WorldWideScience

Sample records for current regulations require

  1. 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...

  2. 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.

  3. Current Regulations and Regulatory Actions

    Science.gov (United States)

    This site will provide basic information on clean air permitting under the title V operating permits program, provide access to state and regional permitting programs, and maintain access to proposed and final regulatory requirements.

  4. Current requirements for polymeric biomaterials in otolaryngology

    Directory of Open Access Journals (Sweden)

    Sternberg, Katrin

    2009-01-01

    Full Text Available In recent years otolaryngology was strongly influenced by newly developed implants which are based on both, innovative biomaterials and novel implant technologies. Since the biomaterials are integrated into biological systems they have to fulfill all technical requirements and accommodate biological interactions. Technical functionality relating to implant specific mechanical properties, a sufficiently high stability in terms of physiological conditions, and good biocompatibility are the demands with regard to suitability of biomaterials. The goal in applying biomaterials for implants is to maintain biofunctionality over extended periods of time. These general demands to biomaterials are equally valid for use in otolaryngology. Different classes of materials can be utilized as biomaterials. Metals belong to the oldest biomaterials. In addition, alloys, ceramics, inorganic glasses and composites have been tested successfully. Furthermore, natural and synthetic polymers are widely used materials, which will be in the focus of the current article with regard to their properties and usage as cochlear implants, osteosynthesis implants, stents, and matrices for tissue engineering. Due to their application as permanent or temporary implants materials are differentiated into biostable and biodegradable polymers. The here identified general and up to date requirements for biomaterials and the illustrated applications in otolaryngology emphasize ongoing research efforts in this area and at the same time demonstrate the high significance of interdisciplinary cooperation between natural sciences, engineering, and medical sciences.

  5. Current perspectives on shoot branching regulation

    Directory of Open Access Journals (Sweden)

    Cunquan YUAN,Lin XI,Yaping KOU,Yu ZHAO,Liangjun ZHAO

    2015-03-01

    Full Text Available Shoot branching is regulated by the complex interactions among hormones, development, and environmental factors. Recent studies into the regulatory mecha-nisms of shoot branching have focused on strigolactones, which is a new area of investigation in shoot branching regulation. Elucidation of the function of the D53 gene has allowed exploration of detailed mechanisms of action of strigolactones in regulating shoot branching. In addition, the recent discovery that sucrose is key for axillary bud release has challenged the established auxin theory, in which auxin is the principal agent in the control of apical dominance. These developments increase our understan-ding of branching control and indicate that regulation of shoot branching involves a complex network. Here, we first summarize advances in the systematic regulatory network of plant shoot branching based on current information. Then we describe recent developments in the synthesis and signal transduction of strigolactones. Based on these considerations, we further summarize the plant shoot branching regulatory network, including long distance systemic signals and local gene activity mediated by strigolactones following perception of external envi-ronmental signals, such as shading, in order to provide a comprehensive overview of plant shoot branching.

  6. 49 CFR 236.562 - Minimum rail current required.

    Science.gov (United States)

    2010-10-01

    ..., INSPECTION, MAINTENANCE, AND REPAIR OF SIGNAL AND TRAIN CONTROL SYSTEMS, DEVICES, AND APPLIANCES Automatic Train Stop, Train Control and Cab Signal Systems Rules and Instructions; Locomotives § 236.562 Minimum rail current required. The minimum rail current required to restore the locomotive equipment...

  7. Energy-conserving development regulations: current practice

    Energy Technology Data Exchange (ETDEWEB)

    1980-05-01

    Almost every aspect of land development has an effect on energy use, from minute architectural details to broad considerations of urban density. Energy-efficiency depends in part on how development is planned and carried out. Conventional development regulations, such as zoning ordinances and subdivision regulations, can be adapted in many ways to promote energy conservation at the community level. This report is about energy-efficient site and neighborhood design. It examines recent experiences of local governments that have adopted new development regulations or amended existing ones to promote energy conservation, more efficient generation and distribution, or a switch to alternative, renewable sources. Although much has been written in recent years about saving energy through community design, actual experience in applying these new ideas is still limited. To date, most communities have focused their efforts on studying the problem, documenting consumption patterns, and writing reports and plans. Only a handful have amended their land-use controls for the express purpose of saving energy. This study identifies 13 of these pioneering communities, after undertaking a survey of over 1400 local, regional, and state planning agencies. It takes a look at their experiences, to learn what has been done, how well it has worked, and what problems have been encountered.

