells ectopically expressing BATF were more efficient at promoting allergic inflammation than control transduced cells. These data indicate that BATF is a central regulator of the Th9 phenotype and contributes to the development of allergic inflammation.

  12. Revocation of General Safety Test Regulations That Are Duplicative of Requirements in Biologics License Applications. Final rule.

    Science.gov (United States)

    2015-07-02

    The Food and Drug Administration (FDA) is amending the biologics regulations by removing the general safety test (GST) requirements for biological products. FDA is finalizing this action because the existing codified GST regulations are duplicative of requirements that are also specified in biologics license applications (BLAs), or are no longer necessary or appropriate to help ensure the safety, purity, and potency of licensed biological products. FDA is taking this action as part of its retrospective review of its regulations to promote improvement and innovation, in response to the Executive order.

  13. The experience of family members of ICU patients who require extensive monitoring: a qualitative study.

    Science.gov (United States)

    Smith, Claudia DiSabatino; Custard, Kristi

    2014-09-01

    A mixed methods study using family research with a phenomenological approach (n = 5 families) was conducted to explore family members' perceptions about the extensive monitoring technology used on their critically ill family member after cardiac surgery, as experienced when family members initially visited the patient in the cardiovascular intensive care unit. Five relevant themes emerged: overwhelmed by all of the machines; feelings of uncertainty; methods of coping; meaning of the numbers on the machines; and need for education.

  14. 40 CFR 98.164 - Monitoring and QA/QC requirements.

    Science.gov (United States)

    2010-07-01

    ... requirements in § 98.3(i) of this part. (2) Determine the carbon content and the molecular weight annually of... analysis is required to determine the carbon content and molecular weight of the fuel and feedstock. (3... methods to determine the carbon content for all fuels and feedstocks, and molecular weight of...

  15. A monomer is the minimum functional unit required for channel and ATPase activity of the cystic fibrosis transmembrane conductance regulator.

    Science.gov (United States)

    Ramjeesingh, M; Li, C; Kogan, I; Wang, Y; Huan, L J; Bear, C E

    2001-09-04

    The cystic fibrosis transmembrane conductance regulator (CFTR) normally functions as a phosphorylation-regulated chloride channel on the apical surface of epithelial cells, and lack of this function is the primary cause for the fatal disease cystic fibrosis (CF). Previous studies showed that purified, reconstituted CFTR can function as a chloride channel and, further, that its intrinsic ATPase activity is required to regulate opening and closing of the channel gate. However, these previous studies did not identify the quaternary structure required to mediate conduction and catalysis. Our present studies show that CFTR molecules may self-associate in CHO and Sf9 membranes, as complexes close to the predicted size of CFTR dimers can be captured by chemical cross-linking reagents and detected using nondissociative PAGE. However, CFTR function does not require a multimeric complex for function as we determined that purified, reconstituted CFTR monomers are sufficient to mediate regulated chloride conduction and ATPase activity.

  16. Profiles of Motivated Self-Regulation in College Computer Science Courses: Differences in Major versus Required Non-Major Courses

    Science.gov (United States)

    Shell, Duane F.; Soh, Leen-Kiat

    2013-01-01

    The goal of the present study was to utilize a profiling approach to understand differences in motivation and strategic self-regulation among post-secondary STEM students in major versus required non-major computer science courses. Participants were 233 students from required introductory computer science courses (194 men; 35 women; 4 unknown) at…

  17. 50 CFR 404.5 - Requirements for a vessel monitoring system.

    Science.gov (United States)

    2010-10-01

    ... ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE); ENDANGERED SPECIES COMMITTEE REGULATIONS SUBCHAPTER A... when voyaging within the Monument. An operating VMS includes an operating mobile transmitting unit on... operable. (e) Access to position data. As a condition of authorized access to the Monument, a vessel owner...

  18. Power Control and Monitoring Requirements for Thermal Vacuum/Thermal Balance Testing of the MAP Observatory

    Science.gov (United States)

    Johnson, Chris; Hinkle, R. Kenneth (Technical Monitor)

    2002-01-01

    The specific heater control requirements for the thermal vacuum and thermal balance testing of the Microwave Anisotropy Probe (MAP) Observatory at the Goddard Space Flight Center (GSFC) in Greenbelt, Maryland are described. The testing was conducted in the 10m wide x 18.3m high Space Environment Simulator (SES) Thermal Vacuum Facility. The MAP thermal testing required accurate quantification of spacecraft and fixture power levels while minimizing heater electrical emissions. The special requirements of the MAP test necessitated construction of five (5) new heater racks.

  19. Performance monitoring and the medial prefrontal cortex: A review of individual differences and context effects as a window on self-regulation

    Directory of Open Access Journals (Sweden)

    Stefon evan Noordt

    2012-07-01

    Full Text Available The medial prefrontal cortex (MPFC is central to self-regulation and has been implicated in generating a cluster of event-related potential components, collectively referred to as medial frontal negativities (MFNs. These MFNs are elicited while individuals monitor behavioural and environmental consequences, and include the error-related negativity, Nogo N2, and the feedback-related negativity. A growing cognitive and affective neuroscience literature indicates that the activation of the anterior cingulate cortex and surrounding medial prefrontal regions during performance monitoring is not only influenced by task context, but that these patterns of activity also vary as a function of individual differences (e.g., personality, temperament, clinical and non-clinical symptomatology, socio-political orientation, and genetic polymorphisms, as well as interactions between individual differences and task context. In this review we survey the neuroscience literature on the relations between performance monitoring, personality, task context, and brain functioning with a focus on the MPFC. We relate these issues to the role of affect in the paradigms used to elicit performance-monitoring neural responses and highlight some of the theoretical and clinical implications of this research. We conclude with a discussion of the complexity of these issues and how some of the basic assumptions required for their interpretation may be clarified with future research.

  20. Sunsynchronous low Earth orbit spacecraft concepts and technology requirements for global change monitoring

    Science.gov (United States)

    Garrett, L. Bernard; Butterfield, Ansel J.; Taback, Israel; Garn, Paul A.; Burrowbridge, Donald R., Jr.

    1991-01-01

    The Global Change Technology Initiative listing of instruments for operation in low Earth, sunsynchronous orbits contain 21 entries, of which 20 are carried aboard multi-instrument spacecraft. This list identifies the temporal requirements for repetition of measurements and also includes groups of instruments that make complementing measurements. Definitions for individual spacecraft follows the temporal and grouping requirements to establish constellations which will provide the measurement data. The definitions of constellations for multi-instrument spacecraft show two alternatives: a constellation of 10 spacecraft, each compatible with launch by a Delta booster; a constellation of 4 spacecraft, each requiring a Titan booster. Operating subsystems for the individual spacecraft can use modular concepts that are adaptations based upon current plans for improving the performance of the NASA-Goddard Multimission Modular units. The descriptions of the spacecraft and constellations begins with a compilation of instrument related requirements that define the principal system performance parameters and operating capabilities.

  1. BDNF activates mTOR to regulate GluR1 expression required for memory formation.

    Directory of Open Access Journals (Sweden)

    Leandro Slipczuk

    Full Text Available BACKGROUND: The mammalian target of Rapamycin (mTOR kinase plays a key role in translational control of a subset of mRNAs through regulation of its initiation step. In neurons, mTOR is present at the synaptic region, where it modulates the activity-dependent expression of locally-translated proteins independently of mRNA synthesis. Indeed, mTOR is necessary for different forms of synaptic plasticity and long-term memory (LTM formation. However, little is known about the time course of mTOR activation and the extracellular signals governing this process or the identity of the proteins whose translation is regulated by this kinase, during mnemonic processing. METHODOLOGY/PRINCIPAL FINDINGS: Here we show that consolidation of inhibitory avoidance (IA LTM entails mTOR activation in the dorsal hippocampus at the moment of and 3 h after training and is associated with a rapid and rapamycin-sensitive increase in AMPA receptor GluR1 subunit expression, which was also blocked by intra-hippocampal delivery of GluR1 antisense oligonucleotides (ASO. In addition, we found that pre- or post-training administration of function-blocking anti-BDNF antibodies into dorsal CA1 hampered IA LTM retention, abolished the learning-induced biphasic activation of mTOR and its readout, p70S6K and blocked GluR1 expression, indicating that BDNF is an upstream factor controlling mTOR signaling during fear-memory consolidation. Interestingly, BDNF ASO hindered LTM retention only when given into dorsal CA1 1 h after but not 2 h before training, suggesting that BDNF controls the biphasic requirement of mTOR during LTM consolidation through different mechanisms: an early one involving BDNF already available at the moment of training, and a late one, happening around 3 h post-training that needs de novo synthesis of this neurotrophin. CONCLUSIONS/SIGNIFICANCE: IN CONCLUSION, OUR FINDINGS DEMONSTRATE THAT: 1 mTOR-mediated mRNA translation is required for memory consolidation during

  2. Experiences in certification of packages for transportation of fresh nuclear fuel in the context of new safety requirements established by IAEA regulations (IAEA-96 regulations, ST-1) for air transportation of nuclear materials (requirements to C-type packages)

    Energy Technology Data Exchange (ETDEWEB)

    Dudai, V.I.; Kovtun, A.D.; Matveev, V.Z.; Morenko, A.I.; Nilulin, V.M.; Shapovalov, V.I.; Yakushev, V.A.; Bobrovsky, V.S.; Rozhkov, V.V.; Agapov, A.M.; Kolesnikov, A.S. [Russian Federal Nuclear Centre - All-Russian Research Inst. of Experimental Physics, Sarov (Russian Federation)]|[JSC ' ' MSZ' ' , Electrostal (Russian Federation)]|[JSC ' ' NPCC' ' , Novosibirsk (Russian Federation)]|[Minatom of Russia, Moscow (Russian Federation)]|[Gosatomnadzor of Russia, Moscow (Russian Federation)

    2004-07-01

    Every year in Russia, a large amount of domestic and international transportation of fresh nuclear fuel (FNF) used in Russian and foreign energy and research atomic reactors and referred to fissile materials based on IAEA Regulations is performed. Here, bulk transportation is performed by air, and it concerns international transportation in particular. According to national ''Main Regulations for Safe Transport and physical Protection of Nuclear Materials (OPBZ- 83)'' and ''Regulations for the Safe Transport of Radioactive Materials'' of the International Atomic Energy Agency (IAEA Regulations), nuclear and radiation security under normal (accident free) and accident conditions of transport must be completely provided by the package design. In this context, high requirements to fissile packages exposed to heat and mechanical loads in transport accidents are imposed. A long-standing experience in accident free transportation of FM has shown that such approach to provide nuclear and radiation security pays for itself completely. Nevertheless, once in 10 years the International Atomic Energy Agency on every revision of the ''Regulations for the Safe Transport of Radioactive Materials'' places more stringent requirements upon the FM and transportation thereof, resulting from the objectively increasing risk associated with constant rise in volume and density of transportation, and also strained social and economical situation in a number of regions in the world. In the new edition of the IAEA Regulations (ST-1), published in 1996 and brought into force in 2001 (IAEA-96 Regulations), the requirements to FM packages conveyed by aircraft were radically changed. These requirements are completely presented in new Russian ''Regulations for the Safe Transport of Radioactive Materials'' (PBTRM- 2004) which will be brought into force in the time ahead.

  3. Regulations and monitoring of the financial part of the electricity market; Regler och tillsyn inom den finansiella delen av el-marknaden

    Energy Technology Data Exchange (ETDEWEB)

    Eriksson, Svante; Eliasson, Torben; Jenssen Aasmund [ECON Center for Economic Analysis A.S, Oslo (Norway)

    2001-11-01

    The electricity derivatives market has grown significantly during the last few years. It refers to all commodity derivatives (options, futures and forwards) based on electricity and traded either on the Nord Pool Exchange or bilaterally between single parties. The growth of the derivatives market has also led to an increasing need for relevant regulation and monitoring. In this report ECON describes how the common financial regulations (e.g. Sweden's Securities Operations Act) affect power sector companies and how the electricity derivatives market is being monitored by the Swedish and the Norwegian financial supervisory authorities. The aim of the report is to give ideas about possible future research projects about the electricity derivatives market. In Sweden commodity derivatives based on electricity are generally considered to be 'financial instruments' according to The Trading in Financial Instruments Act. At least this seems to be the case with contracts traded on Nord Pool and bilateral contracts that can be subject to clearing by Nord Pool. In some cases, companies wanting to offer services regarding financial instruments in the Swedish market need a special licence and it comes from the Swedish Financial Supervisory Authority. The services that require a special permit are: trading financial instruments, in one's own name, on behalf of another party, brokering of contacts between purchasers and sellers, trading in financial instruments on one's own account, management of another party's financial instruments, and underwriting or other participation in issuances of securities or offers to purchase or sell financial instruments directly to the public. A licence to conduct a securities operation brings with it, among other things, certain mandatory capital requirements. Securities operations should also be conducted in such a manner that public confidence is maintained in the securities markets. Regulation should insure that for

  4. Dentate Gyrus Development Requires ERK Activity to Maintain Progenitor Population and MAPK Pathway Feedback Regulation.

    Science.gov (United States)

    Vithayathil, Joseph; Pucilowska, Joanna; Goodnough, L Henry; Atit, Radhika P; Landreth, Gary E

    2015-04-29

    The ERK/MAPK pathway is an important developmental signaling pathway. Mutations in upstream elements of this pathway result in neuro-cardio-facial cutaneous (NCFC) syndromes, which are typified by impaired neurocognitive abilities that are reliant upon hippocampal function. The role of ERK signaling during hippocampal development has not been examined and may provide critical insight into the cause of hippocampal dysfunction in NCFC syndromes. In this study, we have generated ERK1 and conditional ERK2 compound knock-out mice to determine the role of ERK signaling during development of the hippocampal dentate gyrus. We found that loss of both ERK1 and ERK2 resulted in 60% fewer granule cells and near complete absence of neural progenitor pools in the postnatal dentate gyrus. Loss of ERK1/2 impaired maintenance of neural progenitors as they migrate from the dentate ventricular zone to the dentate gyrus proper, resulting in premature depletion of neural progenitor cells beginning at E16.5, which prevented generation of granule cells later in development. Finally, loss of ERK2 alone does not impair development of the dentate gyrus as animals expressing only ERK1 developed a normal hippocampus. These findings establish that ERK signaling regulates maintenance of progenitor cells required for development of the dentate gyrus. Copyright © 2015 the authors 0270-6474/15/356836-13$15.00/0.

  5. CSL protein regulates transcription of genes required to prevent catastrophic mitosis in fission yeast.

    Science.gov (United States)

    Převorovský, Martin; Oravcová, Martina; Zach, Róbert; Jordáková, Anna; Bähler, Jürg; Půta, František; Folk, Petr

    2016-11-16

    For every eukaryotic cell to grow and divide, intricately coordinated action of numerous proteins is required to ensure proper cell-cycle progression. The fission yeast Schizosaccharomyces pombe has been instrumental in elucidating the fundamental principles of cell-cycle control. Mutations in S. pombe 'cut' (cell untimely torn) genes cause failed coordination between cell and nuclear division, resulting in catastrophic mitosis. Deletion of cbf11, a fission yeast CSL transcription factor gene, triggers a 'cut' phenotype, but the precise role of Cbf11 in promoting mitotic fidelity is not known. We report that Cbf11 directly activates the transcription of the acetyl-coenzyme A carboxylase gene cut6, and the biotin uptake/biosynthesis genes vht1 and bio2, with the former 2 implicated in mitotic fidelity. Cbf11 binds to a canonical, metazoan-like CSL response element (GTGGGAA) in the cut6 promoter. Expression of Cbf11 target genes shows apparent oscillations during the cell cycle using temperature-sensitive cdc25-22 and cdc10-M17 block-release experiments, but not with other synchronization methods. The penetrance of catastrophic mitosis in cbf11 and cut6 mutants is nutrient-dependent. We also show that drastic decrease in biotin availability arrests cell proliferation but does not cause mitotic defects. Taken together, our results raise the possibility that CSL proteins play conserved roles in regulating cell-cycle progression, and they could guide experiments into mitotic CSL functions in mammals.

  6. Reciprocal regulation of Wnt and Gpr177/mouse Wntless is required for embryonic axis formation.

    Science.gov (United States)

    Fu, Jiang; Jiang, Ming; Mirando, Anthony J; Yu, Hsiao-Man Ivy; Hsu, Wei

    2009-11-03

    Members of the Wnt family are secreted glycoproteins that trigger cellular signals essential for proper development of organisms. Cellular signaling induced by Wnt proteins is involved in diverse developmental processes and human diseases. Previous studies have generated an enormous wealth of knowledge on the events in signal-receiving cells. However, relatively little is known about the making of Wnt in signal-producing cells. Here, we describe that Gpr177, the mouse orthologue of Drosophila Wls, is expressed during formation of embryonic axes. Embryos with deficient Gpr177 exhibit defects in establishment of the body axis, a phenotype highly reminiscent to the loss of Wnt3. Although many different mammalian Wnt proteins are required for a wide range of developmental processes, the Wnt3 ablation exhibits the earliest developmental abnormality. This suggests that the Gpr177-mediated Wnt production cannot be substituted. As a direct target of Wnt, Gpr177 is activated by beta-catenin and LEF/TCF-dependent transcription. This activation alters the cellular distributions of Gpr177 which binds to Wnt proteins and assists their sorting and secretion in a feedback regulatory mechanism. Our findings demonstrate that the loss of Gpr177 affects Wnt production in the signal-producing cells, leading to alterations of Wnt signaling in the signal-receiving cells. A reciprocal regulation of Wnt and Gpr177 is essential for the patterning of the anterior-posterior axis during mammalian development.

  7. Fgf19 regulated by Hh signaling is required for zebrafish forebrain development.

    Science.gov (United States)

    Miyake, Ayumi; Nakayama, Yoshiaki; Konishi, Morichika; Itoh, Nobuyuki

    2005-12-01

    Fibroblast growth factor (Fgf) signaling plays important roles in brain development. Fgf3 and Fgf8 are crucial for the formation of the forebrain and hindbrain. Fgf8 is also required for the midbrain to form. Here, we identified zebrafish Fgf19 and examined its roles in brain development by knocking down Fgf19 function. We found that Fgf19 expressed in the forebrain, midbrain and hindbrain was involved in cell proliferation and cell survival during embryonic brain development. Fgf19 was also essential for development of the ventral telencephalon and diencephalon. Regional specification is linked to cell type specification. Fgf19 was also essential for the specification of gamma-aminobutyric acid (GABA)ergic interneurons and oligodendrocytes generated in the ventral telencephalon and diencephalon. The cross talk between Fgf and Hh signaling is critical for brain development. In the forebrain, Fgf19 expression was down-regulated on inhibition of Hh but not of Fgf3/Fgf8, and overexpression of Fgf19 rescued partially the phenotype on inhibition of Hh. The present findings indicate that Fgf19 signaling is crucial for forebrain development by interacting with Hh and provide new insights into the roles of Fgf signaling in brain development.

  8. ADAM10 regulates transcription factor expression required for plasma cell function.

    Directory of Open Access Journals (Sweden)

    Natalia S Chaimowitz

    Full Text Available A disintegrin and metalloprotease 10 (ADAM10 is a key regulator of cellular processes by shedding extracellular domains of transmembrane proteins. We have previously demonstrated that deletion of B cell expressed ADAM10 results in changes in lymphoid tissue architecture and impaired germinal center (GC formation. In this study, mice were generated in which ADAM10 is deleted in B cells following class switch recombination (ADAM10(Δ/ΔIgG1-cre(+/- mice. Despite normal GC formation, antibody responses were impaired in ADAM10(Δ/ΔIgG1-cre(+/- mice, implicating ADAM10 in post-GC and extrafollicular B cell terminal differentiation. Surprisingly, plasma cell (PC numbers were normal in ADAM10(Δ/ΔIgG1-cre(+/- mice when compared to controls. However, PCs isolated from ADAM10(Δ/ΔIgG1-cre(+/- mice exhibited decreased expression of transcription factors important for PC function: Prdm1, Xbp1 and Irf4. Bcl6 is a GC transcriptional repressor that inhibits the PC transcriptional program and thus must be downregulated for PC differentiation to occur. Bcl6 expression was increased in PCs isolated from ADAM10(Δ/ΔIgG1-cre(+/- mice at both the mRNA and protein level. These results demonstrate that ADAM10 is required for proper transcription factor expression in PCs and thus, for normal PC function.

  9. 75 FR 20825 - Information Collection Requirement; Defense Federal Acquisition Regulation Supplement; DoD Pilot...

    Science.gov (United States)

    2010-04-21

    ... Regulation Supplement; DoD Pilot Mentor-Protege Program AGENCY: Defense Acquisition Regulations System... Acquisition Regulation Supplement (DFARS) Appendix I, DoD Pilot Mentor-Protege Program; OMB Control Number... Mentor-Protege Program have been met. The purposes of the Program are to (1) provide incentives to...

  10. The European Union's REACH regulation: a review of its history and requirements.

    Science.gov (United States)

    Williams, E Spencer; Panko, Julie; Paustenbach, Dennis J

    2009-01-01

    In 2006, the European Union (EU) promulgated a monumental regulatory initiative for the Registration, Evaluation, Authorization, and Restriction of Chemicals (REACH). To date, several thousand pages of text have been needed to describe the expectations of this regulation. There were numerous reasons for the promulgation of REACH, but, by and large, it is an extension of the global desire to produce fewer industrial chemicals, to understand the possible human and ecological hazards of those that are produced, and to insure that any major threat is anticipated, as well as prevented. Most industry-related groups consider it the most wide-ranging and costly regulatory initiatives related to health risk assessment ever to be promulgated. This review presents a description of REACH that should inform scientists, managers, and others about its objectives and the means to satisfy them. Registration is required for all chemicals manufactured or imported into the EU, unless specifically exempted. Registration is expected to be a collaborative process among companies, which will generate a dossier containing data on physicochemical characteristics, as well as toxicological and ecotoxicological properties. Though the magnitude of the gaps in the data required for registration is uncertain at this point, it is clear that basic toxicology testing will have to be conducted for many chemical substances that have not undergone formal review up to this point. For many chemicals, an examination of hazards and risks arising from the use of these substances will also be required in the form of a chemical safety report (CSR). Beginning with the dual processes of dossier and substance evaluation, the European Chemicals Agency (ECHA), the Member States of the EU, and the European Commission will identify chemicals that may pose unacceptable hazards to human health and/or the environment, and will curtail or restrict their usage. The implementation of REACH will expand and deepen the

  11. SNM gamma-ray fingerprint monitor functional requirements and design specifications

    Energy Technology Data Exchange (ETDEWEB)

    Bieber, A.M. Jr.; Kane, W.R.

    1994-07-01

    A number of DOE facilities need to perform confirmatory inventory measurements on items of special nuclear material (SNM). The DOE Office of Safeguards and Security (OSS) has tasked the Safeguards, Safety and Nonproliferation Division (SSN) of the Department of Advanced Technology at Brookhaven National Laboratory (BNL) to develop a high-resolution gamma-ray-spectroscopy-based instrument for performing confirmatory inventory measurements on such materials, a ``gamma-ray fingerprint monitor`` (GRFM). This document is a conceptual design for the SSN GRFM system. This conceptual design is based on previous experience with measurements of plutonium-bearing materials and comparison of gamma-ray spectrum features, not on actual tests of the procedures or hardware described. As a result, modifications may be necessary when actual prototype hardware and software are tested in realistic circumstances on actual materials of interest.

  12. 40 CFR 63.8600 - What are my monitoring installation, operation, and maintenance requirements?

    Science.gov (United States)

    2010-07-01

    ... Clay Ceramics Manufacturing Testing and Initial Compliance Requirements § 63.8600 What are my....5 inch of water or a transducer with a minimum measurement sensitivity of 1 percent of the pressure... Filter Bag Leak Detection Guidance,” (EPA-454/R-98-015, September 1997). This document is available...

  13. 40 CFR 63.7331 - What are the installation, operation, and maintenance requirements for my monitors?

    Science.gov (United States)

    2010-07-01

    ... pushing emissions, you must install, operate, and maintain CPMS to measure and record the pressure drop... the pressure drop across each multicyclone during each push according to the requirements in... a manner consistent with the guidance document, “Fabric Filter Bag Leak Detection Guidance”...

  14. 40 CFR 63.11092 - What testing and monitoring requirements must I meet?

    Science.gov (United States)

    2010-07-01

    .... Should the Administrator deem the prior test data unacceptable, the facility is still required to meet... suction line, with the measurements displayed on a gauge that can be visually observed. Each carbon bed... valve sequencing, cycle time, gasoline flow, purge air flow, and operating temperatures....

  15. 40 CFR 98.34 - Monitoring and QA/QC requirements.

    Science.gov (United States)

    2010-07-01

    ...) The carbon content and, if applicable, molecular weight of the fuels shall be determined according to... the carbon content and molecular weight of the fuel is required if the necessary equipment is in place... a particular type of fuel, sampling and analysis for carbon content and molecular weight...

  16. Monitoring and control requirement definition study for dispersed storage and generation (DSG). Volume II. Final report, Appendix A: selected DSG technologies and their general control requirements

    Energy Technology Data Exchange (ETDEWEB)

    1980-10-01

    A major aim of the US National Energy Policy, as well as that of the New York State Energy Research and Development Authority, is to conserve energy and to shift from oil to more abundant domestic fuels and renewable energy sources. Dispersed Storage and Generation (DSG) is the term that characterizes the present and future dispersed, relatively small (<30 MW) energy systems, such as solar thermal electric, photovoltaic, wind, fuel cell, storage battery, hydro, and cogeneration, which can help achieve these national energy goals and can be dispersed throughout the distribution portion of an electric utility system. The purpose of this survey and identification of DSG technologies is to present an understanding of the special characteristics of each of these technologies in sufficient detail so that the physical principles of their operation and the internal control of each technology are evident. In this way, a better appreciation can be obtained of the monitoring and control requirements for these DSGs from a remote distribution dispatch center. A consistent approach is being sought for both hardware and software which will handle the monitoring and control necessary to integrate a number of different DSG technologies into a common distribution dispatch network. From this study it appears that the control of each of the DSG technologies is compatible with a supervisory control method of operation that lends itself to remote control from a distribution dispatch center.

  17. The effects of divided attention at study and reporting procedure on regulation and monitoring for episodic recall.

    Science.gov (United States)

    Sauer, James; Hope, Lorraine

    2016-09-01

    Eyewitnesses regulate the level of detail (grain size) reported to balance competing demands for informativeness and accuracy. However, research to date has predominantly examined metacognitive monitoring for semantic memory tasks, and used relatively artificial phased reporting procedures. Further, although the established role of confidence in this regulation process may affect the confidence-accuracy relation for volunteered responses in predictable ways, previous investigations of the confidence-accuracy relation for eyewitness recall have largely overlooked the regulation of response granularity. Using a non-phased paradigm, Experiment 1 compared reporting and monitoring following optimal and sub-optimal (divided attention) encoding conditions. Participants showed evidence of sacrificing accuracy for informativeness, even when memory quality was relatively weak. Participants in the divided (cf. full) attention condition showed reduced accuracy for fine- but not coarse-grained responses. However, indices of discrimination and confidence diagnosticity showed no effect of divided attention. Experiment 2 compared the effects of divided attention at encoding on reporting and monitoring using both non-phased and 2-phase procedures. Divided attention effects were consistent with Experiment 1. However, compared to those in the non-phased condition, participants in the 2-phase condition displayed a more conservative control strategy, and confidence ratings were less diagnostic of accuracy. When memory quality was reduced, although attempts to balance informativeness and accuracy increased the chance of fine-grained response errors, confidence provided an index of the likely accuracy of volunteered fine-grained responses for both condition.

  18. Requirement for safety monitoring for approved multiple sclerosis therapies: an overview.

    Science.gov (United States)

    Rommer, P S; Zettl, U K; Kieseier, B; Hartung, H-P; Menge, T; Frohman, E; Greenberg, B M; Hemmer, B; Stüve, O

    2014-03-01

    During the last two decades, treatment options for patients with multiple sclerosis (MS) have broadened tremendously. All agents that are currently approved for clinical use have potential side effects, and a careful risk-benefit evaluation is part of a decision algorithm to identify the optimal treatment choice for an individual patient. Whereas glatiramer acetate and interferon beta preparations have been used in MS for decades and have a proven safety record, more recently approved drugs appear to be more effective, but potential risks might be more severe. The potential complications of some novel therapies might not even have been identified to their full extent. This review is aimed at the clinical neurologist in that it offers insights into potential adverse events of each of the approved MS therapeutics: interferon beta, glatiramer acetate, mitoxantrone, natalizumab, fingolimod and teriflunomide, as well as recently approved therapeutics such as dimethyl fumarate and alemtuzumab. It also provides recommendations for monitoring the different drugs during therapy in order to avoid common side effects.

  19. Novel insights into iron regulation and requirement in marine medaka Oryzias melastigma

    Science.gov (United States)

    Wang, Jian; Wang, Wen-Xiong

    2016-05-01

    Iron (Fe) is an essential trace element for marine fish. However, our knowledge of Fe requirements at different development stages of marine fish is still limited. Here, we reported the efficient Fe absorption strategies adopted by larval fish under different dietary Fe supplementary levels (i.e., 0-640 mg/kg). Biokinetically, the larval fish controlled their dietary Fe assimilation efficiency (AE, 1.6-18.5%), and enhanced their waterborne Fe uptake (ca. 2.5 fold change of uptake rate constant) once the dietary Fe was deficient (i.e., 27.4 mg Fe/kg feed). Transcriptionally, the expression of hepcidin1 (hep1; Fe regulator; i.e., 2.3-15.7 fold change) in larval fish was positively correlated with the Fe supplementary levels. Comparatively, the female adult fish were poor in assimilating the added Fe source (i.e., ferric form) with similar life-sustainable levels of Fe (i.e., 0.046-0.12 μg/g/d assimilated for Fe supplementary levels of 27.4, 162 and 657 mg Fe/kg feed). The overall feeding experiments suggested that dietary net Fe flux sufficient for the normal growth of larval medaka was 0.71-1.75 μg/g/d (i.e., 83.9 mg Fe/kg feed), consistent with the modeled value (i.e., 1.09-2.16 μg/g/d). In female adults, the estimated essential net Fe flux was 0.88-0.90 μg/g/d.

  20. 25 CFR 518.7 - If a tribe holds a certificate of self-regulation, is it required to report information to the...

    Science.gov (United States)

    2010-04-01

    ... 25 Indians 2 2010-04-01 2010-04-01 false If a tribe holds a certificate of self-regulation, is it... REGULATION OF CLASS II GAMING § 518.7 If a tribe holds a certificate of self-regulation, is it required to... certificate of self-regulation shall be required to submit a self-regulation report annually to the Commission...

  1. [Justification of regulations for the use of computers with LCD monitor during academic studies].

    Science.gov (United States)

    Stepanova, M I; Sazanyuk, Z I; Laponova, E D; Voronova, B Z; Lashneva, I P

    2014-01-01

    In the article there are reported the results of studies of the effect on the functional state of the body of schoolchildren curriculum with the use of personal computers with LCD monitors. The resistance to the development of visual and general fatigue in schoolchildren -users of personal computers with LCD monitors was found to be higher because they provide a more favorable environment for visual work in comparison with monitors based on cathode-ray tube. The use of LED lighting in classrooms contributes to the creation a more favorable light environment than in fluorescent lighting and reduces fatigable impact (both general and visual) of classes with the use of personal computers.

  2. 78 FR 30898 - Information Collection Requirement; Defense Federal Acquisition Regulation Supplement; Rights in...

    Science.gov (United States)

    2013-05-23

    ... Technical Data and Computer Software--Small Business Innovation Research (SBIR) Program. In accordance with... Regulation Supplement; Rights in Technical Data and Computer Software AGENCY: Defense Acquisition Regulations... in Technical Data, and Subpart 227.72, Rights in Computer Software and Computer Software...

  3. Melanocortin 4 receptor is not required for estrogenic regulations on energy homeostasis and reproduction

    Science.gov (United States)

    Brain estrogen receptor-a (ERa) is essential for estrogenic regulation of energy homeostasis and reproduction. We previously showed that ERa expressed by pro-opiomelanocortin (POMC) neurons mediates estrogen's effects on food intake, body weight, negative regulation of hypothalamic–pituitary–gonadal...

  4. 75 FR 8063 - Stakeholder Meeting Regarding Revisions to the Unregulated Contaminant Monitoring Regulation

    Science.gov (United States)

    2010-02-23

    ... From the Federal Register Online via the Government Publishing Office ENVIRONMENTAL PROTECTION AGENCY RIN 2040-AF10 Stakeholder Meeting Regarding Revisions to the Unregulated Contaminant Monitoring... this notice is to announce a public stakeholder meeting to present information to...

  5. Aquatic monitoring programs conducted during environmental impact assessments in Canada: preliminary assessment before and after weakened environmental regulation.

    Science.gov (United States)

    Roach, Brynn; Walker, Tony R

    2017-03-01

    Aquatic monitoring programs are imperative for the functioning of the environmental impact assessment (EIA) process and a cornerstone for industrial compliance in Canada. However, in 2012, several leading pieces of federal environmental legislation (e.g., Canadian Environmental Assessment Act c.19, s. 52, 2012) were drastically altered, effectively weakening levels of environmental protection for aquatic ecosystems during project developments. This paper assesses the impact of CEAA 2012 on aquatic monitoring programs (and subsequent monitoring data reporting) across Canada for ten projects (five completed pre-CEAA 2012 and five completed post-CEAA 2012). Projects included four energy and six mining projects and were selected based on the following criteria: (i) representative of Canada's resource economy; (ii) project information was publicly available; and (iii) strong public interest. Projects pre- and post-CEAA 2012 exhibited few apparent differences before and after environmental regulatory changes. However, wide discrepancies exist in numbers and types of parameters reported, along with a lack of consistency in reporting. Projects pre-CEAA 2012 provided more follow-up monitoring commitments. Although qualitative differences remain inconclusive, this paper highlights requirements for further assessment of aquatic monitoring and follow-up programs in Canada. Recommendations for the government to consider during reviews of the federal environmental assessment processes include (i) improved transparency on the Canadian Environmental Assessment Agency website ( https://www.ceaa-acee.gc.ca/ ); (ii) creation of a legally binding standardized aquatic monitoring program framework to ensure that all Canadian aquatic ecosystems are monitored with equal rigour; and (iii) commitments and justification related to frequency of aquatic monitoring of water quality.

  6. 77 FR 53870 - Availability of a Legal Entity Identifier Meeting the Requirements of the Regulations of the...

    Science.gov (United States)

    2012-09-04

    ... in recordkeeping and swap data reporting. Section 45.6(f)(1) provides that: When a legal entity... COMMISSION Availability of a Legal Entity Identifier Meeting the Requirements of the Regulations of the Commodity Futures Trading Commission and Designation of Provider of Legal Entity Identifiers To Be Used...

  7. 40 CFR Table A-1 to Subpart A of... - Summary of Applicable Requirements for Reference and Equivalent Methods for Air Monitoring of...

    Science.gov (United States)

    2010-07-01

    ... Reference and Equivalent Methods for Air Monitoring of Criteria Pollutants A Table A-1 to Subpart A of Part...) AMBIENT AIR MONITORING REFERENCE AND EQUIVALENT METHODS General Provisions Pt. 53, Subpt. A, Table A-1 Table A-1 to Subpart A of Part 53—Summary of Applicable Requirements for Reference and...

  8. 76 FR 4555 - Authority To Require Supervision and Regulation of Certain Nonbank Financial Companies

    Science.gov (United States)

    2011-01-26

    ... those activities most prone to systemic risk through contagion. To measure interconnectedness, commenters suggested evaluating, among other things, liquidity profile, contagion risk, counterparty credit... prudential regulators or the Office of Financial Research to make its determinations. 11. Should the degree...

  9. 76 FR 78126 - Disclosure Requirements for Depository Institutions Lacking Federal Deposit Insurance (Regulation I)

    Science.gov (United States)

    2011-12-16

    ... INFORMATION CONTACT: Krista Ayoub or Jane Gao, Office of Regulations, at (202) 435-7700. SUPPLEMENTARY... accepts deposits or opens accounts (excluding automated teller machines or point of sale terminals), and...

  10. 78 FR 4788 - Regulated Navigation Area; Reporting Requirements for Barges Loaded With Certain Dangerous...

    Science.gov (United States)

    2013-01-23

    ... under section 4(a) of the Administrative Procedure Act (APA) (5 U.S.C. 553(b)). This provision... follows: PART 165--REGULATED NAVIGATION AREAS AND LIMITED ACCESS AREAS 0 1. The authority citation...

  11. 76 FR 51040 - Food and Drug Administration Clinical Trial Requirements, Regulations, Compliance, and Good...

    Science.gov (United States)

    2011-08-17

    ..., in co-sponsorship with the Society of Clinical Research Associates (SoCRA) is announcing a public... Clinical Trial; (2) Adverse Event Reporting--Science, Regulation, Error, and Safety; (3) Part 11 Compliance...

  12. 77 FR 8886 - Food and Drug Administration Clinical Trial Requirements, Regulations, Compliance, and Good...

    Science.gov (United States)

    2012-02-15

    ...-sponsorship with the Society of Clinical Research Associates (SoCRA) is announcing a public workshop. The... Pharmaceutical Clinical Trial; (2) Adverse Event Reporting--Science, Regulation, Error, and Safety; (3) Part 11...

  13. Histone Methylation Regulator PTIP Is Required for PPARγ and C/EBPα Expression and Adipogenesis

    OpenAIRE

    Cho, Young-Wook; Hong, SunHwa; Jin, Qihuang; Wang, Lifeng; Lee, Ji-Eun; Gavrilova, Oksana; Ge, Kai

    2009-01-01

    PPARγ and C/EBPα cooperate to control preadipocyte differentiation (adipogenesis). However, the factors that regulate PPARγ and C/EBPα expression during adipogenesis remain largely unclear. Here we show PTIP, a protein that associates with histone H3K4 methyltransferases, regulates PPARγ and C/EBPα expression in mouse embryonic fibroblasts (MEFs) and during preadipocyte differentiation. PTIP deletion in MEFs leads to marked decreases of PPARγ expression and PPARγ-stimulated C/EBPα expression....

  14. Continuous Glucose Monitoring Analysis as Predictor of Islet Yield and Insulin Requirements in Autologous Islet Transplantation After Complete Pancreatectomy

    Science.gov (United States)

    Georgiev, George Ivanov; Cercone, Renee; Tiwari, Mukesh; Rilo, Horacio L. R.

    2014-01-01

    We analyzed the pretransplant continuous glucose monitoring (CGM) data of 45 patients that underwent total pancreatectomy followed by autologous islet transplantation (AIT) at the University of Arizona Medical Center. Traditional and novel metrics of CGM time series were correlated to the total islet count (TIC), islet equivalents (IEQs), and weight-normalized IEQs (IEQ/kg). In a subset cohort (n = 26) we analyzed the relationship among the infused number of islets, the CGM indicators, and the first recorded insulin requirement after the procedure. We conclude that receiving a high islet yield is sufficient yet not necessary to achieve low or null insulin requirements within the first 50 days after surgery. Furthermore, CGM inertia and CGM length of curve (2 novel CGM indicators) are shown to be correlated to islet yield, and the CGMs normalized area (Ao) and time ratio above hyperglycemic level (To) are strongly correlated to insulin requirement. A screening test based on To is shown to have 100% sensitivity and 88% specificity discriminating insulin independence upon discharge. PMID:25190081

  15. A mechanism regulating G protein-coupled receptor signaling that requires cycles of protein palmitoylation and depalmitoylation.

    Science.gov (United States)

    Jia, Lixia; Chisari, Mariangela; Maktabi, Mohammad H; Sobieski, Courtney; Zhou, Hao; Konopko, Aaron M; Martin, Brent R; Mennerick, Steven J; Blumer, Kendall J

    2014-02-28

    Reversible attachment and removal of palmitate or other long-chain fatty acids on proteins has been hypothesized, like phosphorylation, to control diverse biological processes. Indeed, palmitate turnover regulates Ras trafficking and signaling. Beyond this example, however, the functions of palmitate turnover on specific proteins remain poorly understood. Here, we show that a mechanism regulating G protein-coupled receptor signaling in neuronal cells requires palmitate turnover. We used hexadecyl fluorophosphonate or palmostatin B to inhibit enzymes in the serine hydrolase family that depalmitoylate proteins, and we studied R7 regulator of G protein signaling (RGS)-binding protein (R7BP), a palmitoylated allosteric modulator of R7 RGS proteins that accelerate deactivation of Gi/o class G proteins. Depalmitoylation inhibition caused R7BP to redistribute from the plasma membrane to endomembrane compartments, dissociated R7BP-bound R7 RGS complexes from Gi/o-gated G protein-regulated inwardly rectifying K(+) (GIRK) channels and delayed GIRK channel closure. In contrast, targeting R7BP to the plasma membrane with a polybasic domain and an irreversibly attached lipid instead of palmitate rendered GIRK channel closure insensitive to depalmitoylation inhibitors. Palmitate turnover therefore is required for localizing R7BP to the plasma membrane and facilitating Gi/o deactivation by R7 RGS proteins on GIRK channels. Our findings broaden the scope of biological processes regulated by palmitate turnover on specific target proteins. Inhibiting R7BP depalmitoylation may provide a means of enhancing GIRK activity in neurological disorders.

  16. Down-regulation of stathmin expression is required for megakaryocyte maturation and platelet production.

    Science.gov (United States)

    Iancu-Rubin, Camelia; Gajzer, David; Tripodi, Joseph; Najfeld, Vesna; Gordon, Ronald E; Hoffman, Ronald; Atweh, George F

    2011-04-28

    The final stages of of megakaryocyte (MK) maturation involve a series of steps, including polyploidization and proplatelet formation. Although these processes are highly dependent on dynamic changes in the microtubule (MT) cytoskeleton, the mechanisms responsible for regulation of MTs in MKs remain poorly defined. Stathmin is a highly conserved MT-regulatory protein that has been suggested to play a role in MK differentiation of human leukemic cell lines. However, previous studies defining this relationship have reached contradictory conclusions. In this study, we addressed this controversy and investigated the role of stathmin in primary human MKs. To explore the importance of stathmin down-regulation during megakaryocytopoiesis, we used a lentiviral-mediated gene delivery system to prevent physiologic down-regulation of stathmin in primary MKs. We demonstrated that sustained expression of constitutively active stathmin delayed cytoplasmic maturation (ie, glycoprotein GPIb and platelet factor 4 expression) and reduced the ability of MKs to achieve high levels of ploidy. Moreover, platelet production was impaired in MKs in which down-regulation of stathmin expression was prevented. These studies indicate that suppression of stathmin is biologically important for MK maturation and platelet production and support the importance of MT regulation during the final stages of thrombopoiesis.

  17. 77 FR 26071 - Revisions to the Unregulated Contaminant Monitoring Regulation (UCMR 3) for Public Water Systems

    Science.gov (United States)

    2012-05-02

    ... fulfills EPA's obligation under SDWA by identifying 29 unregulated contaminants for monitoring during the... or the State. Changes to the rule language clarify that when identifying a representative well, the... Anion using EPA Method 300.1 (IC/Conductivity) \\6\\ or alternate SM \\7\\ or ASTM Methods: \\8\\ chlorate....