  8. Development and regulation of biosimilars: current status and future challenges.

    Science.gov (United States)

    Tsiftsoglou, Asterios S; Ruiz, Sol; Schneider, Christian K

    2013-06-01

    provided through several Committee for Medicinal Products for Human Use (CHMP) guidelines as well as individual scientific advice requested from the European Medicines Agency (EMA) by various companies for the development and regulation of biosimilars. This review is mainly focused on the current status of regulation of biosimilars in the EU as well as on future challenges lying ahead for the improvement of the requirements needed for the marketing authorization of biosimilars. Emphasis is given on the quality requirements concerning these medicinal products (biologics).

  9. Review of the current state of UAV regulations

    NARCIS (Netherlands)

    Stöcker, Elvira Claudia; Bennett, Rohan; Nex, Francesco; Gerke, Markus; Zevenbergen, Jaap

    2017-01-01

    UAVs-unmanned aerial vehicles-facilitate data acquisition at temporal and spatial scales that still remain unachievable for traditional remote sensing platforms. However, current legal frameworks that regulate UAVs present significant barriers to research and development. To highlight the

  10. Audit Risk Assessment in the Light of Current European Regulations

    OpenAIRE

    Ciprian-Costel Munteanu

    2015-01-01

    Recent European reforms on audit regulations have been motivated by efforts to increase audit quality, functioning and performance. We believe the adoption of Directive 2014/56 and Regulation 537/2014 strengthened the role of independent audit and risk committees, which will positively contribute towards audit quality. This paper aims to critically assess the status quo of audit risk assessment in current European standards and regulations, by conducting a theoretical analysis of ...

  11. Current drive at plasma densities required for thermonuclear reactors.

    Science.gov (United States)

    Cesario, R; Amicucci, L; Cardinali, A; Castaldo, C; Marinucci, M; Panaccione, L; Santini, F; Tudisco, O; Apicella, M L; Calabrò, G; Cianfarani, C; Frigione, D; Galli, A; Mazzitelli, G; Mazzotta, C; Pericoli, V; Schettini, G; Tuccillo, A A

    2010-08-10

    Progress in thermonuclear fusion energy research based on deuterium plasmas magnetically confined in toroidal tokamak devices requires the development of efficient current drive methods. Previous experiments have shown that plasma current can be driven effectively by externally launched radio frequency power coupled to lower hybrid plasma waves. However, at the high plasma densities required for fusion power plants, the coupled radio frequency power does not penetrate into the plasma core, possibly because of strong wave interactions with the plasma edge. Here we show experiments performed on FTU (Frascati Tokamak Upgrade) based on theoretical predictions that nonlinear interactions diminish when the peripheral plasma electron temperature is high, allowing significant wave penetration at high density. The results show that the coupled radio frequency power can penetrate into high-density plasmas due to weaker plasma edge effects, thus extending the effective range of lower hybrid current drive towards the domain relevant for fusion reactors.

  12. 26 CFR 1.651(a)-2 - Income required to be distributed currently.

    Science.gov (United States)

    2010-04-01

    ... requirement to distribute currently all the income as defined under section 643(b) and the applicable regulations, the trust shall be treated as having sold the property for its fair market value on the date of distribution. If no amount in excess of the amount of income as defined under section 643(b) and the applicable...

  13. 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...

  14. Utility survey of requirements for a HTS fault current limiter

    DEFF Research Database (Denmark)

    Nielsen, Jan Nygaard; Jørgensen, P.; Østergaard, Jacob;

    2000-01-01

    The application of superconducting fault current limiters (SFCL) in the electric utility sector will clearly dependent on to what extent the needs and requirements of electric utilities can be met by the ongoing development of SFCL technology. This paper considers a questionnaire survey of which ...... needs and expectations the Danish electric utilities have to this new technology. A bus-tie application of SFCL in a distribution substation with three parallel-coupled transformers is discussed......The application of superconducting fault current limiters (SFCL) in the electric utility sector will clearly dependent on to what extent the needs and requirements of electric utilities can be met by the ongoing development of SFCL technology. This paper considers a questionnaire survey of which...

  15. Enhanced current and voltage regulators for stand-alone applications

    DEFF Research Database (Denmark)

    Federico, de Bosio; Pastorelli, Michele; Antonio DeSouza Ribeiro, Luiz

    2016-01-01

    State feedback decoupling permits to achieve a better dynamic response for Voltage Source in stand-alone applications. The design of current and voltage regulators is performed in the discrete-time domain since it provides better accuracy and allows direct pole placement. As the attainable...... bandwidth of the current loop is mainly limited by computational and PWM delays, a lead compensator structure is proposed to overcome this limitation. The design of the voltage regulator is based on the Nyquist criterion, verifying to guarantee a high sensitivity peak. Discrete-time domain implementation...

  16. A summarized discussion of current good manufacturing practice regulations.

    Science.gov (United States)

    Allen, Loyd V

    2013-01-01

    In light of recent events and discussions of compounding pharmacy, it is important to discuss and understand the purpose of good manufacturing practices. This article provides a summary of the current Good Manufacturing Practice Regulations which were established by the U.S. Food and Drug Administration.

  17. Review of the current state of UAV regulations

    NARCIS (Netherlands)

    Stöcker, Elvira Claudia; Bennett, Rohan; Nex, Francesco; Gerke, Markus; Zevenbergen, Jaap

    2017-01-01

    UAVs-unmanned aerial vehicles-facilitate data acquisition at temporal and spatial scales that still remain unachievable for traditional remote sensing platforms. However, current legal frameworks that regulate UAVs present significant barriers to research and development. To highlight the importance

  18. An assessment of joint chat requirements from current usage patterns

    OpenAIRE

    Bryan A. Eovito

    2006-01-01

    This research assesses the impact of synchronous (real-time), text-based chat on military command and control (C2) processes. Chat use among the services, particularly among joint forces, has evolved in ad hoc fashion to fill gaps in currently fielded C2 systems. This growth-by-improvisation inhibits clear definition of the underlying requirements: precisely what C2 deficiencies are being addressed by text-based chat tools? Or, from a bottom-up perspective: what capabilities do text-based cha...

  19. Labelling of electronic cigarettes: regulations and current practice

    Science.gov (United States)

    Buonocore, Federico; Marques Gomes, Ana C N; Nabhani-Gebara, Shereen; Barton, Stephen J; Calabrese, Gianpiero

    2017-01-01

    Background Over the past decade e-cigarettes have established themselves in the global market. E-cigarettes triggered much interest in relation to their content and efficacy as smoking cessation tools, but less attention has been paid to users and environmental safety warnings and guidance. Several regulations have been introduced to promote their safe handling and disposal. From May 2016, liquids and cartridges will be regulated by European Community Directives (ECDs) 2001/83/EC and 93/42/EEC, or 2014/40/EU if marketed as tobacco-related products. Currently, manufacturers and distributors must abide by the Chemical (Hazard Information and Packaging for Supply) Regulations 2009 (CHIP) or Classification, Labelling and Packaging Regulations (CLP), the latter replacing CHIP in June 2015. Objective In this work, the compliance of marketed e-liquids and e-cigarettes with current European Union and UK legislations is assessed. Results E-liquids and e-cigarettes (21 and 9 brands, respectively) were evaluated. Evidence of non-compliance was found in relation to the CHIP/CLP toxic (13%) and environmental (37%) pictograms, tactile warning (23%), nominal amount of solution (30%), supplier contact telephone number and address (40%). None of the evaluated e-cigarettes displayed information on the correct disposal/recycling of batteries in line with the ECD 2006/66/EC. Conclusions More stringent enforcement of regulations is needed to ensure not only the user's safety and awareness, but also the safeguarding of the environment. PMID:26790924

  20. Voltage and Current Regulators Design of Power Converters in Islanded Microgrids based on State Feedback Decoupling

    DEFF Research Database (Denmark)

    Federico, de Bosio; de Sousa Ribeiro, Luiz Antonio; Freijedo Fernandez, Francisco Daniel

    2016-01-01

    In stand-alone microgrids based on voltage source inverters state feedback coupling between the capacitor voltage and inductor current degrades significantly the dynamics performance of voltage and current regulators. The decoupling of the controlled states is proposed, considering the limitations...... introduced by system delays. Moreover, a proportional resonant voltage controller is designed according to Nyquist criterion taking into account application requirements. Experimental tests performed in compliance with the UPS standards verify the theoretical analysis....