  18. Profiles of Motivated Self-Regulation in College Computer Science Courses: Differences in Major versus Required Non-Major Courses

    Science.gov (United States)

    Shell, Duane F.; Soh, Leen-Kiat

    2013-12-01

    The goal of the present study was to utilize a profiling approach to understand differences in motivation and strategic self-regulation among post-secondary STEM students in major versus required non-major computer science courses. Participants were 233 students from required introductory computer science courses (194 men; 35 women; 4 unknown) at a large Midwestern state university. Cluster analysis identified five profiles: (1) a strategic profile of a highly motivated by-any-means good strategy user; (2) a knowledge-building profile of an intrinsically motivated autonomous, mastery-oriented student; (3) a surface learning profile of a utility motivated minimally engaged student; (4) an apathetic profile of an amotivational disengaged student; and (5) a learned helpless profile of a motivated but unable to effectively self-regulate student. Among CS majors and students in courses in their major field, the strategic and knowledge-building profiles were the most prevalent. Among non-CS majors and students in required non-major courses, the learned helpless, surface learning, and apathetic profiles were the most prevalent. Students in the strategic and knowledge-building profiles had significantly higher retention of computational thinking knowledge than students in other profiles. Students in the apathetic and surface learning profiles saw little instrumentality of the course for their future academic and career objectives. Findings show that students in STEM fields taking required computer science courses exhibit the same constellation of motivated strategic self-regulation profiles found in other post-secondary and K-12 settings.

  19. 76 FR 71707 - Revising Underground Storage Tank Regulations-Revisions to Existing Requirements and New...

    Science.gov (United States)

    2011-11-18

    ..., classroom training must be signed by the trainer; computer based training does not require a signature but... Containment and Operator Training; Proposed Rule #0;#0;Federal Register / Vol. 76 , No. 223 / Friday, November... Requirements for Secondary Containment and Operator Training AGENCY: Environmental Protection Agency...

  20. 75 FR 34277 - Federal Acquisition Regulation; FAR Case 2008-007, Additional Requirements for Market Research

    Science.gov (United States)

    2010-06-16

    ... executive agencies. Specifically, the head of the agency must conduct market research before issuing an... million for the procurement of items other than commercial items is required to conduct market research... 10 U.S.C. 2377(c), ``Preliminary Market Research'', to require the head of an agency to...

  1. 76 FR 14548 - Federal Acquisition Regulation; Requirements for Acquisitions Pursuant to Multiple-Award Contracts

    Science.gov (United States)

    2011-03-16

    ...- going competition, when deciding how many BPAs are appropriate and document the decision in the... GSA Advantage! on-line shopping service, by reviewing the catalogs or pricelists of at least three... requirement(s); (B) The benefits of on-going competition and the need to periodically compare multiple...

  2. 76 FR 68027 - Federal Acquisition Regulation; Certification Requirement and Procurement Prohibition Relating to...

    Science.gov (United States)

    2011-11-02

    .... Applicability to Construction Comment: One respondent was concerned that the prescription at FAR 25.1103, which... be interpreted to exclude construction. The respondent suggested changing the prescription to require... require use of the provision in ``all solicitations.'' B. Commercial Database of Persons Doing...

  3. Nonlinear software sensor for monitoring genetic regulation processes with noise and modeling errors

    CERN Document Server

    Ibarra-Junquera, V; Rosu, H C; Arguello, G; Collado-Vides, J

    2004-01-01

    Nonlinear control techniques by means of a software sensor that are commonly used in chemical engineering could be also applied to genetic regulation processes. We provide here a realistic formulation of this procedure by introducing an additive white Gaussian noise, which is usually found in experimental data. Besides, we include model errors, meaning that we assume we do not know the nonlinear regulation function of the process. In order to illustrate this procedure, we employ the Goodwin dynamics of the concentrations (1963) in the simple form recently discussed by De Jong (2002), which involves the dynamics of the mRNA a, given protein A, and metabolite K concentrations. However instead of considering their full dynamics, we use only the data of metabolite K and a designed software sensor. We also show, more generally, that it is possible to rebuild the complete set of n concentrations despite the uncertainties in the regulation function and the perturbation due to the additive white Gaussian noise

  4. 78 FR 72620 - Federal Acquisition Regulation; Higher-Level Contract Quality Requirements

    Science.gov (United States)

    2013-12-03

    ... posted without change to http://www.regulations.gov , including any personal and/or business confidential... counterfeit electronic parts. However, because of the globalization of the marketplace, the problem of... and industry. Globalization in the marketplace increased the risk of counterfeit items in...

  5. Regulation of Hepatic Triacylglycerol Metabolism by CGI-58 Does Not Require ATGL Co-activation

    Directory of Open Access Journals (Sweden)

    Caleb C. Lord

    2016-07-01

    Full Text Available Adipose triglyceride lipase (ATGL and comparative gene identification 58 (CGI-58 are critical regulators of triacylglycerol (TAG turnover. CGI-58 is thought to regulate TAG mobilization by stimulating the enzymatic activity of ATGL. However, it is not known whether this coactivation function of CGI-58 occurs in vivo. Moreover, the phenotype of human CGI-58 mutations suggests ATGL-independent functions. Through direct comparison of mice with single or double deficiency of CGI-58 and ATGL, we show here that CGI-58 knockdown causes hepatic steatosis in both the presence and absence of ATGL. CGI-58 also regulates hepatic diacylglycerol (DAG and inflammation in an ATGL-independent manner. Interestingly, ATGL deficiency, but not CGI-58 deficiency, results in suppression of the hepatic and adipose de novo lipogenic program. Collectively, these findings show that CGI-58 regulates hepatic neutral lipid storage and inflammation in the genetic absence of ATGL, demonstrating that mechanisms driving TAG lipolysis in hepatocytes differ significantly from those in adipocytes.

  6. 78 FR 19467 - Information Collection Requirement; Defense Federal Acquisition Regulation Supplement (DFARS...

    Science.gov (United States)

    2013-04-01

    ... Number: Defense Federal Acquisition Regulation Supplement (DFARS), Appendix I, DoD Pilot Mentor-Protege... purposes of the DoD Pilot Mentor-Protege Program have been met. The purposes of the Program are to (1) provide incentives to major DoD contractors to assist protege firms in enhancing their capabilities...

  7. RovA, a global regulator of Yersinia pestis, specifically required for bubonic plague.

    Science.gov (United States)

    Cathelyn, Jason S; Crosby, Seth D; Lathem, Wyndham W; Goldman, William E; Miller, Virginia L

    2006-09-05

    The pathogenic species of Yersinia contain the transcriptional regulator RovA. In Yersinia pseudotuberculosis and Yersinia enterocolitica, RovA regulates expression of the invasion factor invasin (inv), which mediates translocation across the intestinal epithelium. A Y. enterocolitica rovA mutant has a significant decrease in virulence by LD(50) analysis and an altered rate of dissemination compared with either wild type or an inv mutant, suggesting that RovA regulates multiple virulence factors. Here, we show the involvement of RovA in the virulence of Yersinia pestis, which naturally lacks a functional inv gene. A Y. pestis DeltarovA mutant is attenuated approximately 80-fold by LD(50) and is defective in dissemination/colonization of spleens and lungs after s.c. inoculation. However, the DeltarovA mutant is only slightly attenuated when given via an intranasal or i.p. route, indicating a more important role for RovA in bubonic plague than pneumonic plague or systemic infection. Microarray analysis was used to define the RovA regulon. The psa locus was among the most highly down-regulated loci in the DeltarovA mutant. A DeltapsaA mutant had a significant dissemination defect after s.c. infection but only slight attenuation by the pneumonic-disease model, closely mimicking the virulence defect seen with the DeltarovA mutant. DNA-binding studies revealed that RovA specifically interacts with the psaE and psaA promoter regions, indicating a direct role for RovA in regulating this locus. Thus, RovA appears to be a global transcription factor in Y. pestis and, through its regulatory influence on genes such as psaEFABC, contributes to the virulence of Y. pestis.

  8. Regulated bioluminescence as a tool for bioremediation process monitoring and control of bacterial cultures

    Science.gov (United States)

    Burlage, Robert S.; Heitzer, Armin; Digrazia, Philip M.

    1991-01-01

    An effective on-line monitoring technique for toxic waste bioremediation using bioluminescent microorganisms has shown great potential for the description and optimization of biological processes. The lux genes of the bacterium Vibrio fischeri are used by this species to produce visible light. The lux genes can be genetically fused to the control region of a catabolic gene, with the result that bioluminescence is produced whenever the catabolic gene is induced. Thus the detection of light from a sample indicates that genetic expression from a specific gene is occurring. This technique was used to monitor biodegradation of specific contaminants from waste sites. For these studies, fusions between the lux genes and the operons for naphthalene and toluene/xylene degradation were constructed. Strains carrying one of these fusions respond sensitively and specifically to target substrates. Bioluminescence from these cultures can be rapidly measured in a nondestructive and noninvasive manner. The potential for this technique in this and other biological systems is discussed.

  9. Good for your health? An analysis of the requirements for scientific substantiation in European health claims regulation

    Directory of Open Access Journals (Sweden)

    Oliver Todt

    2016-05-01

    Full Text Available Objective. To identify the various types of evidence, as well as their relative importance in European health claims regulation, in order to analyze the consequences for consumer protection of the requirements for scientific substantiation in this regulation. Materials and methods. Qualitative analysis of various documents relevant to the regulatory process, particularly as to the implications of the standards of proof for the functional food market, as well as consumer behavior. Results. European regulation defines a hierarchy of evidence that turns randomized controlled trials into a necessary and sufficient condition for health claim autho- rizations. Conclusions. Consumer protection can be interpreted in different manners. High standards of proof protect consumers from false information about the health outcomes of functional foods, while lower standards lead to more, albeit less accurate information about such outcomes being available to consumers.

  10. glp-1 is required in the germ line for regulation of the decision between mitosis and meiosis in C. elegans.

    Science.gov (United States)

    Austin, J; Kimble, J

    1987-11-20

    In the wild-type C. elegans germ line there are both mitotic and meiotic germ cells. Mutations in glp-1 cause germ cells that would normally divide mitotically to enter meiosis. This mutant phenotype mimics the effect of killing the distal tip cell, a somatic cell that interacts with the germ line to regulate the mitotic/meiotic decision. In addition, wild-type glp-1 product is required maternally for embryogenesis. Temperature-shift experiments indicate that the temporal requirement for glp-1 activity in the germ line is the same as that for distal tip cell regulation. Mosaic analyses suggest that glp-1 is produced in the germ line. We propose that glp-1 acts as part of the receiving mechanism in the interaction between the distal tip cell and germ line.

  11. Development of technology-neutral safety requirements for the regulation of future nuclear power reactors: Back to basics

    Energy Technology Data Exchange (ETDEWEB)

    Tronea, Madalina, E-mail: madalina.tronea@gmail.co [Faculty of Physics, University of Bucharest (Romania)

    2011-03-15

    This paper explores the current trends as regards the development of technology-neutral safety requirements to be used in the regulation of future nuclear power reactors and the role of the quantitative safety goals in the design of reactor safety systems. The use of the recommendations of the International Commission on Radiological Protection (ICRP) on protection against potential exposure could form the basis of a technology-neutral framework for safety requirements on new reactor designs and could contribute to international harmonisation of nuclear safety assessment practices as part of the licensing processes for future nuclear power plants.

  12. Crop Monitoring Based on SPOT-5 Take-5 and Sentinel-1A Data for the Estimation of Crop Water Requirements

    Directory of Open Access Journals (Sweden)

    Ana Navarro

    2016-06-01

    Full Text Available Optical and microwave images have been combined for land cover monitoring in different agriculture scenarios, providing useful information on qualitative and quantitative land cover changes. This study aims to assess the complementarity and interoperability of optical (SPOT-5 Take-5 and synthetic aperture radar (SAR (Sentinel-1A data for crop parameter (basal crop coefficient (Kcb values and the length of the crop’s development stages retrieval and crop type classification, with a focus on crop water requirements, for an irrigation perimeter in Angola. SPOT-5 Take-5 images are used as a proxy of Sentinel-2 data to evaluate the potential of their enhanced temporal resolution for agricultural applications. In situ data are also used to complement the Earth O=observation (EO data. The Normalized Difference Vegetation Index (NDVI and dual (VV + VH polarization backscattering time series are used to compute the Kcb curve for four crop types (maize, soybean, bean and pasture and to estimate the length of each phenological growth stage. The Kcb values are then used to compute the crop’s evapotranspiration and to subsequently estimate the crop irrigation requirements based on a soil water balance model. A significant R2 correlation between NDVI and backscatter time series was observed for all crops, demonstrating that optical data can be replaced by microwave data in the presence of cloud cover. However, it was not possible to properly identify each stage of the crop cycle due to the lack of EO data for the complete growing season.

  13. Feedback regulation of NEUROG2 activity by MTGR1 is required for progression of neurogenesis.

    Science.gov (United States)

    Aaker, Joshua D; Patineau, Andrea L; Yang, Hyun-Jin; Ewart, David T; Gong, Wuming; Li, Tongbin; Nakagawa, Yasushi; McLoon, Steven C; Koyano-Nakagawa, Naoko

    2009-12-01

    The sequential steps of neurogenesis are characterized by highly choreographed changes in transcription factor activity. In contrast to the well-studied mechanisms of transcription factor activation during neurogenesis, much less is understood regarding how such activity is terminated. We previously showed that MTGR1, a member of the MTG family of transcriptional repressors, is strongly induced by a proneural basic helix-loop-helix transcription factor, NEUROG2 in developing nervous system. In this study, we describe a novel feedback regulation of NEUROG2 activity by MTGR1. We show that MTGR1 physically interacts with NEUROG2 and represses transcriptional activity of NEUROG2. MTGR1 also prevents DNA binding of the NEUROG2/E47 complex. In addition, we provide evidence that proper termination of NEUROG2 activity by MTGR1 is necessary for normal progression of neurogenesis in the developing spinal cord. These results highlight the importance of feedback regulation of proneural gene activity in neurodevelopment.

  14. Tyrosine phosphorylation of Eps15 is required for ligand-regulated, but not constitutive, endocytosis

    DEFF Research Database (Denmark)

    Confalonieri, S; Salcini, A E; Puri, C;

    2000-01-01

    for endocytosis of the epidermal growth factor receptor (EGFR), the prototypical ligand-inducible receptor, but not of the transferrin receptor (TfR), the prototypical constitutively internalized receptor. Eps15, an endocytic protein that is tyrosine phosphorylated by EGFR, is a candidate for such a function...... of the EGFR, but not of the TfR. A phosphopeptide, corresponding to the phosphorylated sequence of Eps15, inhibited EGFR endocytosis, suggesting that phosphotyrosine in Eps15 serves as a docking site for a phosphotyrosine binding protein. Thus, tyrosine phosphorylation of Eps15 represents the first molecular...... determinant, other than those contained in the receptors themselves, which is involved in the differential regulation of constitutive vs. regulated endocytosis....

  15. Substantiating Recommendations on the Choice of an Efficient Strategy of the State Regulation for System of Monitoring Economy Branches

    Directory of Open Access Journals (Sweden)

    Ashchaulov Vitalii V.

    2016-01-01

    Full Text Available The article is concerned with substantiating recommendations on the choice of an effective strategy of the State regulation for system of monitoring economy branches. To display the financial aspect of the enterprise's strategic positioning in the sectoral environment, a modified ADL matrix was used. A matrix of choosing an optimum variant (scenario of the enterprise's financial strategy has been considered taking into account possibilities of its use in the system of monitoring the development of economic sectors. Disadvantages in the use of modified ADL matrices and in choosing an optimum variant (scenario financial strategy of the enterprise's financial strategy have been identified. Factors of internal and external environment, which affect the strategic choice of enterprises, have been determined and analyzed. The SPACE matrix for building the model of choosing a strategy of financing the development of enterprises in the economy branches has been modified in the context of the system of monitoring the development of economy branches. A description of the strategies proposed by author for financing the development of enterprise has been provided, their advantages and disadvantages have been determined. Individual tools of the proposed models of actions can be accepted as a way of improving the strategic positions with a view to achieving a higher quantitative-qualitative situation as well as adjustment of the existing model of behavior.

  16. Netrin-1 up-regulation in inflammatory bowel diseases is required for colorectal cancer progression.

    Science.gov (United States)

    Paradisi, Andrea; Maisse, Carine; Coissieux, Marie-May; Gadot, Nicolas; Lépinasse, Florian; Delloye-Bourgeois, Céline; Delcros, Jean-Guy; Svrcek, Magali; Neufert, Clemens; Fléjou, Jean-François; Scoazec, Jean-Yves; Mehlen, Patrick

    2009-10-06

    Chronic inflammation and cancer are intimately associated. This is particularly true for inflammatory bowel diseases (IBD), such as ulcerative colitis and Crohn's disease, which show a major increased risk for colorectal cancer. While the understanding of the molecular pathogenesis of IBD has recently improved, the mechanisms that link these chronic inflammatory states to colorectal cancer development are in large part unknown. One of these mechanisms is NF-kappaB pathway activation which in turn may contribute to tumor formation by providing anti-apoptotic survival signals to the epithelial cells. Based on the observation that netrin-1, the anti-apoptotic ligand for the dependence receptors DCC and UNC5H is up-regulated in colonic crypts in response to NF-kappaB, we show here that colorectal cancers from inflammatory bowel diseases patients have selected up-regulation of netrin-1. Moreover, we demonstrate that this inflammation-driven netrin-1 up-regulation is causal for colorectal cancer development as interference with netrin-1 autocrine loop in a mouse model for ulcerative colitis-associated colorectal cancer, while showing no effect on inflammation, inhibits colorectal cancer progression.

  17. Multinational Firm’s Production Decisions under Overlapping Free Trade Agreements: Rule of Origin Requirements and Environmental Regulation

    Directory of Open Access Journals (Sweden)

    Sung Hee Lee

    2016-12-01

    Full Text Available In this paper, we study the impact of the rule of origin (ROO requirements accompanied by free trade agreements (FTAs, which specify the minimum portion of supplies that should satisfy the origin requirement, on a multinational firm’s production decisions. We consider a multinational firm who exports its product to multiple countries and analyze its production decision in the presence of multiple free trading agreements (FTAs. The ROO requirements in FTAs not only refer to the origin of supplies but are also involved with an environmental regulation of a country of the supplies. As such, meeting multiple ROO requirements can be costly since it may be involved with an adjustment of production facility and suppliers according to different environmental standards. We investigate a multinational firm’s choice of the ROO level in its production decision under multiple FTAs. It is well known that, in the presence of overlapping FTAs, the firm may strategically choose not to comply with the minimum ROO requirements in the FTAs. Instead, the firm may choose to comply with an ROO level that is higher than required, or pay the tariff instead without enjoying tariff exemption by the FTA in the new country. Such unintended outcomes in the FTAs are called the Spaghetti Bowl Effect. We characterize and quantify two types of such Spaghetti Bowl Effects with the optimal production decisions of a multinational firm under multiple ROO requirements and derive policy implications.

  18. Monitoring and enforcement of environmental regulations. Lessons from a natural field experiment in Norway

    Energy Technology Data Exchange (ETDEWEB)

    Telle, Kjetil

    2012-07-01

    Relying on a small natural field experiment with random assignment of treatments, I estimate effects of three core elements of most monitoring and enforcement practices: self-reporting, audit frequency and specific deterrence. I find evidence of evasive reporting of violations in self-audits, as more violations are detected in on-site audits than in self-audits. Announcing the increased audit frequency has no effect on compliance, but an audit raises the firm's subsequent compliance substantially.(Author)

  19. Nonlinear software sensor for monitoring genetic regulation processes with noise and modeling errors

    Science.gov (United States)

    Ibarra-Junquera, V.; Torres, L. A.; Rosu, H. C.; Argüello, G.; Collado-Vides, J.

    2005-07-01

    Nonlinear control techniques by means of a software sensor that are commonly used in chemical engineering could be also applied to genetic regulation processes. We provide here a realistic formulation of this procedure by introducing an additive white Gaussian noise, which is usually found in experimental data. Besides, we include model errors, meaning that we assume we do not know the nonlinear regulation function of the process. In order to illustrate this procedure, we employ the Goodwin dynamics of the concentrations [B. C. Goodwin, Temporal Oscillations in Cells (Academic, New York, 1963)] in the simple form recently applied to single gene systems and some operon cases [H. De Jong, J. Comput. Biol. 9, 67 (2002)], which involves the dynamics of the mRNA, given protein and metabolite concentrations. Further, we present results for a three gene case in coregulated sets of transcription units as they occur in prokaryotes. However, instead of considering their full dynamics, we use only the data of the metabolites and a designed software sensor. We also show, more generally, that it is possible to rebuild the complete set of nonmeasured concentrations despite the uncertainties in the regulation function or, even more, in the case of not knowing the mRNA dynamics. In addition, the rebuilding of concentrations is not affected by the perturbation due to the additive white Gaussian noise and also we managed to filter the noisy output of the biological system.

  20. Emdogain-regulated gene expression in palatal fibroblasts requires TGF-βRI kinase signaling.

    OpenAIRE

    Stähli, Alexandra Beatrice; Bosshardt, Dieter; Sculean, Anton; Gruber, Reinhard

    2014-01-01

    Genome-wide microarrays have suggested that Emdogain regulates TGF-β target genes in gingival and palatal fibroblasts. However, definitive support for this contention and the extent to which TGF-β signaling contributes to the effects of Emdogain has remained elusive. We therefore studied the role of the TGF-β receptor I (TGF-βRI) kinase to mediate the effect of Emdogain on palatal fibroblasts. Palatal fibroblasts were exposed to Emdogain with and without the inhibitor for TGF-βRI kinase, SB43...

  1. Emdogain-Regulated Gene Expression in Palatal Fibroblasts Requires TGF-βRI Kinase Signaling

    OpenAIRE

    Alexandra Stähli; Dieter Bosshardt; Anton Sculean; Reinhard Gruber

    2014-01-01

    Genome-wide microarrays have suggested that Emdogain regulates TGF-β target genes in gingival and palatal fibroblasts. However, definitive support for this contention and the extent to which TGF-β signaling contributes to the effects of Emdogain has remained elusive. We therefore studied the role of the TGF-β receptor I (TGF-βRI) kinase to mediate the effect of Emdogain on palatal fibroblasts. Palatal fibroblasts were exposed to Emdogain with and without the inhibitor for TGF-βRI kinase, SB43...

  2. Determining the requirements for e-selection in a small recruitment company - using the regulative cycle

    NARCIS (Netherlands)

    Bondarouk, T.V.; Ruel, H.J.M.; Timmermans, P.; Buragga, K.A.; Zaman, N.

    2013-01-01

    The requirements for e-selection technology to be of practical use for Company T have been investigated. Company T’s main business is in identifying and seconding personnel, especially in the technical sector, and it had been anticipating a shortage in the supply of candidates. This served as the in

  3. 78 FR 69267 - Defense Federal Acquisition Regulation Supplement: Requirements Relating to Supply Chain Risk...

    Science.gov (United States)

    2013-11-18

    ... supply chain risk. This rule implements section 806's three supply-chain risk-management approaches as... to improve supply chain risk management (SCRM). In doing so, the DoDI requires, among other things... consider the impact of supply chain risk in specified types of procurements related to national...

  4. 78 FR 54403 - Removal of Transferred OTS Regulations Regarding Recordkeeping and Confirmation Requirements for...

    Science.gov (United States)

    2013-09-04

    ... minor technical nuances,\\23\\ the OTS's rule was otherwise substantively similar to the Agencies... descriptive charts. Additionally, the FDIC recognizes that some State savings associations availed themselves.... Regulatory Analysis and Procedure A. The Paperwork Reduction Act In accordance with the requirements of the...

  5. 77 FR 59139 - Prompt Corrective Action, Requirements for Insurance, and Promulgation of NCUA Rules and Regulations

    Science.gov (United States)

    2012-09-26

    ...'' credit union for determining whether risk-based net worth requirements apply, and 12 CFR 741.3(b)(5)(i... risk to the National Credit Union Share Insurance Fund (NCUSIF). The Board believes the $10 million... the NCUSIF based on recent trends. (iii) Credit Union Complexity and NCUSIF Risk The Board...

  6. 78 FR 63461 - Information Collection Requirement; Defense Federal Acquisition Regulation Supplement; Foreign...

    Science.gov (United States)

    2013-10-24

    ... Canada--Submission with Offer, and 252.225-7004, Report of Intended Performance Outside the United States and Canada-- Submission after Award, as prescribed in 225.7204(a) and (b) respectively, require... list any qualifying country, NAFTA country, or other foreign end product that it intends to furnish...

  7. On-line monitoring of base current and forward emitter current gain of the voltage regulator's serial pnp transistor in a radiation environment

    Directory of Open Access Journals (Sweden)

    Vukić Vladimir Đ.

    2012-01-01

    Full Text Available A method of on-line monitoring of the low-dropout voltage regulator's operation in a radiation environment is developed in this paper. The method had to enable detection of the circuit's degradation during exploitation, without terminating its operation in an ionizing radiation field. Moreover, it had to enable automatic measurement and data collection, as well as the detection of any considerable degradation, well before the monitored voltage regulator's malfunction. The principal parameters of the voltage regulator's operation that were monitored were the serial pnp transistor's base current and the forward emitter current gain. These parameters were procured indirectly, from the data on the voltage regulator's load and quiescent currents. Since the internal consumption current in moderately and heavily loaded devices was used, the quiescent current of a negligibly loaded voltage regulator of the same type served as a reference. Results acquired by on-line monitoring demonstrated marked agreement with the results acquired from examinations of the voltage regulator's maximum output current and minimum dropout voltage in a radiation environment. The results were particularly consistent in tests with heavily loaded devices. Results obtained for moderately loaded voltage regulators and the risks accompanying the application of the presented method, were also analyzed.

  8. Arabidopsis MSI1 Is Required for Negative Regulation of the Response to Drought Stress

    Institute of Scientific and Technical Information of China (English)

    Cristina Alexandre; Yvonne M(o)ller-Steinbach; Nicole Sch(o)nrock; Wilhelm Gruissem; Lars Hennig

    2009-01-01

    Arabidopsis MSI1 has fundamental functions in plant development.MSI1 is a subunit of Polycomb group protein complexes and Chromatin assembly factor 1,and it interacts with the Retinoblastoma-related protein 1.Altered levels of MSI1 result in pleiotropic phenotypes,reflecting the complexity of MSI1 protein functions.In order to uncover additional functions of MSI1,we performed transcriptional profiling of wild-type and plants with highly reduced MSI1 levels (msil-cs).Surprisingly,the known functions of MSI1 could only account for a minor part of the transcriptional changes in msi1-cs plants.One of the most striking unexpected observations was the up-regulation of a subset of ABA-responsive genes eliciting the response to drought and salt stress.We report that MSI1 can bind to the chromatin of the drought-inducible downstream target RD20 and suggest a new role for MSI1 in the negative regulation of the Arabidopsis drought-stress response.

  9. The Ustilago maydis Nit2 homolog regulates nitrogen utilization and is required for efficient induction of filamentous growth.

    Science.gov (United States)

    Horst, Robin J; Zeh, Christine; Saur, Alexandra; Sonnewald, Sophia; Sonnewald, Uwe; Voll, Lars M

    2012-03-01

    Nitrogen catabolite repression (NCR) is a regulatory strategy found in microorganisms that restricts the utilization of complex and unfavored nitrogen sources in the presence of favored nitrogen sources. In fungi, this concept has been best studied in yeasts and filamentous ascomycetes, where the GATA transcription factors Gln3p and Gat1p (in yeasts) and Nit2/AreA (in ascomycetes) constitute the main positive regulators of NCR. The reason why functional Nit2 homologs of some phytopathogenic fungi are required for full virulence in their hosts has remained elusive. We have identified the Nit2 homolog in the basidiomycetous phytopathogen Ustilago maydis and show that it is a major, but not the exclusive, positive regulator of nitrogen utilization. By transcriptome analysis of sporidia grown on artificial media devoid of favored nitrogen sources, we show that only a subset of nitrogen-responsive genes are regulated by Nit2, including the Gal4-like transcription factor Ton1 (a target of Nit2). Ustilagic acid biosynthesis is not under the control of Nit2, while nitrogen starvation-induced filamentous growth is largely dependent on functional Nit2. nit2 deletion mutants show the delayed initiation of filamentous growth on maize leaves and exhibit strongly compromised virulence, demonstrating that Nit2 is required to efficiently initiate the pathogenicity program of U. maydis.

  10. Wdr18 is required for Kupffer's vesicle formation and regulation of body asymmetry in zebrafish.

    Directory of Open Access Journals (Sweden)

    Wei Gao

    Full Text Available Correct specification of the left-right (L-R axis is important for organ morphogenesis. Conserved mechanisms involving cilia rotation inside node-like structures and asymmetric Nodal signaling in the lateral plate mesoderm (LPM, which are important symmetry-breaking events, have been intensively studied. In zebrafish, the clustering and migration of dorsal forerunner cells (DFCs is critical for the formation of the Kuppfer's vesicle (KV. However, molecular events underlying DFC clustering and migration are less understood. The WD-repeat proteins function in a variety of biological processes, including cytoskeleton assembly, intracellular trafficking, mRNA splicing, transcriptional regulation and cell migration. However, little is known about the function of WD-repeat proteins in L-R asymmetry determination. Here, we report the identification and functional analyses of zebrafish wdr18, a novel gene that encodes a WD-repeat protein that is highly conserved among vertebrate species. wdr18 was identified from a Tol2 transposon-mediated enhancer trap screen. Follow-up analysis of wdr18 mRNA expression showed that it was detected in DFCs or the KV progenitor cells and later in the KV at early somitogenesis stages. Morpholino knockdown of wdr18 resulted in laterality defects in the visceral organs, which were preceded by the mis-expression of Nodal-related genes, including spaw and pitx2. Examination of morphants at earlier stages revealed that the KV had fewer and shorter cilia which are immotile and a smaller cavity. We further investigated the organization of DFCs in wdr18 morphant embryos using ntl and sox17 as specific markers and found that the clustering and migration of DFC was altered, leading to a disorganized KV. Finally, through a combination of wdr18 and itgb1b morpholino injections, we provided evidence that wdr18 and itgb1b genetically interact in the laterality determination process. Thus, we reveal a new and essential role for WD

  11. DNMT1 is a Required Genomic Regulator for Murine Liver Histogenesis and Regeneration

    DEFF Research Database (Denmark)

    Kaji, Kosuke; Factor, Valentina M; Andersen, Jesper B

    2016-01-01

    conditional knockout mice (Dnmt1(Δalb) ) by crossing Dnmt1(fl/fl) with Albumin-Cre (Alb-Cre) transgenic mice. Serum, liver tissues and primary hepatocytes were collected from 1-20 week old mice. The Dnmt1(Δalb) phenotype was assessed by histology, confocal and electron microscopy, biochemistry as well...... hepatocytes caused global hypomethylation, enhanced DNA damage response and initiated a senescence state causing a progressive inability to maintain tissue homeostasis and proliferate in response to injury. The liver regenerated via activation and repopulation from progenitors due to lineage...... hepatocytes did not affect liver homeostasis. CONCLUSION: These results establish the indispensable role of DNMT1-mediated epigenetic regulation in postnatal liver growth and regeneration. The Dnmt1(Δalb) mice provide a unique experimental model to study the role of senescence and contribution of progenitor...

  12. Regulation of extracellular matrix genes by arecoline in primary gingival fibroblasts requires epithelial factors.

    Science.gov (United States)

    Thangjam, G S; Agarwal, P; Balapure, A K; Rao, S Girish; Kondaiah, P

    2009-12-01

    Oral submucous fibrosis, a disease of collagen disorder, has been attributed to arecoline present in the saliva of betel quid chewers. However, the molecular basis of the action of arecoline in the pathogenesis of oral submucous fibrosis is poorly understood. The basic aim of our study was to elucidate the mechanism underlying the action of arecoline on the expression of genes in oral fibroblasts. Human keratinocytes (HaCaT cells) and primary human gingival fibroblasts were treated with arecoline in combination with various pathway inhibitors, and the expression of transforming growth factor-beta isoform genes and of collagen isoforms was assessed using reverse transcription-polymerase chain reaction analysis. We observed the induction of transforming growth factor-beta2 by arecoline in HaCaT cells and this induction was found to be caused by activation of the M-3 muscarinic acid receptor via the induction of calcium and the protein kinase C pathway. Most importantly, we showed that transforming growth factor-beta2 was significantly overexpressed in oral submucous fibrosis tissues (p = 0.008), with a median of 2.13 (n = 21) compared with 0.75 (n = 18) in normal buccal mucosal tissues. Furthermore, arecoline down-regulated the expression of collagens 1A1 and 3A1 in human primary gingival fibroblasts; however these collagens were induced by arecoline in the presence of spent medium of cultured human keratinocytes. Treatment with a transforming growth factor-beta blocker, transforming growth factor-beta1 latency-associated peptide, reversed this up-regulation of collagen, suggesting a role for profibrotic cytokines, such as transforming growth factor-beta, in the induction of collagens. Taken together, our data highlight the importance of arecolineinduced epithelial changes in the pathogenesis of oral submucous fibrosis.

  13. The rgg0182 gene encodes a transcriptional regulator required for the full Streptococcus thermophilus LMG18311 thermal adaptation

    Directory of Open Access Journals (Sweden)

    Bertin Stéphane

    2011-10-01

    Full Text Available Abstract Background Streptococcus thermophilus is an important starter strain for the production of yogurt and cheeses. The analysis of sequenced genomes of four strains of S. thermophilus indicates that they contain several genes of the rgg familly potentially encoding transcriptional regulators. Some of the Rgg proteins are known to be involved in bacterial stress adaptation. Results In this study, we demonstrated that Streptococcus thermophilus thermal stress adaptation required the rgg0182 gene which transcription depends on the culture medium and the growth temperature. This gene encoded a protein showing similarity with members of the Rgg family transcriptional regulator. Our data confirmed that Rgg0182 is a transcriptional regulator controlling the expression of its neighboring genes as well as chaperones and proteases encoding genes. Therefore, analysis of a Δrgg0182 mutant revealed that this protein played a role in the heat shock adaptation of Streptococcus thermophilus LMG18311. Conclusions These data showed the importance of the Rgg0182 transcriptional regulator on the survival of S. thermophilus during dairy processes and more specifically during changes in temperature.

  14. Phytochromes Regulate SA and JA Signaling Pathways in Rice and Are Required for Developmentally Controlled Resistance to Magnaporthe grisea

    Institute of Scientific and Technical Information of China (English)

    Xian-Zhi Xie; Yan-Jiu Xue; Jin-Jun Zhoua; Bin Zhang; Hong Chang; Makoto Takano

    2011-01-01

    Old leaves of wild-type rice plants (Oryza sativa L. Cv. Nipponbare)are more resistant to blast fungus (Mag-naporthe grisea)than new leaves. In contrast, both old and new leaves of the rice phytochrome triple mutant (phyAphyB-phyC)are susceptible to blast fungus. We demonstrate that pathogenesis-related class 1 (PR1)proteins are rapidly and strongly induced during M. Grisea infection and following exogenous jasmonate (JA)or salicylic acid (SA)exposure in the old leaves, but not in the new leaves of the wild-type. In contrast, the accumulation of PR1 proteins was significantly attenuated in old and new leaves of the phyAphyBphyC mutant. These results suggest that phytochromes are required for the induction of PR1 proteins in rice. Basal transcription levels of Prla and PRlb were substantially higher in the wild-type as compared to the phyAphyBphyC mutant, suggesting that phytochromes also are required for basal expression of PR1 genes. Moreover, the transcript levels of genes known to function in SA-or JA-dependent defense pathways were regulated by leaf age and functional phytochromes. Taken together, our findings demonstrate that phytochromes are required in rice for age-related resistance to M, grisea and may indirectly increase PR1 gene expression by regulating SA-and JA-dependent defense pathways.

  15. Differential developmental requirement and peripheral regulation for dermal Vγ4 and Vγ6T17 cells in health and inflammation.

    Science.gov (United States)

    Cai, Yihua; Xue, Feng; Fleming, Chris; Yang, Jie; Ding, Chuanlin; Ma, Yunfeng; Liu, Min; Zhang, Huang-ge; Zheng, Jie; Xiong, Na; Yan, Jun

    2014-06-09

    Dermal IL-17-producing γδT cells have a critical role in skin inflammation. However, their development and peripheral regulation have not been fully elucidated. Here we demonstrate that dermal γδT cells develop from the embryonic thymus and undergo homeostatic proliferation after birth with diversified TCR repertoire. Vγ6T cells are bona fide resident, but precursors of dermal Vγ4T cells may require extrathymic environment for imprinting skin-homing properties. Thymic Vγ6T cells are more competitive than Vγ4 for dermal γδT cell reconstitution and TCRδ(-/-) mice reconstituted with Vγ6 develop psoriasis-like inflammation after IMQ-application. Although both IL-23 and IL-1β promote Vγ4 and Vγ6 proliferation, Vγ4 are the main source of IL-17 production that requires IL-1 signalling. Mice with deficiency of IL-1RI signalling have significantly decreased skin inflammation. These studies reveal a differential developmental requirement and peripheral regulation for dermal Vγ6 and Vγ4 γδT cells, implying a new mechanism that may be involved in skin inflammation.

  16. Exploring the structural requirements for jasmonates and related compounds as novel plant growth regulators: a current computational perspective.

    Science.gov (United States)

    Chen, Ke-Xian; Li, Zu-Guang

    2009-11-01

    Jasmonates and related compounds have been highlighted recently in the field of plant physiology and plant molecular biology due to their significant regulatory roles in the signaling pathway for the diverse aspects of plant development and survival. Though a considerable amount of studies concerning their biological effects in different plants have been widely reported, the molecular details of the signaling mechanism are still poorly understood. This review sheds new light on the structural requirements for the bioactivity/property of jasmonic acid derivatives in current computational perspective, which differs from previous research that mainly focus on their biological evaluation, gene and metabolic regulation and the enzymes in their biosynthesis. The computational results may contribute to further understanding the mechanism of drug-receptor interactions in their signaling pathway and designing novel plant growth regulators as high effective ecological pesticides.

  17. ChREBP, but not LXRs, is required for the induction of glucose-regulated genes in mouse liver

    Science.gov (United States)

    Denechaud, Pierre-Damien; Bossard, Pascale; Lobaccaro, Jean-Marc A.; Millatt, Lesley; Staels, Bart; Girard, Jean; Postic, Catherine

    2008-01-01

    The transcription factor carbohydrate-responsive element–binding protein (ChREBP) has emerged as a central regulator of lipid synthesis in liver because it is required for glucose-induced expression of the glycolytic enzyme liver–pyruvate kinase (L-PK) and acts in synergy with SREBP to induce lipogenic genes such as acetyl-CoA carboxylase (ACC) and fatty acid synthase (FAS). Liver X receptors (LXRs) are also important regulators of the lipogenic pathway, and the recent finding that ChREBP is a direct target of LXRs and that glucose itself can bind and activate LXRs prompted us to study the role of LXRs in the induction of glucose-regulated genes in liver. Using an LXR agonist in wild-type mice, we found that LXR stimulation did not promote ChREBP phosphorylation or nuclear localization in the absence of an increased intrahepatic glucose flux. Furthermore, the induction of ChREBP, L-PK, and ACC by glucose or high-carbohydrate diet was similar in LXRα/β knockout compared with wild-type mice, suggesting that the activation of these genes by glucose occurs by an LXR-independent mechanism. We used fluorescence resonance energy transfer analysis to demonstrate that glucose failed to promote the interaction of LXRα/β with specific cofactors. Finally, siRNA silencing of ChREBP in LXRα/β knockout hepatocytes abrogated glucose-induced expression of L-PK and ACC, further demonstrating the central role of ChREBP in glucose signaling. Taken together, our results demonstrate that glucose is required for ChREBP functional activity and that LXRs are not necessary for the induction of glucose-regulated genes in liver. PMID:18292813

  18. Regulation of the ERK pathway in the dentate gyrus by in vivo dopamine D1 receptor stimulation requires glutamatergic transmission.

    Science.gov (United States)

    Gangarossa, Giuseppe; Valjent, Emmanuel

    2012-11-01

    Acute systemic administration of the dopamine D1/D5 receptors (D1Rs) agonist, SKF81297, activates the extracellular signal-regulated protein kinases (ERK) pathway selectively in the granule cells of the dentate gyrus. In this study, we examined the mechanisms involved in this regulation and investigated the molecular components that could promote ERK-dependent transcription and translation. SKF81297 induced phosphorylation of ERK and histone H3 required intact glutamatergic transmission. Blockade of glutamate release achieved by the mGluR2/3 agonist, LY354740 or the selective adenosine A1R agonist, CCPA as well as neurotoxic lesions of lateral entorhinal cortex reduced the ability of SKF81297 to induce ERK activation in the dentate gyrus. This activation required the combined stimulation of NR2B-containing NMDARs, mGluR1 and mGluR5. SKF81297 evoked phosphorylation of the ribosomal protein S6 (rpS6) selectively at the Ser235/236 site while the Ser240/244 site remains unchanged. The SKF81297 induced increased phosphorylation of rpS6 was dependent on PKC and ERK/p90RSK activation. Surprisingly, administration of D1Rs agonist suppressed mTORC1/p70S6K pathway suggesting an mTOR-independent regulation of rpS6 phosphorylation. Taken together, our results show that intact glutamatergic transmission plays a major role in the regulation of ERK-dependent phosphorylation of histone H3 and rpS6 observed in the mouse dentate gyrus after systemic administration of SKF81297.

  19. Optogenetic monitoring identifies phosphatidylthreonine-regulated calcium homeostasis in Toxoplasma gondii

    Directory of Open Access Journals (Sweden)

    Arunakar Kuchipudi

    2016-05-01

    Full Text Available Toxoplasma gondii is an obligate intracellular parasite, which inflicts acute as well as chronic infections in a wide range of warm-blooded vertebrates. Our recent work has demonstrated the natural occurrence and autonomous synthesis of an exclusive lipid phosphatidylthreonine in T. gondii. Targeted gene disruption of phosphatidylthreonine synthase impairs the parasite virulence due to unforeseen attenuation of the consecutive events of motility, egress and invasion. However, the underlying basis of such an intriguing phenotype in the parasite mutant remains unknown. Using an optogenetic sensor (gene-encoded calcium indicator, GCaMP6s, we show that loss of phosphatidylthreonine depletes calcium stores in intracellular tachyzoites, which leads to dysregulation of calcium release into the cytosol during the egress phase of the mutant. Consistently, the parasite motility and egress phenotypes in the mutant can be entirely restored by ionophore-induced mobilization of calcium. Collectively, our results suggest a novel regulatory function of phosphatidylthreonine in calcium signaling of a prevalent parasitic protist. Moreover, our application of an optogenetic sensor to monitor subcellular calcium in a model intracellular pathogen exemplifies its wider utility to other entwined systems.

  20. Vangl2-regulated polarisation of second heart field-derived cells is required for outflow tract lengthening during cardiac development.

    Directory of Open Access Journals (Sweden)

    Simon A Ramsbottom

    2014-12-01

    Full Text Available Planar cell polarity (PCP is the mechanism by which cells orient themselves in the plane of an epithelium or during directed cell migration, and is regulated by a highly conserved signalling pathway. Mutations in the PCP gene Vangl2, as well as in other key components of the pathway, cause a spectrum of cardiac outflow tract defects. However, it is unclear why cells within the mesodermal heart tissue require PCP signalling. Using a new conditionally floxed allele we show that Vangl2 is required solely within the second heart field (SHF to direct normal outflow tract lengthening, a process that is required for septation and normal alignment of the aorta and pulmonary trunk with the ventricular chambers. Analysis of a range of markers of polarised epithelial tissues showed that in the normal heart, undifferentiated SHF cells move from the dorsal pericardial wall into the distal outflow tract where they acquire an epithelial phenotype, before moving proximally where they differentiate into cardiomyocytes. Thus there is a transition zone in the distal outflow tract where SHF cells become more polarised, turn off progenitor markers and start to differentiate to cardiomyocytes. Membrane-bound Vangl2 marks the proximal extent of this transition zone and in the absence of Vangl2, the SHF-derived cells are abnormally polarised and disorganised. The consequent thickening, rather than lengthening, of the outflow wall leads to a shortened outflow tract. Premature down regulation of the SHF-progenitor marker Isl1 in the mutants, and accompanied premature differentiation to cardiomyocytes, suggests that the organisation of the cells within the transition zone is important for maintaining the undifferentiated phenotype. Thus, Vangl2-regulated polarisation and subsequent acquisition of an epithelial phenotype is essential to lengthen the tubular outflow vessel, a process that is essential for on-going cardiac morphogenesis.