  1. Audit Risk Assessment in the Light of Current European Regulations

    Directory of Open Access Journals (Sweden)

    Ciprian-Costel Munteanu

    2015-06-01

    Full Text Available Recent European reforms on audit regulations have been motivated by efforts to increase audit quality, functioning and performance. We believe the adoption of Directive 2014/56 and Regulation 537/2014 strengthened the role of independent audit and risk committees, which will positively contribute towards audit quality. This paper aims to critically assess the status quo of audit risk assessment in current European standards and regulations, by conducting a theoretical analysis of different aspects of audit risk. Our main objective is to stress the importance of detecting inherent and control risk, which lead to material misstatement at the assertion level. They need to be assessed so as to determine the nature, timing and extent of further audit procedures necessary to obtain sufficient appropriate audit evidence. These pieces of evidence enable the auditor to express an opinion on the financial statements at an acceptably low level of audit risk. Therefore, we point to the fact that researchers as well as practitioners and policymakers have to be careful when using audit tools and assessing risk levels, as their conclusions continuously shape the regulations.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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...

  7. 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...

  8. Current state of OLED technology relative to military avionics requirements

    Science.gov (United States)

    Tchon, Joseph L.; Barnidge, Tracy J.; Hufnagel, Bruce D.; Bahadur, Birendra

    2014-06-01

    The paper will review optical and environmental performance thresholds required for OLED technology to be used on various military platforms. Life study results will be summarized to highlight trends while identifying remaining performance gaps to make this technology viable for future military avionics platforms.

  9. Parallel Computational Fluid Dynamics: Current Status and Future Requirements

    Science.gov (United States)

    Simon, Horst D.; VanDalsem, William R.; Dagum, Leonardo; Kutler, Paul (Technical Monitor)

    1994-01-01

    One or the key objectives of the Applied Research Branch in the Numerical Aerodynamic Simulation (NAS) Systems Division at NASA Allies Research Center is the accelerated introduction of highly parallel machines into a full operational environment. In this report we discuss the performance results obtained from the implementation of some computational fluid dynamics (CFD) applications on the Connection Machine CM-2 and the Intel iPSC/860. We summarize some of the experiences made so far with the parallel testbed machines at the NAS Applied Research Branch. Then we discuss the long term computational requirements for accomplishing some of the grand challenge problems in computational aerosciences. We argue that only massively parallel machines will be able to meet these grand challenge requirements, and we outline the computer science and algorithm research challenges ahead.

  10. Comparison of resonant current regulators for DFIG during grid voltage distortion

    Institute of Scientific and Technical Information of China (English)

    Yi-peng SONG; Heng NIAN

    2013-01-01

    We investigate two different kinds of resonant current regulators for a doubly fed induction generator (DFIG) under distorted grid voltage conditions:proportional integral resonant (PIR) regulator with traditional resonant part and vector propor-tional integral (VPI) regulator with VPI resonant part. Based on the mathematical model of DFIG under distorted grid voltage, the transfer function and frequency response characteristics of the two current regulators are analyzed and compared. The superiority of the VPI current regulator over the PIR regulator is pointed out, and the influence of discretization methods on the performance of the resonant current regulator is studied. All the results are validated by MATLAB simulation and experiments.

  11. Effectiveness of reserve requirements on current account imbalances

    OpenAIRE

    Dalkıran, Dilşat Tugba

    2012-01-01

    Ankara : The Department of Economics, İhsan Doğramacı Bilkent University, 2012. Thesis (Master's) -- Bilkent University, 2012. Includes bibliographical references leaves 35. Following the recent financial crisis, reserve requirements have become a policy instrument preferred in many emerging markets such as China, Brazil and Turkey for various purposes. Therefore, the formulating a theoretical framework to study the policy effectiveness remains an important issue. In thi...

  12. Geosensors to support crop production: current applications and user requirements.

    Science.gov (United States)

    Thessler, Sirpa; Kooistra, Lammert; Teye, Frederick; Huitu, Hanna; Bregt, Arnold K

    2011-01-01

    Sensor technology, which benefits from high temporal measuring resolution, real-time data transfer and high spatial resolution of sensor data that shows in-field variations, has the potential to provide added value for crop production. The present paper explores how sensors and sensor networks have been utilised in the crop production process and what their added-value and the main bottlenecks are from the perspective of users. The focus is on sensor based applications and on requirements that users pose for them. Literature and two use cases were reviewed and applications were classified according to the crop production process: sensing of growth conditions, fertilising, irrigation, plant protection, harvesting and fleet control. The potential of sensor technology was widely acknowledged along the crop production chain. Users of the sensors require easy-to-use and reliable applications that are actionable in crop production at reasonable costs. The challenges are to develop sensor technology, data interoperability and management tools as well as data and measurement services in a way that requirements can be met, and potential benefits and added value can be realized in the farms in terms of higher yields, improved quality of yields, decreased input costs and production risks, and less work time and load.