  1. Vangl2-regulated polarisation of second heart field-derived cells is required for outflow tract lengthening during cardiac development.

    Science.gov (United States)

    Ramsbottom, Simon A; Sharma, Vipul; Rhee, Hong Jun; Eley, Lorraine; Phillips, Helen M; Rigby, Hannah F; Dean, Charlotte; Chaudhry, Bill; Henderson, Deborah J

    2014-12-01

    Planar cell polarity (PCP) is the mechanism by which cells orient themselves in the plane of an epithelium or during directed cell migration, and is regulated by a highly conserved signalling pathway. Mutations in the PCP gene Vangl2, as well as in other key components of the pathway, cause a spectrum of cardiac outflow tract defects. However, it is unclear why cells within the mesodermal heart tissue require PCP signalling. Using a new conditionally floxed allele we show that Vangl2 is required solely within the second heart field (SHF) to direct normal outflow tract lengthening, a process that is required for septation and normal alignment of the aorta and pulmonary trunk with the ventricular chambers. Analysis of a range of markers of polarised epithelial tissues showed that in the normal heart, undifferentiated SHF cells move from the dorsal pericardial wall into the distal outflow tract where they acquire an epithelial phenotype, before moving proximally where they differentiate into cardiomyocytes. Thus there is a transition zone in the distal outflow tract where SHF cells become more polarised, turn off progenitor markers and start to differentiate to cardiomyocytes. Membrane-bound Vangl2 marks the proximal extent of this transition zone and in the absence of Vangl2, the SHF-derived cells are abnormally polarised and disorganised. The consequent thickening, rather than lengthening, of the outflow wall leads to a shortened outflow tract. Premature down regulation of the SHF-progenitor marker Isl1 in the mutants, and accompanied premature differentiation to cardiomyocytes, suggests that the organisation of the cells within the transition zone is important for maintaining the undifferentiated phenotype. Thus, Vangl2-regulated polarisation and subsequent acquisition of an epithelial phenotype is essential to lengthen the tubular outflow vessel, a process that is essential for on-going cardiac morphogenesis.

  2. cAMP-Signalling Regulates Gametocyte-Infected Erythrocyte Deformability Required for Malaria Parasite Transmission.

    Directory of Open Access Journals (Sweden)

    Ghania Ramdani

    2015-05-01

    Full Text Available Blocking Plasmodium falciparum transmission to mosquitoes has been designated a strategic objective in the global agenda of malaria elimination. Transmission is ensured by gametocyte-infected erythrocytes (GIE that sequester in the bone marrow and at maturation are released into peripheral blood from where they are taken up during a mosquito blood meal. Release into the blood circulation is accompanied by an increase in GIE deformability that allows them to pass through the spleen. Here, we used a microsphere matrix to mimic splenic filtration and investigated the role of cAMP-signalling in regulating GIE deformability. We demonstrated that mature GIE deformability is dependent on reduced cAMP-signalling and on increased phosphodiesterase expression in stage V gametocytes, and that parasite cAMP-dependent kinase activity contributes to the stiffness of immature gametocytes. Importantly, pharmacological agents that raise cAMP levels in transmissible stage V gametocytes render them less deformable and hence less likely to circulate through the spleen. Therefore, phosphodiesterase inhibitors that raise cAMP levels in P. falciparum infected erythrocytes, such as sildenafil, represent new candidate drugs to block transmission of malaria parasites.

  3. Down-regulation of Stathmin Is Required for the Phenotypic Changes and Classical Activation of Macrophages.

    Science.gov (United States)

    Xu, Kewei; Harrison, Rene E

    2015-07-31

    Macrophages are important cells of innate immunity with specialized capacity for recognition and elimination of pathogens and presentation of antigens to lymphocytes for adaptive immunity. Macrophages become activated upon exposure to pro-inflammatory cytokines and pathogenic stimuli. Classical activation of macrophages with interferon-γ (IFNγ) and lipopolysaccharide (LPS) triggers a wide range of signaling events and morphological changes to induce the immune response. Our previous microtubule (MT) proteomic work revealed that the stathmin association with MTs is considerably reduced in activated macrophages, which contain significantly more stabilized MTs. Here, we show that there is a global decrease in stathmin levels, an MT catastrophe protein, in activated macrophages using both immunoblotting and immunofluorescent microscopy. This is an LPS-specific response that induces proteasome-mediated degradation of stathmin. We explored the functions of stathmin down-regulation in activated macrophages by generating a stable cell line overexpressing stathmin-GFP. We show that stathmin-GFP overexpression impacts MT stability, impairs cell spreading, and reduces activation-associated phenotypes. Furthermore, overexpressing stathmin reduces complement receptor 3-mediated phagocytosis and cellular activation, implicating a pivotal inhibitory role for stathmin in classically activated macrophages.

  4. Redox regulation of SUMO enzymes is required for ATM activity and survival in oxidative stress.

    Science.gov (United States)

    Stankovic-Valentin, Nicolas; Drzewicka, Katarzyna; König, Cornelia; Schiebel, Elmar; Melchior, Frauke

    2016-06-15

    To sense and defend against oxidative stress, cells depend on signal transduction cascades involving redox-sensitive proteins. We previously identified SUMO (small ubiquitin-related modifier) enzymes as downstream effectors of reactive oxygen species (ROS). Hydrogen peroxide transiently inactivates SUMO E1 and E2 enzymes by inducing a disulfide bond between their catalytic cysteines. How important their oxidation is in light of many other redox-regulated proteins has however been unclear. To selectively disrupt this redox switch, we identified a catalytically fully active SUMO E2 enzyme variant (Ubc9 D100A) with strongly reduced propensity to maintain a disulfide with the E1 enzyme in vitro and in cells. Replacement of Ubc9 by this variant impairs cell survival both under acute and mild chronic oxidative stresses. Intriguingly, Ubc9 D100A cells fail to maintain activity of the ATM-Chk2 DNA damage response pathway that is induced by hydrogen peroxide. In line with this, these cells are also more sensitive to the ROS-producing chemotherapeutic drugs etoposide/Vp16 and Ara-C. These findings reveal that SUMO E1~E2 oxidation is an essential redox switch in oxidative stress.

  5. Emdogain-regulated gene expression in palatal fibroblasts requires TGF-βRI kinase signaling.

    Directory of Open Access Journals (Sweden)

    Alexandra Stähli

    Full Text Available Genome-wide microarrays have suggested that Emdogain regulates TGF-β target genes in gingival and palatal fibroblasts. However, definitive support for this contention and the extent to which TGF-β signaling contributes to the effects of Emdogain has remained elusive. We therefore studied the role of the TGF-β receptor I (TGF-βRI kinase to mediate the effect of Emdogain on palatal fibroblasts. Palatal fibroblasts were exposed to Emdogain with and without the inhibitor for TGF-βRI kinase, SB431542. Emdogain caused 39 coding genes to be differentially expressed in palatal fibroblasts by microarray analysis (p10-fold. Importantly, in the presence of the TGF-βRI kinase inhibitor SB431542, Emdogain failed to cause any significant changes in gene expression. Consistent with this mechanism, three independent TGF-βRI kinase inhibitors and a TGF-β neutralizing antibody abrogated the increased expression of IL-11, a selected Emdogain target gene. The MAPK inhibitors SB203580 and U0126 lowered the impact of Emdogain on IL-11 expression. The data support that TGF-βRI kinase activity is necessary to mediate the effects of Emdogain on gene expression in vitro.

  6. Emdogain-regulated gene expression in palatal fibroblasts requires TGF-βRI kinase signaling.

    Science.gov (United States)

    Stähli, Alexandra; Bosshardt, Dieter; Sculean, Anton; Gruber, Reinhard

    2014-01-01

    Genome-wide microarrays have suggested that Emdogain regulates TGF-β target genes in gingival and palatal fibroblasts. However, definitive support for this contention and the extent to which TGF-β signaling contributes to the effects of Emdogain has remained elusive. We therefore studied the role of the TGF-β receptor I (TGF-βRI) kinase to mediate the effect of Emdogain on palatal fibroblasts. Palatal fibroblasts were exposed to Emdogain with and without the inhibitor for TGF-βRI kinase, SB431542. Emdogain caused 39 coding genes to be differentially expressed in palatal fibroblasts by microarray analysis (p10-fold). Importantly, in the presence of the TGF-βRI kinase inhibitor SB431542, Emdogain failed to cause any significant changes in gene expression. Consistent with this mechanism, three independent TGF-βRI kinase inhibitors and a TGF-β neutralizing antibody abrogated the increased expression of IL-11, a selected Emdogain target gene. The MAPK inhibitors SB203580 and U0126 lowered the impact of Emdogain on IL-11 expression. The data support that TGF-βRI kinase activity is necessary to mediate the effects of Emdogain on gene expression in vitro.

  7. Mitotic destruction of the cell cycle regulated NIMA protein kinase of Aspergillus nidulans is required for mitotic exit.

    OpenAIRE

    Pu, R T; Osmani, S A

    1995-01-01

    NIMA is a cell cycle regulated protein kinase required, in addition to p34cdc2/cyclin B, for initiation of mitosis in Aspergillus nidulans. Like cyclin B, NIMA accumulates when cells are arrested in G2 and is degraded as cells traverse mitosis. However, it is stable in cells arrested in mitosis. NIMA, and related kinases, have an N-terminal kinase domain and a C-terminal extension. Deletion of the C-terminus does not completely inactivate NIMA kinase activity but does prevent functional compl...

  8. High Silicon Accumulation in the Shoot is Required for Down-Regulating the Expression of Si Transporter Genes in Rice.

    Science.gov (United States)

    Mitani-Ueno, Namiki; Yamaji, Naoki; Ma, Jian Feng

    2016-12-01

    Rice requires high silicon (Si) for its high and sustainable yield. The efficient uptake of Si in rice is mediated by two transporters OsLsi1 and OsLsi2, which function as influx and efflux transporters, respectively. Our previous studies showed that the mRNA expression levels of these transporter genes were down-regulated by Si. Herein we investigated the mechanism underlying regulation of OsLsi1 and OsLsi2 expression. There was a negative correlation between the expression level of OsLsi1 and OsLsi2 and shoot Si accumulation when the rice seedlings were exposed to different Si supply conditions. A split root experiment showed that the expression of both OsLsi1 and OsLsi2 was also down-regulated in half the roots without direct Si exposure when the other half of the roots were exposed to Si. Analysis with transgenic rice carrying different lengths of OsLsi1 promoter regions fused with green fluorescent protein (GFP) as a reporter gene revealed that the region responsible for the Si response of OsLsi1 expression is present between -327 to -292 in the promoter. However, this region was not associated with the tissue and cellular localization of OsLsi1. In conclusion, the Si-induced down-regulation of Si transporter genes is controlled by shoot Si, not root Si, and the region between -327 and -292 in the OsLsi1 promoter is involved in this regulation of OsLsi1 expression in rice.

  9. Using reporting requirements to improve employer wellness incentives and their regulation.

    Science.gov (United States)

    Madison, Kristin; Schmidt, Harald; Volpp, Kevin G

    2014-10-01

    Employer interest in offering financial incentives for healthy behaviors has been increasing. Some employers have begun to tie health plan-based rewards or penalties to standards involving tobacco use or biometric measures such as body mass index. The Patient Protection and Affordable Care Act attempts to strike a balance between the potential benefits and risks of wellness incentive programs by permitting these incentives but simultaneously limiting their use. Evidence about the implications of the newest generation of incentive programs for health, health costs, and burdens on individual employees will be critical for informing both private and public decision makers. After describing the many pieces of information that would be valuable for assessing these programs, this article proposes more narrowly targeted reporting requirements that could facilitate incentive program development, evaluation, and oversight. Copyright © 2014 by Duke University Press.

  10. Basic Requirements Of The Basel Committee On Regulating Capital Adequacy And Liquidity Of Commercial Banks

    Directory of Open Access Journals (Sweden)

    Bahriddin Berdiyarov

    2012-03-01

    Full Text Available The current paper highlights theBaselI, Basel II & Basel III requirements on capital adequacy and liquidity of commercial banks.  In the paper, Basel II structure, methods of loan risk assessment, coefficients of loan risk assessment, credit risk measurement for counterparty banks are discussed.  Moreover, assessments of Basel III on bank chances against crisis driven from financial and economic crunches, risk management, performance quality and bank transparency improvement measures are presented.  At the end, the author gives his conclusions on the essence and necessity of new regulatory standards of the Basel Committee on bank’s supervision in the structure of the supervision of credit institutions.

  11. Heterochromatic genome stability requires regulators of histone H3 K9 methylation.

    Directory of Open Access Journals (Sweden)

    Jamy C Peng

    2009-03-01

    Full Text Available Heterochromatin contains many repetitive DNA elements and few protein-encoding genes, yet it is essential for chromosome organization and inheritance. Here, we show that Drosophila that lack the Su(var3-9 H3K9 methyltransferase display significantly elevated frequencies of spontaneous DNA damage in heterochromatin, in both somatic and germ-line cells. Accumulated DNA damage in these mutants correlates with chromosomal defects, such as translocations and loss of heterozygosity. DNA repair and mitotic checkpoints are also activated in mutant animals and are required for their viability. Similar effects of lower magnitude were observed in animals that lack the RNA interference pathway component Dcr2. These results suggest that the H3K9 methylation and RNAi pathways ensure heterochromatin stability.

  12. Heterochromatic genome stability requires regulators of histone H3 K9 methylation.

    Science.gov (United States)

    Peng, Jamy C; Karpen, Gary H

    2009-03-01

    Heterochromatin contains many repetitive DNA elements and few protein-encoding genes, yet it is essential for chromosome organization and inheritance. Here, we show that Drosophila that lack the Su(var)3-9 H3K9 methyltransferase display significantly elevated frequencies of spontaneous DNA damage in heterochromatin, in both somatic and germ-line cells. Accumulated DNA damage in these mutants correlates with chromosomal defects, such as translocations and loss of heterozygosity. DNA repair and mitotic checkpoints are also activated in mutant animals and are required for their viability. Similar effects of lower magnitude were observed in animals that lack the RNA interference pathway component Dcr2. These results suggest that the H3K9 methylation and RNAi pathways ensure heterochromatin stability.

  13. Survival of Listeria monocytogenes in Soil Requires AgrA-Mediated Regulation.

    Science.gov (United States)

    Vivant, Anne-Laure; Garmyn, Dominique; Gal, Laurent; Hartmann, Alain; Piveteau, Pascal

    2015-08-01

    In a recent paper, we demonstrated that inactivation of the Agr system affects the patterns of survival of Listeria monocytogenes (A.-L. Vivant, D. Garmyn, L. Gal, and P. Piveteau, Front Cell Infect Microbiol 4:160, http://dx.doi.org/10.3389/fcimb.2014.00160). In this study, we investigated whether the Agr-mediated response is triggered during adaptation in soil, and we compared survival patterns in a set of 10 soils. The fate of the parental strain L. monocytogenes L9 (a rifampin-resistant mutant of L. monocytogenes EGD-e) and that of a ΔagrA deletion mutant were compared in a collection of 10 soil microcosms. The ΔagrA mutant displayed significantly reduced survival in these biotic soil microcosms, and differential transcriptome analyses showed large alterations of the transcriptome when AgrA was not functional, while the variations in the transcriptomes between the wild type and the ΔagrA deletion mutant were modest under abiotic conditions. Indeed, in biotic soil environments, 578 protein-coding genes and an extensive repertoire of noncoding RNAs (ncRNAs) were differentially transcribed. The transcription of genes coding for proteins involved in cell envelope and cellular processes, including the phosphotransferase system and ABC transporters, and proteins involved in resistance to antimicrobial peptides was affected. Under sterilized soil conditions, the differences were limited to 86 genes and 29 ncRNAs. These results suggest that the response regulator AgrA of the Agr communication system plays important roles during the saprophytic life of L. monocytogenes in soil.

  14. N-cadherin regulates signaling mechanisms required for lens fiber cell elongation and lens morphogenesis.

    Science.gov (United States)

    Logan, Caitlin M; Rajakaruna, Suren; Bowen, Caitlin; Radice, Glenn L; Robinson, Michael L; Menko, A Sue

    2017-08-01

    Tissue development and regeneration involve high-ordered morphogenetic processes that are governed by elements of the cytoskeleton in conjunction with cell adhesion molecules. Such processes are particularly important in the lens whose structure dictates its function. Studies of our lens-specific N-cadherin conditional knockout mouse (N-cadcKO) revealed an essential role for N-cadherin in the migration of the apical tips of differentiating lens fiber cells along the apical surfaces of the epithelium, a region termed the Epithelial Fiber Interface (EFI), that is necessary for normal fiber cell elongation and the morphogenesis. Studies of the N-cadcKO lens suggest that N-cadherin function in fiber cell morphogenesis is linked to the activation of Rac1 and myosin II, both signaling pathways central to the regulation of cell motility including determining the directionality of cellular movement. The absence of N-cadherin did not disrupt lateral contacts between fiber cells during development, and the maintenance of Aquaporin-0 and increased expression of EphA2 at cell-cell interfaces suggests that these molecules may function in this role. E-cadherin was maintained in newly differentiating fiber cells without interfering with expression of lens-specific differentiation proteins but was not able to replace N-cadherin function in these cells. The dependence of migration of the fiber cell apical domains along the EFI for lens morphogenesis on N-cadherin provides new insight into the process of tissue development. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. A Trypanosomatid Iron Transporter that Regulates Mitochondrial Function Is Required for Leishmania amazonensis Virulence.

    Directory of Open Access Journals (Sweden)

    Bidyottam Mittra

    2016-01-01

    Full Text Available Iron, an essential co-factor of respiratory chain proteins, is critical for mitochondrial function and maintenance of its redox balance. We previously reported a role for iron uptake in differentiation of Leishmania amazonensis into virulent amastigotes, by a mechanism that involves reactive oxygen species (ROS production and is independent of the classical pH and temperature cues. Iron import into mitochondria was proposed to be essential for this process, but evidence supporting this hypothesis was lacking because the Leishmania mitochondrial iron transporter was unknown. Here we describe MIT1, a homolog of the mitochondrial iron importer genes mrs3 (yeast and mitoferrin-1 (human that is highly conserved among trypanosomatids. MIT1 expression was essential for the survival of Trypanosoma brucei procyclic but not bloodstream forms, which lack functional respiratory complexes. L. amazonensis LMIT1 null mutants could not be generated, suggesting that this mitochondrial iron importer is essential for promastigote viability. Promastigotes lacking one LMIT1 allele (LMIT1/Δlmit1 showed growth defects and were more susceptible to ROS toxicity, consistent with the role of iron as the essential co-factor of trypanosomatid mitochondrial superoxide dismutases. LMIT1/Δlmit1 metacyclic promastigotes were unable to replicate as intracellular amastigotes after infecting macrophages or cause cutaneous lesions in mice. When induced to differentiate axenically into amastigotes, LMIT1/Δlmit1 showed strong defects in iron content and function of mitochondria, were unable to upregulate the ROS-regulatory enzyme FeSOD, and showed mitochondrial changes suggestive of redox imbalance. Our results demonstrate the importance of mitochondrial iron uptake in trypanosomatid parasites, and highlight the role of LMIT1 in the iron-regulated process that orchestrates differentiation of L. amazonensis into infective amastigotes.

  16. The Drosophila cell adhesion molecule klingon is required for long-term memory formation and is regulated by Notch.

    Science.gov (United States)

    Matsuno, Motomi; Horiuchi, Junjiro; Tully, Tim; Saitoe, Minoru

    2009-01-06

    The ruslan (rus) mutant was previously identified in a behavioral screen for mutants defective in long-lasting memory, which consists of two consolidated memory types, anesthesia-resistant memory, and protein synthesis-dependent long-term memory (LTM). We demonstrate here that rus is a new allele of klingon (klg), which encodes a homophilic cell adhesion molecule. Klg is acutely required for LTM but not anesthesia-resistant memory formation, and Klg expression increases upon LTM induction. LTM formation also requires activity of the Notch cell-surface receptor. Although defects in Notch have been implicated in memory loss because of Alzheimer's disease, downstream signaling linking Notch to memory have not been determined. Strikingly, we found that Notch activity increases upon LTM induction and regulates Klg expression. Furthermore, Notch-induced enhancement of LTM is disrupted by a klg mutation. We propose that Klg is a downstream effector of Notch signaling that links Notch activity to memory.

  17. Blood volume-monitored regulation of ultrafiltration in fluid-overloaded hemodialysis patients: study protocol for a randomized controlled trial

    Directory of Open Access Journals (Sweden)

    Hecking Manfred

    2012-06-01

    Full Text Available Abstract Background Data generated with the body composition monitor (BCM, Fresenius show, based on bioimpedance technology, that chronic fluid overload in hemodialysis patients is associated with poor survival. However, removing excess fluid by lowering dry weight can be accompanied by intradialytic and postdialytic complications. Here, we aim at testing the hypothesis that, in comparison to conventional hemodialysis, blood volume-monitored regulation of ultrafiltration and dialysate conductivity (UCR and/or regulation of ultrafiltration and temperature (UTR will decrease complications when ultrafiltration volumes are systematically increased in fluid-overloaded hemodialysis patients. Methods/design BCM measurements yield results on fluid overload (in liters, relative to extracellular water (ECW. In this prospective, multicenter, triple-arm, parallel-group, crossover, randomized, controlled clinical trial, we use BCM measurements, routinely introduced in our three maintenance hemodialysis centers shortly prior to the start of the study, to recruit sixty hemodialysis patients with fluid overload (defined as ≥15% ECW. Patients are randomized 1:1:1 into UCR, UTR and conventional hemodialysis groups. BCM-determined, ‘final’ dry weight is set to normohydration weight −7% of ECW postdialysis, and reached by reducing the previous dry weight, in steps of 0.1 kg per 10 kg body weight, during 12 hemodialysis sessions (one study phase. In case of intradialytic complications, dry weight reduction is decreased, according to a prespecified algorithm. A comparison of intra- and post-dialytic complications among study groups constitutes the primary endpoint. In addition, we will assess relative weight reduction, changes in residual renal function, quality of life measures, and predialysis levels of various laboratory parameters including C-reactive protein, troponin T, and N-terminal pro-B-type natriuretic peptide, before and after the first study

  18. Depletion suspends the comparator mechanism in monitoring: The role of chronic self-consciousness in sequential self-regulation.

    Science.gov (United States)

    Jia, Lile; Hirt, Edward R

    2016-08-01

    In 4 studies, we accumulated support for the prediction that depletion suspends the comparator mechanism of self-regulatory monitoring. We adopted an individual difference approach and designated chronic self-consciousness as a signature variable for the comparator mechanism. In the nondepletion condition, we found that self-consciousness predicted self-regulation by itself (Study 1), or by interacting synergistically with other motivational factors such as online goal focus (Study 2) and task motivation (Study 3). In the depletion condition, self-consciousness ceased to predict task performance, which suggested that the self-focused comparator mechanism is suspended by depletion. Instead, depleted participants' self-control was predicted by their implicit goal to rest (Study 3), indicating that depletion does not indiscriminately suspend all self-regulatory processes. In Study 4, we showed that when an effective comparator mechanism is counterproductive to task performance, depletion can actually increase task performance. Implications of our findings for the underlying process of depletion as well as models positing ultimate explanations for sequential self-regulation are discussed. (PsycINFO Database Record

  19. 40 CFR 63.3968 - What are the requirements for continuous parameter monitoring system installation, operation, and...

    Science.gov (United States)

    2010-07-01

    .... Monitoring failures that are caused in part by poor maintenance or careless operation are not malfunctions... install a device to monitor pressure drop across the zeolite wheel or rotary carbon bed. The pressure... signal simulations or via relative accuracy testing. (v) Conduct an accuracy audit every quarter and...

  20. 40 CFR 63.4168 - What are the requirements for continuous parameter monitoring system installation, operation, and...

    Science.gov (United States)

    2010-07-01

    ... Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NATIONAL... monitor pressure drop across the zeolite wheel or rotary carbon bed. The pressure monitoring device must... pressure sensor(s) in or as close to a position that provides a representative measurement of the pressure...

  1. Fyn tyrosine kinase regulates oligodendroglial cell development but is not required for morphological differentiation of oligodendrocytes.

    Science.gov (United States)

    Sperber, B R; McMorris, F A

    2001-02-15

    The non-receptor protein tyrosine kinase Fyn, which is a member of the Src family of kinases, has been shown to be essential for normal myelination and has been suggested to play a role in oligodendrocyte development. However, oligodendrocyte development has not been studied directly in cells lacking Fyn. Additionally, because Fyn is expressed in neurons as well as oligodendrocytes, it is possible that normal myelination requires Fyn expression in neurons but not in oligodendrocytes. To address these issues, we analyzed the development of oligodendrocytes in neuron-free glial cell cultures from fyn(-/-) mice that express no Fyn protein. We observed that oligodendrocytes develop to the stage where they elaborate an extensive network of membranous processes and express the antigenic components of mature oligodendrocytes in the complete absence of Fyn. However, as compared with fyn(+/+) controls, fewer oligodendroglia developed in fyn(-/-) cell cultures, and a smaller proportion of them matured to the stage characterized by a high degree of morphological complexity. In addition, we found that insulin-like growth factor-I, a potent stimulator of oligodendrocyte development, failed to stimulate morphological maturation of fyn(-/-) oligodendroglia. The pyrazolopyrimidine PP2, believed to be a selective inhibitor of Fyn, did not prevent the development of morphologically complex oligodendrocytes. Unexpectedly, however, it was toxic to both fyn(+/+) and fyn(-/-) glial cells, indicating that this class of inhibitors can have significant effects that are independent of Fyn.

  2. Distinct Akt phosphorylation states are required for insulin regulated Glut4 and Glut1-mediated glucose uptake.

    Science.gov (United States)

    Beg, Muheeb; Abdullah, Nazish; Thowfeik, Fathima Shazna; Altorki, Nasser K; McGraw, Timothy E

    2017-06-07

    Insulin, downstream of Akt activation, promotes glucose uptake into fat and muscle cells to lower postprandial blood glucose, an enforced change in cellular metabolism to maintain glucose homeostasis. This effect is mediated by the Glut4 glucose transporter. Growth factors also enhance glucose uptake to fuel an anabolic metabolism required for tissue growth and repair. This activity is predominantly mediated by the Glut1. Akt is activated by phosphorylation of its kinase and hydrophobic motif (HM) domains. We show that insulin-stimulated Glut4-mediated glucose uptake requires PDPK1 phosphorylation of the kinase domain but not mTORC2 phosphorylation of the HM domain. Nonetheless, an intact HM domain is required for Glut4-mediated glucose uptake. Whereas, Glut1-mediated glucose uptake also requires mTORC2 phosphorylation of the HM domain, demonstrating both phosphorylation-dependent and independent roles of the HM domain in regulating glucose uptake. Thus, mTORC2 links Akt to the distinct physiologic programs related to Glut4 and Glut1-mediated glucose uptake.

  3. Surface swarming motility by Pectobacterium atrosepticum is a latent phenotype that requires O antigen and is regulated by quorum sensing.

    Science.gov (United States)

    Bowden, Steven D; Hale, Nicola; Chung, Jade C S; Hodgkinson, James T; Spring, David R; Welch, Martin

    2013-11-01

    We describe a previously cryptic phenotype associated with the opportunistic phytopathogen Pectobacterium atrosepticum (Pca): surface swarming. We found that when Pca was spotted onto plates containing <0.5% (w/v) agar, the culture produced copious amounts of extracellular matrix material containing highly motile cells. Once produced, this 'slime layer' spread rapidly across the plate either as an advancing front or as tendrils. Transposon mutagenesis was used to identify mutants that were affected in swarming. Hypo-swarmer mutants mostly carried insertions in a horizontally acquired island (HAI5), which encodes a cluster of genes involved in O antigen biosynthesis. Hyper-swarmer mutants mostly carried insertions in hexY, a known antagonist of the class I flagellar master regulator, FlhD4C2. In addition, we found that the nucleoid protein, histone-like nuclear structuring protein 2 (H-NS2), also regulated swarming behaviour. A mutant in which hns2 was overexpressed displayed a hyper-swarming phenotype, whereas a mutant in which the hns2 ORF was inactivated had a hypo-swarming phenotype. Swarming was also regulated by quorum sensing (QS) and by the carbon source being utilized. We show, using a range of epistasis experiments, that optimal swarming requires both motility and O antigen biosynthesis, and that H-NS2 and QS both promote swarming through their effects on motility.

  4. Differential requirement for the CD45 splicing regulator hnRNPLL for accumulation of NKT and conventional T cells.

    Directory of Open Access Journals (Sweden)

    Mehmet Yabas

    Full Text Available Natural killer T (NKT cells represent an important regulatory T cell subset that develops in the thymus and contains immature (NK1.1(lo and mature (NK1.1(hi cell subsets. Here we show in mice that an inherited mutation in heterogeneous ribonucleoprotein L-like protein (hnRNPLL(thunder, that shortens the survival of conventional T cells, has no discernible effect on NKT cell development, homeostasis or effector function. Thus, Hnrpll deficiency effectively increases the NKT∶T cell ratio in the periphery. However, Hnrpll mutation disrupts CD45RA, RB and RC exon silencing of the Ptprc mRNA in both NKT and conventional T cells, and leads to a comparably dramatic shift to high molecular weight CD45 isoforms. In addition, Hnrpll mutation has a cell intrinsic effect on the expression of the developmentally regulated cell surface marker NK1.1 on NKT cells in the thymus and periphery but does not affect cell numbers. Therefore our results highlight both overlapping and divergent roles for hnRNPLL between conventional T cells and NKT cells. In both cell subsets it is required as a trans-acting factor to regulate alternative splicing of the Ptprc mRNA, but it is only required for survival of conventional T cells.

  5. Ezrin is required for the functional regulation of the epithelial sodium proton exchanger, NHE3.

    Directory of Open Access Journals (Sweden)

    Hisayoshi Hayashi

    Full Text Available The sodium hydrogen exchanger isoform 3 (NHE3 mediates absorption of sodium, bicarbonate and water from renal and intestinal lumina. This activity is fundamental to the maintenance of a physiological plasma pH and blood pressure. To perform this function NHE3 must be present in the apical membrane of renal tubular and intestinal epithelia. The molecular determinants of this localization have not been conclusively determined, although linkage to the apical actin cytoskeleton through ezrin has been proposed. We set out to evaluate this hypothesis. Functional studies of NHE3 activity were performed on ezrin knockdown mice (Vil2(kd/kd and NHE3 activity similar to wild-type animals detected. Interpretation of this finding was difficult as other ERM (ezrin/radixin/moesin proteins were present. We therefore generated an epithelial cell culture model where ezrin was the only detectable ERM. After knockdown of ezrin expression with siRNA, radixin and moesin expression remained undetectable. Consistent with the animal ultrastructural data, cells lacking ezrin retained an epithelial phenotype but had shortened and thicker microvilli. NHE3 localization was identical to cells transfected with non-targeting siRNA. The attachment of NHE3 to the apical cytoskeleton was unaltered as assessed by fluorescent recovery after photobleaching (FRAP and the solubility of NHE3 in Triton X-100. Baseline NHE3 activity was unaltered, however, cAMP-dependent inhibition of NHE3 was largely lost even though NHE3 was phosphorylated at serines 552 and 605. Thus, ezrin is not necessary for the apical localization, attachment to the cytoskeleton, baseline activity or cAMP induced phosphrylation of NHE3, but instead is required for cAMP mediated inhibition.

  6. 40 CFR Table 4 to Subpart G of... - Process Vents-Monitoring, Recordkeeping, and Reporting Requirements For Maintaining a TRE Index...

    Science.gov (United States)

    2010-07-01

    ..., and Reporting Requirements For Maintaining a TRE Index Value >1.0 and. â¤4.0 4 Table 4 to Subpart G of... for Organic Hazardous Air Pollutants From the Synthetic Organic Chemical Manufacturing Industry for... TRE Index Value >1.0 and. ≤4.0 Final recovery device Parameters to be monitored a Recordkeeping...

  7. 40 CFR 60.4237 - What are the monitoring requirements if I am an owner or operator of an emergency stationary SI...

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 6 2010-07-01 2010-07-01 false What are the monitoring requirements if I am an owner or operator of an emergency stationary SI internal combustion engine? 60.4237 Section 60.4237 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW...

  8. CovR and VicRK regulate cell surface biogenesis genes required for biofilm formation in Streptococcus mutans.

    Directory of Open Access Journals (Sweden)

    Rafael N Stipp

    Full Text Available The two-component system VicRK and the orphan regulator CovR of Streptococcus mutans co-regulate a group of virulence genes associated with the synthesis of and interaction with extracellular polysaccharides of the biofilm matrix. Knockout mutants of vicK and covR display abnormal cell division and morphology phenotypes, although the gene function defects involved are as yet unknown. Using transcriptomic comparisons between parent strain UA159 with vicK (UAvic or covR (UAcov deletion mutants together with electrophoretic motility shift assays (EMSA, we identified genes directly regulated by both VicR and CovR with putative functions in cell wall/surface biogenesis, including gbpB, wapE, smaA, SMU.2146c, and lysM. Deletion mutants of genes regulated by VicR and CovR (wapE, lysM, smaA, or regulated only by VicR (SMU.2146c or CovR (epsC promoted significant alterations in biofilm initiation, including increased fragility, defects in microcolony formation, and atypical cell morphology and/or chaining. Significant reductions in mureinolytic activity and/or increases in DNA release during growth were observed in knockout mutants of smaA, wapE, lysM, SMU.2146c and epsC, implying roles in cell wall biogenesis. WapE and lysM mutations also affected cell hydrophobicity and sensitivity to osmotic or oxidative stress. Finally, vicR, covR and VicRK/CovR-targets (gbpB, wapE, smaA, SMU.2146c, lysM, epsC are up-regulated in UA159 during biofilm initiation, in a sucrose-dependent manner. These data support a model in which VicRK and CovR coordinate cell division and surface biogenesis with the extracellular synthesis of polysaccharides, a process apparently required for formation of structurally stable biofilms in the presence of sucrose.

  9. Ecological Monitoring and Compliance Program 2012 Report

    Energy Technology Data Exchange (ETDEWEB)

    Hall, Derek B.; Anderson, David C.; Greger, Paul D.; Ostler, W. Kent; Hansen, Dennis J.

    2013-07-03

    The Ecological Monitoring and Compliance Program (EMAC), funded through the U.S. Department of Energy, National Nuclear Security Administration Nevada Field Office (NNSA/NFO, formerly Nevada Site Office), monitors the ecosystem of the Nevada National Security Site (NNSS) and ensures compliance with laws and regulations pertaining to NNSS biota. This report summarizes the program’s activities conducted by National Security Technologies, LLC (NSTec), during calendar year 2012. Program activities included (a) biological surveys at proposed construction sites, (b) desert tortoise compliance, (c) ecosystem monitoring, (d) sensitive plant species monitoring, (e) sensitive and protected/regulated animal monitoring, (f) habitat restoration monitoring, and (g) monitoring of the Nonproliferation Test and Evaluation Complex (NPTEC). During 2012, all applicable laws, regulations, and permit requirements were met, enabling EMAC to achieve its intended goals and objectives.

  10. Ecological Monitoring and Compliance Program 2011 Report

    Energy Technology Data Exchange (ETDEWEB)

    Hansen, D. J.; Anderson, D. C.; Hall, D. B.; Greger, P. D.; Ostler, W. K.

    2012-06-13

    The Ecological Monitoring and Compliance (EMAC) Program, funded through the U.S. Department of Energy, National Nuclear Security Administration Nevada Site Office, monitors the ecosystem of the Nevada National Security Site and ensures compliance with laws and regulations pertaining to NNSS biota. This report summarizes the program's activities conducted by National Security Technologies, LLC, during calendar year 2011. Program activities included (a) biological surveys at proposed construction sites, (b) desert tortoise compliance, (c) ecosystem monitoring, (d) sensitive plant species monitoring, (e) sensitive and protected/regulated animal monitoring, (f) habitat restoration monitoring, and (g) monitoring of the Nonproliferation Test and Evaluation Complex. During 2011, all applicable laws, regulations, and permit requirements were met, enabling EMAC to achieve its intended goals and objectives.

  11. Ecological Monitoring and Compliance Program 2009 Report

    Energy Technology Data Exchange (ETDEWEB)

    Hansen, J. Dennis; Anderson, David C.; Hall, Derek B.; Greger, Paul D.; Ostler, W. Kent

    2010-07-13

    The Ecological Monitoring and Compliance Program (EMAC), funded through the U.S. Department of Energy, National Nuclear Security Administration Nevada Site Office, monitors the ecosystem of the Nevada Test Site and ensures compliance with laws and regulations pertaining to NTS biota. This report summarizes the program’s activities conducted by National Security Technologies, LLC, during calendar year 2009. Program activities included (a) biological surveys at proposed construction sites, (b) desert tortoise compliance, (c) ecosystem mapping and data management, (d) sensitive plant species monitoring, (e) sensitive and protected/regulated animal monitoring, (f) habitat monitoring, (g) habitat restoration monitoring, and (h) monitoring of the Nonproliferation Test and Evaluation Complex. During 2009, all applicable laws, regulations, and permit requirements were met, enabling EMAC to achieve its intended goals and objectives.

  12. Ecological Monitoring and Compliance Program 2010 Report

    Energy Technology Data Exchange (ETDEWEB)

    Hansen, D.J.; Anderson, D.C.; Hall, D.B.; Greger, P.D.; Ostler, W.K.

    2011-07-01

    The Ecological Monitoring and Compliance (EMAC) Program, funded through the U.S. Department of Energy, National Nuclear Security Administration Nevada Site Office (NNSA/NSO), monitors the ecosystem of the Nevada National Security Site (NNSS) and ensures compliance with laws and regulations pertaining to NNSS biota. This report summarizes the program’s activities conducted by National Security Technologies, LLC (NSTec), during calendar year 2010. Program activities included (a) biological surveys at proposed construction sites, (b) desert tortoise compliance, (c) ecosystem monitoring, (d) sensitive plant species monitoring, (e) sensitive and protected/regulated animal monitoring, (f) habitat restoration monitoring, and (g) monitoring of the Nonproliferation Test and Evaluation Complex (NPTEC). During 2010, all applicable laws, regulations, and permit requirements were met, enabling EMAC to achieve its intended goals and objectives.

  13. The new DIN EN 50379. Requirements and testing regulated throughout Europe for portable exhaust gas measuring devices; Die neue DIN EN 50379. Anforderungen und Pruefung an tragbare Abgasmessgeraete europaweit geregelt

    Energy Technology Data Exchange (ETDEWEB)

    Ester, S. [Woehler Messgeraete Kehrgeraete GmbH, Bad Wuennenberg (Germany); Schrameyer, A. [TUEV Sued Industrie Service GmbH, Muenchen (Germany)

    2006-03-15

    The requirements for measuring devices for monitoring of small combustion systems have, up to now, been regulated in Germany in the context of the Federal Prevention of Pollution Ordinance and the individual states' Street Hygiene and Inspection Ordinances. This article describes the new DIN EN 50 379 series of standards, which, for the first time ever, now also covers devices for determination of combustion parameters for installation and maintenance of heating equipment and thus goes well beyond the previous scope of applicability. The new series of standards will, in future, assure a high quality level throughout Europe. (orig.)

  14. 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.

  15. Notification: Hotline Complaint – Region 7 Compliance with Bid, Payment, and Performance Bond Requirements Outlined in Federal Acquisition Regulation (FAR) Part 28

    Science.gov (United States)

    June 1, 2012. In response to an anonymous hotline complaint, the OIG plans to conduct a review of Region 7’s compliance with bid, payment, and performance bond requirements outlined in Federal Acquisition Regulation (FAR) Part 28.

  16. Application of wide selected-ion monitoring data-independent acquisition to identify tomato fruit proteins regulated by the CUTIN DEFICIENT2 transcription factor

    Science.gov (United States)

    We describe here the use of label-free wide selected-ion monitoring data-independent acquisition (WiSIM-DIA) to identify proteins that are involved in the formation of tomato (Solanum lycopersicum) fruit cuticles and that are regulated by the transcription factor CUTIN DEFICIENT2 (CD2). A spectral l...

  17. Protein kinase A regulation of P2X(4) receptors: requirement for a specific motif in the C-terminus.

    Science.gov (United States)

    Brown, David A; Yule, David I

    2010-02-01

    The P2X purinergic receptor sub-family of ligand-gated ion channels are subject to protein kinase modulation. We have previously demonstrated that P2X(4)R signaling can be positively regulated by increasing intracellular cAMP levels. The molecular mechanism underlying this effect was, however, unknown. The present study initially addressed whether protein kinase A (PKA) activation was required. Subsequently a mutational approach was utilized to determine which region of the receptor was required for this potentiation. In both DT-40 3KO and HEK-293 cells transiently expressing P2X(4)R, forskolin treatment enhanced ATP-mediated signaling. Specific PKA inhibitors prevented the forskolin-induced enhancement of ATP-mediated inward currents in P2X(4)R expressing HEK-293 cells. To define which region of the P2X(4)R was required for the potentiation, mutations were generated in the cytoplasmic C-terminal tail. It was determined that a limited region of the C-terminus, consisting of a non-canonical tyrosine based sorting motif, was required for the effects of PKA. Of note, this region does not harbor any recognizable PKA phosphorylation motifs, and no direct phosphorylation of P2X(4)R was detected, suggesting that PKA phosphorylation of an accessory protein interacts with the endocytosis motif in the C-terminus of the P2X(4)R. In support of this notion, using Total Internal Reflection Fluorescence Microscopy (TIRF)\\ P2X(4)-EGFP was shown to accumulate at/near the plasma membrane following forskolin treatment. In addition, disrupting the endocytosis machinery using a dominant-negative dynamin construct also prevented the PKA-mediated enhancement of ATP-stimulated Ca(2+) signals. Our results are consistent with a novel mechanism of P2XR regulation, whereby PKA activity, without directly phosphorylating P2X(4)R, markedly enhances ATP-stimulated P2X(4)R currents and hence cytosolic Ca(2+) signals. This may occur at least in part, by altering the trafficking of a population of

  18. The ciliogenic transcription factor RFX3 regulates early midline distribution of guidepost neurons required for corpus callosum development.

    Directory of Open Access Journals (Sweden)

    Carine Benadiba

    Full Text Available The corpus callosum (CC is the major commissure that bridges the cerebral hemispheres. Agenesis of the CC is associated with human ciliopathies, but the origin of this default is unclear. Regulatory Factor X3 (RFX3 is a transcription factor involved in the control of ciliogenesis, and Rfx3-deficient mice show several hallmarks of ciliopathies including left-right asymmetry defects and hydrocephalus. Here we show that Rfx3-deficient mice suffer from CC agenesis associated with a marked disorganisation of guidepost neurons required for axon pathfinding across the midline. Using transplantation assays, we demonstrate that abnormalities of the mutant midline region are primarily responsible for the CC malformation. Conditional genetic inactivation shows that RFX3 is not required in guidepost cells for proper CC formation, but is required before E12.5 for proper patterning of the cortical septal boundary and hence accurate distribution of guidepost neurons at later stages. We observe focused but consistent ectopic expression of Fibroblast growth factor 8 (Fgf8 at the rostro commissural plate associated with a reduced ratio of GLIoma-associated oncogene family zinc finger 3 (GLI3 repressor to activator forms. We demonstrate on brain explant cultures that ectopic FGF8 reproduces the guidepost neuronal defects observed in Rfx3 mutants. This study unravels a crucial role of RFX3 during early brain development by indirectly regulating GLI3 activity, which leads to FGF8 upregulation and ultimately to disturbed distribution of guidepost neurons required for CC morphogenesis. Hence, the RFX3 mutant mouse model brings novel understandings of the mechanisms that underlie CC agenesis in ciliopathies.

  19. Fragile X protein FMRP is required for homeostatic plasticity and regulation of synaptic strength by retinoic acid.

    Science.gov (United States)

    Soden, Marta E; Chen, Lu

    2010-12-15

    Homeostatic synaptic plasticity adjusts the strength of synapses during global changes in neural activity, thereby stabilizing the overall activity of neural networks. Suppression of synaptic activity increases synaptic strength by inducing synthesis of retinoic acid (RA), which activates postsynaptic synthesis of AMPA-type glutamate receptors (AMPARs) in dendrites and promotes synaptic insertion of newly synthesized AMPARs. Here, we show that fragile X mental retardation protein (FMRP), an RNA-binding protein that regulates dendritic protein synthesis, is essential for increases in synaptic strength induced by RA or by blockade of neural activity in the mouse hippocampus. Although activity-dependent RA synthesis is maintained in Fmr1 knock-out neurons, RA-dependent dendritic translation of GluR1-type AMPA receptors is impaired. Intriguingly, FMRP is only required for the form of homeostatic plasticity that is dependent on both RA signaling and local protein synthesis. Postsynaptic expression of wild-type or mutant FMRP(I304N) in knock-out neurons reduced the total, surface, and synaptic levels of AMPARs, implying a role for FMRP in regulating AMPAR abundance. Expression of FMRP lacking the RGG box RNA-binding domain had no effect on AMPAR levels. Importantly, postsynaptic expression of wild-type FMRP, but not FMRP(I304N) or FMRPΔRGG, restored synaptic scaling when expressed in knock-out neurons. Together, these findings identify an unanticipated role for FMRP in regulating homeostatic synaptic plasticity downstream of RA. Our results raise the possibility that at least some of the symptoms of fragile X syndrome reflect impaired homeostatic plasticity and impaired RA signaling.

  20. Mycobacterium tuberculosis universal stress protein Rv2623 regulates bacillary growth by ATP-Binding: requirement for establishing chronic persistent infection.

    Directory of Open Access Journals (Sweden)

    Joshua E Drumm

    2009-05-01

    Full Text Available Tuberculous latency and reactivation play a significant role in the pathogenesis of tuberculosis, yet the mechanisms that regulate these processes remain unclear. The Mycobacterium tuberculosisuniversal stress protein (USP homolog, rv2623, is among the most highly induced genes when the tubercle bacillus is subjected to hypoxia and nitrosative stress, conditions thought to promote latency. Induction of rv2623 also occurs when M. tuberculosis encounters conditions associated with growth arrest, such as the intracellular milieu of macrophages and in the lungs of mice with chronic tuberculosis. Therefore, we tested the hypothesis that Rv2623 regulates tuberculosis latency. We observed that an Rv2623-deficient mutant fails to establish chronic tuberculous infection in guinea pigs and mice, exhibiting a hypervirulence phenotype associated with increased bacterial burden and mortality. Consistent with this in vivo growth-regulatory role, constitutive overexpression of rv2623 attenuates mycobacterial growth in vitro. Biochemical analysis of purified Rv2623 suggested that this mycobacterial USP binds ATP, and the 2.9-A-resolution crystal structure revealed that Rv2623 engages ATP in a novel nucleotide-binding pocket. Structure-guided mutagenesis yielded Rv2623 mutants with reduced ATP-binding capacity. Analysis of mycobacteria overexpressing these mutants revealed that the in vitro growth-inhibitory property of Rv2623 correlates with its ability to bind ATP. Together, the results indicate that i M. tuberculosis Rv2623 regulates mycobacterial growth in vitro and in vivo, and ii Rv2623 is required for the entry of the tubercle bacillus into the chronic phase of infection in the host; in addition, iii Rv2623 binds ATP; and iv the growth-regulatory attribute of this USP is dependent on its ATP-binding activity. We propose that Rv2623 may function as an ATP-dependent signaling intermediate in a pathway that promotes persistent infection.

  1. ST2 negatively regulates TLR2 signaling, but is not required for bacterial lipoprotein-induced tolerance.

    LENUS (Irish Health Repository)

    Liu, Jinghua

    2010-05-15

    Activation of TLR signaling is critical for host innate immunity against bacterial infection. Previous studies reported that the ST2 receptor, a member of the Toll\\/IL-1 receptor superfamily, functions as a negative regulator of TLR4 signaling and maintains LPS tolerance. However, it is undetermined whether ST2 negatively regulates TLR2 signaling and furthermore, whether a TLR2 agonist, bacterial lipoprotein (BLP)-induced tolerance is dependent on ST2. In this study, we show that BLP stimulation-induced production of proinflammatory cytokines and immunocomplex formation of TLR2-MyD88 and MyD88-IL-1R-associated kinase (IRAK) were significantly enhanced in ST2-deficient macrophages compared with those in wild-type controls. Furthermore, overexpression of ST2 dose-dependently attenuated BLP-induced NF-kappaB activation, suggesting a negative regulatory role of ST2 in TLR2 signaling. A moderate but significantly attenuated production of TNF-alpha and IL-6 on a second BLP stimulation was observed in BLP-pretreated, ST2-deficient macrophages, which is associated with substantially reduced IRAK-1 protein expression and downregulated TLR2-MyD88 and MyD88-IRAK immunocomplex formation. ST2-deficient mice, when pretreated with a nonlethal dose of BLP, benefitted from an improved survival against a subsequent lethal BLP challenge, indicating BLP tolerance develops in the absence of the ST2 receptor. Taken together, our results demonstrate that ST2 acts as a negative regulator of TLR2 signaling, but is not required for BLP-induced tolerance.

  2. Nfil3 is a glucocorticoid-regulated gene required for glucocorticoid-induced apoptosis in male murine T cells.

    Science.gov (United States)

    Carey, Kirstyn T; Tan, Kheng H; Ng, Judy; Liddicoat, Douglas R; Godfrey, Dale I; Cole, Timothy J

    2013-04-01

    Glucocorticoids (GCs) have essential roles in the regulation of development, integrated metabolism, and immune and neurological responses, and act primarily via the glucocorticoid receptor (GR). In most cells, GC treatment results in down-regulation of GR mRNA and protein levels via negative feedback mechanisms. However, in GC-treated thymocytes, GR protein levels are maintained at a high level, increasing sensitivity of thymocytes to GCs, resulting in apoptosis termed glucocorticoid-induced cell death (GICD). CD4(+)CD8(+) double-positive thymocytes and thymic natural killer T cells in particular are highly sensitive to GICD. Although GICD is exploited via the use of synthetic GC analogues in the treatment of hematopoietic malignancies, the intracellular molecular pathway of GICD is not well understood. To explore GICD in thymocytes, the authors performed whole genome expression microarray analysis in mouse GR exon 2 null vs wild-type thymus RNA 3 hours after dexamethasone treatment. Identified and validated direct GR targets included P21 and Bim, in addition to an important transcriptional regulator Nfil3, which previously has been associated with GICD and is essential for natural killer cell development in vivo. Immunostaining of NFIL3 in whole thymus localized NFIL3 primarily to the medullary region, and double labeling colocalized NFIL3 to apoptotic cells. In silico analysis revealed a putative GC response element 5 kb upstream of the Nfil3 promoter that is strongly conserved in the rat genome and was confirmed to bind GR by chromatin immunoprecipitation. The knockdown of Nfil3 mRNA levels to 20% of normal using specific small interfering RNAs abrogated GICD, indicating that NFIL3 is required for normal GICD in CTLL-2 T cells.

  3. Transcriptome Reprogramming by Plasmid-Encoded Transcriptional Regulators Is Required for Host Niche Adaption of a Macrophage Pathogen

    Science.gov (United States)

    Coulson, Garry B.; Miranda-CasoLuengo, Aleksandra A.; Miranda-CasoLuengo, Raúl; Wang, Xiaoguang; Oliver, Jenna; Willingham-Lane, Jennifer M.

    2015-01-01

    Rhodococcus equi is a facultative intracellular pathogen of macrophages, relying on the presence of a conjugative virulence plasmid harboring a 21-kb pathogenicity island (PAI) for growth in host macrophages. The PAI encodes a family of 6 virulence-associated proteins (Vaps) in addition to 20 other proteins. The contribution of these to virulence has remained unclear. We show that the presence of only 3 virulence plasmid genes (of 73 in total) is required and sufficient for intracellular growth. These include a single vap family member, vapA, and two PAI-located transcriptional regulators, virR and virS. Both transcriptional regulators are essential for wild-type-level expression of vapA, yet vapA expression alone is not sufficient to allow intracellular growth. A whole-genome microarray analysis revealed that VirR and VirS substantially integrate themselves into the chromosomal regulatory network, significantly altering the transcription of 18% of all chromosomal genes. This pathoadaptation involved significant enrichment of select gene ontologies, in particular, enrichment of genes involved in transport processes, energy production, and cellular metabolism, suggesting a major change in cell physiology allowing the bacterium to grow in the hostile environment of the host cell. The results suggest that following the acquisition of the virulence plasmid by an avirulent ancestor of R. equi, coevolution between the plasmid and the chromosome took place, allowing VirR and VirS to regulate the transcription of chromosomal genes in a process that ultimately promoted intracellular growth. Our findings suggest a mechanism for cooption of existing chromosomal traits during the evolution of a pathogenic bacterium from an avirulent saprophyte. PMID:26015480

  4. Mycobacterium tuberculosis Universal Stress Protein Rv2623 Regulates Bacillary Growth by ATP Binding: Requirement for Establishing Chronic Persistent Infection

    Energy Technology Data Exchange (ETDEWEB)

    Drumm, J.; Mi, K; Bilder, P; Sun, M; Lim, J; Bielefeldt-Ohmann, H; Basaraba, R; So, M; Zhu, G; et. al.

    2009-01-01

    Tuberculous latency and reactivation play a significant role in the pathogenesis of tuberculosis, yet the mechanisms that regulate these processes remain unclear. The Mycobacterium tuberculosisuniversal stress protein (USP) homolog, rv2623, is among the most highly induced genes when the tubercle bacillus is subjected to hypoxia and nitrosative stress, conditions thought to promote latency. Induction of rv2623 also occurs when M. tuberculosis encounters conditions associated with growth arrest, such as the intracellular milieu of macrophages and in the lungs of mice with chronic tuberculosis. Therefore, we tested the hypothesis that Rv2623 regulates tuberculosis latency. We observed that an Rv2623-deficient mutant fails to establish chronic tuberculous infection in guinea pigs and mice, exhibiting a hypervirulence phenotype associated with increased bacterial burden and mortality. Consistent with this in vivo growth-regulatory role, constitutive overexpression of rv2623 attenuates mycobacterial growth in vitro. Biochemical analysis of purified Rv2623 suggested that this mycobacterial USP binds ATP, and the 2.9-A-resolution crystal structure revealed that Rv2623 engages ATP in a novel nucleotide-binding pocket. Structure-guided mutagenesis yielded Rv2623 mutants with reduced ATP-binding capacity. Analysis of mycobacteria overexpressing these mutants revealed that the in vitro growth-inhibitory property of Rv2623 correlates with its ability to bind ATP. Together, the results indicate that i M. tuberculosis Rv2623 regulates mycobacterial growth in vitro and in vivo, and ii Rv2623 is required for the entry of the tubercle bacillus into the chronic phase of infection in the host; in addition, iii Rv2623 binds ATP; and iv the growth-regulatory attribute of this USP is dependent on its ATP-binding activity. We propose that Rv2623 may function as an ATP-dependent signaling intermediate in a pathway that promotes persistent infection.

  5. Intracellular pH regulation in unstimulated Calliphora salivary glands is Na+ dependent and requires V-ATPase activity.

    Science.gov (United States)

    Schewe, Bettina; Blenau, Wolfgang; Walz, Bernd

    2012-04-15

    Salivary gland cells of the blowfly Calliphora vicina have a vacuolar-type H(+)-ATPase (V-ATPase) that lies in their apical membrane and energizes the secretion of a KCl-rich primary saliva upon stimulation with serotonin (5-hydroxytryptamine). Whether and to what extent V-ATPase contributes to intracellular pH (pH(i)) regulation in unstimulated gland cells is unknown. We used the fluorescent dye BCECF to study intracellular pH(i) regulation microfluorometrically and show that: (1) under resting conditions, the application of Na(+)-free physiological saline induces an intracellular alkalinization attributable to the inhibition of the activity of a Na(+)-dependent glutamate transporter; (2) the maintenance of resting pH(i) is Na(+), Cl(-), concanamycin A and DIDS sensitive; (3) recovery from an intracellular acid load is Na(+) sensitive and requires V-ATPase activity; (4) the Na(+)/H(+) antiporter is not involved in pH(i) recovery after a NH(4)Cl prepulse; and (5) at least one Na(+)-dependent transporter and the V-ATPase maintain recovery from an intracellular acid load. Thus, under resting conditions, the V-ATPase and at least one Na(+)-dependent transporter maintain normal pH(i) values of pH 7.5. We have also detected the presence of a Na(+)-dependent glutamate transporter, which seems to act as an acid loader. Despite this not being a common pH(i)-regulating transporter, its activity affects steady-state pH(i) in C. vicina salivary gland cells.

  6. S1P-Yap1 signaling regulates endoderm formation required for cardiac precursor cell migration in zebrafish.

    Science.gov (United States)

    Fukui, Hajime; Terai, Kenta; Nakajima, Hiroyuki; Chiba, Ayano; Fukuhara, Shigetomo; Mochizuki, Naoki

    2014-10-13

    To form the primary heart tube in zebrafish, bilateral cardiac precursor cells (CPCs) migrate toward the midline beneath the endoderm. Mutants lacking endoderm and fish with defective sphingosine 1-phosphate (S1P) signaling exhibit cardia bifida. Endoderm defects lead to the lack of foothold for the CPCs, whereas the cause of cardia bifida in S1P signaling mutants remains unclear. Here we show that S1P signaling regulates CPC migration through Yes-associated protein 1 (Yap1)-dependent endoderm survival. Cardia bifida seen in spns2 (S1P transporter) morphants and s1pr2 (S1P receptor-2) morphants could be rescued by endodermal expression of nuclear localized form of yap1. yap1 morphants had decreased expression of the Yap1/Tead target connective tissue growth factor a (Ctgfa) and consequently increased endodermal cell apoptosis. Consistently, ctgfa morphants showed defects of the endodermal sheet and cardia bifida. Collectively, we show that S1pr2/Yap1-regulated ctgfa expression is essential for the proper endoderm formation required for CPC migration.

  7. Regulation of the New Arabidopsis Imprinted Gene AtBMI1C Requires the Interplay of Different Epigenetic Mechanisms

    Institute of Scientific and Technical Information of China (English)

    Fabian Bratzel; ChaoYang; Alexandra Ancelova; Gema López-Torrejón; Marcus Koch; Juan Carlos del Pozo; Myriam Calonje

    2012-01-01

    Recently,it has been shown that plants contain homologs to the animal Polycomb repressive complex 1 (PRC1)components BMI1 and RING1A/B.In Arabidopsis,there are three BMI1-like genes,two of which,AtBMI1A and B,are required during post-embryonic plant growth to repress embryonic traits and allow cell differentiation.However,little is known about the third BMI1-like gene,AtBMI1C.In this work,we show that AtBMI1C is only expressed during endosperm and stamen development.AtBMI1C is an imprinted gene expressed from the maternal allele in the endosperm but biallelically expressed in stamen.We found that the characteristic expression pattern of AtBMI1C is the result of a complex epigenetic regulation that involves CG DNA methylation,RNA-directed non-CG DNA methylation (RdDM),and PcG activity.Our results show the orchestrated interplay of different epigenetic mechanisms in regulating gene expression throughout development,shedding light on the current hypotheses for the origin and mechanism of imprinting in plant endosperm.

  8. Phytophthora capsici homologue of the cell cycle regulator SDA1 is required for sporangial morphology, mycelial growth and plant infection.

    Science.gov (United States)

    Zhu, Chunyuan; Yang, Xiaoyan; Lv, Rongfei; Li, Zhuang; Ding, Xiaomeng; Tyler, Brett M; Zhang, Xiuguo

    2016-04-01

    SDA1 encodes a highly conserved protein that is widely distributed in eukaryotic organisms. SDA1 is essential for cell cycle progression and organization of the actin cytoskeleton in yeasts and humans. In this study, we identified a Phytophthora capsici orthologue of yeast SDA1, named PcSDA1. In P. capsici, PcSDA1 is strongly expressed in three asexual developmental states (mycelium, sporangia and germinating cysts), as well as late in infection. Silencing or overexpression of PcSDA1 in P. capsici transformants affected the growth of hyphae and sporangiophores, sporangial development, cyst germination and zoospore release. Phalloidin staining confirmed that PcSDA1 is required for organization of the actin cytoskeleton. Moreover, 4',6-diamidino-2-phenylindole (DAPI) staining and PcSDA1-green fluorescent protein (GFP) fusions revealed that PcSDA1 is involved in the regulation of nuclear distribution in hyphae and sporangia. Both silenced and overexpression transformants showed severely diminished virulence. Thus, our results suggest that PcSDA1 plays a similar role in the regulation of the actin cytoskeleton and nuclear division in this filamentous organism as in non-filamentous yeasts and human cells.

  9. HDAC1 negatively regulates Bdnf and Pvalb required for parvalbumin interneuron maturation in an experience-dependent manner.

    Science.gov (United States)

    Koh, Dawn X P; Sng, Judy C G

    2016-11-01

    During early postnatal development, neuronal circuits are sculpted by sensory experience provided by the external environment. This experience-dependent regulation of circuitry development consolidates the balance of excitatory-inhibitory (E/I) neurons in the brain. The cortical barrel-column that innervates a single principal whisker is used to provide a clear reference frame for studying the consolidation of E/I circuitry. Sensory deprivation of S1 at birth disrupts the consolidation of excitatory-inhibitory balance by decreasing inhibitory transmission of parvalbumin interneurons. The molecular mechanisms underlying this decrease in inhibition are not completely understood. Our findings show that epigenetic mechanisms, in particular histone deacetylation by histone deacetylases, negatively regulate the expression of brain-derived neurotrophic factor (Bdnf) and parvalbumin (Pvalb) genes during development, which are required for the maturation of parvalbumin interneurons. After whisker deprivation, increased histone deacetylase 1 expression and activity led to increased histone deacetylase 1 binding and decreased histone acetylation at Bdnf promoters I-IV and Pvalb promoter, resulting in the repression of Bdnf and Pvalb gene transcription. The decrease in Bdnf expression further affected parvalbumin interneuron maturation at layer II/III in S1, demonstrated by decreased parvalbumin expression, a marker for parvalbumin interneuron maturation. Knockdown of HDAC1 recovered Bdnf and Pvalb gene transcription and also prevented the decrease of inhibitory synapses accompanying whisker deprivation.

  10. FgFlbD regulates hyphal differentiation required for sexual and asexual reproduction in the ascomycete fungus Fusarium graminearum.

    Science.gov (United States)

    Son, Hokyoung; Kim, Myung-Gu; Chae, Suhn-Kee; Lee, Yin-Won

    2014-11-01

    Fusarium graminearum is a filamentous fungal plant pathogen that infects major cereal crops. The fungus produces both sexual and asexual spores in order to endure unfavorable environmental conditions and increase their numbers and distribution across plants. In a model filamentous fungus, Aspergillus nidulans, early induction of conidiogenesis is orchestrated by the fluffy genes. The objectives of this study were to characterize fluffy gene homologs involved in conidiogenesis and their mechanism of action in F. graminearum. We characterized five fluffy gene homologs in F. graminearum and found that FlbD is the only conserved regulator for conidiogenesis in A. nidulans and F. graminearum. Deletion of fgflbD prevented hyphal differentiation and the formation of perithecia. Successful interspecies complementation using A. nidulans flbD demonstrated that the molecular mechanisms responsible for FlbD functions are conserved in F. graminearum. Moreover, abaA-wetA pathway is positively regulated by FgFlbD during conidiogenesis in F. graminearum. Deleting fgflbD abolished morphological effects of abaA overexpression, which suggests that additional factors for FgFlbD or an AbaA-independent pathway for conidiogenesis are required for F. graminearum conidiation. Importantly, this study led to the construction of a genetic pathway of F. graminearum conidiogenesis and provides new insights into the genetics of conidiogenesis in fungi.

  11. The bifunctional abiotic stress signalling regulator and endogenous RNA silencing suppressor FIERY1 is required for lateral root formation

    KAUST Repository

    Chen, Hao

    2010-09-28

    The Arabidopsis FIERY1 (FRY1) locus was originally identified as a negative regulator of stress-responsive gene expression and later shown to be required for suppression of RNA silencing. In this study we discovered that the FRY1 locus also regulates lateral root formation. Compared with the wild type, fry1 mutant seedlings generated significantly fewer lateral roots under normal growth conditions and also exhibited a dramatically reduced sensitivity to auxin in inducing lateral root initiation. Using transgenic plants that overexpress a yeast homolog of FRY1 that possesses only the 3\\', 5\\'-bisphosphate nucleotidase activity but not the inositol 1-phosphatase activity, we demonstrated that the lateral root phenotypes in fry1 result from loss of the nucleotidase activity. Furthermore, a T-DNA insertion mutant of another RNA silencing suppressor, XRN4 (but not XRN2 or XRN3), which is an exoribonuclease that is inhibited by the substrate of the FRY1 3\\', 5\\'-bisphosphate nucleotidase, exhibits similar lateral root defects. Although fry1 and xrn4 exhibited reduced sensitivity to ethylene, our experiments demonstrated that restoration of ethylene sensitivity in the fry1 mutant is not sufficient to rescue the lateral root phenotypes of fry1. Our results indicate that RNA silencing modulated by FRY1 and XRN4 plays an important role in shaping root architecture. © 2010 Blackwell Publishing Ltd.

  12. Greater-than-Class C low-level radioactive waste transportation regulations and requirements study. National Low-Level Waste Management Program

    Energy Technology Data Exchange (ETDEWEB)

    Tyacke, M.; Schmitt, R.

    1993-07-01

    The purpose of this report is to identify the regulations and requirements for transporting greater-than-Class C (GTCC) low-level radioactive waste (LLW) and to identify planning activities that need to be accomplished in preparation for transporting GTCC LLW. The regulations and requirements for transporting hazardous materials, of which GTCC LLW is included, are complex and include several Federal agencies, state and local governments, and Indian tribes. This report is divided into five sections and three appendices. Section 1 introduces the report. Section 2 identifies and discusses the transportation regulations and requirements. The regulations and requirements are divided into Federal, state, local government, and Indian tribes subsections. This report does not identify the regulations or requirements of specific state, local government, and Indian tribes, since the storage, treatment, and disposal facility locations and transportation routes have not been specifically identified. Section 3 identifies the planning needed to ensure that all transportation activities are in compliance with the regulations and requirements. It is divided into (a) transportation packaging; (b) transportation operations; (c) system safety and risk analysis, (d) route selection; (e) emergency preparedness and response; and (f) safeguards and security. This section does not provide actual planning since the details of the Department of Energy (DOE) GTCC LLW Program have not been finalized, e.g., waste characterization and quantity, storage, treatment and disposal facility locations, and acceptance criteria. Sections 4 and 5 provide conclusions and referenced documents, respectively.

  13. A DOE contractor`s perspective of environmental monitoring requirements at a low-level waste facility

    Energy Technology Data Exchange (ETDEWEB)

    Ferns, T.W. [Idaho National Engineering Lab., ID (United States)

    1989-11-01

    Environmental monitoring at a low-level waste disposal facility (LLWDF) should, (1) demonstrate compliance with environmental laws; (2) detect any spatial or temporal environmental changes; and (3) provide information on the potential or actual exposure of humans and/or the environment to disposed waste and/or waste by-products. Under the DOE Order system the LLWDF site manager has more freedom of implementation for a monitoring program than either the semi-prescriptive NRC, or the prescriptive EPA hazardous waste programs. This paper will attempt to compare and contrast environmental monitoring under the different systems (DOE, NRC, and EPA), and determine if the DOE might benefit from a more prescriptive system.

  14. 46 CFR 197.575 - Observation of monitoring.

    Science.gov (United States)

    2010-10-01

    ... 46 Shipping 7 2010-10-01 2010-10-01 false Observation of monitoring. 197.575 Section 197.575... STANDARDS GENERAL PROVISIONS Benzene § 197.575 Observation of monitoring. (a) Persons involved in benzene.... (b) When observation of monitoring requires entry into regulated areas, the observers shall use...

  15. Regulation of carotenoid and bacteriochlorophyll biosynthesis genes and identification of an evolutionarily conserved gene required for bacteriochlorophyll accumulation.

    Science.gov (United States)

    Armstrong, G A; Cook, D N; Ma, D; Alberti, M; Burke, D H; Hearst, J E

    1993-05-01

    The temporal expression of ten clustered genes required for carotenoid (crt) and bacteriochlorophyll (bch) biosynthesis was examined during the transition from aerobic respiration to anaerobiosis requisite for the development of the photosynthetic membrane in the bacterium Rhodobacter capsulatus. Accumulation of crtA, crtC, crtD, crtE, crtF, crtK, bchC and bchD mRNAs increased transiently and coordinately, up to 12-fold following removal of oxygen from the growth medium, paralleling increases in mRNAs encoding pigment-binding polypeptides of the photosynthetic apparatus. The crtB and crtI genes, in contrast, were expressed similarly in the presence or absence of oxygen. The regulation patterns of promoters for the crtA and crtI genes and the bchCXYZ operon were characterized using lacZ transcriptional fusion and qualitatively reflected the corresponding mRNA accumulation patterns. We also report that the bchI gene product, encoded by a DNA sequence previously considered to be a portion of crtA, shares 49% sequence identity with the nuclear-encoded Arabidopsis thaliana Cs chloroplast protein required for normal pigmentation in plants.

  16. Up-regulation of insulin-like growth factor 2 by ketamine requires glycogen synthase kinase-3 inhibition.

    Science.gov (United States)

    Grieco, Steven F; Cheng, Yuyan; Eldar-Finkelman, Hagit; Jope, Richard S; Beurel, Eléonore

    2017-01-04

    An antidepressant dose of the rapidly-acting ketamine inhibits glycogen synthase kinase-3 (GSK3) in mouse hippocampus, and this inhibition is required for the antidepressant effect of ketamine in learned helplessness depression-like behavior. Here we report that treatment with an antidepressant dose of ketamine (10mg/kg) increased expression of insulin-like growth factor 2 (IGF2) in mouse hippocampus, an effect that required ketamine-induced inhibition of GSK3. Ketamine also inhibited hippocampal GSK3 and increased expression of hippocampal IGF2 in mice when administered after the induction of learned helplessness. Treatment with the specific GSK3 inhibitor L803-mts was sufficient to up-regulate hippocampal IGF2 expression. Administration of IGF2 siRNA reduced ketamine's antidepressant effect in the learned helplessness paradigm. Mice subjected to the learned helplessness paradigm were separated into two groups, those that were resilient (non-depressed) and those that were susceptible (depressed). Non-depressed resilient mice displayed higher expression of IGF2 than susceptible mice. These results indicate that IGF2 contributes to ketamine's antidepressant effect and that IGF2 may confer resilience to depression-like behavior. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. 40 CFR Table 5 to Subpart Ppp of... - Process Vents From Batch Unit Operations-Monitoring, Recordkeeping, and Reporting Requirements

    Science.gov (United States)

    2010-07-01

    ... Hazardous Air Pollutant Emissions for Polyether Polyols Production Pt. 63, Subpt. PPP, Table 5 Table 5 to... all instances when monitoring data are not collected—PR. d,e If a base absorbent is used, report all pH values that are below the minimum operating values. If an acid absorbent is used, report all...

  18. 40 CFR 63.4568 - What are the requirements for continuous parameter monitoring system installation, operation, and...

    Science.gov (United States)

    2010-07-01

    ... caused in part by poor maintenance or careless operation are not malfunctions. Any period for which the... install a device to monitor pressure drop across the zeolite wheel or rotary carbon bed. The pressure... simulations or via relative accuracy testing. (v) Conduct an accuracy audit every quarter and after every...

  19. Department of Labor Needs to Give CETA Prime Sponsors More Guidance and Assistance for Implementing Monitoring Requirements.

    Science.gov (United States)

    1981-09-03

    minimun the IMUs’ yearly monitoring plan must include 15 areas of emphasis. (See enc. I.) The letter recommended a minimum of one comprehensive...Financial management 8. Allowance payment system 9. Participant wages and benefits 10. Client tracking system 11. Complaint/grievance procedure * 12

  20. Validation of limited sampling models (LSM) for estimating AUC in therapeutic drug monitoring - is a separate validation group required?

    NARCIS (Netherlands)

    Proost, J. H.

    2007-01-01

    Objective: Limited sampling models (LSM) for estimating AUC in therapeutic drug monitoring are usually validated in a separate group of patients, according to published guidelines. The aim of this study is to evaluate the validation of LSM by comparing independent validation with cross-validation us

  1. 40 CFR Table 11 to Subpart G of... - Wastewater-Inspection and Monitoring Requirements for Waste Management Units

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 9 2010-07-01 2010-07-01 false Wastewater-Inspection and Monitoring... Operations, and Wastewater Pt. 63, Subpt. G, Table 11 Table 11 to Subpart G of Part 63—Wastewater—Inspection... wastewater tank for control equipment failures and improper work practices Initially Semi-annually...

  2. Osh proteins regulate membrane sterol organization but are not required for sterol movement between the ER and PM

    Science.gov (United States)

    Georgiev, Alexander; Sullivan, David P.; Kersting, Michael C.; Dittman, Jeremy S.; Beh, Christopher T.; Menon, Anant K.

    2011-01-01

    Sterol transport between the endoplasmic reticulum (ER) and plasma membrane (PM) occurs by an ATP-dependent, non-vesicular mechanism that is presumed to require sterol transport proteins (STPs). In Saccharomyces cerevisiae, homologues of the mammalian oxysterol-binding protein (Osh1–7) have been proposed to function as STPs. To evaluate this proposal we took two approaches. First we used dehydroergosterol (DHE) to visualize sterol movement in living cells by fluorescence microscopy. DHE was introduced into the PM under hypoxic conditions and observed to redistribute to lipid droplets on growing the cells aerobically. Redistribution required ATP and the sterol acyltransferase Are2, but did not require PM-derived transport vesicles. DHE redistribution occurred robustly in a conditional yeast mutant (oshΔ osh4-1ts) that lacks all functional Osh proteins at 37°C. In a second approach we used a pulse-chase protocol to analyze the movement of metabolically radiolabeled ergosterol from the ER to the PM. Arrival of radiolabeled ergosterol at the PM was assessed in isolated PM-enriched fractions as well by extracting sterols from intact cells with methyl-β-cyclodextrin. These experiments revealed that whereas ergosterol is transported effectively from the ER to the PM in Osh-deficient cells, the rate at which it moves within the PM to equilibrate with the methyl-β-cyclodextrin extractable sterol pool is slowed. We conclude (i) that the role of Osh proteins in nonvesicular sterol transport between the PM, ER and lipid droplets is either minimal, or subsumed by other mechanisms and (ii) that Osh proteins regulate the organization of sterols at the PM. PMID:21689253

  3. Half Pint/Puf68 is required for negative regulation of splicing by the SR factor Transformer2

    Science.gov (United States)

    Wang, Shanzhi; Wagner, Eric J; Mattox, William

    2013-01-01

    The SR family of proteins plays important regulatory roles in the control of alternative splicing in a wide range of organisms. These factors affect splicing through both positive and negative controls of splice site recognition by pre-spliceosomal factors. Recent studies indicate that the Drosophila SR factor Transformer 2 (Tra2) activates and represses splicing through distinct and separable effector regions of the protein. While the interactions of its Arg-Ser-rich activator region have been well studied, cofactors involved in splicing repression have yet to be found. Here we use a luciferase-based splicing reporter assay to screen for novel proteins necessary for Tra2-dependent repression of splicing. This approach identified Half pint, also known as Puf68, as a co-repressor required for Tra2-mediated autoregulation of the M1 intron. In vivo, Half pint is required for Tra2-dependent repression of M1 splicing but is not necessary for Tra2-dependent activation of doublesex splicing. Further experiments indicate that the effect of Hfp is sequence-specific and that it associates with these target transcripts in cells. Importantly, known M1 splicing regulatory elements are sufficient to sensitize a heterologous intron to Hfp regulation. Two alternative proteins deriving from Hfp transcripts, Hfp68, and Hfp58, were found to be expressed in vivo but differed dramatically in their effect on M1 splicing. Comparison of the cellular localization of these forms in S2 cells revealed that Hfp68 is predominantly localized to the nucleus while Hfp58 is distributed across both the nucleus and cytoplasm. This accords with their observed effects on splicing and suggests that differential compartmentalization may contribute to the specificity of these isoforms. Together, these studies reveal a function for Half pint in splicing repression and demonstrate it to be specifically required for Tra2-dependent intron inclusion. PMID:23880637

  4. Half pint/Puf68 is required for negative regulation of splicing by the SR splicing factor Transformer2.

    Science.gov (United States)

    Wang, Shanzhi; Wagner, Eric J; Mattox, William

    2013-08-01

    The SR family of proteins plays important regulatory roles in the control of alternative splicing in a wide range of organisms. These factors affect splicing through both positive and negative controls of splice site recognition by pre-spliceosomal factors. Recent studies indicate that the Drosophila SR factor Transformer 2 (Tra2) activates and represses splicing through distinct and separable effector regions of the protein. While the interactions of its Arg-Ser-rich activator region have been well studied, cofactors involved in splicing repression have yet to be found. Here we use a luciferase-based splicing reporter assay to screen for novel proteins necessary for Tra2-dependent repression of splicing. This approach identified Half pint, also known as Puf68, as a co-repressor required for Tra2-mediated autoregulation of the M1 intron. In vivo, Half pint is required for Tra2-dependent repression of M1 splicing but is not necessary for Tra2-dependent activation of doublesex splicing. Further experiments indicate that the effect of Hfp is sequence-specific and that it associates with these target transcripts in cells. Importantly, known M1 splicing regulatory elements are sufficient to sensitize a heterologous intron to Hfp regulation. Two alternative proteins deriving from Hfp transcripts, Hfp68, and Hfp58, were found to be expressed in vivo but differed dramatically in their effect on M1 splicing. Comparison of the cellular localization of these forms in S2 cells revealed that Hfp68 is predominantly localized to the nucleus while Hfp58 is distributed across both the nucleus and cytoplasm. This accords with their observed effects on splicing and suggests that differential compartmentalization may contribute to the specificity of these isoforms. Together, these studies reveal a function for Half pint in splicing repression and demonstrate it to be specifically required for Tra2-dependent intron inclusion.

  5. Ecological Monitoring and Compliance Program 2013 Report

    Energy Technology Data Exchange (ETDEWEB)

    Hall, Derek B.; Anderson, David C.; Greger, Paul D.

    2014-06-05

    The Ecological Monitoring and Compliance Program (EMAC), funded through the U.S. Department of Energy, National Nuclear Security Administration Nevada Field Office (NNSA/NFO, formerly Nevada Site Office), monitors the ecosystem of the Nevada National Security Site (NNSS) and ensures compliance with laws and regulations pertaining to NNSS biota. This report summarizes the program’s activities conducted by National Security Technologies, LLC (NSTec), during calendar year 2013. Program activities included (a) biological surveys at proposed activity sites, (b) desert tortoise compliance, (c) ecosystem monitoring, (d) sensitive plant species monitoring, (e) sensitive and protected/regulated animal monitoring, and (f) habitat restoration monitoring. During 2013, all applicable laws, regulations, and permit requirements were met, enabling EMAC to achieve its intended goals and objectives.

  6. SafeLand guidelines for landslide monitoring and early warning systems in Europe - Design and required technology

    Science.gov (United States)

    Bazin, S.

    2012-04-01

    Landslide monitoring means the comparison of landslide characteristics like areal extent, speed of movement, surface topography and soil humidity from different periods in order to assess landslide activity. An ultimate "universal" methodology for this purpose does not exist; every technology has its own advantages and disadvantages. End-users should carefully consider each one to select the methodologies that represent the best compromise between pros and cons, and are best suited for their needs. Besides monitoring technology, there are many factors governing the choice of an Early Warning System (EWS). A people-centred EWS necessarily comprises five key elements: (1) knowledge of the risks; (2) identification, monitoring, analysis and forecasting of the hazards; (3) operational centre; (4) communication or dissemination of alerts and warnings; and (5) local capabilities to respond to the warnings received. The expression "end-to-end warning system" is also used to emphasize that EWSs need to span all steps from hazard detection through to community response. The aim of the present work is to provide guidelines for establishing the different components for landslide EWSs. One of the main deliverables of the EC-FP7 SafeLand project addresses the technical and practical issues related to monitoring and early warning for landslides, and identifies the best technologies available in the context of both hazard assessment and design of EWSs. This deliverable targets the end-users and aims to facilitate the decision process by providing guidelines. For the purpose of sharing the globally accumulated expertise, a screening study was done on 14 EWSs from 8 different countries. On these bases, the report presents a synoptic view of existing monitoring methodologies and early-warning strategies and their applicability for different landslide types, scales and risk management steps. Several comprehensive checklists and toolboxes are also included to support informed

  7. From the Accounting Treatment Required by the Accounting Regulations Corresponding with European Directives to the Accounting Treatment Required by IFRS - Legislative Steps Taken by Romanian Credit Institutions

    OpenAIRE

    Doroº Alexandra

    2013-01-01

    Legislative stages in accordance with IFRS accounting treatment began to OMF no. 907/2005- approving categories of entities applying accounting rules in accordance with International Financial Reporting Standards, accounting regulations that comply with European directives, as amended, continued with NBR Order no. 13/2008, NBR Order no. 15/2009, NBR Order no. 9/2010 and ends with NBR Order No. 27/16.12.2010 - for approval of Accounting Regulations in accordance with International Financial Re...

  8. CXCR3 Requirement for the Interleukin-13-Mediated Up-Regulation of Interleukin-13Rα2 in Pulmonary Fibroblasts.

    Science.gov (United States)

    Barnes, Jennifer C; Lumsden, Robert V; Worrell, Julie; Counihan, Ian P; O'Beirne, Sarah L; Belperio, John A; Fabre, Aurelie; Donnelly, Seamas C; Boylan, Denise; Kane, Rosemary; Keane, Michael P

    2015-08-01

    Idiopathic pulmonary fibrosis (IPF) is a progressive disease characterized by fibrosis and abnormal vascularity. IL-13, a profibrotic cytokine that plays a role in IPF, functions through the Jak/STAT pathway after binding to the IL-13 receptor α1 (IL-13Rα1)/IL-4Rα complex. IL-13 also binds to IL-13Rα2, which has been thought to function as a nonsignaling decoy receptor, although possible signaling roles of this receptor have been proposed. CXCR3 and its IFN-inducible ligands-CXCL9, CXCL10, and CXCL11-have been implicated in vascular remodeling and fibroblast motility during the development of IPF. In this study, CXCR3 expression was demonstrated in cultured pulmonary fibroblasts from wild-type BALB/c mice and was found to be necessary for the IL-13-mediated gene and protein up-regulation of IL-13Rα2. In fibroblasts from CXCR3-deficient mice, STAT6 activation was prolonged. This study is the first to demonstrate the expression of CXCR3 in fibroblasts and its association with the expression of IL-13Rα2. Taken together, the results from this study point strongly to a requirement for CXCR3 for IL-13-mediated IL-13Rα2 gene expression. Understanding the function of CXCR3 in IL-13-mediated lung injury may lead to novel approaches to combat the development of pulmonary fibrosis, whether by limiting the effects of IL-13 or by manipulation of angiostatic pathways. The elucidation of the complex relationship between these antifibrotic receptors and manipulation of the CXCR3-mediated regulation of IL-13Rα2 may represent a novel therapeutic modality in cases of acute lung injury or chronic inflammation that may progress to fibrosis.

  9. Fyn requires HnRNPA2B1 and Sam68 to synergistically regulate apoptosis in pancreatic cancer.

    Science.gov (United States)

    Chen, Zhi-Yu; Cai, Lei; Zhu, Jin; Chen, Min; Chen, Jian; Li, Zhi-Hua; Liu, Xiang-De; Wang, Shu-Guang; Bie, Ping; Jiang, Peng; Dong, Jia-Hong; Li, Xiao-Wu

    2011-10-01

    The Src family kinase Fyn, heterogenous nuclear ribonucleoprotein (HnRNP) A2B1 and Sam68 are thought to be associated with the metastasis of tumors, but their roles in the regulation of apoptosis remain unclear. This study investigated the role of Fyn and its potential relationship with HnRNPA2B1 and Sam68 in the regulation of apoptosis in pancreatic cancer. Experimental design. We examined both the activity of Fyn and the expression of HnRNPA2B1 in human pancreatic cancer tissues and systematically investigated the apoptotic mechanisms induced by Fyn activity using multiple experimental approaches. We found that Fyn activity was increased in metastatic pancreatic cancer tissues. In the pancreatic cancer BxPc3 cell line, the inhibition of Fyn activity by kinase-dead Fyn downregulated HnRNPA2B1 expression. Further analysis showed that HnRNPA2B1 expression was associated with pancreatic cancer progression. In BxPc3 cells, HnRNPA2B1 bound to Bcl-x messenger RNA (mRNA), which affected splicing and therefore, the formation of Bcl-x(s). Downregulation of HnRNPA2B1 by RNA interference (RNAi) resulted in the increased formation of the pro-apoptotic Bcl-x(s) and promoted apoptosis of BxPc3 cells. In addition, deactivation of Fyn in BxPc3 cells reduced Sam68 phosphorylation. This resulted in increased binding between Sam68 and Bcl-x mRNA, promoting the formation of the anti-apoptotic Bcl-x(L). The knockdown of Sam68 by RNAi also increased the formation of Bcl-x(L). Finally, HnRNPA2B1 overexpression or Sam68 knockdown could rescue pancreatic cancer cells from apoptosis. Our results suggest a mechanism by which Fyn requires HnRNPA2B1 and Sam68 to coordinate and regulate apoptosis, thus promoting the proliferation and metastasis of pancreatic cancer.

  10. Zinc finger protein Loz1 is required for zinc-responsive regulation of gene expression in fission yeast

    Science.gov (United States)

    Corkins, Mark E.; May, Margot; Ehrensberger, Kate M.; Hu, Ya-Mei; Liu, Yi-Hsuan; Bloor, Sean D.; Jenkins, Blair; Runge, Kurt W.; Bird, Amanda J.