  13. Geosensors to Support Crop Production: Current Applications and User Requirements

    Directory of Open Access Journals (Sweden)

    Lammert Kooistra

    2011-06-01

    Full Text Available Sensor technology, which benefits from high temporal measuring resolution, real-time data transfer and high spatial resolution of sensor data that shows in-field variations, has the potential to provide added value for crop production. The present paper explores how sensors and sensor networks have been utilised in the crop production process and what their added-value and the main bottlenecks are from the perspective of users. The focus is on sensor based applications and on requirements that users pose for them. Literature and two use cases were reviewed and applications were classified according to the crop production process: sensing of growth conditions, fertilising, irrigation, plant protection, harvesting and fleet control. The potential of sensor technology was widely acknowledged along the crop production chain. Users of the sensors require easy-to-use and reliable applications that are actionable in crop production at reasonable costs. The challenges are to develop sensor technology, data interoperability and management tools as well as data and measurement services in a way that requirements can be met, and potential benefits and added value can be realized in the farms in terms of higher yields, improved quality of yields, decreased input costs and production risks, and less work time and load.

  14. Redox regulation in malaria: current concepts and pharmacotherapeutic implications.

    Science.gov (United States)

    Goyal, M; Alam, A; Bandyopadhyay, U

    2012-01-01

    Malaria imposes a serious threat to human and becomes more prevalent due to the emergence of drug resistant parasite. Understanding of the underlying mechanism of drug resistance and identification of novel drug targets are key effective processes for the management of malaria. Malaria parasite is highly susceptible to oxidative stress but lives in a pro-oxidant rich environment containing oxygen and iron, which produce a large amount of reactive oxygen species. Management of oxidative stress in malaria parasite is tightly regulated through active redox and antioxidant defense systems. The elevation of oxidative stress as a result of inhibition of any component of this defense system leads to redox imbalance and ultimately parasite death. Therefore, redox system plays an indispensable role for the survival of parasite within the host. Identification of key molecules, which disrupt parasite redox balance by altering key redox reactions and promote oxidative stress in parasites, would be an effective approach to develop novel antimalarial drugs. During the last few decades, contributions by researchers around the globe provide even better understanding of redox biology of malaria parasite. Here, in this review, we are highlighting the knowledge gathered so far regarding the essential redox-active processes and their components in malaria parasite to overcome elevated oxidative insults. We have also given maximum efforts to enlist currently used redox-active antimalarials, their mode of action and pharmacotherapeutic implications.

  15. Factors affecting the regulation of pacing: current perspectives

    Directory of Open Access Journals (Sweden)

    Mauger AR

    2014-09-01

    Full Text Available Alexis R Mauger Endurance Research Group, School of Sport and Exercise Sciences, Faculty of Science, University of Kent, Chatham, UK Abstract: During prolonged dynamic and rhythmic exercise, muscular pain and discomfort arises as a result of an increased concentration of deleterious metabolites. Sensed by peripheral nociceptors and transmitted via afferent feedback to the brain, this provides important information regarding the physiological state of the muscle. These sensations ultimately contribute to what is termed "exercise-induced pain". Despite being well recognized by athletes and coaches, and suggested to be integral to exercise performance, this construct has largely escaped attention in experimental work. This perspective article highlights the current understanding of pacing in endurance performance, and the causes of exercise-induced pain. A new perspective is described, which proposes how exercise-induced pain may be a contributing factor in helping individuals to regulate their work rate during exercise and thus provides an important construct in pacing. Keywords: pain, exercise-induced pain, discomfort, exercise performance, self-paced

  16. 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.

  17. 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.

  18. 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...

  19. 46 CFR 11.713 - Requirements for maintaining current knowledge of waters to be navigated.

    Science.gov (United States)

    2010-10-01

    ... 46 Shipping 1 2010-10-01 2010-10-01 false Requirements for maintaining current knowledge of waters... § 11.713 Requirements for maintaining current knowledge of waters to be navigated. (a) If a first class... current knowledge of the route. Persons using this method of re-familiarization shall certify,...

  20. 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...

  1. 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...

  2. 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...

  3. How Should Risk-Based Regulation Reflect Current Public Opinion?

    Science.gov (United States)

    Pollock, Christopher John

    2016-08-01

    Risk-based regulation of novel agricultural products with public choice manifest via traceability and labelling is a more effective approach than the use of regulatory processes to reflect public concerns, which may not always be supported by evidence.