    2013-01-01

    In Schizosaccharomyces pombe, alcohol dehydrogenase 1 (Adh1) is an abundant zinc-requiring enzyme that catalyses the conversion of acetaldehyde to ethanol during fermentation. In a zinc-replete cell, adh1 is highly expressed. However, in zinc-limited cells, adh1 gene expression is repressed, and cells induce the expression of an alternative alcohol dehydrogenase encoded by the adh4 gene. In our studies examining this zinc-dependent switch in alcohol dehydrogenase gene expression, we isolated an adh1Δ strain containing a partial loss of function mutation that resulted in higher levels of adh4 transcripts in zinc-replete cells. This mutation also led to the aberrant expression of other genes that are typically regulated by zinc. Using linkage analysis, we have mapped the position of this mutation to a single gene called Loss Of Zinc sensing 1 (loz1). Loz1 is a 55-kDa protein that contains a double C2H2-type zinc finger domain. The mapped mutation that disrupts Loz1 function leads to an arginine to glycine substitution in the second zinc finger domain, suggesting that the double zinc finger domain is important for Loz1 function. We show that loz1Δ cells hyperaccumulate zinc and that Loz1 is required for gene repression in zinc-replete cells. We also have found that Loz1 negatively autoregulates its own expression. We propose that Loz1 is a unique metalloregulatory factor that plays a central role in zinc homeostasis in S. pombe. PMID:24003116

  11. The Epstein-Barr virus (EBV) glycoprotein B cytoplasmic C-terminal tail domain regulates the energy requirement for EBV-induced membrane fusion.

    Science.gov (United States)

    Chen, Jia; Zhang, Xianming; Jardetzky, Theodore S; Longnecker, Richard

    2014-10-01

    The entry of enveloped viruses into host cells is preceded by membrane fusion, which in Epstein-Barr virus (EBV) is thought to be mediated by the refolding of glycoprotein B (gB) from a prefusion to a postfusion state. In our current studies, we characterized a gB C-terminal tail domain (CTD) mutant truncated at amino acid 843 (gB843). This truncation mutant is hyperfusogenic as monitored by syncytium formation and in a quantitative fusion assay and is dependent on gH/gL for fusion activity. gB843 can rescue the fusion function of other glycoprotein mutants that have null or decreased fusion activity in epithelial and B cells. In addition, gB843 requires less gp42 and gH/gL for fusion, and can function in fusion at a lower temperature than wild-type gB, indicating a lower energy requirement for fusion activation. Since a key step in fusion is the conversion of gB from a prefusion to an active postfusion state by gH/gL, gB843 may access this activated gB state more readily. Our studies indicate that the gB CTD may participate in the fusion function by maintaining gB in an inactive prefusion form prior to activation by receptor binding. Importance: Diseases resulting from Epstein-Barr virus (EBV) infection in humans range from the fairly benign disease infectious mononucleosis to life-threatening cancer. As an enveloped virus, EBV must fuse with a host cell membrane for entry and infection by using glycoproteins gH/gL, gB, and gp42. Among these glycoproteins, gB is thought to be the protein that executes fusion. To further characterize the function of the EBV gB cytoplasmic C-terminal tail domain (CTD) in fusion, we used a previously constructed CTD truncation mutant and studied its fusion activity in the context of other EBV glycoprotein mutants. From these studies, we find that the gB CTD regulates fusion by altering the energy requirements for the triggering of fusion mediated by gH/gL or gp42. Overall, our studies may lead to a better understanding of EBV fusion

  12. 42 CFR 495.316 - State monitoring and reporting regarding activities required to receive an incentive payment.

    Science.gov (United States)

    2010-10-01

    ...) Provider adoption, implementation, or upgrade of certified EHR technology activities and payments; and (2... upgraded certified EHR technology. (ii) Aggregated data tables representing the provider adoption... STANDARDS FOR THE ELECTRONIC HEALTH RECORD TECHNOLOGY INCENTIVE PROGRAM Requirements Specific to...

  13. Induction of interleukin-8 by Naegleria fowleri lysates requires activation of extracellular signal-regulated kinase in human astroglial cells.

    Science.gov (United States)

    Kim, Jong-Hyun; Sohn, Hae-Jin; Lee, Sang-Hee; Kwon, Daeho; Shin, Ho-Joon

    2012-08-01

    Naegleria fowleri is a pathogenic free-living amoeba which causes primary amoebic meningoencephalitis in humans and experimental animals. To investigate the mechanisms of such inflammatory diseases, potential chemokine gene activation in human astroglial cells was investigated following treatment with N. fowleri lysates. We demonstrated that N. fowleri are potent inducers for the expression of interleukin-8 (IL-8) genes in human astroglial cells which was preceded by activation of extracellular signal-regulated kinase (ERK). In addition, N. fowleri lysates induces the DNA binding activity of activator protein-1 (AP-1), an important transcription factor for IL-8 induction. The specific mitogen-activated protein kinase kinase/ERK inhibitor, U0126, blocks N. fowleri-mediated AP-1 activation and subsequent IL-8 induction. N. fowleri-induced IL-8 expression requires activation of ERK in human astroglial cells. These findings indicate that treatment of N. fowleri on human astroglial cells leads to the activation of AP-1 and subsequent expression of IL-8 which are dependent on ERK activation. These results may help understand the N. fowleri-mediated upregulation of chemokine and cytokine expression in the astroglial cells.

  14. International Regulations of Propolis Quality: Required Assays do not Necessarily Reflect their Polyphenolic-Related In Vitro Activities.

    Science.gov (United States)

    Bridi, Raquel; Montenegro, Gloria; Nuñez-Quijada, Gabriel; Giordano, Ady; Fernanda Morán-Romero, Maria; Jara-Pezoa, Isaac; Speisky, Hernán; Atala, Elias; López-Alarcón, Camilo

    2015-06-01

    Propolis has been proposed as a polyphenolic-rich natural product potentially able to be used for human consumption or even for medicinal proposes. To guarantee a minimum phenolic and flavonoid content and as consequence of their related-biological activities, international requirements of propolis quality are commonly applied. In this work we assessed phenolic and flavonoid contents of propolis; the antioxidant capacity (toward peroxyl radicals and hypochlorous acid); the ability to generate nitric oxide (NO); and, finally the antimicrobial activity of 6 propolis samples from the VI region of Chile. Our results show that the total phenolic and flavonoid content of propolis samples are not always in agreement with their polyphenolic-associated in vitro activities. For example, P03 and P06 samples showed the lowest (25 ± 4 GAE/g propolis) and the highest (105 ± 3 GAE/g propolis) total phenolic content, respectively. This was in agreement with flavonoid content and their Oxygen Radical Absorbance Capacity (ORAC) activity. However, this dependence was not observed toward HOCl, NO release and antimicrobial activity. Based on our results, we consider that, in order to guarantee the antioxidant or antimicrobial in vitro effects, the international regulations of propolis quality should contemplate the convenience of incorporating other simple analytical test such as ORAC or antimicrobial tests. © 2015 Institute of Food Technologists®

  15. The Second Subunit of DNA Polymerase Delta Is Required for Genomic Stability and Epigenetic Regulation1[OPEN

    Science.gov (United States)

    Cheng, Jinkui; Lai, Jinsheng; Gong, Zhizhong

    2016-01-01

    DNA polymerase δ plays crucial roles in DNA repair and replication as well as maintaining genomic stability. However, the function of POLD2, the second small subunit of DNA polymerase δ, has not been characterized yet in Arabidopsis (Arabidopsis thaliana). During a genetic screen for release of transcriptional gene silencing, we identified a mutation in POLD2. Whole-genome bisulfite sequencing indicated that POLD2 is not involved in the regulation of DNA methylation. POLD2 genetically interacts with Ataxia Telangiectasia-mutated and Rad3-related and DNA polymerase α. The pold2-1 mutant exhibits genomic instability with a high frequency of homologous recombination. It also exhibits hypersensitivity to DNA-damaging reagents and short telomere length. Whole-genome chromatin immunoprecipitation sequencing and RNA sequencing analyses suggest that pold2-1 changes H3K27me3 and H3K4me3 modifications, and these changes are correlated with the gene expression levels. Our study suggests that POLD2 is required for maintaining genome integrity and properly establishing the epigenetic markers during DNA replication to modulate gene expression. PMID:27208288

  16. CD81 is a central regulator of cellular events required for hepatitis C virus infection of human hepatocytes.

    Science.gov (United States)

    Brazzoli, Michela; Bianchi, Alessia; Filippini, Sara; Weiner, Amy; Zhu, Qing; Pizza, Mariagrazia; Crotta, Stefania

    2008-09-01

    Infection with hepatitis C virus (HCV) is still a major public health problem, and the events leading to hepatocyte infection are not yet fully understood. Combining confocal microscopy with biochemical analysis and studies of infection requirements using pharmacological inhibitors and small interfering RNAs, we show here that engagement of CD81 activates the Rho GTPase family members Rac, Rho, and Cdc42 and that the block of these signaling pathways drastically reduces HCV infectivity. Activation of Rho GTPases mediates actin-dependent relocalization of the HCV E2/CD81 complex to cell-cell contact areas where CD81 comes into contact with the tight-junction proteins occludin, ZO-1, and claudin-1, which was recently described as an HCV coreceptor. Finally, we show that CD81 engagement activates the Raf/MEK/ERK signaling cascade and that this pathway affects postentry events of the virus life cycle. In conclusion, we describe a range of cellular events that are manipulated by HCV to coordinate interactions with its multiple coreceptors and to establish productive infections and find that CD81 is a central regulator of these events.

  17. Surface water mineralization of isopyrazam according to OECD 309: observations on implementation of the new data requirement within agrochemical regulation.

    Science.gov (United States)

    Hand, Laurence H; Moreland, Harriet J

    2014-03-01

    A surface water mineralization study (according to the Organisation for Economic Co-operation and Development [OECD] guideline OECD 309) is a new requirement in European Union agrochemical regulations; therefore, industry has little experience with this test. The guideline allows for a number of options within the test design, notably the options to conduct the study under diffuse light and to include an inoculum of suspended sediment. The present study was designed to investigate the potential impact of these options on the degradation rate of a representative compound. The fungicide, isopyrazam, was chosen as it was previously shown to be susceptible to metabolism by phototrophic organisms under a fluorescent light-dark cycle. The impact of diffuse light was investigated at light intensities representative of those at depth in large, open water bodies (test compound [DT50] < 50 d), whereas degradation in continuous darkness was negligible. Furthermore, investigation at 2 different light intensities resulted in similar degradation rates, indicating that this transformation mechanism was not proportional to light intensity, provided that there was sufficient light for photosynthesis to occur. Inclusion of suspended sediment did not have a significant impact on the degradation rate of isopyrazam, except at extremely high sediment concentrations, which were not considered representative of conditions in large, open water bodies.

  18. Unattended Monitoring System Design Methodology

    Energy Technology Data Exchange (ETDEWEB)

    Drayer, D.D.; DeLand, S.M.; Harmon, C.D.; Matter, J.C.; Martinez, R.L.; Smith, J.D.

    1999-07-08

    A methodology for designing Unattended Monitoring Systems starting at a systems level has been developed at Sandia National Laboratories. This proven methodology provides a template that describes the process for selecting and applying appropriate technologies to meet unattended system requirements, as well as providing a framework for development of both training courses and workshops associated with unattended monitoring. The design and implementation of unattended monitoring systems is generally intended to respond to some form of policy based requirements resulting from international agreements or domestic regulations. Once the monitoring requirements are established, a review of the associated process and its related facilities enables identification of strategic monitoring locations and development of a conceptual system design. The detailed design effort results in the definition of detection components as well as the supporting communications network and data management scheme. The data analyses then enables a coherent display of the knowledge generated during the monitoring effort. The resultant knowledge is then compared to the original system objectives to ensure that the design adequately addresses the fundamental principles stated in the policy agreements. Implementation of this design methodology will ensure that comprehensive unattended monitoring system designs provide appropriate answers to those critical questions imposed by specific agreements or regulations. This paper describes the main features of the methodology and discusses how it can be applied in real world situations.

  19. Guide to NRC reporting and recordkeeping requirements. Compiled from requirements in Title 10 of the U.S. Code of Federal Regulations as codified on December 31, 1993; Revision 1

    Energy Technology Data Exchange (ETDEWEB)

    Collins, M.; Shelton, B.

    1994-07-01

    This compilation includes in the first two sections the reporting and recordkeeping requirements applicable to US Nuclear Regulatory Commission (NRC) licensees and applicants and to members of the public. It includes those requirements codified in Title 10 of the code of Federal Regulations, Chapter 1, on December 31, 1993. It also includes, in a separate section, any of those requirements that were superseded or discontinued between January 1992 and December 1993. Finally, the appendix lists mailing and delivery addresses for NRC Headquarters and Regional Offices mentioned in the compilation. The Office of Information Resources Management staff compiled this listing of reporting and recordkeeping requirements to briefly describe each in a single document primarily to help licensees readily identify the requirements. The compilation is not a substitute for the regulations, and is not intended to impose any new requirements or technical positions. It is part of NRC`s continuing efforts to comply with the Paperwork Reduction Act of 1980 and the Office of Management and Budget regulations that mandate effective and efficient Federal information resources management programs.

  20. 40 CFR 60.4345 - What are the requirements for the continuous emission monitoring system equipment, if I choose to...

    Science.gov (United States)

    2010-07-01

    ... choose to use this option? If the option to use a NOX CEMS is chosen: (a) Each NOX diluent CEMS must be..., Procedure 1 in appendix F to this part is not required. Alternatively, a NOX diluent CEMS that is installed... subpart. The relative accuracy test audit (RATA) of the CEMS shall be performed on a lb/MMBtu basis....

  1. Nuclear localization of the transcriptional regulator MIER1α requires interaction with HDAC1/2 in breast cancer cells.

    Directory of Open Access Journals (Sweden)

    Shengnan Li

    Full Text Available MIER1α is a transcriptional regulator that functions in gene repression through its ability to interact with various chromatin modifiers and transcription factors. We have also shown that MIER1α interacts with ERα and inhibits estrogen-stimulated growth. While MIER1α is localized in the nucleus of MCF7 cells, previous studies have shown that it does not contain a nuclear localization signal. In this report, we investigate the mechanism involved in transporting MIER1α into the nucleus. We explored the possibility that MIER1α is transported into the nucleus through a 'piggyback' mechanism. One obvious choice is via interaction with ERα, however we demonstrate that nuclear targeting of MIER1α does not require ERα. Knockdown of ERα reduced protein expression to 22% of control, but did not alter the percentage of cells with nuclear MIER1α (98% nuclear with scrambled shRNA vs. 95% with ERα shRNA. Further evidence was obtained using two stable transfectants derived from the ER-negative MDA231 cell line: MC2 (ERα+ and VC5 (ERα-. Confocal analysis showed no difference in MIER1α localization (86% nuclear in MC2 vs. 89% in VC5. These data demonstrate that ERα is not involved in nuclear localization of MIER1α. To identify the critical MIER1α sequence, we performed a deletion analysis and determined that the ELM2 domain was necessary and sufficient for nuclear localization. This domain binds HDAC1 & 2, therefore we investigated their role. Confocal analysis of an MIER1α containing an ELM2 point mutation previously shown to abolish HDAC binding revealed that this mutation results in almost complete loss of nuclear targeting: 10% nuclear vs. 97% with WT-MIER1α. Moreover, double knockdown of HDAC1 and 2 caused a reduction in percent nuclear from 86% to 44%. The results of this study demonstrate that nuclear targeting of MIER1α requires an intact ELM2 domain and is dependent on interaction with HDAC1/2.

  2. RCN1-regulated phosphatase activity and EIN2 modulate hypocotyl gravitropism by a mechanism that does not require ethylene signaling.

    Science.gov (United States)

    Muday, Gloria K; Brady, Shari R; Argueso, Cristiana; Deruère, Jean; Kieber, Joseph J; DeLong, Alison

    2006-08-01

    The roots curl in naphthylphthalamic acid1 (rcn1) mutant of Arabidopsis (Arabidopsis thaliana) has altered auxin transport, gravitropism, and ethylene response, providing an opportunity to analyze the interplay between ethylene and auxin in control of seedling growth. Roots of rcn1 seedlings were previously shown to have altered auxin transport, growth, and gravitropism, while rcn1 hypocotyl elongation exhibited enhanced ethylene response. We have characterized auxin transport and gravitropism phenotypes of rcn1 hypocotyls and have explored the roles of auxin and ethylene in controlling these phenotypes. As in roots, auxin transport is increased in etiolated rcn1 hypocotyls. Hypocotyl gravity response is accelerated, although overall elongation is reduced, in etiolated rcn1 hypocotyls. Etiolated, but not light grown, rcn1 seedlings also overproduce ethylene, and mutations conferring ethylene insensitivity restore normal hypocotyl elongation to rcn1. Auxin transport is unaffected by treatment with the ethylene precursor 1-aminocyclopropane carboxylic acid in etiolated hypocotyls of wild-type and rcn1 seedlings. Surprisingly, the ethylene insensitive2-1 (ein2-1) and ein2-5 mutations dramatically reduce gravitropic bending in hypocotyls. However, the ethylene resistant1-3 (etr1-3) mutation does not significantly affect hypocotyl gravity response. Furthermore, neither the etr1 nor the ein2 mutation abrogates the accelerated gravitropism observed in rcn1 hypocotyls, indicating that both wild-type gravity response and enhanced gravity response in rcn1 do not require an intact ethylene-signaling pathway. We therefore conclude that the RCN1 protein affects overall hypocotyl elongation via negative regulation of ethylene synthesis in etiolated seedlings, and that RCN1 and EIN2 modulate hypocotyl gravitropism and ethylene responses through independent pathways.

  3. Sterol regulation of human fatty acid synthase promoter I requires nuclear factor-Y- and Sp-1-binding sites.

    Science.gov (United States)

    Xiong, S; Chirala, S S; Wakil, S J

    2000-04-11

    To understand cholesterol-mediated regulation of human fatty acid synthase promoter I, we tested various 5'-deletion constructs of promoter I-luciferase reporter gene constructs in HepG2 cells. The reporter gene constructs that contained only the Sp-1-binding site (nucleotides -82 to -74) and the two tandem sterol regulatory elements (SREs; nucleotides -63 to -46) did not respond to cholesterol. Only the reporter gene constructs containing a nuclear factor-Y (NF-Y) sequence, the CCAAT sequence (nucleotides -90 to -86), an Sp-1 sequence, and the two tandem SREs responded to cholesterol. The NF-Y-binding site, therefore, is essential for cholesterol response. Mutating the SREs or the NF-Y site and inserting 4 bp between the Sp-1- and NF-Y-binding sites both resulted in a minimal cholesterol response of the reporter genes. Electrophoretic mobility-shift assays using anti-SRE-binding protein (SREBP) and anti-NF-Ya antibodies confirmed that these SREs and the NF-Y site bind the respective factors. We also identified a second Sp-1 site located between nucleotides -40 and -30 that can substitute for the mutated Sp-1 site located between nucleotides -82 and -74. The reporter gene expression of the wild-type promoter and the Sp-1 site (nucleotides -82 to -74) mutant promoter was similar when SREBP1a [the N-terminal domain of SREBP (amino acids 1-520)] was constitutively overexpressed, suggesting that Sp-1 recruits SREBP to the SREs. Under the same conditions, an NF-Y site mutation resulted in significant loss of reporter gene expression, suggesting that NF-Y is required to activate the cholesterol response.

  4. Cellular zinc is required for intestinal epithelial barrier maintenance via the regulation of claudin-3 and occludin expression.

    Science.gov (United States)

    Miyoshi, Yuka; Tanabe, Soichi; Suzuki, Takuya

    2016-07-01

    Intracellular zinc is required for a variety of cell functions, but its precise roles in the maintenance of the intestinal tight junction (TJ) barrier remain unclear. The present study investigated the essential roles of intracellular zinc in the preservation of intestinal TJ integrity and the underlying molecular mechanisms. Depletion of intracellular zinc in both intestinal Caco-2 cells and mouse colons through the application of a cell-permeable zinc chelator N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) induced a disruption of the TJ barrier, as indicated by increased FITC-labeled dextran flux and decreased transepithelial electrical resistance. The TPEN-induced TJ disruption is associated with downregulation of two TJ proteins, occludin and claudin-3. Biotinylation of cell surface proteins revealed that the zinc depletion induced the proteolysis of occludin but not claudin-3. Occludin proteolysis was sensitive to the inhibition of calpain activity, and increased calpain activity was observed in the zinc-depleted cells. Although quantitative PCR analysis and promoter reporter assay have demonstrated that the zinc depletion-induced claudin-3 downregulation occurred at transcriptional levels, a site-directed mutation in the egr1 binding site in the claudin-3 promoter sequence induced loss of both the basal promoter activity and the TPEN-induced decreases. Reduced egr1 expression by a specific siRNA also inhibited claudin-3 expression and transepithelial electrical resistance maintenance in cells. This study shows that intracellular zinc has an essential role in the maintenance of the intestinal epithelial TJ barrier through regulation of occludin proteolysis and claudin-3 transcription.

  5. Requirements and Implementation Feasibility for a CubeSat Thermal Infrared Imaging System to Monitor the Structure of Volcanic Ash Clouds

    Science.gov (United States)

    Thorsen, D.; Carroll, R.; Webley, P.; Hawkins, J.

    2014-12-01

    The 2010 eruption of the Eyjafjallajökull volcano in Iceland caused the cancellation of approximately 108,000 flights over an 8-day period, disrupted air traffic worldwide, and cost the airline industry more than $400 million per day. The inconvenience and economic impact of this and similar events, such as Puyehue-Cordon-Caulle in 2011, have heightened the interest in developing improved satellite remote sensing techniques for monitoring volcanic plumes and drifting clouds. For aviation safety, the operational/research community has started to move towards classifying the concentrations within volcanic plumes and clouds. Additionally, volcanic ash transport and dispersion (VATD) models are often used for forecasting ash cloud locations and they require knowledge of the structure of the erupting column to improve their ash simulations and also downwind 3-D maps of the ash cloud to calibrate/validate their modeling output. Existing remote sensing satellites utilize a brightness temperature method with thermal infrared (TIR) measurements from 10 - 12 μm to determine mass loading of volcanic ash along a single line of sight, but they have infrequent revisit times and they cannot resolve the three-dimensional structure of the ash clouds. A cluster of CubeSats dedicated to the monitoring of volcanic ash and plumes could provide both more frequent updates and the multi-aspect images needed to resolve the density structure of volcanic ash clouds and plumes. In this presentation, we discuss the feasibility and requirements for a CubeSat TIR imaging system and the associated on-board image processing that would be required to monitor the structure of volcanic ash clouds from Low Earth Orbit.

  6. Remote sensing in precision farming: real-time monitoring of water and fertilizer requirements of agricultural crops

    Science.gov (United States)

    Zilberman, Arkadi; Ben Asher, Jiftah; Kopeika, Norman S.

    2016-10-01

    The advancements in remote sensing in combination with sensor technology (both passive and active) enable growers to analyze an entire crop field as well as its local features. In particular, changes of actual evapo-transpiration (ET) as a function of water availability can be measured remotely with infrared radiometers. Detection of crop water stress and ET and combining it with the soil water flow model enable rational irrigation timing and application amounts. Nutrient deficiency, and in particular nitrogen deficiency, causes substantial crop losses. This deficiency needs to be identified immediately. A faster the detection and correction, a lesser the damage to the crop yield. In the present work, to retrieve ET a novel deterministic approach was used which is based on the remote sensing data. The algorithm can automatically provide timely valuable information on plant and soil water status, which can improve the management of irrigated crops. The solution is capable of bridging between Penman-Monteith ET model and Richards soil water flow model. This bridging can serve as a preliminary tool for expert irrigation system. To support decisions regarding fertilizers the greenness of plant canopies is assessed and quantified by using the spectral reflectance sensors and digital color imaging. Fertilization management can be provided on the basis of sampling and monitoring of crop nitrogen conditions using RS technique and translating measured N concentration in crop to kg/ha N application in the field.

  7. Around-the-clock ambulatory blood pressure monitoring is required to properly diagnose resistant hypertension and assess associated vascular risk.

    Science.gov (United States)

    Hermida, Ramón C; Ayala, Diana E; Ríos, María T; Fernández, José R; Mojón, Artemio; Smolensky, Michael H

    2014-07-01

    Diagnosis of resistant hypertension (RH) is currently based upon awake-time office blood pressure (BP). An increasing number of studies have documented abnormally elevated sleep-time BP in most RH patients, indicating that diagnosis of true RH cannot be determined solely by comparison of office BP with either patient awake-time BP self-measurements or awake-BP mean from ambulatory monitoring (ABPM), as is customary in the published literature. Moreover, the ABPM-determined sleep-time BP mean is an independent and stronger predictor of cardiovascular and cerebrovascular disease (CVD) risk than either daytime office/ABPM-derived awake or 24-hour means. Results of the recently completed MAPEC (Monitorización Ambulatoria para Predicción de Eventos Cardiovasculares) prospective outcomes study, which included a large cohort of RH patients, established that time of treatment relative to circadian rhythms constituted a critically important yet often neglected variable with respect to BP control. The study found that bedtime versus morning ingestion of the full dose of ≥1 BP-lowering medications resulted in both better therapeutic normalization of sleep-time BP and reduced CVD morbidity and mortality, including in RH patients. Accordingly, ABPM is highly recommended to properly diagnose and manage true RH, with a bedtime hypertension medication regimen as the therapeutic scheme of choice.

  8. Endothelial-derived tissue factor pathway inhibitor regulates arterial thrombosis but is not required for development or hemostasis.

    NARCIS (Netherlands)

    White, T.A.; Johnson, T.; Zarzhevsky, N.; Tom, C.; Delacroix, S.; Holroyd, E.W.; Maroney, S.A.; Singh, R.; Pan, S.; Fay, W.P.; Deursen, J.M.A. van; Mast, A.E.; Sandhu, G.S.; Simari, R.D.

    2010-01-01

    The antithrombotic surface of endothelium is regulated in a coordinated manner. Tissue factor pathway inhibitor (TFPI) localized at the endothelial cell surface regulates the production of FXa by inhibiting the TF/VIIa complex. Systemic homozygotic deletion of the first Kunitz (K1) domain of TFPI

  9. The chemotaxis regulator pilG of Xylella fastidiosa is required for virulence in Vitis vinifera grapevines

    Science.gov (United States)

    Xylella fastidiosa is a Gram-negative, xylem-limited pathogenic bacterium that causes Pierce’s disease of grapevines. Type IV pili of X. fastidiosa are regulated by pilG, a chemotaxis regulator in Pil-Chp operon involving signal transduction pathways. To elucidate the role of pilG in twitching motil...

  10. ArgR and AhrC are both required for regulation of arginine metabolism in Lactococcus lactis

    NARCIS (Netherlands)

    Larsen, R; Buist, G; Kuipers, OP; Kok, J

    2004-01-01

    The DNA binding proteins ArgR and AhrC are essential for regulation of arginine metabolism in Escherichia Coli and Bacillus subtilis, respectively. A unique property of these regulators is that they form hexameric protein complexes, mediating repression of arginine biosynthetic pathways as well as a

  11. The chemotaxis regulator pilG of Xylella fastidiosa is required for virulence in Vitis vinifera grapevines

    Science.gov (United States)

    Type IV pili of X. fastidiosa are regulated by pilG, a response regulator protein putatively involved in chemotaxis-like operon sensing stimuli through signal transduction pathways. To elucidate roles of pilG in pathogenicity of X. fastidiosa, the pilG-deletion mutant and complementary strain contai...

  12. Monitoring the inputs required to extend and sustain hygiene promotion: findings from the GLAAS 2013/2014 survey.

    Science.gov (United States)

    Moreland, Leslie D; Gore, Fiona M; Andre, Nathalie; Cairncross, Sandy; Ensink, Jeroen H J

    2016-08-01

    There are significant gaps in information about the inputs required to effectively extend and sustain hygiene promotion activities to improve people's health outcomes through water, sanitation and hygiene (WASH) interventions. We sought to analyse current country and global trends in the use of key inputs required for effective and sustainable implementation of hygiene promotion to help guide hygiene promotion policy and decision-making after 2015. Data collected in response to the GLAAS 2013/2014 survey from 93 countries of 94 were included, and responses were analysed for 12 questions assessing the inputs and enabling environment for hygiene promotion under four thematic areas. Data were included and analysed from 20 External Support Agencies (ESA) of 23 collected through self-administered surveys. Firstly, the data showed a large variation in the way in which hygiene promotion is defined and what constitutes key activities in this area. Secondly, challenges to implement hygiene promotion are considerable: include poor implementation of policies and plans, weak coordination mechanisms, human resource limitations and a lack of available hygiene promotion budget data. Despite the proven benefits of hand washing with soap, a critical hygiene-related factor in minimising infection, GLAAS 2013/2014 survey data showed that hygiene promotion remains a neglected component of WASH. Additional research to identify the context-specific strategies and inputs required to enhance the effectiveness of hygiene promotion at scale are needed. Improved data collection methods are also necessary to advance the availability and reliability of hygiene-specific information. © 2016 John Wiley & Sons Ltd.

  13. Conforming STOP Violence Against Women Formula Grant Program Regulations to Statutory Change; Definitions and Confidentiality Requirements Applicable to All OVW Grant Programs. Final rule.

    Science.gov (United States)

    2016-11-29

    This rule amends the regulations for the STOP (ServicesTrainingOfficersProsecutors) Violence Against Women Formula Grant Program (STOP Program) and the general provisions governing Office on Violence Against Women (OVW) programs to comply with statutory changes and reduce repetition of statutory language. Also, this rule implements statutory requirements for nondisclosure of confidential or private information relating to all OVW grant programs.

  14. FUNCTIONS AND REQUIREMENTS FOR RUSSIAN PULSATING MONITOR DEPLOYMENT IN THE GUNITE AND ASSOCIATED TANKS AT OAK RIDGE NATIONAL LABORATORY

    Energy Technology Data Exchange (ETDEWEB)

    Thomas Albert

    1999-01-01

    This document provides functions and requirements to support deployment of pulsating mixer pump technology in the Oak Ridge National Laboratory (ORNL) Gunite and Associated Tanks to mobilize and mix the settled sludge and solids in these tanks. In FY 1998 pulsating mixer pump technology, a jet mixer powered by a reciprocating air supply, was selected for FY 1999 deployment in one of the GAAT tanks to mobilize settled solids. Pulsating mixer pump technology was identified in FY 1996 during technical exchanges between the US Department of Energy (DOE) Tanks Focus Area Retrieval and Closure program, the DOE Environmental Management International Programs, and delegates from Russia as a promising technology that could be implemented in the US. The pulsating mixer pump technology, provided by the Russian Integrated Mining Chemical Company, was tested at Pacific Northwest National Laboratory (PNNL) to observe its ability to mobilize settled solids. Based on the results of this demonstration, ORNL and DOE staff determined that a modified pulsating mixer pump would meet project needs for bulk mobilization of Gunite tank sludge prior to deployment of other retrieval systems. The deployment of this device is expected to significantly reduce the costs of operation and maintenance of more expensive retrieval systems. The functions and requirements presented here were developed by evaluating the results and recommendations that resulted from the pulsating mixer pump demonstration at PNNL, and by coupling this with the remediation needs identified by staff at ORNL involved with the remediation of the Gunite and Associated Tanks.

  15. VirS, an OmpR/PhoB subfamily response regulator, is required for activation of vapA gene expression in Rhodococcus equi.

    Science.gov (United States)

    Kakuda, Tsutomu; Hirota, Takuya; Takeuchi, Tatsuya; Hagiuda, Hirofumi; Miyazaki, Shiko; Takai, Shinji

    2014-10-03

    Rhodococcus equi is an important pulmonary pathogen in foals and in immunocompromised individuals. Virulent R. equi strains carry an 80-90 kb virulence plasmid that expresses the virulence-associated protein A (VapA). VapA expression is regulated by temperature and pH. The LysR-type transcriptional regulator, VirR, is involved in the regulation of the vapA gene. To examine the mechanism underlying transcriptional regulation of vapA, we characterized an R. equi mutant in which another putative transcriptional regulator encoded on the virulence plasmid, VirS, was deleted. Deletion of virS reduced vapA promoter activity to non-inducible levels. Complementary expression of VirS in the virS deletion mutant restored transcription at the PvapA promoter, even under non-inducing conditions (30°C and pH 8.0). In addition, VirS expression increased PvapA promoter activity in the absence of functional VirR. Further, transcription of the icgA operon containing virS was regulated by pH and temperature in the same manner as vapA. This study suggests that VirS is required for VapA expression and that regulation of PvapA-promoter activity may be achieved by controlling VirS expression levels.

  16. Requirement of regulated endocrine-specific protein-18 for development and expression of regulated endocrine-specific protein-18 isoform c in mice.

    Science.gov (United States)

    Liang, Min; Yang, Jian Ling; Bian, Min Juan; Liu, Jie; Hong, Xiao Qi; Wang, Yan Cong; Huang, Yu Fang; Gu, Shu Ping; Yu, Mei; Huang, Fang; Fei, Jian

    2011-04-01

    Regulated endocrine-specific protein-18 (RESP18) is distributed mainly in the peripheral endocrine and neuroendocrine tissues. The expression of RESP18 protein is regulated by physiological factors, such as blood glucose or dopaminergic drugs, but its functions remain unclear. In this study, to explore the biological functions of RESP18 in vivo, we generated RESP18 heterozygous deficient mice, and further found RESP18 was essential for embryonic development. In addition, we cloned a new isoform of mouse RESP18 by reverse transcription-polymerase chain reaction (RT-PCR), and denominated it as RESP18-c. Mouse RESP18-c, by skipping exon4 (43 bp in length), encodes a shorter protein of 120 amino acid residues. The distribution of RESP18-c mRNA is similar with that of RESP18 mRNA in the peripheral tissues and brains of mice.

  17. Growth Arrest Specific 2 Is Up-Regulated in Chronic Myeloid Leukemia Cells and Required for Their Growth

    OpenAIRE

    Haixia Zhou; Yue Ge; Lili Sun; Wenjuan Ma; Jie Wu; Xiuyan Zhang; Xiaohui Hu; Eaves, Connie J; Depei Wu; Yun Zhao

    2014-01-01

    Although the generation of BCR-ABL is the molecular hallmark of chronic myeloid leukemia (CML), the comprehensive molecular mechanisms of the disease remain unclear yet. Growth arrest specific 2 (GAS2) regulates multiple cellular functions including cell cycle, apoptosis and calpain activities. In the present study, we found GAS2 was up-regulated in CML cells including CD34+ progenitor cells compared to their normal counterparts. We utilized RNAi and the expression of dominant negative form o...

  18. Balancing spatially regulated β-actin translation and dynamin-mediated endocytosis is required to assemble functional epithelial monolayers.

    Science.gov (United States)

    Cruz, Lissette A; Vedula, Pavan; Gutierrez, Natasha; Shah, Neel; Rodriguez, Steven; Ayee, Brian; Davis, Justin; Rodriguez, Alexis J

    2015-12-01

    Regulating adherens junction complex assembly/disassembly is critical to maintaining epithelial homeostasis in healthy epithelial tissues. Consequently, adherens junction structure and function is often perturbed in clinically advanced tumors of epithelial origin. Some of the most studied factors driving adherens junction complex perturbation in epithelial cancers are transcriptional and epigenetic down-regulation of E-cadherin expression. However, numerous reports demonstrate that post-translational regulatory mechanisms such as endocytosis also regulate early phases of epithelial-mesenchymal transition and metastatic progression. In already assembled healthy epithelia, E-cadherin endocytosis recycles cadherin-catenin complexes to regulate the number of mature adherens junctions found at cell-cell contact sites. However, following de novo epithelial cell-cell contact, endocytosis negatively regulates adherens junction assembly by removing E-cadherin from the cell surface. By contrast, following de novo epithelial cell-cell contact, spatially localized β-actin translation drives cytoskeletal remodeling and consequently E-cadherin clustering at cell-cell contact sites and therefore positively regulates adherens junction assembly. In this report we demonstrate that dynamin-mediated endocytosis and β-actin translation-dependent cadherin-catenin complex anchoring oppose each other following epithelial cell-cell contact. Consequently, the final extent of adherens junction assembly depends on which of these processes is dominant following epithelial cell-cell contact. We expressed β-actin transcripts impaired in their ability to properly localize monomer synthesis (Δ3'UTR) in MDCK cells to perturb actin filament remodeling and anchoring, and demonstrate the resulting defect in adherens junction structure and function is rescued by inhibiting dynamin mediated endocytosis. Therefore, we demonstrate balancing spatially regulated β-actin translation and dynamin

  19. Establishing Bedding Requirements during Transport and Monitoring Skin Temperature during Cold and Mild Seasons after Transport for Finishing Pigs

    Directory of Open Access Journals (Sweden)

    John McGlone

    2014-05-01

    Full Text Available The broad aim of this study was to determine whether bedding level in the transport trailer influenced pig performance and welfare. Specifically, the objective was to define the bedding requirements of pigs during transportation in commercial settings during cold and mild weather. Animals (n = 112,078 pigs on 572 trailers used were raised in commercial finishing sites and transported in trailers to commercial processing plants. Dead on arrival (DOA, non-ambulatory (NA, and total dead and down (D&D data were collected and skin surface temperatures of the pigs were measured by infrared thermography. Data were collected during winter (Experiment 1 and fall/spring (Experiment 2. Total D&D percent showed no interaction between bedding level and outside air temperature in any experiments. Average skin surface temperature during unloading increased with outside air temperature linearly in both experiments (P < 0.01. In conclusion, over-use of bedding may be economically inefficient. Pig skin surface temperature could be a useful measure of pig welfare during or after transport.

  20. Technical requirements for implementation of an individual monitoring service for evaluation of operational quantity HP(10) using thermoluminescent dosimetry; Requisitos tecnicos para a implantacao de um servico de monitoracao individual externa de corpo inteiro para fotons utilizando dosimetria termoluminescente

    Energy Technology Data Exchange (ETDEWEB)

    Francisco, Adelaide Benedita Armando

    2016-11-01

    This work aims to establish technical requirements for the development of a TLDs system for the assessment of operational quantity H{sub P}(10), in order to implement an external individual monitoring service in countries who do not have. This allows a better understanding of the technic and the thermoluminescent dosimetry system, thus contributing to identify the technical criteria to be followed by a dosimetry laboratory and evaluation of the dosimetric system performance. For this, the review of the specific literature of the dosimetry field was conducted and later the type and performance tests that must be followed by a dosimetric system were reproduced in practice. In additional was made a analysis of internationals standards norms and the technical regulation used in Brazil, to define the essentials type testes to a dosimetric system. To check the performance of a dosimetry system, a performance analysis of the Brazilian TLDs system was carried out over the past 6 years using the trumpet curve, where it was observed that most of TLDs system, in this review period, were approved and have excellent performance. The technical requirements for the development of a thermoluminescent dosimetry system ensure that the system provides technically reliable results and allow demonstration of compliance with the standard criteria established by national and international standards, and the implementation of the dosimetry system, is verified the compliance of the annual doses limits set for occupationally exposed. (author)

  1. Adjustment of Energy requirements in TEK; TEK= Technical Regulations under the Norwegian Planning and Building Act; Justering av energikrav i TEK

    Energy Technology Data Exchange (ETDEWEB)

    Thyholt, Marit; Dokka, Tor Helge; Schild, Peter; Grini, Catherine; Mysen, Mads; Sartori, Igor

    2008-07-01

    The National Office of Building Technology and Administration (BE) desired to review the consequences of different levels of ambition for requirements for heat gaining from vent air, as well as a possible requirement for energy efficient design of building fronts. In addition the energy scope in the regulation (TEK2007) should be adjusted according to the final establishment of a new calculation standard (Norwegian Standard - NS 3031:2007). A statement on these subjects has been carried out at SINTEF Byggforsk, and is described in this report. Adjustments of framework regulations.There are only minor differences between adjusted calculations according to NS 3031 and the original energy framework calculations, i.e. the difference for net energy need amounts to the size of 0 to 6 percent. Heat gain.The report shows that it is possible - both from techical and financial considerations - to increase the requirement level for heat gain from vent air for most categories of buildings. This implies a sharpening of the annual median temperature efficiency from 70 % to 80 %, for all building categories, except from hospitals, institutions and light industry/workshops. A possible sharpening of regulations for heat gain in houses has not been evaluated. Depending on building category a sharpening of regulations for heat gain from vent air will imply that net energy need will be reduced on a scale of 20 to 30 kWh/m2 per annum. The report demonstrates that despite a possible sharpening of the requirements on energy efficiency for heat recovery devices does not prevent the use of large areas of windows and window panes. Vulnerability analyses show that deviations from the prerequisites in the basis for the energy framework concerning air quantities and air temperatures give the possibility of weakening the building's heating characteristics. Building fronts. Different methods for added requirements for building fronts have been examined. The aim has been to find methods and

  2. Requirements for an Advanced Low Earth Orbit (LEO) Sounder (ALS) for Improved Regional Weather Prediction and Monitoring of Greenhouse Gases

    Science.gov (United States)

    Pagano, Thomas S.; Chahine, Moustafa T.; Susskind, Joel

    2008-01-01

    Hyperspectral infrared atmospheric sounders (e.g., the Atmospheric Infrared Sounder (AIRS) on Aqua and the Infrared Atmospheric Sounding Interferometer (IASI) on Met Op) provide highly accurate temperature and water vapor profiles in the lower to upper troposphere. These systems are vital operational components of our National Weather Prediction system and the AIRS has demonstrated over 6 hrs of forecast improvement on the 5 day operational forecast. Despite the success in the mid troposphere to lower stratosphere, a reduction in sensitivity and accuracy has been seen in these systems in the boundary layer over land. In this paper we demonstrate the potential improvement associated with higher spatial resolution (1 km vs currently 13.5 km) on the accuracy of boundary layer products with an added consequence of higher yield of cloud free scenes. This latter feature is related to the number of samples that can be assimilated and has also shown to have a significant impact on improving forecast accuracy. We also present a set of frequencies and resolutions that will improve vertical resolution of temperature and water vapor and trace gas species throughout the atmosphere. Development of an Advanced Low Earth Orbit (LEO) Sounder (ALS) with these improvements will improve weather forecast at the regional scale and of tropical storms and hurricanes. Improvements are also expected in the accuracy of the water vapor and cloud properties products, enhancing process studies and providing a better match to the resolution of future climate models. The improvements of technology required for the ALS are consistent with the current state of technology as demonstrated in NASA Instrument Incubator Program and NOAA's Hyperspectral Environmental Suite (HES) formulation phase development programs.

  3. Requirements for an Advanced Low Earth Orbit (LEO) Sounder (ALS) for improved regional weather prediction and monitoring of greenhouse gases

    Science.gov (United States)

    Pagano, Thomas S.; Chahine, Moustafa T.; Susskind, Joel

    2008-12-01

    Hyperspectral infrared atmospheric sounders (e.g. the Atmospheric Infrared Sounder (AIRS) on Aqua and the Infrared Atmospheric Sounding Interferometer (IASI) on MetOp) provide highly accurate temperature and water vapor profiles in the lower to upper troposphere. These systems are vital operational components of our National Weather Prediction system and the AIRS has demonstrated over 6 hrs of forecast improvement on the 5 day operational forecast1. Despite the success in the mid troposphere to lower stratosphere, a reduction in sensitivity and accuracy has been seen in these systems in the boundary layer over land. In this paper we demonstrate the potential improvement associated with higher spatial resolution (1km vs currently 13.5 km) on the accuracy of boundary layer products with an added consequence of higher yield of cloud free scenes. This latter feature is related to the number of samples that can be assimilated and has also shown to have a significant impact on improving forecast accuracy. We also present a set of frequencies and resolutions that will improve vertical resolution of temperature and water vapor and trace gas species throughout the atmosphere. Development of an Advanced Low Earth Orbit (LEO) Sounder (ALS) with these improvements will improve weather forecast at the regional scale and of tropical storms and hurricanes. Improvements are also expected in the accuracy of the water vapor and cloud properties products, enhancing process studies and providing a better match to the resolution of future climate models. The improvements of technology required for the ALS are consistent with the current state of technology as demonstrated in NASA Instrument Incubator Program and NOAA's Hyperspectral Environmental Suite (HES) formulation phase development programs.

  4. Monitoring madness

    Energy Technology Data Exchange (ETDEWEB)

    Blankinship, S.

    2006-01-15

    High quality continuous emission monitoring capability can be as essential as high quality emission control equipment. Future mercury monitoring and control requirements add to the justification for better CEMS. The article discusses two prominent mercury measurement methods - the cold vapour atomic absorptive spectrometer (CVAAs) and the atomic absorptive spectrometer (AFS). It stresses the importance of maintaining a CEMS. 1 photo.

  5. Clavibacter michiganensis subsp. michiganensis Vatr1 and Vatr2 transcriptional regulators are required for virulence in tomato.

    Science.gov (United States)

    Savidor, Alon; Chalupowicz, Laura; Teper, Doron; Gartemann, Karl-Heinz; Eichenlaub, Rudolf; Manulis-Sasson, Shulamit; Barash, Isaac; Sessa, Guido

    2014-10-01

    The plant pathogen Clavibacter michiganensis subsp. michiganensis is a gram-positive bacterium responsible for wilt and canker disease of tomato. Although disease development is well characterized and diagnosed, molecular mechanisms of C. michiganensis subsp. michiganensis virulence are poorly understood. Here, we identified and characterized two C. michiganensis subsp. michiganensis transcriptional regulators, Vatr1 and Vatr2, that are involved in pathogenicity of C. michiganensis subsp. michiganensis. Vatr1 and Vatr2 belong to TetR and MocR families of transcriptional regulators, respectively. Mutations in their corresponding genes caused attenuated virulence, with the Δvatr2 mutant showing a more dramatic effect than Δvatr1. Although both mutants grew well in vitro and reached a high titer in planta, they caused reduced wilting and canker development in infected plants compared with the wild-type bacterium. They also led to a reduced expression of the ethylene-synthesizing tomato enzyme ACC-oxidase compared with wild-type C. michiganensis subsp. michiganensis and to reduced ethylene production in the plant. Transcriptomic analysis of wild-type C. michiganensis subsp. michiganensis and the two mutants under infection-mimicking conditions revealed that Vatr1 and Vatr2 regulate expression of virulence factors, membrane and secreted proteins, and signal-transducing proteins. A 70% overlap between the sets of genes positively regulated by Vatr1 and Vatr2 suggests that these transcriptional regulators are on the same molecular pathway responsible for C. michiganensis subsp. michiganensis virulence.

  6. JMJD-5/KDM8 regulates H3K36me2 and is required for late steps of homologous recombination and genome integrity

    DEFF Research Database (Denmark)

    Amendola, Pier Giorgio; Zaghet, Nico; Ramalho, João J

    2017-01-01

    The eukaryotic genome is organized in a three-dimensional structure called chromatin, constituted by DNA and associated proteins, the majority of which are histones. Post-translational modifications of histone proteins greatly influence chromatin structure and regulate many DNA-based biological...... processes. Methylation of lysine 36 of histone 3 (H3K36) is a post-translational modification functionally relevant during early steps of DNA damage repair. Here, we show that the JMJD-5 regulates H3K36 di-methylation and it is required at late stages of double strand break repair mediated by homologous...

  7. Wdpcp, a PCP protein required for ciliogenesis, regulates directional cell migration and cell polarity by direct modulation of the actin cytoskeleton.

    Science.gov (United States)

    Cui, Cheng; Chatterjee, Bishwanath; Lozito, Thomas P; Zhang, Zhen; Francis, Richard J; Yagi, Hisato; Swanhart, Lisa M; Sanker, Subramaniam; Francis, Deanne; Yu, Qing; San Agustin, Jovenal T; Puligilla, Chandrakala; Chatterjee, Tania; Tansey, Terry; Liu, Xiaoqin; Kelley, Matthew W; Spiliotis, Elias T; Kwiatkowski, Adam V; Tuan, Rocky; Pazour, Gregory J; Hukriede, Neil A; Lo, Cecilia W

    2013-11-01

    Planar cell polarity (PCP) regulates cell alignment required for collective cell movement during embryonic development. This requires PCP/PCP effector proteins, some of which also play essential roles in ciliogenesis, highlighting the long-standing question of the role of the cilium in PCP. Wdpcp, a PCP effector, was recently shown to regulate both ciliogenesis and collective cell movement, but the underlying mechanism is unknown. Here we show Wdpcp can regulate PCP by direct modulation of the actin cytoskeleton. These studies were made possible by recovery of a Wdpcp mutant mouse model. Wdpcp-deficient mice exhibit phenotypes reminiscent of Bardet-Biedl/Meckel-Gruber ciliopathy syndromes, including cardiac outflow tract and cochlea defects associated with PCP perturbation. We observed Wdpcp is localized to the transition zone, and in Wdpcp-deficient cells, Sept2, Nphp1, and Mks1 were lost from the transition zone, indicating Wdpcp is required for recruitment of proteins essential for ciliogenesis. Wdpcp is also found in the cytoplasm, where it is localized in the actin cytoskeleton and in focal adhesions. Wdpcp interacts with Sept2 and is colocalized with Sept2 in actin filaments, but in Wdpcp-deficient cells, Sept2 was lost from the actin cytoskeleton, suggesting Wdpcp is required for Sept2 recruitment to actin filaments. Significantly, organization of the actin filaments and focal contacts were markedly changed in Wdpcp-deficient cells. This was associated with decreased membrane ruffling, failure to establish cell polarity, and loss of directional cell migration. These results suggest the PCP defects in Wdpcp mutants are not caused by loss of cilia, but by direct disruption of the actin cytoskeleton. Consistent with this, Wdpcp mutant cochlea has normal kinocilia and yet exhibits PCP defects. Together, these findings provide the first evidence, to our knowledge, that a PCP component required for ciliogenesis can directly modulate the actin cytoskeleton to

  8. Wdpcp, a PCP protein required for ciliogenesis, regulates directional cell migration and cell polarity by direct modulation of the actin cytoskeleton.

    Directory of Open Access Journals (Sweden)

    Cheng Cui

    2013-11-01

    Full Text Available Planar cell polarity (PCP regulates cell alignment required for collective cell movement during embryonic development. This requires PCP/PCP effector proteins, some of which also play essential roles in ciliogenesis, highlighting the long-standing question of the role of the cilium in PCP. Wdpcp, a PCP effector, was recently shown to regulate both ciliogenesis and collective cell movement, but the underlying mechanism is unknown. Here we show Wdpcp can regulate PCP by direct modulation of the actin cytoskeleton. These studies were made possible by recovery of a Wdpcp mutant mouse model. Wdpcp-deficient mice exhibit phenotypes reminiscent of Bardet-Biedl/Meckel-Gruber ciliopathy syndromes, including cardiac outflow tract and cochlea defects associated with PCP perturbation. We observed Wdpcp is localized to the transition zone, and in Wdpcp-deficient cells, Sept2, Nphp1, and Mks1 were lost from the transition zone, indicating Wdpcp is required for recruitment of proteins essential for ciliogenesis. Wdpcp is also found in the cytoplasm, where it is localized in the actin cytoskeleton and in focal adhesions. Wdpcp interacts with Sept2 and is colocalized with Sept2 in actin filaments, but in Wdpcp-deficient cells, Sept2 was lost from the actin cytoskeleton, suggesting Wdpcp is required for Sept2 recruitment to actin filaments. Significantly, organization of the actin filaments and focal contacts were markedly changed in Wdpcp-deficient cells. This was associated with decreased membrane ruffling, failure to establish cell polarity, and loss of directional cell migration. These results suggest the PCP defects in Wdpcp mutants are not caused by loss of cilia, but by direct disruption of the actin cytoskeleton. Consistent with this, Wdpcp mutant cochlea has normal kinocilia and yet exhibits PCP defects. Together, these findings provide the first evidence, to our knowledge, that a PCP component required for ciliogenesis can directly modulate the actin

  9. Associations between statewide prescription drug monitoring program (PDMP) requirement and physician patterns of prescribing opioid analgesics for patients with non-cancer chronic pain.

    Science.gov (United States)

    Lin, Hsien-Chang; Wang, Zhi; Boyd, Carol; Simoni-Wastila, Linda; Buu, Anne

    2018-01-01

    State-level prescription drug monitoring programs (PDMPs) have been implemented in most states. PDMPs enable registered prescribers to obtain real-time information on patients' prescription history to reduce non-medical use of controlled drugs. This study examined whether PDMP implementation and different levels of PDMP requirements were associated with physicians' patterns of prescribing opioid analgesics for patients with non-cancer chronic pain. This is a secondary analysis study using cross-sectional national data. Patients with non-cancer chronic pain from the 2012 National Ambulatory Medical Care Survey were included (weighted N=81,018,131; unweighted N=3295). Heckman two-step selection procedure employing two logistic regressions was used to explore the associations between PDMP requirements and physicians' prescribing behaviors, controlling for physician characteristics, patient characteristics, physician-healthcare system interaction, and physician-patient relationship, guided by the Eisenberg's model of physician decision making. State PDMP implementation status and requirement levels were not associated with physician opioid prescribing for non-cancer chronic pain treatment (p's ranged 0.30-0.32). Patients with Medicare coverage were more likely to be prescribed opioid analgesics than those with private health insurance (OR=1.55, prequirements and enforcement for prescribers and related stakeholders. Future studies also are needed to identify characteristics contributing to PDMP effectiveness in reducing non-medical use of prescription opioids. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. BolA Is Required for the Accurate Regulation of c-di-GMP, a Central Player in Biofilm Formation.

    Science.gov (United States)

    Moreira, Ricardo N; Dressaire, Clémentine; Barahona, Susana; Galego, Lisete; Kaever, Volkhard; Jenal, Urs; Arraiano, Cecília M

    2017-09-19

    The bacterial second messenger cyclic dimeric GMP (c-di-GMP) is a nearly ubiquitous intracellular signaling molecule involved in the transition from the motile to the sessile/biofilm state in bacteria. C-di-GMP regulates various cellular processes, including biofilm formation, motility, and virulence. BolA is a transcription factor that promotes survival in different stresses and is also involved in biofilm formation. Both BolA and c-di-GMP participate in the regulation of motility mechanisms leading to similar phenotypes. Here, we establish the importance of the balance between these two factors for accurate regulation of the transition between the planktonic and sessile lifestyles. This balance is achieved by negative-feedback regulation of BolA and c-di-GMP. BolA not only contributes directly to the motility of bacteria but also regulates the expression of diguanylate cyclases and phosphodiesterases. This expression modulation influences the synthesis and degradation of c-di-GMP, while this signaling metabolite has a negative influence in bolA mRNA transcription. Finally, we present evidence of the dominant role of BolA in biofilm, showing that, even in the presence of elevated c-di-GMP levels, biofilm formation is reduced in the absence of BolA. C-di-GMP is one of the most important bacterial second messengers involved in several cellular processes, including virulence, cell cycle regulation, biofilm formation, and flagellar synthesis. In this study, we unravelled a direct connection between the bolA morphogene and the c-di-GMP signaling molecule. We show the important cross-talk that occurs between these two molecular regulators during the transition between the motile/planktonic and adhesive/sessile lifestyles in Escherichia coli This work provides important clues that can be helpful in the development of new strategies, and the results can be applied to other organisms with relevance for human health.IMPORTANCE Bacterial cells have evolved several mechanisms

  11. Monitoring water stress and fruit quality in an orange orchard under regulated deficit irrigation using narrow-band structural and physiological remote sensing indices

    Science.gov (United States)

    Stagakis, S.; González-Dugo, V.; Cid, P.; Guillén-Climent, M. L.; Zarco-Tejada, P. J.

    2012-07-01

    This paper deals with the monitoring of water status and the assessment of the effect of stress on citrus fruit quality using structural and physiological remote sensing indices. Four flights were conducted over a citrus orchard in 2009 using an unmanned aerial vehicle (UAV) carrying a multispectral camera with six narrow spectral bands in the visible and near infrared. Physiological indices such as the Photochemical Reflectance Index (PRI570), a new structurally robust PRI formulation that uses the 515 nm as the reference band (PRI515), and a chlorophyll ratio (R700/R670) were compared against the Normalized Difference Vegetation Index (NDVI), Renormalized Difference Vegetation Index (RDVI) and Modified Triangular Vegetation Index (MTVI) canopy structural indices for their performance in tracking water status and the effects of sustained water stress on fruit quality at harvest. The irrigation setup in the commercial orchard was compared against a treatment scheduled to satisfy full requirements (based on estimated crop evapotranspiration) using two regulated deficit irrigation (RDI) strategies. The water status of the trees throughout the experiment was monitored with frequent field measurements of stem water potential (Ψx), while titratable acidity (TA) and total soluble solids (TSS) were measured at harvest on selected trees from each irrigation treatment. The high spatial resolution of the multispectral imagery (30 cm pixel size) enabled identification of pure tree crown components, extracting the tree reflectance from shaded, sunlit and aggregated pixels. The physiological and structural indices were then calculated from each tree at the following levels: (i) pure sunlit tree crown, (ii) entire crown, aggregating the within-crown shadows, and (iii) simulating a lower resolution pixel, including tree crown, sunlit and shaded soil pixels. The resulting analysis demonstrated that both PRI formulations were able to track water status, except when water stress

  12. Regulation of Telomere Length Requires a Conserved N-Terminal Domain of Rif2 in Saccharomyces cerevisiae.

    Science.gov (United States)

    Kaizer, Hannah; Connelly, Carla J; Bettridge, Kelsey; Viggiani, Christopher; Greider, Carol W

    2015-10-01

    The regulation of telomere length equilibrium is essential for cell growth and survival since critically short telomeres signal DNA damage and cell cycle arrest. While the broad principles of length regulation are well established, the molecular mechanism of how these steps occur is not fully understood. We mutagenized the RIF2 gene in Saccharomyces cerevisiae to understand how this protein blocks excess telomere elongation. We identified an N-terminal domain in Rif2 that is essential for length regulation, which we have termed BAT domain for Blocks Addition of Telomeres. Tethering this BAT domain to Rap1 blocked telomere elongation not only in rif2Δ mutants but also in rif1Δ and rap1C-terminal deletion mutants. Mutation of a single amino acid in the BAT domain, phenylalanine at position 8 to alanine, recapitulated the rif2Δ mutant phenotype. Substitution of F8 with tryptophan mimicked the wild-type phenylalanine, suggesting the aromatic amino acid represents a protein interaction site that is essential for telomere length regulation.

  13. Three fim genes required for the regulation of length and mediation of adhesion of Escherichia coli type 1 fimbriae

    DEFF Research Database (Denmark)

    Klemm, P; Christiansen, Gunna

    1987-01-01

    Three novel fim genes of Escherichia coli, fimF, fimG and fimH, were characterized. These genes were not necessary for the production of fimbriae but were shown to be involved in the adhesive property and longitudinal regulation of these structures. Complementation experiments indicated that both...

  14. 77 FR 33635 - Amendment to the Bank Secrecy Act Regulations-Requirement That Clerks of Court Report Certain...

    Science.gov (United States)

    2012-06-07

    ... Revenue Code of 1986'' file reports with FinCEN in the time and manner prescribed by regulation. Section... because of the applicable restrictions on disclosure in the U.S. tax code. During calendar year 2010... statement under section 202. List of Subjects in 31 CFR Part 1010 Administrative practice and procedure...

  15. 40 CFR 141.29 - Monitoring of consecutive public water systems.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 22 2010-07-01 2010-07-01 false Monitoring of consecutive public water... PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Monitoring and Analytical Requirements § 141.29 Monitoring of consecutive public water systems. When a public water system supplies water to...

  16. 34 CFR 303.171 - Supervision and monitoring of programs.

    Science.gov (United States)

    2010-07-01

    ... 34 Education 2 2010-07-01 2010-07-01 false Supervision and monitoring of programs. 303.171 Section 303.171 Education Regulations of the Offices of the Department of Education (Continued) OFFICE OF...-Application Requirements § 303.171 Supervision and monitoring of programs. Each application must...

  17. The H-loop in the Second Nucleotide-binding Domain of the Cystic Fibrosis Transmembrane Conductance Regulator is Required for Efficient Chloride Channel Closing

    OpenAIRE

    Kloch, Monika; Milewski, Michał; Nurowska, Ewa; Dworakowska, Beata; Cutting, Garry R.; Dołowy, Krzysztof

    2010-01-01

    The cystic fibrosis transmembrane conductance regulator (CFTR) is an ATP-binding cassette (ABC) transporter that functions as a cAMP-activated chloride channel. The recent model of CFTR gating predicts that the ATP binding to both nucleotide-binding domains (NBD1 and NBD2) of CFTR is required for the opening of the channel, while the ATP hydrolysis at NBD2 induces subsequent channel closing. In most ABC proteins, efficient hydrolysis of ATP requires the presence of the invariant histidine res...

  18. Binding of a 100-kDa ubiquitous factor to the human prolactin promoter is required for its basal and hormone-regulated activity.

    Science.gov (United States)

    Peers, B; Nalda, A M; Monget, P; Voz, M L; Belayew, A; Martial, J A

    1992-11-15

    cAMP strongly stimulates the activity of the human prolactin (hPRL) promoter. We have previously shown that two types of cis-element are required for this cAMP regulation; binding sites for the pituitary-specific factor Pit-1, and the sequence spanning nucleotides -115 to -85 (named sequence A). Sequence A contains the TGACG motif found in the consensus sequence of the cAMP-responsive element (CRE). In this study, we show that a mutation in the TGACG motif of sequence A strongly reduces not only the cAMP regulation but also the Ca2+ regulation and basal activity of the hPRL promoter. Furthermore, gel-shift assays indicate that the mutation prevents binding of a ubiquitous factor which is not the CRE-binding protein. Southwestern experiments suggest that this ubiquitous factor's molecular mass is approximately 100 kDa. We conclude that binding of a 100-kDa ubiquitous factor to sequence A is required for full basal and hormonal regulation of hPRL-promoter activity.

  19. Pyrrolizidine alkaloids (PAs) in honey and pollen-legal regulation of PA levels in food and animal feed required.

    Science.gov (United States)

    Kempf, Michael; Reinhard, Annika; Beuerle, Till

    2010-01-01

    Pyrrolizidine alkaloids (PAs) are secondary plant constituents that comprise about 400 different structures and occur in two major forms, a tertiary form and the corresponding N-oxide. PAs containing a 1,2-double bond are pre-toxins and metabolically activated by the action of hepatic P-450 enzymes to toxic pyrroles. Besides the acute toxic effects, the genotoxic and tumorigenicity potential of PAs was demonstrated in some eukaryotic model systems. Recently, the potential PA contamination of food and feeding stuff attracted recurrent great deals of attention. Humans are exposed to these toxins by consumption of herbal medicine, herbal teas, dietary supplements or food containing PA plant material. In numerous studies the potential threat to human health by PAs is stated. In pharmaceuticals, the use of these plants is regulated. Considering the PA concentrations observed especially in authentic honey from PA producing plants and pollen products, the results provoke an international regulation of PAs in food.

  20. RTN3 Regulates the Expression Level of Chemokine Receptor CXCR4 and is Required for Migration of Primordial Germ Cells

    Directory of Open Access Journals (Sweden)

    Haitao Li

    2016-04-01

    Full Text Available CXCR4 is a crucial chemokine receptor that plays key roles in primordial germ cell (PGC homing. To further characterize the CXCR4-mediated migration of PGCs, we screened CXCR4-interacting proteins using yeast two-hybrid screening. We identified reticulon3 (RTN3, a member of the reticulon family, and considered an apoptotic signal transducer, as able to interact directly with CXCR4. Furthermore, we discovered that the mRNA and protein expression levels of CXCR4 could be regulated by RTN3. We also found that RTN3 altered CXCR4 translocation and localization. Moreover, increasing the signaling of either CXCR4b or RTN3 produced similar PGC mislocalization phenotypes in zebrafish. These results suggested that RTN3 modulates PGC migration through interaction with, and regulation of, CXCR4.

  1. Statutory Management Requirements on "Animal Identification and Registration" (Act A7, Act A8: monitoring methods for compliance and related costs in four Italian farms

    Directory of Open Access Journals (Sweden)

    Marisanna Speroni

    2015-11-01

    Full Text Available The European and national laws concerning the identification and registration of livestock are meant to make possible their traceability and facilitate food safety and animal health, especially in case of a disease outbreak; such rules have also become prerequisites that farmers must meet to obtain single farm payments under the common agricultural policy. Failure to comply with these obligations entails the reduction or exclusion from direct payments. Act A7, reports the obligations imposed by the EC Regulation 1760/2000 establishing a system for the identification and registration of bovine animals and regarding the labelling of beef and beef products; Act A8, report the obligations established by the EC Reg. 21/2004 establishing a system for the identification and registration of sheep and goats. The project MO.NA.CO. monitored the application of rules for identification and registration of cattle, sheep and goats and their costs. The system of identification and registration of dairy cows resulted well organized with a good level of coordination between the involved actors in both farms. The activities necessary for compliance with rules of identification and registration of cattle are distributed throughout the year, but vary from day to day. The average total cost for annual obligations amounted to € 533.34 year-1 while the average cost for individual fulfilment in the monitoring period amounted to € 4.10. Even in the case of sheep and goats, the monitoring showed a good cooperation between farms and technicians. However, some difficulties were detected, mainly due to the size of the rumen boluses and the limited effectiveness of the ear tags. The operators suggested using smaller rumen boluses and tattoo instead of ear tags; they also suggest to extend the period within which the animals have to be labelled from the current 6 months to 9 months. The cost of compliance amounted to € 5.27 head-1 for sheep and € 4.90 head-1 for

  2. Arabidopsis thaliana VOZ (Vascular plant One-Zinc finger) transcription factors are required for proper regulation of flowering time.

    Science.gov (United States)

    Celesnik, Helena; Ali, Gul S; Robison, Faith M; Reddy, Anireddy S N

    2013-04-15

    Transition to flowering in plants is tightly controlled by environmental cues, which regulate the photoperiod and vernalization pathways, and endogenous signals, which mediate the autonomous and gibberellin pathways. In this work, we investigated the role of two Zn(2+)-finger transcription factors, the paralogues AtVOZ1 and AtVOZ2, in Arabidopsis thaliana flowering. Single atvoz1-1 and atvoz2-1 mutants showed no significant phenotypes as compared to wild type. However, atvoz1-1 atvoz2-1 double mutant plants exhibited several phenotypes characteristic of flowering-time mutants. The double mutant displayed a severe delay in flowering, together with additional pleiotropic phenotypes. Late flowering correlated with elevated expression of FLOWERING LOCUS C (FLC), which encodes a potent floral repressor, and decreased expression of its target, the floral promoter FD. Vernalization rescued delayed flowering of atvoz1-1 atvoz2-1 and reversed elevated FLC levels. Accumulation of FLC transcripts in atvoz1-1 atvoz2-1 correlated with increased expression of several FLC activators, including components of the PAF1 and SWR1 chromatin-modifying complexes. Additionally, AtVOZs were shown to bind the promoter of MOS3/SAR3 and directly regulate expression of this nuclear pore protein, which is known to participate in the regulation of flowering time, suggesting that AtVOZs exert at least some of their flowering regulation by influencing the nuclear pore function. Complementation of atvoz1-1 atvoz2-1 with AtVOZ2 reversed all double mutant phenotypes, confirming that the observed morphological and molecular changes arise from the absence of functional AtVOZ proteins, and validating the functional redundancy between AtVOZ1 and AtVOZ2.

  3. TyrR, the regulator of aromatic amino acid metabolism, is required for mice infection of Yersinia pestis

    Directory of Open Access Journals (Sweden)

    Zhongliang eDeng

    2015-02-01

    Full Text Available Yersinia pestis, the causative agent of plague, poses a serious health threat to rodents and human beings. TyrR is a transcriptional regulator that controls the metabolism of aromatic amino acids in Escherichia coli. In this paper, TyrR played an important role in Y. pestis virulence. Inactivation of tyrR did not seem to affect the in vitro growth of this organism, but resulted in at least 10,000-fold attenuation compared with the wild-type (WT strain upon subcutaneous infection to mice. In addition, loads of tyrR mutant within mice livers and spleens significantly decreased compared with the WT strain. Transcriptome analysis revealed that TyrR, directly or indirectly, regulated 29 genes encoded on Y. pestis chromosome or plasmids under in vitro growth condition. Similar to the regulatory function of this protein in E. coli, five aromatic-pathway genes (aroF-tyrA, aroP, aroL, and tyrP were significantly reduced upon deletion of the tyrR gene. Two genes (glnL and glnG that encode sensory histidine kinase and regulator in a two-component regulatory system involved in nitrogen assimilation were downregulated in the tyrR mutant. Several genes encoding type III secretion proteins were transcribed by 2.0- to 4.2-fold in a tyrR mutant relative to the WT strain. Interestingly, the acid-stressed genes, hdeB and hdeD, were downregulated, and such downregulation partly accounted for the decrease in tolerance of the tyrR mutant under acidic conditions. In conclusion, regulation of TyrR in Y. pestis is similar to, but distinct from, that in E. coli. TyrR is a metabolic virulence determinant in Y. pestis that is important for extracellular survival and/or proliferation.

  4. Savannah River Site Environmental Monitoring Plan. Volume 1, Section 1000 Addendum: Revision 3

    Energy Technology Data Exchange (ETDEWEB)

    Jannik, G.T.

    1994-10-01

    This document -- the Savannah River Site Environmental Monitoring Plan (SRS EM Plan) -- has been prepared according to guidance contained in the DOE 5400 Series orders, in 10 CFR 834, and in DOE/EH-0173T, Environmental Regulatory Guide for Radiological Effluent Monitoring and environmental Surveillance [DOE, 1991]. The SRS EM Plan`s purpose is to define the criteria, regulations, and guideline requirements with which SRS will comply. These criteria and requirements are applicable to environmental monitoring activities performed in support of the SRS Environmental Monitoring Program (SRS EM Program), WSRC-3Q1-2, Volume 1, Section 1100. They are not applicable to monitoring activities utilized exclusively for process monitoring/control. The environmental monitoring program requirements documented in the SRS EM Plan incorporate all applicable should requirements of DOE/EH-0173T and expand upon them to include nonradiological environmental monitoring program requirements.

  5. Cartilage development requires the function of Estrogen-related receptor alpha that directly regulates sox9 expression in zebrafish.

    Science.gov (United States)

    Kim, Yong-Il; No Lee, Joon; Bhandari, Sushil; Nam, In-Koo; Yoo, Kyeong-Won; Kim, Se-Jin; Oh, Gi-Su; Kim, Hyung-Jin; So, Hong-Seob; Choe, Seong-Kyu; Park, Raekil

    2015-12-10

    Estrogen-related receptor alpha (ESRRa) regulates a number of cellular processes including development of bone and muscles. However, direct evidence regarding its involvement in cartilage development remains elusive. In this report, we establish an in vivo role of Esrra in cartilage development during embryogenesis in zebrafish. Gene expression analysis indicates that esrra is expressed in developing pharyngeal arches where genes necessary for cartilage development are also expressed. Loss of function analysis shows that knockdown of esrra impairs expression of genes including sox9, col2a1, sox5, sox6, runx2 and col10a1 thus induces abnormally formed cartilage in pharyngeal arches. Importantly, we identify putative ESRRa binding elements in upstream regions of sox9 to which ESRRa can directly bind, indicating that Esrra may directly regulate sox9 expression. Accordingly, ectopic expression of sox9 rescues defective formation of cartilage induced by the knockdown of esrra. Taken together, our results indicate for the first time that ESRRa is essential for cartilage development by regulating sox9 expression during vertebrate development.

  6. TALEN Gene Knockouts Reveal No Requirement for the Conserved Human Shelterin Protein Rap1 in Telomere Protection and Length Regulation

    Directory of Open Access Journals (Sweden)

    Shaheen Kabir

    2014-11-01

    Full Text Available The conserved protein Rap1 functions at telomeres in fungi, protozoa, and vertebrates. Like yeast Rap1, human Rap1 has been implicated in telomere length regulation and repression of nonhomologous end-joining (NHEJ at telomeres. However, mouse telomeres lacking Rap1 do not succumb to NHEJ. To determine the functions of human Rap1, we generated several transcription activator-like effector nuclease (TALEN-mediated human cell lines lacking Rap1. Loss of Rap1 did not affect the other components of shelterin, the modification of telomeric histones, the subnuclear position of telomeres, or the 3′ telomeric overhang. Telomeres lacking Rap1 did not show a DNA damage response, NHEJ, or consistent changes in their length, indicating that Rap1 does not have an important function in protection or length regulation of human telomeres. As human Rap1, like its mouse and unicellular orthologs, affects gene expression, we propose that the conservation of Rap1 reflects its role in transcriptional regulation rather than a function at telomeres.

  7. Suppressor of fusion, a Fusarium oxysporum homolog of Ndt80, is required for nutrient-dependent regulation of anastomosis.

    Science.gov (United States)

    Shahi, Shermineh; Fokkens, Like; Houterman, Petra M; Rep, Martijn

    2016-10-01

    Heterokaryon formation is an essential step in asexual recombination in Fusarium oxysporum. Filamentous fungi have an elaborate nonself recognition machinery to prevent formation and proliferation of heterokaryotic cells, called heterokaryon incompatibility (HI). In F. oxysporum the regulation of this machinery is not well understood. In Neurospora crassa, Vib-1, a putative transcription factor of the p53-like Ndt80 family of transcription factors, has been identified as global regulator of HI. In this study we investigated the role of the F. oxysporum homolog of Vib-1, called Suf, in vegetative hyphal and conidial anastomosis tube (CAT) fusion and HI. We identified a novel function for an Ndt80 homolog as a nutrient-dependent regulator of anastomosis. Strains carrying the SUF deletion mutation display a hyper-fusion phenotype during vegetative growth as well as germling development. In addition, conidial paring of incompatible SUF deletion strains led to more heterokaryon formation, which is independent of suppression of HI. Our data provides further proof for the divergence in the functions of different members Ndt80 family. We propose that Ndt80 homologs mediate responses to nutrient quality and quantity, with specific responses varying between species.

  8. Cdc42 is a key regulator of B cell differentiation and is required for antiviral humoral immunity

    DEFF Research Database (Denmark)

    Burbage, Marianne; Keppler, Selina J; Gasparrini, Francesca

    2015-01-01

    The small Rho GTPase Cdc42, known to interact with Wiskott-Aldrich syndrome (WAS) protein, is an important regulator of actin remodeling. Here, we show that genetic ablation of Cdc42 exclusively in the B cell lineage is sufficient to render mice unable to mount antibody responses. Indeed Cdc42-de...... and actin remodeling; defective antigen presentation and in vivo interaction with T cells; and a severe B cell-intrinsic block in plasma cell differentiation. Thus, our study presents a new perspective on Cdc42 as key regulator of B cell physiology.......The small Rho GTPase Cdc42, known to interact with Wiskott-Aldrich syndrome (WAS) protein, is an important regulator of actin remodeling. Here, we show that genetic ablation of Cdc42 exclusively in the B cell lineage is sufficient to render mice unable to mount antibody responses. Indeed Cdc42......-deficient mice are incapable of forming germinal centers or generating plasma B cells upon either viral infection or immunization. Such severe immune deficiency is caused by multiple and profound B cell abnormalities, including early blocks during B cell development; impaired antigen-driven BCR signaling...

  9. Issues with monitoring the safety of psychoactive products under a legal regulated market for new psychoactive substances ('legal highs') in New Zealand.

    Science.gov (United States)

    Rychert, Marta; Wilkins, Chris; Witten, Karen

    2017-02-23

    New Zealand's Psychoactive Substances Act (2013) established the world's first regulated market for 'low risk' psychoactive products ('legal highs'). Under an interim PSA regime, 47 existing products were permitted to be continued to be sold. To explore issues with the implementation of regulatory systems to monitor the safety of products on the legal market under the interim Psychoactive Substances Act regime. Semi-structured interviews with 30 key stakeholders, including industry, government agency, health and drug service professionals were conducted, transcribed and analysed thematically. In retrospect stakeholders questioned the decision to approve strong synthetic cannabinoid smoking products, noting their health risks because of product formulation, inconsistent manufacturing practices and smoking as the means of administration. Industry actors claimed the decision to approve synthetic cannabinoid smokeable products prevented potentially safer products from gaining market share. The system for withdrawing approved products which were subsequently found to be harmful was criticised for the poor quality of data available, limited engagement with health professionals and the slowness of product withdrawal. Many of the problems with the regime were attributed to the urgency under which the legal market under the interim Psychoactive Substances Act was established and implemented. The selection of 'safer' products, implementation of the product monitoring system, and engagement with health professionals may have benefited from more time and resources. An incremental approach to establishing the new market may have made the regulatory management of the new regime more workable. [Rychert M, Wilkins C, Witten K. Issues with monitoring the safety of psychoactive products under a legal regulated market for new psychoactive substances ('legal highs') in New Zealand. Drug Alcohol Rev 2017;00:000-000]. © 2017 Australasian Professional Society on Alcohol and other Drugs.

  10. Growth arrest specific 2 is up-regulated in chronic myeloid leukemia cells and required for their growth.

    Directory of Open Access Journals (Sweden)

    Haixia Zhou

    Full Text Available Although the generation of BCR-ABL is the molecular hallmark of chronic myeloid leukemia (CML, the comprehensive molecular mechanisms of the disease remain unclear yet. Growth arrest specific 2 (GAS2 regulates multiple cellular functions including cell cycle, apoptosis and calpain activities. In the present study, we found GAS2 was up-regulated in CML cells including CD34+ progenitor cells compared to their normal counterparts. We utilized RNAi and the expression of dominant negative form of GAS2 (GAS2DN to target GAS2, which resulted in calpain activity enhancement and growth inhibition of both K562 and MEG-01 cells. Targeting GAS2 also sensitized K562 cells to Imatinib mesylate (IM. GAS2DN suppressed the tumorigenic ability of MEG-01 cells and impaired the tumour growth as well. Moreover, the CD34+ cells from CML patients and healthy donors were transduced with control and GAS2DN lentiviral vectors, and the CD34+ transduced (YFP+ progeny cells (CD34+YFP+ were plated for colony-forming cell (CFC assay. The results showed that GAS2DN inhibited the CFC production of CML cells by 57±3% (n = 3, while affected those of normal hematopoietic cells by 31±1% (n = 2. Next, we found the inhibition of CML cells by GAS2DN was dependent on calpain activity but not the degradation of beta-catenin. Lastly, we generated microarray data to identify the differentially expressed genes upon GAS2DN and validated that the expression of HNRPDL, PTK7 and UCHL5 was suppressed by GAS2DN. These 3 genes were up-regulated in CML cells compared to normal control cells and the growth of K562 cells was inhibited upon HNRPDL silence. Taken together, we have demonstrated that GAS2 is up-regulated in CML cells and the inhibition of GAS2 impairs the growth of CML cells, which indicates GAS2 is a novel regulator of CML cells and a potential therapeutic target of this disease.

  11. Assessing Drinking Water Quality and Water Safety Management in Sub-Saharan Africa Using Regulated Monitoring Data.

    Science.gov (United States)

    Kumpel, Emily; Peletz, Rachel; Bonham, Mateyo; Khush, Ranjiv

    2016-10-18

    Universal access to safe drinking water is prioritized in the post-2015 Sustainable Development Goals. Collecting reliable and actionable water quality information in low-resource settings, however, is challenging, and little is known about the correspondence between water quality data collected by local monitoring agencies and global frameworks for water safety. Using 42 926 microbial water quality test results from 32 surveillance agencies and water suppliers in seven sub-Saharan African countries, we determined the degree to which water sources were monitored, how water quality varied by source type, and institutional responses to results. Sixty-four percent of the water samples were collected from piped supplies, although the majority of Africans rely on nonpiped sources. Piped supplies had the lowest levels of fecal indicator bacteria (FIB) compared to any other source type: only 4% of samples of water piped to plots and 2% of samples from water piped to public taps/standpipes were positive for FIB (n = 14 948 and n = 12 278, respectively). Among other types of improved sources, samples from harvested rainwater and boreholes were less often positive for FIB (22%, n = 167 and 31%, n = 3329, respectively) than protected springs or protected dug wells (39%, n = 472 and 65%, n = 505). When data from different settings were aggregated, the FIB levels in different source types broadly reflected the source-type water safety framework used by the Joint Monitoring Programme. However, the insufficient testing of nonpiped sources relative to their use indicates important gaps in current assessments. Our results emphasize the importance of local data collection for water safety management and measurement of progress toward universal safe drinking water access.

  12. The Regulation and Monitor of Metacognitive Knowledge in SFL Writing%二语写作中元认知知识的调节与监控

    Institute of Scientific and Technical Information of China (English)

    何春霞

    2012-01-01

    The present studies both home and abroad have affirmed the importance of metacognitive knowledge to SFL writing,which,therefore,can get better reaction in turn provided that the writer's metacognitive awareness is consciously improved and their application of metacognitive knowledge is greatly enhanced.Centred on the cognitive subject,the paper discusses how the metacognitive knowledge is regulated and monitored effectively during SFL writing.Finally it is concluded that only in the dynamic process when self-montior and out-monitor,macro-monitor and micro-monitor interact each other can the writer fully exert his metacognitive knowledge and further serve the SFL writing.%国内外相关研究已证实元认知知识在二语写作中的重要性。因此,自觉提高写作者的元认知意识,加强元认知知识的运用,无疑能更好反作用于二语写作。文章从认知主体出发,探讨了如何在二语写作中有效地对元认知知识加以调节和监控。最后得出只有在自我监控与外部监控、宏观监控与微观监控相互作用的动态过程中才能最大限度地发挥写作者的元认知知识,更好地服务于二语写作。

  13. Monitoring human factor risk characteristics at nuclear legacy sites in northwest Russia in support of radiation safety regulation.

    Science.gov (United States)

    Scheblanov, V Y; Sneve, M K; Bobrov, A F

    2012-12-01

    This paper describes research aimed at improving regulatory supervision of radiation safety during work associated with the management of spent nuclear fuel and radioactive waste at legacy sites in northwest Russia through timely identification of employees presenting unfavourable human factor risk characteristics. The legacy sites of interest include sites of temporary storage now operated by SevRAO on behalf of Rosatom. The sites were previously operational bases for servicing nuclear powered submarines and are now subject to major remediation activities. These activities include hazardous operations for recovery of spent nuclear fuel and radioactive waste from sub-optimal storage conditions. The paper describes the results of analysis of methods, procedures, techniques and informational issues leading to the development of an expert-diagnostic information system for monitoring of workers involved in carrying out the most hazardous operations. The system serves as a tool for human factor and professional reliability risk monitoring and has been tested in practical working environments and implemented as part of regulatory supervision. The work has been carried out by the Burnasyan Federal Medical Biophysical Center, within the framework of the regulatory cooperation programme between the Federal Medical-Biological Agency of Russia and the Norwegian Radiation Protection Authority.

  14. The H-loop in the second nucleotide-binding domain of the cystic fibrosis transmembrane conductance regulator is required for efficient chloride channel closing.

    Science.gov (United States)

    Kloch, Monika; Milewski, Michał; Nurowska, Ewa; Dworakowska, Beata; Cutting, Garry R; Dołowy, Krzysztof

    2010-01-01

    The cystic fibrosis transmembrane conductance regulator (CFTR) is an ATP-binding cassette (ABC) transporter that functions as a cAMP-activated chloride channel. The recent model of CFTR gating predicts that the ATP binding to both nucleotide-binding domains (NBD1 and NBD2) of CFTR is required for the opening of the channel, while the ATP hydrolysis at NBD2 induces subsequent channel closing. In most ABC proteins, efficient hydrolysis of ATP requires the presence of the invariant histidine residue within the H-loop located in the C-terminal part of the NBD. However, the contribution of the corresponding region (H-loop) of NBD2 to the CFTR channel gating has not been examined so far. Here we report that the alanine substitution of the conserved dipeptide HR motif (HR-->AA) in the H-loop of NBD2 leads to prolonged open states of CFTR channel, indicating that the H-loop is required for efficient channel closing. On the other hand, the HR-->AA substitution lead to the substantial decrease of CFTR-mediated current density (pA/pF) in transfected HEK 293 cells, as recorded in the whole-cell patch-clamp analysis. These results suggest that the H-loop of NBD2, apart from being required for CFTR channel closing, may be involved in regulating CFTR trafficking to the cell surface.

  15. Ripening-regulated susceptibility of tomato fruit to Botrytis cinerea requires NOR but not RIN or ethylene.

    Science.gov (United States)

    Cantu, Dario; Blanco-Ulate, Barbara; Yang, Liya; Labavitch, John M; Bennett, Alan B; Powell, Ann L T

    2009-07-01

    Fruit ripening is a developmental process that is associated with increased susceptibility to the necrotrophic pathogen Botrytis cinerea. Histochemical observations demonstrate that unripe tomato (Solanum lycopersicum) fruit activate pathogen defense responses, but these responses are attenuated in ripe fruit infected by B. cinerea. Tomato fruit ripening is regulated independently and cooperatively by ethylene and transcription factors, including NON-RIPENING (NOR) and RIPENING-INHIBITOR (RIN). Mutations in NOR or RIN or interference with ethylene perception prevent fruit from ripening and, thereby, would be expected to influence susceptibility. We show, however, that the susceptibility of ripe fruit is dependent on NOR but not on RIN and only partially on ethylene perception, leading to the conclusion that not all of the pathways and events that constitute ripening render fruit susceptible. Additionally, on unripe fruit, B. cinerea induces the expression of genes also expressed as uninfected fruit ripen. Among the ripening-associated genes induced by B. cinerea are LePG (for polygalacturonase) and LeExp1 (for expansin), which encode cell wall-modifying proteins and have been shown to facilitate susceptibility. LePG and LeExp1 are induced only in susceptible rin fruit and not in resistant nor fruit. Thus, to infect fruit, B. cinerea relies on some of the processes and events that occur during ripening, and the fungus induces these pathways in unripe fruit, suggesting that the pathogen itself can initiate the induction of susceptibility by exploiting endogenous developmental programs. These results demonstrate the developmental plasticity of plant responses to the fungus and indicate how known regulators of fruit ripening participate in regulating ripening-associated pathogen susceptibility.

  16. 12 CFR 27.6 - Substitute monitoring program.

    Science.gov (United States)

    2010-01-01

    ....6 Banks and Banking COMPTROLLER OF THE CURRENCY, DEPARTMENT OF THE TREASURY FAIR HOUSING HOME LOAN... substitute monitoring program as authorized under § 202.13(d) of Regulation B of the Federal Reserve Board... compliance with the requirements of § 202.13 of Regulation B. ...

  17. The catalytic activity of the CD45 membrane-proximal phosphatase domain is required for TCR signaling and regulation

    DEFF Research Database (Denmark)

    Desai, D M; Sap, J; Silvennoinen, O;

    1994-01-01

    Cell surface expression of CD45, a receptor-like protein tyrosine phosphatase (PTPase), is required for T cell antigen receptor (TCR)-mediated signal transduction. Like the majority of transmembrane PTPases, CD45 contains two cytoplasmic phosphatase domains, whose relative in vivo function is not...

  18. 77 FR 30473 - Notice of Intent To Revise Stormwater Regulations To Specify That an NPDES Permit Is Not Required...

    Science.gov (United States)

    2012-05-23

    ... programs, including forest stewardship and sustainability initiatives. Under these required or voluntary... performance standards are encouraged to be abandoned. Abandonment strategies may involve the removal of stream... order by forest industry land, corporate non-industrial land, and private non-industrial land (Prud...

  19. 7 CFR 319.5 - Requirements for submitting requests to change the regulations in 7 CFR part 319.

    Science.gov (United States)

    2010-01-01

    ... required in order to complete a pest risk analysis in accordance with international standards for pest risk... implementation. (8) Existing documentation: Relevant pest risk analyses, environmental assessment(s), biological... Analysis and Operations, PPQ, APHIS, 4700 River Road Unit 140, Riverdale, MD 20737. (d) Information....