  4. 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.

  5. Methods, systems and apparatus for synchronous current regulation of a five-phase machine

    Science.gov (United States)

    Gallegos-Lopez, Gabriel; Perisic, Milun

    2012-10-09

    Methods, systems and apparatus are provided for controlling operation of and regulating current provided to a five-phase machine when one or more phases has experienced a fault or has failed. In one implementation, the disclosed embodiments can be used to synchronously regulate current in a vector controlled motor drive system that includes a five-phase AC machine, a five-phase inverter module coupled to the five-phase AC machine, and a synchronous current regulator.

  6. Design and implementation of a bidirectional current-controlled voltage-regulated DC-DC switched-mode converter

    CSIR Research Space (South Africa)

    Coetzer, A

    2016-01-01

    Full Text Available The design and implementation of a bidirectional current-controlled voltage-regulated DC-DC converter is presented. The converter is required to connect a battery of electrochemical cells (the battery) to an asynchronous motor-drive unit via a...

  7. 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.

  8. Current good manufacturing practices, quality control procedures, quality factors, notification requirements, and records and reports, for infant formula. Final rule.

    Science.gov (United States)

    2014-06-10

    The Food and Drug Administration (FDA or we) is issuing a final rule that adopts, with some modifications, the interim final rule (IFR) entitled "Current Good Manufacturing Practices, Quality Control Procedures, Quality Factors, Notification Requirements, and Records and Reports, for Infant Formula'' (February 10, 2014). This final rule affirms the IFR's changes to FDA's regulations and provides additional modifications and clarifications. The final rule also responds to certain comments submitted in response to the request for comments in the IFR.

  9. Regulation of persistent Na current by interactions between beta subunits of voltage-gated Na channels.

    Science.gov (United States)

    Aman, Teresa K; Grieco-Calub, Tina M; Chen, Chunling; Rusconi, Raffaella; Slat, Emily A; Isom, Lori L; Raman, Indira M

    2009-02-18

    The beta subunits of voltage-gated Na channels (Scnxb) regulate the gating of pore-forming alpha subunits, as well as their trafficking and localization. In heterologous expression systems, beta1, beta2, and beta3 subunits influence inactivation and persistent current in different ways. To test how the beta4 protein regulates Na channel gating, we transfected beta4 into HEK (human embryonic kidney) cells stably expressing Na(V)1.1. Unlike a free peptide with a sequence from the beta4 cytoplasmic domain, the full-length beta4 protein did not block open channels. Instead, beta4 expression favored open states by shifting activation curves negative, decreasing the slope of the inactivation curve, and increasing the percentage of noninactivating current. Consequently, persistent current tripled in amplitude. Expression of beta1 or chimeric subunits including the beta1 extracellular domain, however, favored inactivation. Coexpressing Na(V)1.1 and beta4 with beta1 produced tiny persistent currents, indicating that beta1 overcomes the effects of beta4 in heterotrimeric channels. In contrast, beta1(C121W), which contains an extracellular epilepsy-associated mutation, did not counteract the destabilization of inactivation by beta4 and also required unusually large depolarizations for channel opening. In cultured hippocampal neurons transfected with beta4, persistent current was slightly but significantly increased. Moreover, in beta4-expressing neurons from Scn1b and Scn1b/Scn2b null mice, entry into inactivated states was slowed. These data suggest that beta1 and beta4 have antagonistic roles, the former favoring inactivation, and the latter favoring activation. Because increased Na channel availability may facilitate action potential firing, these results suggest a mechanism for seizure susceptibility of both mice and humans with disrupted beta1 subunits.

  10. GABAB receptor phosphorylation regulates KCTD12-induced K+ current desensitization

    DEFF Research Database (Denmark)

    Adelfinger, L; Turecek, R; Ivankova, K

    2014-01-01

    released from the G-protein. Receptor-activated K+ currents desensitize in the sustained presence of agonist to avoid excessive effects on neuronal activity. Desensitization of K+ currents integrates distinct mechanistic underpinnings. GABAB receptor activity reduces protein kinase-A activity, which...... reduces phosphorylation of serine-892 in GABAB2 and promotes receptor degradation. This form of desensitization operates on the time scale of several minutes to hours. A faster form of desensitization is induced by the auxiliary subunit KCTD12, which interferes with channel activation by binding to the G......-protein βγ subunits. Here we show that the two mechanisms of desensitization influence each other. Serine-892 phosphorylation in heterologous cells rearranges KCTD12 at the receptor and slows KCTD12-induced desensitization. Likewise, protein kinase-A activation in hippocampal neurons slows fast...