  20. 76 FR 14812 - Final Regulation Extending the Reporting Deadline for Year 2010 Data Elements Required Under the...

    Science.gov (United States)

    2011-03-18

    ... Review B. Paperwork Reduction Act C. Regulatory Flexibility Act (RFA) D. Unfunded Mandates Reform Act... facilities. 331492 Zinc dust reclaiming facilities, recovering from scrap and/or alloying purchased metals... addition of facility- and parent-level reporting requirements (75 FR 57669, September 22, 2010); technical...

  1. Inhibition of seed germination and induction of systemic disease resistance by Pseudomonas chlororaphis O6 requires phenazine production regulated by the global regulator, gacS.

    Science.gov (United States)

    Kang, Beom Ryong; Han, Song Hee; Zdor, Rob E; Anderson, Anne J; Spencer, Matt; Yang, Kwang Yeol; Kim, Yong Hwan; Lee, Myung Chul; Cho, Baik Ho; Kim, Young Cheol

    2007-04-01

    Seed coating by a phenazine-producing bacterium, Pseudomonas chlororaphis O6, induced dose-dependent inhibition of germination in wheat and barley seeds, but did not inhibit germination of rice or cucumber seeds. In wheat seedlings grown from inoculated seeds, phenazine production levels near the seed were higher than in the roots. Deletion of the gacS gene reduced transcription from the genes required for phenazine synthesis, the regulatory phzI gene and the biosynthetic phzA gene. The inhibition of seed germination and the induction of systemic disease resistance against a bacterial soft-rot pathogen, Erwinia carotovora subsp. carotovora, were impaired in the gacS and phzA mutants of P chlororaphis O6. Culture filtrates of the gacS and phzA mutants of P chlororaphis 06 did not inhibit seed germination of wheat, whereas that of the wild-type was inhibitory. Our results showed that the production of phenazines by P chlororaphis O6 was correlated with reduced germination of barley and wheat seeds, and the level of systemic resistance in tobacco against E. carotovora.

  2. Identification of mRNAs bound and regulated by human LIN28 proteins and molecular requirements for RNA recognition

    Science.gov (United States)

    Hafner, Markus; Max, Klaas E.A.; Bandaru, Pradeep; Morozov, Pavel; Gerstberger, Stefanie; Brown, Miguel; Molina, Henrik; Tuschl, Thomas

    2013-01-01

    Human LIN28A and LIN28B are RNA-binding proteins (RBPs) conserved in animals with important roles during development and stem cell reprogramming. We used Photoactivatable-Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation (PAR-CLIP) in HEK293 cells and identified a largely overlapping set of ∼3000 mRNAs at ∼9500 sites located in the 3′ UTR and CDS. In vitro and in vivo, LIN28 preferentially bound single-stranded RNA containing a uridine-rich element and one or more flanking guanosines and appeared to be able to disrupt base-pairing to access these elements when embedded in predicted secondary structure. In HEK293 cells, LIN28 protein binding mildly stabilized target mRNAs and increased protein abundance. The top targets were its own mRNAs and those of other RBPs and cell cycle regulators. Alteration of LIN28 protein levels also negatively regulated the abundance of some but not all let-7 miRNA family members, indicating sequence-specific binding of let-7 precursors to LIN28 proteins and competition with cytoplasmic miRNA biogenesis factors. PMID:23481595

  3. Regulation of cell divisions and differentiation by MALE STERILITY32 is required for anther development in maize.

    Science.gov (United States)

    Moon, Jihyun; Skibbe, David; Timofejeva, Ljudmilla; Wang, Chung-Ju Rachel; Kelliher, Timothy; Kremling, Karl; Walbot, Virginia; Cande, William Zacheus

    2013-11-01

    Male fertility in flowering plants relies on proper division and differentiation of cells in the anther, a process that gives rise to four somatic layers surrounding central germinal cells. The maize gene male sterility32 (ms32) encodes a basic helix-loop-helix (bHLH) transcription factor, which functions as an important regulator of both division and differentiation during anther development. After the four somatic cell layers are generated properly through successive periclinal divisions, in the ms32 mutant, tapetal precursor cells fail to differentiate, and, instead, undergo additional periclinal divisions to form extra layers of cells. These cells become vacuolated and expand, and lead to failure in pollen mother cell development. ms32 expression is specific to the pre-meiotic anthers and is distributed initially broadly in the four lobes, but as the anther develops, its expression becomes restricted to the innermost somatic layer, the tapetum. The ms32-ref mac1-1 double mutant is unable to form tapetal precursors and also exhibits excessive somatic proliferation leading to numerous, disorganized cell layers, suggesting a synergistic interaction between ms32 and mac1. Altogether, our results show that MS32 is a major regulator in maize anther development that promotes tapetum differentiation and inhibits periclinal division once a tapetal cell is specified. © 2013 The Authors The Plant Journal © 2013 John Wiley & Sons Ltd.

  4. Opposite-sex attraction in male mice requires testosterone-dependent regulation of adult olfactory bulb neurogenesis

    Science.gov (United States)

    Schellino, Roberta; Trova, Sara; Cimino, Irene; Farinetti, Alice; Jongbloets, Bart C.; Pasterkamp, R. Jeroen; Panzica, Giancarlo; Giacobini, Paolo; De Marchis, Silvia; Peretto, Paolo

    2016-01-01

    Opposite-sex attraction in most mammals depends on the fine-tuned integration of pheromonal stimuli with gonadal hormones in the brain circuits underlying sexual behaviour. Neural activity in these circuits is regulated by sensory processing in the accessory olfactory bulb (AOB), the first central station of the vomeronasal system. Recent evidence indicates adult neurogenesis in the AOB is involved in sex behaviour; however, the mechanisms underlying this function are unknown. By using Semaphorin 7A knockout (Sema7A ko) mice, which show a reduced number of gonadotropin-releasing-hormone neurons, small testicles and subfertility, and wild-type males castrated during adulthood, we demonstrate that the level of circulating testosterone regulates the sex-specific control of AOB neurogenesis and the vomeronasal system activation, which influences opposite-sex cue preference/attraction in mice. Overall, these data highlight adult neurogenesis as a hub for the integration of pheromonal and hormonal cues that control sex-specific responses in brain circuits. PMID:27782186

  5. Serum inducible kinase is a positive regulator of cortical dendrite development and is required for BDNF-promoted dendritic arborization

    Institute of Scientific and Technical Information of China (English)

    Shun-Ling Guo; Guo-He Tan; Shuai Li; Xue-Wen Cheng; Ya Zhou; Yun-Fang Jia; Hui Xiong; Jiong Tao; Zhi-Qi Xiong

    2012-01-01

    Serum inducible kinase (SNK),also known as (p)olo-(l)ike (k)inase 2 (PLK2),is a known regulator of mitosis,synaptogenesis and synaptic homeostasis.However,its role in early cortical development is unknown.Herein,we show that snk is expressed in the cortical plate from embryonic day 14,but not in the ventricular/subventricular zones (VZ/SVZ),and SNK protein localizes to the soma and dendrites of cultured immature cortical neurons.Loss of SNK impaired dendritic but not axonal arborization in a dose-dependent manner and overexpression had opposite effects,both in vitro and in vivo.Overexpression of SNK also caused abnormal branching of the leading process of migrating cortical neurons in electroporated cortices.The kinase activity was necessary for these effects.Extracellular signalregulated kinase (ERK) pathway activity downstream of brain-derived neurotrophic factor (BDNF) stimulation led to increases in SNK protein expression via transcriptional regulation,and this upregulation was necessary for the growth-promoting effect of BDNF on dendritic arborization.Taken together,our results indicate that SNK is essential for dendrite morphogenesis in cortical neurons.

  6. A DNA2 Homolog Is Required for DNA Damage Repair, Cell Cycle Regulation, and Meristem Maintenance in Plants.

    Science.gov (United States)

    Jia, Ning; Liu, Xiaomin; Gao, Hongbo

    2016-05-01

    Plant meristem cells divide and differentiate in a spatially and temporally regulated manner, ultimately giving rise to organs. In this study, we isolated the Arabidopsis jing he sheng 1 (jhs1) mutant, which exhibited retarded growth, an abnormal pattern of meristem cell division and differentiation, and morphological defects such as fasciation, an irregular arrangement of siliques, and short roots. We identified JHS1 as a homolog of human and yeast DNA Replication Helicase/Nuclease2, which is known to be involved in DNA replication and damage repair. JHS1 is strongly expressed in the meristem of Arabidopsis. The jhs1 mutant was sensitive to DNA damage stress and had an increased DNA damage response, including increased expression of genes involved in DNA damage repair and cell cycle regulation, and a higher frequency of homologous recombination. In the meristem of the mutant plants, cell cycle progression was delayed at the G2 or late S phase and genes essential for meristem maintenance were misregulated. These results suggest that JHS1 plays an important role in DNA replication and damage repair, meristem maintenance, and development in plants. © 2016 American Society of Plant Biologists. All Rights Reserved.

  7. 24 CFR 761.21 - Plan requirement.

    Science.gov (United States)

    2010-04-01

    ...-supported activities and describe the system for monitoring and evaluating these activities. Measurable... 24 Housing and Urban Development 4 2010-04-01 2010-04-01 false Plan requirement. 761.21 Section 761.21 Housing and Urban Development Regulations Relating to Housing and Urban Development (Continued...

  8. JMJD-5/KDM8 regulates H3K36me2 and is required for late steps of homologous recombination and genome integrity

    Science.gov (United States)

    Zaghet, Nico; Ramalho, João J.; Vilstrup Johansen, Jens; Salcini, Anna Elisabetta

    2017-01-01

    The eukaryotic genome is organized in a three-dimensional structure called chromatin, constituted by DNA and associated proteins, the majority of which are histones. Post-translational modifications of histone proteins greatly influence chromatin structure and regulate many DNA-based biological processes. Methylation of lysine 36 of histone 3 (H3K36) is a post-translational modification functionally relevant during early steps of DNA damage repair. Here, we show that the JMJD-5 regulates H3K36 di-methylation and it is required at late stages of double strand break repair mediated by homologous recombination. Loss of jmjd-5 results in hypersensitivity to ionizing radiation and in meiotic defects, and it is associated with aberrant retention of RAD-51 at sites of double strand breaks. Analyses of jmjd-5 genetic interactions with genes required for resolving recombination intermediates (rtel-1) or promoting the resolution of RAD-51 double stranded DNA filaments (rfs-1 and helq-1) suggest that jmjd-5 prevents the formation of stalled postsynaptic recombination intermediates and favors RAD-51 removal. As these phenotypes are all recapitulated by a catalytically inactive jmjd-5 mutant, we propose a novel role for H3K36me2 regulation during late steps of homologous recombination critical to preserve genome integrity. PMID:28207814

  9. CD81 Is a Central Regulator of Cellular Events Required for Hepatitis C Virus Infection of Human Hepatocytes▿ †

    OpenAIRE

    Brazzoli, Michela; Bianchi, Alessia; Filippini, Sara; Weiner, Amy; Zhu, Qing; Pizza, Mariagrazia; Crotta, Stefania

    2008-01-01

    Infection with hepatitis C virus (HCV) is still a major public health problem, and the events leading to hepatocyte infection are not yet fully understood. Combining confocal microscopy with biochemical analysis and studies of infection requirements using pharmacological inhibitors and small interfering RNAs, we show here that engagement of CD81 activates the Rho GTPase family members Rac, Rho, and Cdc42 and that the block of these signaling pathways drastically reduces HCV infectivity. Activ...

  10. Engineering requirements due to the ESP/ESPN regulation apply at the port plug for ITER diagnostic system

    Energy Technology Data Exchange (ETDEWEB)

    Giacomin, T., E-mail: Thibaud.Giacomin@iter.org [ITER Organization, Route de Vinon sur Verdon, 13115 St Paul-lez-Durance (France); Delhom, D.; Drevon, J.-M. [Bertin Technologies, Aix en Provence (France); Guirao, J.; Iglesias, S. [NATEC Ingenieros (Spain); Jourdan, T. [ITER Organization, Route de Vinon sur Verdon, 13115 St Paul-lez-Durance (France); Loesser, D. [Princeton Plasma Physics Laboratory (United States); Maquet, P. [ITER Organization, Route de Vinon sur Verdon, 13115 St Paul-lez-Durance (France); Ordieres, J. [NATEC Ingenieros (Spain); Pak, S. [ITER Organization, Route de Vinon sur Verdon, 13115 St Paul-lez-Durance (France); Proust, M. [CEA Cadarache, St. Paul-lez-Durance (France); Smith, M. [Princeton Plasma Physics Laboratory (United States); Udintsev, V.S. [ITER Organization, Route de Vinon sur Verdon, 13115 St Paul-lez-Durance (France); Vacas, C. [NATEC Ingenieros (Spain); Walsh, M.J. [ITER Organization, Route de Vinon sur Verdon, 13115 St Paul-lez-Durance (France); Zhai, Y. [Princeton Plasma Physics Laboratory (United States)

    2015-10-15

    Highlights: • The ESP/ESPN regulation is applied to all equatorial and upper port-based diagnostic systems. • The EPP and UPP structure provides a flexible platform for a variety of diagnostics. • The EPP and UPP structure provide a support the diagnostic shielding module. - Abstract: Due to this position close to the plasma, the port plug structure and the diagnostic first wall (DFW) contain water to allow cooling during operation and for heating during bake-out. To remove the heat coming from the plasma due to radiation and neutrons, the pressure inside these structures should be up to 44 bars. On the other hand, the dominant load expected to drive the design of these structures is of electromagnetic origin during the plasma disruption. Description of the loads acting on DFWs and generic port plug structures and the significance of the load due to the water pressure, with implications on the design and inspection, are discussed in this paper.

  11. CLNR1, the AREA/NIT2-like global nitrogen regulator of the plant fungal pathogen Colletotrichum lindemuthianum is required for the infection cycle.

    Science.gov (United States)

    Pellier, Anne-Laure; Laugé, Richard; Veneault-Fourrey, Claire; Langin, Thierry

    2003-05-01

    Nitrogen starvation is generally assumed to be encountered by biotrophic and hemibiotrophic plant fungal pathogens at the beginning of their infection cycle. We tested whether nitrogen starvation constitutes a cue regulating genes that are required for pathogenicity of Colletotrichum lindemuthianum, a fungal pathogen of common bean. The clnr1 (C. lindemuthianumnitrogen regulator 1) gene, the areA/nit-2 orthologue of C. lindemuthianum, was isolated. The predicted CLNR1 protein exhibits high amino acid sequence similarities with the AREA and NIT2 global fungal nitrogen regulators. Targeted clnr1- mutants are unable to use a wide array of nitrogen sources, indicating that clnr1 is the C. lindemuthianum major nitrogen regulatory gene. The clnr1- mutants are non-pathogenic, although few anthracnose lesions seldom occur on whole plantlets. Surprisingly, cytological analysis reveals that the clnr1- mutants are not disturbed from the penetration stage until the end of the biotrophic phase, but that they are impaired during the setting up of the necrotrophic phase. Thus, through CLNR1, nitrogen starvation constitutes a cue for the regulation of genes that are compulsory for this stage of the C. lindemuthianum infection process. Additionally, clnr1- mutants complemented with the Aspergillus nidulans areA gene are fully pathogenic, indicating that areA is able to activate the C. lindemuthianum suited genes, normally under the control of clnr1.

  12. Idaho National Laboratory Site Environmental Monitoring Plan

    Energy Technology Data Exchange (ETDEWEB)

    Jenifer Nordstrom

    2014-02-01

    This plan provides a high-level summary of environmental monitoring performed by various organizations within and around the Idaho National Laboratory (INL) Site as required by U.S. Department of Energy (DOE) Order 435.1, Radioactive Waste Management, and DOE Order 458.1, Radiation Protection of the Public and the Environment, Guide DOE/EH-0173T, Environmental Regulatory Guide for Radiological Effluent Monitoring and Environmental Surveillance, and in accordance with 40 Code of Federal Regulations (CFR) 61, National Emission Standards for Hazardous Air Pollutants. The purpose of these orders is to 1) implement sound stewardship practices that protect the air, water, land, and other natural and cultural resources that may be impacted by DOE operations, and 2) to establish standards and requirements for the operations of DOE and DOE contractors with respect to protection of the environment and members of the public against undue risk from radiation. This plan describes the organizations responsible for conducting environmental monitoring across the INL Site, the rationale for monitoring, the types of media being monitored, where the monitoring is conducted, and where monitoring results can be obtained. Detailed monitoring procedures, program plans, or other governing documents used by contractors or agencies to implement requirements are referenced in this plan. This plan covers all planned monitoring and environmental surveillance. Nonroutine activities such as special research studies and characterization of individual sites for environmental restoration are outside the scope of this plan.

  13. Requirement for the eIF4E binding proteins for the synergistic down-regulation of protein synthesis by hypertonic conditions and mTOR inhibition.

    Science.gov (United States)

    Clemens, Michael J; Elia, Androulla; Morley, Simon J

    2013-01-01

    The protein kinase mammalian target of rapamycin (mTOR) regulates the phosphorylation and activity of several proteins that have the potential to control translation, including p70S6 kinase and the eIF4E binding proteins 4E-BP1 and 4E-BP2. In spite of this, in exponentially growing cells overall protein synthesis is often resistant to mTOR inhibitors. We report here that sensitivity of wild-type mouse embryonic fibroblasts (MEFs) to mTOR inhibitors can be greatly increased when the cells are subjected to the physiological stress imposed by hypertonic conditions. In contrast, protein synthesis in MEFs with a double knockout of 4E-BP1 and 4E-BP2 remains resistant to mTOR inhibitors under these conditions. Phosphorylation of p70S6 kinase and protein kinase B (Akt) is blocked by the mTOR inhibitor Ku0063794 equally well in both wild-type and 4E-BP knockout cells, under both normal and hypertonic conditions. The response of protein synthesis to hypertonic stress itself does not require the 4E-BPs. These data suggest that under certain stress conditions: (i) translation has a greater requirement for mTOR activity and (ii) there is an absolute requirement for the 4E-BPs for regulation by mTOR. Importantly, dephosphorylation of p70S6 kinase and Akt is not sufficient to affect protein synthesis acutely.

  14. Growth regulator requirement for in vitro embryogenic cultures of snowdrop (Galanthus nivalis L.) suitable for germplasm preservation.

    Science.gov (United States)

    Resetár, Anna; Demeter, Zita; Ficsor, Emese; Balázs, Andrea; Mosolygó, Agnes; Szőke, Eva; Gonda, S; Papp, L; Surányi, G; Máthé, C

    2014-06-01

    In this study, we report on the production of bulb scale-derived tissue cultures capable of efficient shoot and plant regeneration in three genotypes of snowdrop (Galanthus nivalis L., Amaryllidaceae), a protected ornamental plant. For culture line A, high auxin and low cytokinin concentration is required for callus production and plant regeneration. The type of auxin is of key importance: α-naphthaleneacetic acid (NAA) in combination with indole-3-acetic acid (IAA) at concentrations of 2 mg L-1 or 2-10 mg L-1 NAA with 1 mg L-1 N6-benzyladenine (BA), a cytokinin on full-strength media are required for regeneration. Cultures showing regeneration were embryogenic. When lines B and C were induced and maintained with 2 mg L-1 NAA and 1 mg L-1 BA, they produced mature bulblets with shoots, without roots. Line A produced immature bulblets with shoots under the above culture condition. Amplified Fragment Length Polymorphism (AFLP) analysis showed that (i) genetic differences between line A and its bulb explants were not significant, therefore these tissue cultures are suitable for germplasm preservation, and (ii) different morphogenetic responses of lines A, B and C originated from genetic differences. Culture line A is suitable for field-growing, cultivation and germplasm preservation of G. nivalis and for the production of Amaryllidaceae alkaloids.

  15. When Are Mobile Phones Useful for Water Quality Data Collection? An Analysis of Data Flows and ICT Applications among Regulated Monitoring Institutions in Sub-Saharan Africa.

    Science.gov (United States)

    Kumpel, Emily; Peletz, Rachel; Bonham, Mateyo; Fay, Annette; Cock-Esteb, Alicea; Khush, Ranjiv

    2015-09-02

    Water quality monitoring is important for identifying public health risks and ensuring water safety. However, even when water sources are tested, many institutions struggle to access data for immediate action or long-term decision-making. We analyzed water testing structures among 26 regulated water suppliers and public health surveillance agencies across six African countries and identified four water quality data management typologies. Within each typology, we then analyzed the potential for information and communication technology (ICT) tools to facilitate water quality information flows. A consistent feature of all four typologies was that testing activities occurred in laboratories or offices, not at water sources; therefore, mobile phone-based data management may be most beneficial for institutions that collect data from multiple remote laboratories. We implemented a mobile phone application to facilitate water quality data collection within the national public health agency in Senegal, Service National de l'Hygiène. Our results indicate that using the phones to transmit more than just water quality data will likely improve the effectiveness and sustainability of this type of intervention. We conclude that an assessment of program structure, particularly its data flows, provides a sound starting point for understanding the extent to which ICTs might strengthen water quality monitoring efforts.

  16. When Are Mobile Phones Useful for Water Quality Data Collection? An Analysis of Data Flows and ICT Applications among Regulated Monitoring Institutions in Sub-Saharan Africa

    Directory of Open Access Journals (Sweden)

    Emily Kumpel

    2015-09-01

    Full Text Available Water quality monitoring is important for identifying public health risks and ensuring water safety. However, even when water sources are tested, many institutions struggle to access data for immediate action or long-term decision-making. We analyzed water testing structures among 26 regulated water suppliers and public health surveillance agencies across six African countries and identified four water quality data management typologies. Within each typology, we then analyzed the potential for information and communication technology (ICT tools to facilitate water quality information flows. A consistent feature of all four typologies was that testing activities occurred in laboratories or offices, not at water sources; therefore, mobile phone-based data management may be most beneficial for institutions that collect data from multiple remote laboratories. We implemented a mobile phone application to facilitate water quality data collection within the national public health agency in Senegal, Service National de l’Hygiène. Our results indicate that using the phones to transmit more than just water quality data will likely improve the effectiveness and sustainability of this type of intervention. We conclude that an assessment of program structure, particularly its data flows, provides a sound starting point for understanding the extent to which ICTs might strengthen water quality monitoring efforts.

  17. Shape-induced terminal differentiation of human epidermal stem cells requires p38 and is regulated by histone acetylation.

    Directory of Open Access Journals (Sweden)

    John T Connelly

    Full Text Available Engineered model substrates are powerful tools for examining interactions between stem cells and their microenvironment. Using this approach, we have previously shown that restricted cell adhesion promotes terminal differentiation of human epidermal stem cells via activation of serum response factor (SRF and transcription of AP-1 genes. Here we investigate the roles of p38 MAPK and histone acetylation. Inhibition of p38 activity impaired SRF transcriptional activity and shape-induced terminal differentiation of human keratinocytes. In addition, inhibiting p38 reduced histone H3 acetylation at the promoters of SRF target genes, FOS and JUNB. Although histone acetylation correlated with SRF transcriptional activity and target gene expression, treatment with the histone de-acetylase inhibitor, trichostatin A (TSA blocked terminal differentiation on micro-patterned substrates and in suspension. TSA treatment simultaneously maintained expression of LRIG1, TP63, and ITGB1. Therefore, global histone de-acetylation represses stem cell maintenance genes independent of SRF. Our studies establish a novel role for extrinsic physical cues in the regulation of chromatin remodeling, transcription, and differentiation of human epidermal stem cells.

  18. Mammalian diaphanous-related formin 1 regulates GSK3β-dependent microtubule dynamics required for T cell migratory polarization.

    Directory of Open Access Journals (Sweden)

    Baoxia Dong

    Full Text Available The mammalian diaphanous-related formin (mDia1, a Rho-regulated cytoskeletal modulator, has been shown to promote T lymphocyte chemotaxis and interaction with antigen presenting cells, but the mechanisms underpinning mDia1 roles in these processes have not been defined. Here we show that mDia1(-/- T cells exhibit impaired lymphocyte function-associated antigen 1 (LFA-1-mediated T cell adhesion, migration and in vivo trafficking. These defects are associated with impaired microtubule (MT polarization and stabilization, altered MT dynamics and reduced peripheral clustering of the MT plus-end-protein, adenomatous polyposis coli (APC in migrating T cells following LFA-1-engagement. Loss of mDia1 also leads to impaired inducible inactivation of the glycogen synthase kinase (GSK 3β as well as hyperphosphorylation and reduced levels of APC in migrating T cells. These findings identify essential roles for the mDia1 formin in modulating GSK3β-dependent MT contributions to induction of T-cell polarity, adhesion and motility.

  19. Mammalian diaphanous-related formin 1 regulates GSK3β-dependent microtubule dynamics required for T cell migratory polarization.

    Science.gov (United States)

    Dong, Baoxia; Zhang, Steven S; Gao, Wen; Su, Haichun; Chen, Jun; Jin, Fuzi; Bhargava, Ajay; Chen, Xiequn; Jorgensen, Lars; Alberts, Arthur S; Zhang, Jinyi; Siminovitch, Katherine A

    2013-01-01

    The mammalian diaphanous-related formin (mDia1), a Rho-regulated cytoskeletal modulator, has been shown to promote T lymphocyte chemotaxis and interaction with antigen presenting cells, but the mechanisms underpinning mDia1 roles in these processes have not been defined. Here we show that mDia1(-/-) T cells exhibit impaired lymphocyte function-associated antigen 1 (LFA-1)-mediated T cell adhesion, migration and in vivo trafficking. These defects are associated with impaired microtubule (MT) polarization and stabilization, altered MT dynamics and reduced peripheral clustering of the MT plus-end-protein, adenomatous polyposis coli (APC) in migrating T cells following LFA-1-engagement. Loss of mDia1 also leads to impaired inducible inactivation of the glycogen synthase kinase (GSK) 3β as well as hyperphosphorylation and reduced levels of APC in migrating T cells. These findings identify essential roles for the mDia1 formin in modulating GSK3β-dependent MT contributions to induction of T-cell polarity, adhesion and motility.

  20. The plant cuticle is required for osmotic stress regulation of abscisic acid biosynthesis and osmotic stress tolerance in Arabidopsis

    KAUST Repository

    Wang, Zhenyu

    2011-05-01

    Osmotic stress activates the biosynthesis of abscisic acid (ABA). One major step in ABA biosynthesis is the carotenoid cleavage catalyzed by a 9-cis epoxycarotenoid dioxygenase (NCED). To understand the mechanism for osmotic stress activation of ABA biosynthesis, we screened for Arabidopsis thaliana mutants that failed to induce the NCED3 genee xpression in response to osmotic stress treatments. The ced1 (for 9-cis epoxycarotenoid dioxy genase defective 1) mutant isolated in this study showed markedly reduced expression of NCED3 in response to osmotic stress (polyethylene glycol)treatments compared with the wild type. Other ABA biosynthesis genes are also greatly reduced in ced1 under osmotic stress. ced1 mutant plants are very sensitive to even mild osmotic stress. Map-based cloning revealed unexpectedly thatCED1 encodes a putative a/b hydrolase domain-containing protein and is allelic to the BODYGUARD gene that was recently shown to be essential for cuticle biogenesis. Further studies discovered that other cut in biosynthesis mutants are also impaired in osmotic stress induction of ABA biosynthesis genes and are sensitive to osmotic stress. Our work demonstrates that the cuticle functions not merely as a physical barrier to minimize water loss but also mediates osmotic stress signaling and tolerance by regulating ABA biosynthesis and signaling. © 2011 American Society of Plant Biologists. All rights reserved.

  1. The imitation switch ATPase Snf2l is required for superovulation and regulates Fgl2 in differentiating mouse granulosa cells.

    Science.gov (United States)

    Pépin, David; Paradis, François; Perez-Iratxeta, Carol; Picketts, David J; Vanderhyden, Barbara C

    2013-06-01

    Imitation switch (ISWI) proteins are catalytic subunits of chromatin remodeling complexes that alter nucleosome positioning by hydrolyzing ATP to regulate access to DNA. In mice, there are two paralogs, SNF2-homolog (SNF2H) and SNF2-like (SNF2L), which participate in different complexes and have contrasting patterns of expression. Here we investigate the role of SNF2L in ovaries by characterizing a mouse bearing an inactivating deletion of exon 6 that disrupts the ATPase domain. Snf2l mutant mice produce significantly fewer eggs than control mice when superovulated. Gonadotropin stimulation leads to a significant deficit in secondary follicles and an increase in abnormal antral follicles. Mutant females also failed to induce fibrinogen-like 2 (Fgl2) in response to human chorionic gonadotropin (hCG) stimulation, while overexpression of SNF2L was sufficient to drive its expression in granulosa cells. SNF2L was also shown to directly interact with the nuclear receptor co-activator flightless I (FLI-I) as shown by immunoprecipitation. These results begin to establish a role for SNF2L in the precise coordination of gene expression in granulosa cells during folliculogenesis and its broader implications in fertility.

  2. The bristle patterning genes hairy and extramacrochaetae regulate the development of structures required for flight in Diptera☆

    Science.gov (United States)

    Costa, Marta; Calleja, Manuel; Alonso, Claudio R.; Simpson, Pat

    2014-01-01

    The distribution of sensory bristles on the thorax of Diptera (true flies) provides a useful model for the study of the evolution of spatial patterns. Large bristles called macrochaetes are arranged into species-specific stereotypical patterns determined via spatially discrete expression of the proneural genes achaete–scute (ac–sc). In Drosophila ac-sc expression is regulated by transcriptional activation at sites where bristle precursors develop and by repression outside of these sites. Three genes, extramacrochaetae (emc), hairy (h) and stripe (sr), involved in repression have been documented. Here we demonstrate that in Drosophila, the repressor genes emc and h, like sr, play an essential role in the development of structures forming part of the flight apparatus. In addition we find that, in Calliphora vicina a species diverged from D. melanogaster by about 100 Myr, spatial expression of emc, h and sr is conserved at the location of development of those structures. Based on these findings we argue, first, that the role emc, h and sr in development of the flight apparatus preceded their activities for macrochaete patterning; second, that species-specific variation in activation and repression of ac-sc expression is evolving in parallel to establish a unique distribution of macrochaetes in each species. PMID:24384389

  3. Suggested guidelines for gas emission monitoring at danish landfills

    DEFF Research Database (Denmark)

    Kjeldsen, Peter; Scheutz, Charlotte

    2015-01-01

    Landfill gas is produced on waste disposal sites receiving organic waste resulting in emission of methane. Regulation requires that the landfill gas is managed in order to reduce emissions, but very few suggestions exist to how the landfill gas management activities are monitored, what requirements...... measures to determine the efficiency of the performed emission mitigation is defined. Finally, several principles are presented for how criteria can be developed for when a monitoring program can be terminated....

  4. Nrk2b-mediated NAD+ production regulates cell adhesion and is required for muscle morphogenesis in vivo

    Science.gov (United States)

    Goody, Michelle F.; Kelly, Meghan W.; Lessard, Kevin N.; Khalil, Andre; Henry, Clarissa A.

    2010-01-01

    Cell-matrix adhesion complexes (CMACs) play fundamental roles during morphogenesis. Given the ubiquitous nature of CMACs and their roles in many cellular processes, one question is how specificity of CMAC function is modulated. The clearly defined cell behaviors that generate segmentally reiterated axial skeletal muscle during zebrafish development comprise an ideal system with which to investigate CMAC function during morphogenesis. We found that Nicotinamide riboside kinase 2b (Nrk2b) cell autonomously modulates the molecular composition of CMACs in vivo. Nrk2b is required for normal Laminin polymerization at the myotendinous junction (MTJ). In Nrk2b-deficient embryos, at MTJ loci where Laminin is not properly polymerized, muscle fibers elongate into adjacent myotomes and are abnormally long. In yeast and human cells, Nrk2 phosphorylates Nicotinamide Riboside and generates NAD+ through an alternative salvage pathway. Exogenous NAD+ treatment rescues MTJ development in Nrk2b-deficient embryos, but not in laminin mutant embryos. Both Nrk2b and Laminin are required for localization of Paxillin, but not β-Dystroglycan, to CMACs at the MTJ. Overexpression of Paxillin in Nrk2b-deficient embryos is sufficient to rescue MTJ integrity. Taken together, these data show that Nrk2b plays a specific role in modulating subcellular localization of discrete CMAC components that in turn play roles in musculoskeletal development. Furthermore, these data suggest that Nrk2b-mediated synthesis of NAD+ is functionally upstream of Laminin adhesion and Paxillin subcellular localization during MTJ development. These results indicate a previously unrecognized complexity to CMAC assembly in vivo and also elucidate a novel role for NAD+ during morphogenesis. PMID:20566368

  5. HCN channel dendritic targeting requires bipartite interaction with TRIP8b and regulates antidepressant-like behavioral effects

    Science.gov (United States)

    Han, Ye; Heuermann, Robert J.; Lyman, Kyle A.; Fisher, Daniel; Ismail, Quratul-Ain; Chetkovich, Dane M.

    2016-01-01

    Major Depressive Disorder is a prevalent psychiatric condition with limited therapeutic options beyond monoaminergic therapies. Although effective in some individuals, many patients fail to respond adequately to existing treatments and new pharmacologic targets are needed. HCN channels regulate excitability in neurons and blocking HCN channel function has been proposed as a novel antidepressant strategy. However, systemic blockade of HCN channels produces cardiac effects that limit this approach. Knockout (KO) of the brain-specific HCN channel auxiliary subunit TRIP8b also produces antidepressant-like behavioral effects and suggests that inhibiting TRIP8b function could produce antidepressant-like effects without affecting the heart. We examined the structural basis of TRIP8b-mediated HCN channel trafficking and its relationship to antidepressant-like behavior using a viral rescue approach in TRIP8b KO mice. We found that restoring TRIP8b to the hippocampus was sufficient to reverse the impaired HCN channel trafficking and antidepressant-like behavioral effects caused by TRIP8b KO. Moreover, we found that hippocampal expression of a mutated version of TRIP8b further impaired HCN channel trafficking and increased the antidepressant-like behavioral phenotype of TRIP8b KO mice. Thus, modulating the TRIP8b-HCN interaction bidirectionally influences channel trafficking and antidepressant-like behavior. Overall, our work suggests that small molecule inhibitors of the interaction between TRIP8b and HCN should produce antidepressant-like behaviors and could represent a new paradigm for the treatment of Major Depressive Disorder. PMID:27400855

  6. Palmitoylation on the carboxyl terminus tail is required for the selective regulation of dopamine D2 versus D3 receptors.

    Science.gov (United States)

    Zhang, Xiaowei; Le, Hang Thi; Zhang, Xiaohan; Zheng, Mei; Choi, Bo-Gil; Kim, Kyeong-Man

    2016-09-01

    Dopamine D2 receptor (D2R) and D3 receptor (D3R) possess highly conserved amino acid sequences but this study showed that D3R was more extensively palmitoylated than D2R. Based on this finding, the molecular basis of this selective palmitoylation of D3R was determined and the roles of palmitoylation in the regulation of D3R functions were investigated. D3R was palmitoylated on the cysteine residue on its carboxyl terminus tail, the last amino acid residue of D3R, and an exchange of the carboxyl terminus tail between D2R and D3R (D2R-D3C and D3R-D2C) resulted in the switching of the palmitoylation phenotype. When the consensus site for palmitoylation was mutated or the palmitoylation of D3R was inhibited by treatment with 2-bromopalmitate (2BP), a palmitoylation blocker, cell-surface expression, PKC-mediated endocytosis, agonist affinity, and agonist-induced tolerance of D3R were all inhibited. However, these changes were not observed when D3R palmitoylation was inhibited by replacing its carboxyl tail with that of D2R (D3R-D2C) or when the palmitoylation of D2R-D3C was inhibited by treatment with 2BP. Overall, this study shows that D3R is palmitoylated more extensively than D2R even though the carboxyl terminus tails of D2R and D3R are highly homologous, and thus provides a new clue regarding the consensus sequence for palmitoylation. This study also shows that palmitoylation controls various functionalities of D3R only when the receptor is in the intact D3R configuration. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Cue-elicited reward-seeking requires extracellular signal-regulated kinase activation in the nucleus accumbens.

    Science.gov (United States)

    Shiflett, Michael W; Martini, Ross P; Mauna, Jocelyn C; Foster, Rebecca L; Peet, Eloise; Thiels, Edda

    2008-02-01

    The motivation to seek out rewards can come under the control of stimuli associated with reward delivery. The ability of cues to motivate reward-seeking behavior depends on the nucleus accumbens (NAcc). The molecular mechanisms in the NAcc that underlie the ability of a cue to motivate reward-seeking are not well understood. We examined whether extracellular signal-regulated kinase (ERK), an important intracellular signaling pathway in learning and memory, has a role in these motivational processes. We first examined p42 ERK (ERK2) activation in the NAcc after rats were trained to associate an auditory stimulus with food delivery and found that, as a consequence of training, presentation of the auditory cue itself was sufficient to increase ERK2 activation in the NAcc. To examine whether inhibition of ERK in the NAcc prevents cue-induced reward-seeking, we infused an inhibitor of ERK, U0126, into the NAcc before assessing rats' instrumental responding in the presence versus absence of the conditioned cue. We found that, whereas vehicle-infused rats showed increased instrumental responding during cue presentation, rats infused with U0126 showed a profound impairment in cue-induced instrumental responding. In contrast, intra-NAcc U0126 infusion had no effect on rats' food-reinforced instrumental responding or their ability to execute conditioned approach behavior. Our results demonstrate learning-related changes in ERK signaling in the NAcc, and that disruption of ERK activation in this structure interferes with the incentive-motivational effects of conditioned stimuli. The molecular mechanisms described here may have implications for cue-elicited drug craving after repeated exposure to drugs of abuse.

  8. HCN-channel dendritic targeting requires bipartite interaction with TRIP8b and regulates antidepressant-like behavioral effects.

    Science.gov (United States)

    Han, Y; Heuermann, R J; Lyman, K A; Fisher, D; Ismail, Q-A; Chetkovich, D M

    2017-03-01

    Major depressive disorder (MDD) is a prevalent psychiatric condition with limited therapeutic options beyond monoaminergic therapies. Although effective in some individuals, many patients fail to respond adequately to existing treatments, and new pharmacologic targets are needed. Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels regulate excitability in neurons, and blocking HCN channel function has been proposed as a novel antidepressant strategy. However, systemic blockade of HCN channels produces cardiac effects that limit this approach. Knockout (KO) of the brain-specific HCN-channel auxiliary subunit tetratricopeptide repeat-containing Rab8b-interacting protein (TRIP8b) also produces antidepressant-like behavioral effects and suggests that inhibiting TRIP8b function could produce antidepressant-like effects without affecting the heart. We examined the structural basis of TRIP8b-mediated HCN-channel trafficking and its relationship with antidepressant-like behavior using a viral rescue approach in TRIP8b KO mice. We found that restoring TRIP8b to the hippocampus was sufficient to reverse the impaired HCN-channel trafficking and antidepressant-like behavioral effects caused by TRIP8b KO. Moreover, we found that hippocampal expression of a mutated version of TRIP8b further impaired HCN-channel trafficking and increased the antidepressant-like behavioral phenotype of TRIP8b KO mice. Thus, modulating the TRIP8b-HCN interaction bidirectionally influences channel trafficking and antidepressant-like behavior. Overall, our work suggests that small-molecule inhibitors of the interaction between TRIP8b and HCN should produce antidepressant-like behaviors and could represent a new paradigm for the treatment of MDD.

  9. Development and application of a permit information system for shale oil (PERMISSO). Final report appendix: summary sheets of regulations required for oil shale development, June 1978--May 1979

    Energy Technology Data Exchange (ETDEWEB)

    1979-07-01

    This appendix is comprised of summaries of various governmental permits, licenses and other approvals required for oil shale development. The summaries were completed during the period June--October 1978, and are current as of July 1, 1978, although more recent authority was cited in some cases. One of the major purposes of Phase II of the project will be to update these summaries as statutes and regulations are added, changed or eliminated. This updating will be particularly important in the case of environmental permits and approvals. Many legislative and regulatory changes affecting environmental requirements are pending at this time and will alter many of the summaries herein. In addition, many regulatory proposals have been or likely will be challenged in the courts. When such conflicts are resolved further changes may be in order.

  10. Application of wide selected-ion monitoring data-independent acquisition to identify tomato fruit proteins regulated by the CUTIN DEFICIENT2 transcription factor.

    Science.gov (United States)

    Martin, Laetitia B B; Sherwood, Robert W; Nicklay, Joshua J; Yang, Yong; Muratore-Schroeder, Tara L; Anderson, Elizabeth T; Thannhauser, Theodore W; Rose, Jocelyn K C; Zhang, Sheng

    2016-08-01

    We describe here the use of label-free wide selected-ion monitoring data-independent acquisition (WiSIM-DIA) to identify proteins that are involved in the formation of tomato (Solanum lycopersicum) fruit cuticles and that are regulated by the transcription factor CUTIN DEFICIENT2 (CD2). A spectral library consisting of 11 753 unique peptides, corresponding to 2338 tomato protein groups, was used and the DIA analysis was performed at the MS1 level utilizing narrow mass windows for extraction with Skyline 2.6 software. We identified a total of 1140 proteins, 67 of which had expression levels that differed significantly between the cd2 tomato mutant and the wild-type cultivar M82. Differentially expressed proteins including a key protein involved in cutin biosynthesis, were selected for validation by target SRM/MRM and by Western blot analysis. In addition to confirming a role for CD2 in regulating cuticle formation, the results also revealed that CD2 influences pathways associated with cell wall biology, anthocyanin biosynthesis, plant development, and responses to stress, which complements findings of earlier RNA-Seq experiments. Our results provide new insights into molecular processes and aspects of fruit biology associated with CD2 function, and demonstrate that the WiSIM-DIA is an effective quantitative approach for global protein identifications.

  11. Quality Assurance Program Plan for radionuclide airborne emissions monitoring

    Energy Technology Data Exchange (ETDEWEB)

    Vance, L.M.

    1993-07-01

    This Quality Assurance Program Plan (QAPP) describes the quality assurance requirements and responsibilities for radioactive airborne emissions measurements activities from regulated stacks are controlled at the Hanford Site. Detailed monitoring requirements apply to stacks exceeding 1% of the standard of 10 mrem annual effective dose equivalent to the maximally exposed individual from operations of the Hanford Site.

  12. The Carboxy-terminus of BAK1 regulates kinase activity and is required for normal growth of Arabidopsis

    Directory of Open Access Journals (Sweden)

    Man-Ho eOh

    2014-02-01

    Full Text Available Binding of brassinolide to the BRASSINOSTEROID-INSENSTIVE 1 (BRI1 receptor kinase promotes interaction with its co-receptor, BRI1-ASSOCIATED RECEPTOR KINASE 1 (BAK1. Juxtaposition of the kinase domains that occurs then allows reciprocal transphosphorylation and activation of both kinases, but details of that process are not entirely clear. In the present study we show that the carboxy (C - terminal polypeptide of BAK1 may play a role. First, we demonstrate that the C-terminal domain is a strong inhibitor of the transphosphorylation activity of the recombinant BAK1 cytoplasmic domain protein. However, recombinant BAK1 lacking the C-terminal domain is unable to transactivate the peptide kinase activity of BRI1 in vitro. Thus, the C-terminal domain may play both a positive and negative role. Interestingly, a synthetic peptide corresponding to the full C-terminal domain (residues 576 to 615 of BAK1 interacted with recombinant BRI1 in vitro, and that interaction was enhanced by phosphorylation at the Tyr-610 site. Expression of a BAK1 C-terminal domain truncation (designated BAK1-ΔCT-Flag in transgenic Arabidopsis plants lacking endogenous bak1 and its functional paralog, bkk1, produced plants that were wild type in appearance but much smaller than plants expressing full-length BAK1-Flag. The reduction in growth may be attributed to a partial inhibition of BR signaling in vivo as reflected in root growth assays but other factors are likely involved as well. Our working model is that in vivo, the inhibitory action of the C-terminal domain of BAK1 is relieved by binding to BRI1. However, that interaction is not essential for BR signaling, but other aspects of cellular signaling are impacted when the C-terminal domain is truncated and result in inhibition of growth. These results increase the molecular understanding of the C-terminal domain of BAK1 as a regulator of kinase activity that may serve as a model for other receptor kinases.