  11. 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....

  12. 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.

  13. 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.

  14. Brushless DC motor Drive during Speed regulation with Current Controller

    Directory of Open Access Journals (Sweden)

    Bhikshalu Manchala

    2015-04-01

    Full Text Available Brushless DC Motor (BLDC is one of the best electrical drives that have increasing popularity, due to their high efficiency, reliability, good dynamic response and very low maintenance. Due to the increasing demand for compact & reliable motors and the evolution of low cost power semiconductor switches and permanent magnet (PM materials, brushless DC motors become popular in every application from home appliances to aerospace industry. The conventional techniques for controlling the stator phase current in a brushless DC drive are practically effective in low speed and cannot reduce the commutation torque ripple in high speed range. This paper presents the PI controller for speed control of BLDC motor. The output of the PI controllers is summed and is given as the input to the current controller. The BLDC motor is fed from the inverter where the rotor position and current controller is the input. The complete model of the proposed drive system is developed and simulated using MATLAB/Simulink software. The operation principle of using component is analysed and the simulation results are presented in this to verify the theoretical analysis.

  15. 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

  16. Current regulations and state investment in the wind power projects

    Directory of Open Access Journals (Sweden)

    Danijela Simeunović

    2015-09-01

    Full Text Available It is the objective of this paper to present the path which the wind farm projects have undergone in Croatia from their introduction and expansion since the year 2007 up to the today's date when their development is temporarily stayed. The paper first describes the political and legal framework for renewable energy sources in Croatia, and specifically for wind power plants, as well as the current status of investment in building of wind farms in Croatia. The paper further analyses why the Croatian system of promotion of eligible producers has proven to attract investment, albeit unsustainable for a longer period of time, and which regulatory measures have later contributed to the suspension of wind farm investment in 2014. Legal and factual consequences of such suspension are discussed at the end of the paper, as well as some potential new business models for further exploitation of wind energy in Croatia.

  17. 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

  18. Former Soviet Regulations for seismic design of NPPs and comparison with current international practice

    Energy Technology Data Exchange (ETDEWEB)

    Kostarev, V.; Schukin, A.; Berkovski, A. [CKTI-Vibroseism Co. Ltd. (Cape Verde)

    1997-03-01

    This paper presents a summary of current earthquake design criteria used in former Soviet Regulations for equipment and piping systems of nuclear power plants in light of those used in United States and Japan. The detailed comparative seismic analysis of PWR (WWER) Primary Coolant Loop System (PCLS) according to Former Soviet (Russian) PNAE Code and ASME BPV Code with some comments regarding to Japan Code JEAG - 4601 was undertaken for better understanding of the differences and coincidences of seismic design criteria and requirements. The selection of these three guides for the study has very simple explanation: according to ASME BVPC, JEAG and PNAE the huge majority of existing NPPs has been designed. (J.P.N.)

  19. 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.

  20. 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.

  1. 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),...

  2. 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...

  3. REACTIVE CURRENT OF AN INDUCTION ELECTRIC DRIVES WITH THYRISTOR VOLTAGE REGULATOR

    Directory of Open Access Journals (Sweden)

    J.V. Kovalova

    2014-12-01

    Full Text Available A model for a separation of reactive constituent from current of idling of an induction motor at its feed from a thyristor voltage regulator in the dependences on the control angle of thyristors is developed. As a result of modeling, dependence of relative reactive current which is approximated by formula for calculation of effective current of reactive constituent of nonsinusoidal current is obtained.

  4. Low-level radioactive waste disposal in the United States: An overview of current commercial regulations and concepts

    Energy Technology Data Exchange (ETDEWEB)

    Kennedy, W.E. Jr.

    1993-08-01

    Commercial low-level radioactive waste disposal in the United States is regulated by the US Nuclear Regulatory Commission (NRC) under 10 CFR 61 (1991). This regulation was issued in 1981 after a lengthy and thorough development process that considered the radionuclide concentrations and characteristics associated with commercial low-level radioactive waste streams; alternatives for waste classification; alternative technologies for low-level radioactive waste disposal; and data, modeling, and scenario analyses. The development process also included the publication of both draft and final environmental impact statements. The final regulation describes the general provisions; licenses; performance objectives; technical requirements for land disposal; financial assurances; participation by state governments and Indian tribes; and records, reports, tests, and inspections. This paper provides an overview of, and tutorial on, current commercial low-level radioactive waste disposal regulations in the United States.