  13. Transcriptional regulation by triiodothyronine requires synergistic action of the thyroid receptor with another trans-acting factor.

    Science.gov (United States)

    Voz, M L; Peers, B; Wiedig, M J; Jacquemin, P; Belayew, A; Martial, J A

    1992-09-01

    Human placental lactogen B (hCS-B) promoter activity is strongly stimulated by triiodothyronine (T3) in pituitary GC cells through interaction between the thyroid receptor and a thyroid receptor-binding element (TBE) spanning coordinates -67 to -41. This TBE is adjacent to the binding site for pituitary factor GHF1 (-95 to -68) which seems necessary for T3 stimulation of hCS-B promoter activity (M. L. Voz, B. Peers, A. Belayew, and J. A. Martial, J. Biol. Chem. 266:13397-13404, 1991). We here demonstrate actual synergy between the thyroid receptor and GHF1. Indeed, in placental JEG-3 cells devoid of factor GHF1, hCS promoter activity is barely stimulated by T3, while a strong response is observed in pituitary GC cells. In the latter, furthermore, neither the TBE nor the GHF1-binding site alone is sufficient to render the thymidine kinase promoter responsive to T3, while in combination they promote strong T3 stimulation. Close proximity between these sites is required for optimal synergy: T3 stimulation globally decreases with increased spacing. Furthermore, synergy occurs not only with a GHF1-binding site but also with all other factor recognition sequences tested (Sp1, NF1, CP1, Oct1, and CACCC boxes) and even with two other copies of the TBE. Nor is it specific to hCS TBE, since the palindromic sequence TCAGGTCA TGACCTGA (TREpal) also exhibits cooperativity.

  14. A Myb transcription factor of Phytophthora sojae, regulated by MAP kinase PsSAK1, is required for zoospore development.

    Directory of Open Access Journals (Sweden)

    Meng Zhang

    Full Text Available PsSAK1, a mitogen-activated protein (MAP kinase from Phytophthora sojae, plays an important role in host infection and zoospore viability. However, the downstream mechanism of PsSAK1 remains unclear. In this study, the 3'-tag digital gene expression (DGE profiling method was applied to sequence the global transcriptional sequence of PsSAK1-silenced mutants during the cysts stage and 1.5 h after inoculation onto susceptible soybean leaf tissues. Compared with the gene expression levels of the recipient P. sojae strain, several candidates of Myb family were differentially expressed (up or down in response to the loss of PsSAK1, including of a R2R3-type Myb transcription factor, PsMYB1. qRT-PCR indicated that the transcriptional level of PsMYB1 decreased due to PsSAK1 silencing. The transcriptional level of PsMYB1 increased during sporulating hyphae, in germinated cysts, and early infection. Silencing of PsMYB1 results in three phenotypes: a no cleavage of the cytoplasm into uninucleate zoospores or release of normal zoospores, b direct germination of sporangia, and c afunction in zoospore-mediated plant infection. Our data indicate that the PsMYB1 transcription factor functions downstream of MAP kinase PsSAK1 and is required for zoospore development of P. sojae.

  15. A Conserved C-Terminal Domain of the Aspergillus fumigatus Developmental Regulator MedA Is Required for Nuclear Localization, Adhesion and Virulence

    Science.gov (United States)

    Al Abdallah, Qusai; Choe, Se-In; Campoli, Paolo; Baptista, Stefanie; Gravelat, Fabrice N.; Lee, Mark J.; Sheppard, Donald C.

    2012-01-01

    MedA is a developmental regulator that is conserved in the genome of most filamentous fungi. In the pathogenic fungus Aspergillus fumigatus MedA regulates conidiogenesis, adherence to host cells, and pathogenicity. The mechanism by which MedA governs these phenotypes remains unknown. Although the nuclear import of MedA orthologues has been reported in other fungi, no nuclear localization signal, DNA-binding domain or other conserved motifs have been identified within MedA. In this work, we performed a deletion analysis of MedA and identified a novel domain within the C-terminal region of the protein, designated MedA346–557, that is necessary and sufficient for nuclear localization of MedA. We further demonstrate that MedA nuclear localization is required for the function of MedA. Surprisingly, expression of the minimal nuclear localization fragment MedA346–557 alone was sufficient to restore conidogenesis, biofilm formation and virulence to the medA mutant strain. Collectively these results suggest that MedA functions in the regulation of transcription, and that the MedA346–557 domain is both necessary and sufficient to mediate MedA function. PMID:23185496

  16. NF1 regulation of RAS/ERK signaling is required for appropriate granule neuron progenitor expansion and migration in cerebellar development.

    Science.gov (United States)

    Sanchez-Ortiz, Efrain; Cho, Woosung; Nazarenko, Inga; Mo, Wei; Chen, Jian; Parada, Luis F

    2014-11-01

    Cerebellar development is regulated by a coordinated spatiotemporal interplay between granule neuron progenitors (GNPs), Purkinje neurons, and glia. Abnormal development can trigger motor deficits, and more recent data indicate important roles in aspects of memory, behavior, and autism spectrum disorders (ASDs). Germline mutation in the NF1 tumor suppressor gene underlies Neurofibromatosis type 1, a complex disease that enhances susceptibility to certain cancers and neurological disorders, including intellectual deficits and ASD. The NF1 gene encodes for neurofibromin, a RAS GTPase-activating protein, and thus negatively regulates the RAS signaling pathway. Here, using mouse models to direct conditional NF1 ablation in either embryonic cerebellar progenitors or neonatal GNPs, we show that neurofibromin is required for appropriate development of cerebellar folia layering and structure. Remarkably, neonatal administration of inhibitors of the ERK pathway reversed the morphological defects. Thus, our findings establish a critical cell-autonomous role for the NF1-RAS-ERK pathway in the appropriate regulation of cerebellar development and provide a basis for using neonatal ERK inhibitor-based therapies to treat NF1-induced cerebellar disorders.

  17. A LysR-family transcriptional regulator required for virulence in Brucella abortus is highly conserved among the α-proteobacteria.

    Science.gov (United States)

    Sheehan, Lauren M; Budnick, James A; Blanchard, Catlyn; Dunman, Paul M; Caswell, Clayton C

    2015-10-01

    Small RNAs are principal elements of bacterial gene regulation and physiology. Two small RNAs in Brucella abortus, AbcR1 and AbcR2, are required for wild-type virulence. Examination of the abcR loci revealed the presence of a gene encoding a LysR-type transcriptional regulator flanking abcR2 on chromosome 1. Deletion of this lysR gene (bab1_1517) resulted in the complete loss of abcR2 expression while no difference in abcR1 expression was observed. The B. abortus bab1_1517 mutant strain was significantly attenuated in macrophages and mice, and bab1_1517 was subsequently named vtlR for virulence-associated transcriptional LysR-family regulator. Microarray analysis revealed three additional genes encoding small hypothetical proteins also under the control of VtlR. Electrophoretic mobility shift assays demonstrated that VtlR binds directly to the promoter regions of abcR2 and the three hypothetical protein-encoding genes, and DNase I footprint analysis identified the specific nucleotide sequence in these promoters that VtlR binds to and drives gene expression. Strikingly, orthologs of VtlR are encoded in a wide range of host-associated α-proteobacteria, and it is likely that the VtlR genetic system represents a common regulatory circuit critical for host-bacterium interactions.

  18. Requirements for Foreign and Domestic Establishment Registration and Listing for Human Drugs, Including Drugs That Are Regulated Under a Biologics License Application, and Animal Drugs. Final rule.

    Science.gov (United States)

    2016-08-31

    The Food and Drug Administration (FDA) is amending its regulations governing drug establishment registration and drug listing. These amendments reorganize, modify, and clarify current regulations concerning who must register establishments and list human drugs, human drugs that are also biological products, and animal drugs. The final rule requires electronic submission, unless waived in certain circumstances, of registration and listing information. This rulemaking pertains to finished drug products and to active pharmaceutical ingredients (APIs) alone or together with one or more other ingredients. The final rule describes how and when owners or operators of establishments at which drugs are manufactured or processed must register their establishments with FDA and list the drugs they manufacture or process. In addition, the rule makes certain changes to the National Drug Code (NDC) system. We are taking this action to improve management of drug establishment registration and drug listing requirements and make these processes more efficient and effective for industry and for us. This action also supports implementation of the electronic prescribing provisions of the Medicare Prescription Drug, Improvement, and Modernization Act of 2003 (MMA) and the availability of current drug labeling information through DailyMed, a computerized repository of drug information maintained by the National Library of Medicine.

  19. The Drosophila deoxyhypusine hydroxylase homologue nero and its target eIF5A are required for cell growth and the regulation of autophagy.

    Science.gov (United States)

    Patel, Prajal H; Costa-Mattioli, Mauro; Schulze, Karen L; Bellen, Hugo J

    2009-06-29

    Hypusination is a unique posttranslational modification by which lysine is transformed into the atypical amino acid hypusine. eIF5A (eukaryotic initiation factor 5A) is the only known protein to contain hypusine. In this study, we describe the identification and characterization of nero, the Drosophila melanogaster deoxyhypusine hydroxylase (DOHH) homologue. nero mutations affect cell and organ size, bromodeoxyuridine incorporation, and autophagy. Knockdown of the hypusination target eIF5A via RNA interference causes phenotypes similar to nero mutations. However, loss of nero appears to cause milder phenotypes than loss of eIF5A. This is partially explained through a potential compensatory mechanism by which nero mutant cells up-regulate eIF5A levels. The failure of eIF5A up-regulation to rescue nero mutant phenotypes suggests that hypusination is required for eIF5A function. Furthermore, expression of enzymatically impaired forms of DOHH fails to rescue nero clones, indicating that hypusination activity is important for nero function. Our data also indicate that nero and eIF5A are required for cell growth and affect autophagy and protein synthesis.

  20. Novel promoter sequence required for manganese regulation of manganese peroxidase isozyme 1 gene expression in Phanerochaete chrysosporium.

    Science.gov (United States)

    Ma, Biao; Mayfield, Mary B; Godfrey, Bruce J; Gold, Michael H

    2004-06-01

    Manganese peroxidase (MnP) is a major, extracellular component of the lignin-degrading system produced by the wood-rotting basidiomycetous fungus Phanerochaete chrysosporium. The transcription of MnP-encoding genes (mnps) in P. chrysosporium occurs as a secondary metabolic event, triggered by nutrient-nitrogen limitation. In addition, mnp expression occurs only under Mn2+ supplementation. Using a reporter system based on the enhanced green fluorescent protein gene (egfp), we have characterized the P. chrysosporium mnp1 promoter by examining the effects of deletion, replacement, and translocation mutations on mnp1 promoter-directed egfp expression. The 1,528-bp mnp1 promoter fragment drives egfp expression only under Mn2+-sufficient, nitrogen-limiting conditions, as required for endogenous MnP production. However, deletion of a 48-bp fragment, residing 521 bp upstream of the translation start codon in the mnp1 promoter, or replacement of this fragment with an unrelated sequence resulted in egfp expression under nitrogen limitation, both in the absence and presence of exogenous Mn2+. Translocation of the 48-bp fragment to a site 120 bp downstream of its original location resulted in Mn2+-dependent egfp expression under conditions similar to those observed with the wild-type mnp1 promoter. These results suggest that the 48-bp fragment contains at least one Mn2+-responsive cis element. Additional promoter-deletion experiments suggested that the Mn2+ element(s) is located within the 33-bp sequence at the 3' end of the 48-bp fragment. This is the first promoter sequence containing a Mn2+-responsive element(s) to be characterized in any eukaryotic organism. Copyright 2004 American Society for Microbiology

  1. Personal-UV-monitoring in outdoor workplaces - a base for well-balanced regulations in heath prevention and care; UV-Monitoring an Arbeitsplaetzen im Freien - eine Basis fuer ausgewogene Richtlinien zum Gesundheitsschutz

    Energy Technology Data Exchange (ETDEWEB)

    Unverricht, I.; Knuschke, P. [Klinik und Poliklinik fuer Dermatologie, Technische Universitaet Dresden (Germany); Janssen, M.; Ott, G. [Bundesanstalt fuer Arbeitsschutz und Arbeitsmedizin Dortmund (Germany)

    2004-07-01

    The effects of UV-radiation on the skin and the eyes are the same - independent if the UV-exposure was during work time or during spare time. The actual individual UVexposure depends on one hand on the special profession activities on the other hand on the spare time and holiday behaviour. Therefore, the relations between the different outdoor professions in times being outside and further objective and subjective influences on the individual UV-exposure should be investigated by personal-UV-monitoring in the course of the year (BAuA(Federal Institute for Occupational Safety and Health) research project F1777). The investigation should be compared with former results of an analogous personal-UVmonitoring in indoor workers. Guidelines on occupational safety and health for outdoor workplaces cannot be based on the ICNIRP-Guidelines for UV-exposed indoor workplaces. These guidelines refer to unexposed skin e.g. in Wintertime. The skin of outdoor workers builts up an internal self-protection (increased cornea thickness; pigmentation). Therefore, well-balanced regulations for outdoor workplaces should be based on an acceptable UV-exposure level above that of indoor workers. (orig.)

  2. Ecological Monitoring and Compliance Program 2007 Report

    Energy Technology Data Exchange (ETDEWEB)

    Hansen, Dennis; Anderson, David; Derek, Hall; Greger, Paul; Ostler, W. Kent

    2008-03-01

    In accordance with U.S. Department of Energy (DOE) Order 450.1, 'Environmental Protection Program', the Office of the Assistant Manager for Environmental Management of the DOE, National Nuclear Security Administration Nevada Site Office (NNSA/NSO) requires ecological monitoring and biological compliance support for activities and programs conducted at the Nevada Test Site (NTS). National Security Technologies, LLC (NSTec), Ecological Services has implemented the Ecological Monitoring and Compliance (EMAC) Program to provide this support. EMAC is designed to ensure compliance with applicable laws and regulations, delineate and define NTS ecosystems, and provide ecological information that can be used to predict and evaluate the potential impacts of proposed projects and programs on those ecosystems. This report summarizes the EMAC activities conducted by NSTec during calendar year 2007. Monitoring tasks during 2007 included eight program areas: (a) biological surveys, (b) desert tortoise compliance, (c) ecosystem mapping and data management, (d) sensitive plant monitoring, (e) sensitive and protected/regulated animal monitoring, (f) habitat monitoring, (g) habitat restoration monitoring, and (h) biological monitoring at the Nonproliferation Test and Evaluation Complex (NPTEC). The following sections of this report describe work performed under these eight areas.

  3. MST kinases monitor actin cytoskeletal integrity and signal via c-Jun N-terminal kinase stress-activated kinase to regulate p21Waf1/Cip1 stability.

    Science.gov (United States)

    Densham, Ruth M; O'Neill, Eric; Munro, June; König, Ireen; Anderson, Kurt; Kolch, Walter; Olson, Michael F

    2009-12-01

    As well as providing a structural framework, the actin cytoskeleton plays integral roles in cell death, survival, and proliferation. The disruption of the actin cytoskeleton results in the activation of the c-Jun N-terminal kinase (JNK) stress-activated protein kinase (SAPK) pathway; however, the sensor of actin integrity that couples to the JNK pathway has not been characterized in mammalian cells. We now report that the mammalian Ste20-like (MST) kinases mediate the activation of the JNK pathway in response to the disruption of the actin cytoskeleton. One consequence of actin disruption is the JNK-mediated stabilization of p21(Waf1/Cip1) (p21) via the phosphorylation of Thr57. The expression of MST1 or MST2 was sufficient to stabilize p21 in a JNK- and Thr57-dependent manner, while the stabilization of p21 by actin disruption required MST activity. These data indicate that, in addition to being components of the Salvador-Warts-Hippo tumor suppressor network and binding partners of c-Raf and the RASSF1A tumor suppressor, MST kinases serve to monitor cytoskeletal integrity and couple via the JNK SAPK pathway to the regulation of a key cell cycle regulatory protein.

  4. Monitoring of surface and airborne contamination

    Energy Technology Data Exchange (ETDEWEB)

    Pradeep Kumar, K.S. [Bhabha Atomic Research Centre, Bombay (India)

    1997-06-01

    Indian nuclear energy programme aims at total safety in all activities involved in the entire fuel cycle for the occupational workers, members of the public and the environment as a whole. Routine radiation monitoring with clearly laid out procedures are followed for ensuring the safety of workers and public. Radiation monitoring carried out for the nuclear installations comprises of process monitoring, monitoring of effluent releases and also of the radiation protection monitoring of the individuals, work place and environment. Regulations like banning of smoking and consumption of food and drink etc. reduces the risk of direct ingestion even if inadvertent spread of contamination takes place. Though limit of transportable surface contamination is prescribed, the health physicists always follow a ``clean on swipe`` philosophy which compensates any error in the measurement of surface contamination. In this paper, the following items are contained: Necessity of contamination monitoring, accuracy required in the calibration of surface contamination monitors, methodology for contamination monitoring, air monitoring, guidelines for unrestricted release of scrap materials, and problems in contamination monitoring. (G.K.)

  5. A dietary polyphenol resveratrol acts to provide neuroprotection in recurrent stroke models by regulating AMPK and SIRT1 signaling, thereby reducing energy requirements during ischemia.

    Science.gov (United States)

    2016-04-01

    The above article from European Journal of Neuroscience, published online on 5 March 2013 in Wiley Online Library (http://onlinelibrary.wiley.com/doi/10.1111/ejn.12162/full), has been retracted by agreement between the Editors-in-Chief, Paul Bolam and John Foxe, the authors and John Wiley & Sons Ltd. The retraction has been agreed as Dr Phillip Barber has informed the publisher that he had seen neither the original data nor any version of the manuscript, and had not been involved in the work reported. A subsequent Institutional investigation found evidence of misconduct on the part of the submitting author. Reference Wang, L.-M., Wang, Y.-J., Cui, M., Luo, W.-J., Wang, X.-J., Barber, P.A. & Chen, Z.-Y. (2013) A dietary polyphenol resveratrol acts to provide neuroprotection in recurrent stroke models by regulating AMPK and SIRT1 signaling, thereby reducing energy requirements during ischemia.

  6. The calcium-sensing receptor-dependent regulation of cell-cell adhesion and keratinocyte differentiation requires Rho and filamin A.

    Science.gov (United States)

    Tu, Chia-Ling; Chang, Wenhan; Bikle, Daniel D

    2011-05-01

    Extracellular Ca(2+) (Ca(2+)(o)) functioning through the calcium-sensing receptor (CaR) induces E-cadherin-mediated cell-cell adhesion and cellular signals mediating cell differentiation in epidermal keratinocytes. Previous studies indicate that CaR regulates cell-cell adhesion through Fyn/Src tyrosine kinases. In this study, we investigate whether Rho GTPase is a part of the CaR-mediated signaling cascade regulating cell adhesion and differentiation. Suppressing endogenous Rho A expression by small interfering RNA (siRNA)-mediated gene silencing blocked the Ca(2+)(o)-induced association of Fyn with E-cadherin and suppressed the Ca(2+)(o)-induced tyrosine phosphorylation of β-, γ-, and p120-catenin and formation of intercellular adherens junctions. Rho A silencing also decreased the Ca(2+)(o)-stimulated expression of terminal differentiation markers. Elevating the Ca(2+)(o) level induced interactions among CaR, Rho A, E-cadherin, and the scaffolding protein filamin A at the cell membrane. Inactivation of CaR expression by adenoviral expression of a CaR antisense complementary DNA inhibited Ca(2+)(o)-induced activation of endogenous Rho. Ca(2+)(o) activation of Rho required a direct interaction between CaR and filamin A. Interference of CaR-filamin interaction inhibited Ca(2+)(o)-induced Rho activation and the formation of cell-cell junctions. These results indicate that Rho is a downstream mediator of CaR in the regulation of Ca(2+)(o)-induced E-cadherin-mediated cell-cell adhesion and keratinocyte differentiation.

  7. Proper actin ring formation and septum constriction requires coordinated regulation of SIN and MOR pathways through the germinal centre kinase MST-1.

    Science.gov (United States)

    Heilig, Yvonne; Dettmann, Anne; Mouriño-Pérez, Rosa R; Schmitt, Kerstin; Valerius, Oliver; Seiler, Stephan

    2014-04-01

    Nuclear DBF2p-related (NDR) kinases constitute a functionally conserved protein family of eukaryotic regulators that control cell division and polarity. In fungi, they function as effector kinases of the morphogenesis (MOR) and septation initiation (SIN) networks and are activated by pathway-specific germinal centre (GC) kinases. We characterized a third GC kinase, MST-1, that connects both kinase cascades. Genetic and biochemical interactions with SIN components and life cell imaging identify MST-1 as SIN-associated kinase that functions in parallel with the GC kinase SID-1 to activate the SIN-effector kinase DBF-2. SID-1 and MST-1 are both regulated by the upstream SIN kinase CDC-7, yet in an opposite manner. Aberrant cortical actomyosin rings are formed in Δmst-1, which resulted in mis-positioned septa and irregular spirals, indicating that MST-1-dependent regulation of the SIN is required for proper formation and constriction of the septal actomyosin ring. However, MST-1 also interacts with several components of the MOR network and modulates MOR activity at multiple levels. MST-1 functions as promiscuous enzyme and also activates the MOR effector kinase COT-1 through hydrophobic motif phosphorylation. In addition, MST-1 physically interacts with the MOR kinase POD-6, and dimerization of both proteins inactivates the GC kinase hetero-complex. These data specify an antagonistic relationship between the SIN and MOR during septum formation in the filamentous ascomycete model Neurospora crassa that is, at least in part, coordinated through the GC kinase MST-1. The similarity of the SIN and MOR pathways to the animal Hippo and Ndr pathways, respectively, suggests that intensive cross-communication between distinct NDR kinase modules may also be relevant for the homologous NDR kinases of higher eukaryotes.

  8. Proper actin ring formation and septum constriction requires coordinated regulation of SIN and MOR pathways through the germinal centre kinase MST-1.

    Directory of Open Access Journals (Sweden)

    Yvonne Heilig

    2014-04-01

    Full Text Available Nuclear DBF2p-related (NDR kinases constitute a functionally conserved protein family of eukaryotic regulators that control cell division and polarity. In fungi, they function as effector kinases of the morphogenesis (MOR and septation initiation (SIN networks and are activated by pathway-specific germinal centre (GC kinases. We characterized a third GC kinase, MST-1, that connects both kinase cascades. Genetic and biochemical interactions with SIN components and life cell imaging identify MST-1 as SIN-associated kinase that functions in parallel with the GC kinase SID-1 to activate the SIN-effector kinase DBF-2. SID-1 and MST-1 are both regulated by the upstream SIN kinase CDC-7, yet in an opposite manner. Aberrant cortical actomyosin rings are formed in Δmst-1, which resulted in mis-positioned septa and irregular spirals, indicating that MST-1-dependent regulation of the SIN is required for proper formation and constriction of the septal actomyosin ring. However, MST-1 also interacts with several components of the MOR network and modulates MOR activity at multiple levels. MST-1 functions as promiscuous enzyme and also activates the MOR effector kinase COT-1 through hydrophobic motif phosphorylation. In addition, MST-1 physically interacts with the MOR kinase POD-6, and dimerization of both proteins inactivates the GC kinase hetero-complex. These data specify an antagonistic relationship between the SIN and MOR during septum formation in the filamentous ascomycete model Neurospora crassa that is, at least in part, coordinated through the GC kinase MST-1. The similarity of the SIN and MOR pathways to the animal Hippo and Ndr pathways, respectively, suggests that intensive cross-communication between distinct NDR kinase modules may also be relevant for the homologous NDR kinases of higher eukaryotes.

  9. Regulation of abiotic stress signalling by Arabidopsis C-terminal domain phosphatase-like 1 requires interaction with a k-homology domain-containing protein.

    Directory of Open Access Journals (Sweden)

    In Sil Jeong

    Full Text Available Arabidopsis thaliana CARBOXYL-TERMINAL DOMAIN (CTD PHOSPHATASE-LIKE 1 (CPL1 regulates plant transcriptional responses to diverse stress signals. Unlike typical CTD phosphatases, CPL1 contains two double-stranded (ds RNA binding motifs (dsRBMs at its C-terminus. Some dsRBMs can bind to dsRNA and/or other proteins, but the function of the CPL1 dsRBMs has remained obscure. Here, we report identification of REGULATOR OF CBF GENE EXPRESSION 3 (RCF3 as a CPL1-interacting protein. RCF3 co-purified with tandem-affinity-tagged CPL1 from cultured Arabidopsis cells and contains multiple K-homology (KH domains, which were predicted to be important for binding to single-stranded DNA/RNA. Yeast two-hybrid, luciferase complementation imaging, and bimolecular fluorescence complementation analyses established that CPL1 and RCF3 strongly associate in vivo, an interaction mediated by the dsRBM1 of CPL1 and the KH3/KH4 domains of RCF3. Mapping of functional regions of CPL1 indicated that CPL1 in vivo function requires the dsRBM1, catalytic activity, and nuclear targeting of CPL1. Gene expression profiles of rcf3 and cpl1 mutants were similar during iron deficiency, but were distinct during the cold response. These results suggest that tethering CPL1 to RCF3 via dsRBM1 is part of the mechanism that confers specificity to CPL1-mediated transcriptional regulation.

  10. Regulation of Monocarboxylic Acid Transporter 1 Trafficking by the Canonical Wnt/β-Catenin Pathway in Rat Brain Endothelial Cells Requires Cross-talk with Notch Signaling.

    Science.gov (United States)

    Liu, Zejian; Sneve, Mary; Haroldson, Thomas A; Smith, Jeffrey P; Drewes, Lester R

    2016-04-01

    The transport of monocarboxylate fuels such as lactate, pyruvate, and ketone bodies across brain endothelial cells is mediated by monocarboxylic acid transporter 1 (MCT1). Although the canonical Wnt/β-catenin pathway is required for rodent blood-brain barrier development and for the expression of associated nutrient transporters, the role of this pathway in the regulation of brain endothelial MCT1 is unknown. Here we report expression of nine members of the frizzled receptor family by the RBE4 rat brain endothelial cell line. Furthermore, activation of the canonical Wnt/β-catenin pathway in RBE4 cells via nuclear β-catenin signaling with LiCl does not alter brain endothelialMct1mRNA but increases the amount of MCT1 transporter protein. Plasma membrane biotinylation studies and confocal microscopic examination of mCherry-tagged MCT1 indicate that increased transporter results from reduced MCT1 trafficking from the plasma membrane via the endosomal/lysosomal pathway and is facilitated by decreased MCT1 ubiquitination following LiCl treatment. Inhibition of the Notch pathway by the γ-secretase inhibitorN-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycinet-butyl ester negated the up-regulation of MCT1 by LiCl, demonstrating a cross-talk between the canonical Wnt/β-catenin and Notch pathways. Our results are important because they show, for the first time, the regulation of MCT1 in cerebrovascular endothelial cells by the multifunctional canonical Wnt/β-catenin and Notch signaling pathways.

  11. The master regulator for biofilm formation in Bacillus subtilis governs the expression of an operon encoding secreted proteins required for the assembly of complex multicellular communities.

    Energy Technology Data Exchange (ETDEWEB)

    Branda, Steven S. (Harvard Medical School, Cambridge, MA); Losick, Richard (Harvard University, Cambridge, MA); Kolter, Roberto (Harvard Medical School, Cambridge, MA); Kearns, Daniel B. (Harvard University, Cambridge, MA); Chu, Frances (Harvard University, Cambridge, MA)

    2005-08-01

    Wild strains of Bacillus subtilis are capable of forming architecturally complex communities of cells known as biofilms. Critical to biofilm formation is the eps operon, which is believed to be responsible for the biosynthesis of an exopolysaccharide that binds chains of cells together in bundles. We report that transcription of eps is under the negative regulation of SinR, a repressor that was found to bind to multiple sites in the regulatory region of the operon. Mutations in sinR bypassed the requirement in biofilm formation of two genes of unknown function, ylbF and ymcA, and sinI, which is known to encode an antagonist of SinR. We propose that these genes are members of a pathway that is responsible for counteracting SinR-mediated repression. We further propose that SinR is a master regulator that governs the transition between a planktonic state in which the bacteria swim as single cells in liquid or swarm in small groups over surfaces, and a sessile state in which the bacteria adhere to each other to form bundled chains and assemble into multicellular communities.

  12. Casein kinase II is required for proper cell division and acts as a negative regulator of centrosome duplication in Caenorhabditis elegans embryos

    Science.gov (United States)

    Medley, Jeffrey C.; Kabara, Megan M.; Stubenvoll, Michael D.; DeMeyer, Lauren E.

    2017-01-01

    ABSTRACT Centrosomes are the primary microtubule-organizing centers that orchestrate microtubule dynamics during the cell cycle. The correct number of centrosomes is pivotal for establishing bipolar mitotic spindles that ensure accurate segregation of chromosomes. Thus, centrioles must duplicate once per cell cycle, one daughter per mother centriole, the process of which requires highly coordinated actions among core factors and modulators. Protein phosphorylation is shown to regulate the stability, localization and activity of centrosome proteins. Here, we report the function of Casein kinase II (CK2) in early Caenorhabditis elegans embryos. The catalytic subunit (KIN-3/CK2α) of CK2 localizes to nuclei, centrosomes and midbodies. Inactivating CK2 leads to cell division defects, including chromosome missegregation, cytokinesis failure and aberrant centrosome behavior. Furthermore, depletion or inhibiting kinase activity of CK2 results in elevated ZYG-1 levels at centrosomes, restoring centrosome duplication and embryonic viability to zyg-1 mutants. Our data suggest that CK2 functions in cell division and negatively regulates centrosome duplication in a kinase-dependent manner. PMID:27881437

  13. Sgs1's roles in DNA end resection, HJ dissolution, and crossover suppression require a two-step SUMO regulation dependent on Smc5/6.

    Science.gov (United States)

    Bermúdez-López, Marcelino; Villoria, María Teresa; Esteras, Miguel; Jarmuz, Adam; Torres-Rosell, Jordi; Clemente-Blanco, Andres; Aragon, Luis

    2016-06-01

    The RecQ helicase Sgs1 plays critical roles during DNA repair by homologous recombination, from end resection to Holliday junction (HJ) dissolution. Sgs1 has both pro- and anti-recombinogenic roles, and therefore its activity must be tightly regulated. However, the controls involved in recruitment and activation of Sgs1 at damaged sites are unknown. Here we show a two-step role for Smc5/6 in recruiting and activating Sgs1 through SUMOylation. First, auto-SUMOylation of Smc5/6 subunits leads to recruitment of Sgs1 as part of the STR (Sgs1-Top3-Rmi1) complex, mediated by two SUMO-interacting motifs (SIMs) on Sgs1 that specifically recognize SUMOylated Smc5/6. Second, Smc5/6-dependent SUMOylation of Sgs1 and Top3 is required for the efficient function of STR. Sgs1 mutants impaired in recognition of SUMOylated Smc5/6 (sgs1-SIMΔ) or SUMO-dead alleles (sgs1-KR) exhibit unprocessed HJs at damaged replication forks, increased crossover frequencies during double-strand break repair, and severe impairment in DNA end resection. Smc5/6 is a key regulator of Sgs1's recombination functions.

  14. The Acinetobacter baumannii Two-Component System AdeRS Regulates Genes Required for Multidrug Efflux, Biofilm Formation, and Virulence in a Strain-Specific Manner

    Directory of Open Access Journals (Sweden)

    Grace E. Richmond

    2016-04-01

    Full Text Available The opportunistic pathogen Acinetobacter baumannii is able to persist in the environment and is often multidrug resistant (MDR, causing difficulties in the treatment of infections. Here, we show that the two-component system AdeRS, which regulates the production of the AdeABC multidrug resistance efflux pump, is required for the formation of a protective biofilm in an ex vivo porcine mucosal model, which mimics a natural infection of the human epithelium. Interestingly, deletion of adeB impacted only on the ability of strain AYE to form a biofilm on plastic and only on the virulence of strain Singapore 1 for Galleria mellonella. RNA-Seq revealed that loss of AdeRS or AdeB significantly altered the transcriptional landscape, resulting in the changed expression of many genes, notably those associated with antimicrobial resistance and virulence interactions. For example, A. baumannii lacking AdeRS displayed decreased expression of adeABC, pil genes, com genes, and a pgaC-like gene, whereas loss of AdeB resulted in increased expression of pil and com genes and decreased expression of ferric acinetobactin transport system genes. These data define the scope of AdeRS-mediated regulation, show that changes in the production of AdeABC mediate important phenotypes controlled by AdeRS, and suggest that AdeABC is a viable target for antimicrobial drug and antibiofilm discovery.

  15. The Acinetobacter baumannii Two-Component System AdeRS Regulates Genes Required for Multidrug Efflux, Biofilm Formation, and Virulence in a Strain-Specific Manner

    Science.gov (United States)

    Richmond, Grace E.; Evans, Laura P.; Anderson, Michele J.; Wand, Matthew E.; Bonney, Laura C.; Ivens, Alasdair; Chua, Kim Lee; Webber, Mark A.; Sutton, J. Mark; Peterson, Marnie L.

    2016-01-01

    ABSTRACT The opportunistic pathogen Acinetobacter baumannii is able to persist in the environment and is often multidrug resistant (MDR), causing difficulties in the treatment of infections. Here, we show that the two-component system AdeRS, which regulates the production of the AdeABC multidrug resistance efflux pump, is required for the formation of a protective biofilm in an ex vivo porcine mucosal model, which mimics a natural infection of the human epithelium. Interestingly, deletion of adeB impacted only on the ability of strain AYE to form a biofilm on plastic and only on the virulence of strain Singapore 1 for Galleria mellonella. RNA-Seq revealed that loss of AdeRS or AdeB significantly altered the transcriptional landscape, resulting in the changed expression of many genes, notably those associated with antimicrobial resistance and virulence interactions. For example, A. baumannii lacking AdeRS displayed decreased expression of adeABC, pil genes, com genes, and a pgaC-like gene, whereas loss of AdeB resulted in increased expression of pil and com genes and decreased expression of ferric acinetobactin transport system genes. These data define the scope of AdeRS-mediated regulation, show that changes in the production of AdeABC mediate important phenotypes controlled by AdeRS, and suggest that AdeABC is a viable target for antimicrobial drug and antibiofilm discovery. PMID:27094331

  16. Zebrafish cyclin Dx is required for development of motor neuron progenitors, and its expression is regulated by hypoxia-inducible factor 2α.

    Science.gov (United States)

    Lien, Huang-Wei; Yuan, Rey-Yue; Chou, Chih-Ming; Chen, Yi-Chung; Hung, Chin-Chun; Hu, Chin-Hwa; Hwang, Sheng-Ping L; Hwang, Pung-Pung; Shen, Chia-Ning; Chen, Chih-Lung; Cheng, Chia-Hsiung; Huang, Chang-Jen

    2016-06-21

    Cyclins play a central role in cell-cycle regulation; in mammals, the D family of cyclins consists of cyclin D1, D2, and D3. In Xenopus, only homologs of cyclins D1 and D2 have been reported, while a novel cyclin, cyclin Dx (ccndx), was found to be required for the maintenance of motor neuron progenitors during embryogenesis. It remains unknown whether zebrafish possess cyclin D3 or cyclin Dx. In this study, we identified a zebrafish ccndx gene encoding a protein which can form a complex with Cdk4. Through whole-mount in situ hybridization, we observed that zccndx mRNA is expressed in the motor neurons of hindbrain and spinal cord during development. Analysis of a 4-kb promoter sequence of the zccndx gene revealed the presence of HRE sites, which can be regulated by HIF2α. Morpholino knockdown of zebrafish Hif2α and cyclin Dx resulted in the abolishment of isl1 and oligo2 expression in the precursors of motor neurons, and also disrupted axon growth. Overexpression of cyclin Dx mRNA in Hif2α morphants partially rescued zccndx expression. Taken together, our data indicate that zebrafish cyclin Dx plays a role in maintaining the precursors of motor neurons.

  17. AKT induces erythroid-cell maturation of JAK2-deficient fetal liver progenitor cells and is required for Epo regulation of erythroid-cell differentiation.

    Science.gov (United States)

    Ghaffari, Saghi; Kitidis, Claire; Zhao, Wei; Marinkovic, Dragan; Fleming, Mark D; Luo, Biao; Marszalek, Joseph; Lodish, Harvey F

    2006-03-01

    AKT serine threonine kinase of the protein kinase B (PKB) family plays essential roles in cell survival, growth, metabolism, and differentiation. In the erythroid system, AKT is known to be rapidly phosphorylated and activated in response to erythropoietin (Epo) engagement of Epo receptor (EpoR) and to sustain survival signals in cultured erythroid cells. Here we demonstrate that activated AKT complements EpoR signaling and supports erythroid-cell differentiation in wild-type and JAK2-deficient fetal liver cells. We show that erythroid maturation of AKT-transduced cells is not solely dependent on AKT-induced cell survival or proliferation signals, suggesting that AKT transduces also a differentiation-specific signal downstream of EpoR in erythroid cells. Down-regulation of expression of AKT kinase by RNA interference, or AKT activity by expression of dominant negative forms, inhibits significantly fetal liver-derived erythroid-cell colony formation and gene expression, demonstrating that AKT is required for Epo regulation of erythroid-cell maturation.

  18. Fibroblast growth factor 21 regulates lipolysis in white adipose tissue but is not required for ketogenesis and triglyceride clearance in liver.

    Science.gov (United States)

    Hotta, Yuhei; Nakamura, Hirotoshi; Konishi, Morichika; Murata, Yusuke; Takagi, Hiroyuki; Matsumura, Shigenobu; Inoue, Kazuo; Fushiki, Tohru; Itoh, Nobuyuki

    2009-10-01

    Fibroblast growth factors (Fgfs) are polypeptide growth factors with diverse functions. Fgf21, a unique member of the Fgf family, is expected to function as a metabolic regulator in an endocrine manner. Hepatic Fgf21 expression was increased by fasting. The phenotypes of hepatic Fgf21 transgenic or knockdown mice and high-fat, low-carbohydrate ketogenic diet-fed mice suggests that Fgf21 stimulates lipolysis in the white adipose tissue during normal feeding and is required for ketogenesis and triglyceride clearance in the liver during fasting. However, the physiological roles of Fgf21 remain unclear. To elucidate the physiological roles of Fgf21, we generated Fgf21 knockout (KO) mice by targeted disruption. Fgf21 KO mice were viable, fertile, and seemingly normal. Food intake, oxygen consumption, and energy expenditure were also essentially unchanged in Fgf21 KO mice. However, hypertrophy of adipocytes, decreased lipolysis in adipocytes, and decreased blood nonesterified fatty acid levels were observed when Fgf21 KO mice were fed normally. In contrast, increased lipolysis in adipocytes and increased blood nonesterified fatty acid levels were observed in Fgf21 KO mice by fasting for 24 h, indicating that Fgf21 stimulates lipolysis in the white adipose tissue during feeding but inhibits it during fasting. In contrast, unexpectedly, hepatic triglyceride levels were essentially unchanged in Fgf21 KO mice. In addition, ketogenesis in Fgf21 KO mice was not impaired by fasting for 24 h. The present results indicate that Fgf21 regulates lipolysis in adipocytes in response to the metabolic state but is not required for ketogenesis and triglyceride clearance in the liver.

  19. Calcium-regulation of mitochondrial respiration maintains ATP homeostasis and requires ARALAR/AGC1-malate aspartate shuttle in intact cortical neurons.

    Science.gov (United States)

    Llorente-Folch, Irene; Rueda, Carlos B; Amigo, Ignacio; del Arco, Araceli; Saheki, Takeyori; Pardo, Beatriz; Satrústegui, Jorgina

    2013-08-28

    Neuronal respiration is controlled by ATP demand and Ca2+ but the roles played by each are unknown, as any Ca2+ signal also impacts on ATP demand. Ca2+ can control mitochondrial function through Ca2+-regulated mitochondrial carriers, the aspartate-glutamate and ATP-Mg/Pi carriers, ARALAR/AGC1 and SCaMC-3, respectively, or in the matrix after Ca2+ transport through the Ca2+ uniporter. We have studied the role of Ca2+ signaling in the regulation of mitochondrial respiration in intact mouse cortical neurons in basal conditions and in response to increased workload caused by increases in [Na+]cyt (veratridine, high-K+ depolarization) and/or [Ca2+]cyt (carbachol). Respiration in nonstimulated neurons on 2.5-5 mm glucose depends on ARALAR-malate aspartate shuttle (MAS), with a 46% drop in aralar KO neurons. All stimulation conditions induced increased OCR (oxygen consumption rate) in the presence of Ca2+, which was prevented by BAPTA-AM loading (to preserve the workload), or in Ca2+-free medium (which also lowers cell workload). SCaMC-3 limits respiration only in response to high workloads and robust Ca2+ signals. In every condition tested Ca2+ activation of ARALAR-MAS was required to fully stimulate coupled respiration by promoting pyruvate entry into mitochondria. In aralar KO neurons, respiration was stimulated by veratridine, but not by KCl or carbachol, indicating that the Ca2+ uniporter pathway played a role in the first, but not in the second condition, even though KCl caused an increase in [Ca2+]mit. The results suggest a requirement for ARALAR-MAS in priming pyruvate entry in mitochondria as a step needed to activate respiration by Ca2+ in response to moderate workloads.

  20. Tomato LeTHIC is an Fe-requiring HMP-P synthase involved in thiamine synthesis and regulated by multiple factors.

    Science.gov (United States)

    Zhao, Weina; Cheng, Xudong; Huang, Zongan; Fan, Huajie; Wu, Huilan; Ling, Hong-Qing

    2011-06-01

    Thiamine is a key primary metabolite which is necessary for the viability of all organisms. It is a dietary requirement for mammals because only prokaryotes, fungi and plants are thiamine prototrophs. In contrast to the well documented biosynthetic mechanism in bacteria, much remains to be deciphered in plants. In this work, a tomato thiamine-auxotrophic (thiamineless, tl) mutant was characterized. The tl mutant occurs due to inactivation of LeTHIC transcription as a result of insertion of a large unknown DNA fragment in its 5'-untranslated region. Expression of wild-type LeTHIC in tl plants was able to complement the mutant to wild type. LeTHIC possessed the same function as E.cTHIC [an Escherichia coli 4-amino-5-hydroxymethyl-2-methylpyrimidine phosphate (HMP-P) synthase involved in synthesis of the pyrimidine moiety of thiamine] because expression of LeTHIC rescued THIC-deficient strains of E. coli under culture conditions without thiamine supplementation, suggesting that plants employ a bacteria-like route of pyrimidine moiety synthesis. LeTHIC is an Fe-S cluster protein localized in chloroplasts, and Fe is required for maintenance of its enzyme activity because Fe deficiency resulted in a significant reduction of thiamine content in tomato leaves. Further, we also showed that the expression of LeTHIC is tightly regulated at the transcriptional and post-transcriptional level by multiple factors, such as light, Fe status and thiamine pyrophosphate (TPP)-riboswitch. The results clearly demonstrated that a feedback regulation mechanism is involved in synthesis of the pyrimidine moiety for controlling thiamine synthesis in tomato. Our results provide a new insight into understanding the molecular mechanism of thiamine biosynthesis in plants.