  5. Self-regulation of the reconnecting current layer in relativistic pair plasma reconnection

    CERN Document Server

    Zenitani, S

    2008-01-01

    We investigate properties of the reconnecting current layer in relativistic pair plasma reconnection. We found that the current layer self-regulates its thickness when the current layer runs out current carriers and so relativistic reconnection retains a fast reconnection rate. Constructing a steady state Sweet-Parker model, we discuss conditions for the current sheet expansion. Based on the energy argument we conclude that the incompressible assumption is invalid in relativistic Sweet-Parker reconnection. The guide field cases are more incompressible than the anti-parallel cases, and we find a more significant current sheet expansion.

  6. Requirements on localized current drive for the suppression of neoclassical tearing modes

    Science.gov (United States)

    Bertelli, N.; De Lazzari, D.; Westerhof, E.

    2011-10-01

    A heuristic criterion for the full suppression of an NTM was formulated as ηNTM ≡ jCD,max/jBS >= 1.2 (Zohm et al 2005 J. Phys. Conf. Ser. 25 234), where jCD,max is the maximum in the driven current density profile applied to stabilize the mode and jBS is the local bootstrap current density. In this work we subject this criterion to a systematic theoretical analysis on the basis of the generalized Rutherford equation. Taking into account only the effect of jCD inside the island, a new criterion for full suppression by a minimum applied total current is obtained in the form of a maximum allowed value for the width of the driven current, wdep, combined with a required minimum for the total driven current in the form of wdepηNTM, where both limits depend on the marginal and saturated island sizes. These requirements can be relaxed when additional effects are taken into account, such as a change in the stability parameter Δ' from the current driven outside the island, power modulation, the accompanying heating inside the island or when the current drive is applied preemptively. When applied to ITER scenario 2, the requirement for full suppression of either the 3/2 or 2/1 NTM becomes wdep ~ 5 cm in agreement with (Sauter et al 2010 Plasma Phys. Control. Fusion 52 025002). Optimization of the ITER ECRH Upper Port Launcher design towards minimum required power for full NTM suppression requires an increase in the toroidal injection angle of the lower steering mirror of several degrees compared with its present design value, while for the upper steering mirror the present design value is close to the optimum.

  7. Current advances and strategies towards fully automated sample preparation for regulated LC-MS/MS bioanalysis.

    Science.gov (United States)

    Zheng, Naiyu; Jiang, Hao; Zeng, Jianing

    2014-09-01

    Robotic liquid handlers (RLHs) have been widely used in automated sample preparation for liquid chromatography-tandem mass spectrometry (LC-MS/MS) bioanalysis. Automated sample preparation for regulated bioanalysis offers significantly higher assay efficiency, better data quality and potential bioanalytical cost-savings. For RLHs that are used for regulated bioanalysis, there are additional requirements, including 21 CFR Part 11 compliance, software validation, system qualification, calibration verification and proper maintenance. This article reviews recent advances in automated sample preparation for regulated bioanalysis in the last 5 years. Specifically, it covers the following aspects: regulated bioanalysis requirements, recent advances in automation hardware and software development, sample extraction workflow simplification, strategies towards fully automated sample extraction, and best practices in automated sample preparation for regulated bioanalysis.

  8. 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...

  9. 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...

  10. 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...

  11. 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...

  12. The relationship between chiropractor required and current level of business knowledge.

    Science.gov (United States)

    Ciolfi, Michael Anthony; Kasen, Patsy Anne

    2017-01-01

    Chiropractors frequently practice within health care systems requiring the business acumen of an entrepreneur. However, some chiropractors do not know the relationship between the level of business knowledge required for practice success and their current level of business knowledge. The purpose of this quantitative study was to examine the relationship between chiropractors' perceived level of business knowledge required and their perceived level of current business knowledge. Two hundred and seventy-four participants completed an online survey (Health Care Training and Education Needs Survey) which included eight key business items. Participants rated the level of perceived business knowledge required (Part I) and their current perceived level of knowledge (Part II) for the same eight items. Data was collected from November 27, 2013 to December 18, 2013. Data were analyzed using Spearman's ranked correlation to determine the statistically significant relationships for the perceived level of knowledge required and the perceived current level of knowledge for each of the paired eight items from Parts I and II of the survey. Wilcoxon Signed Ranks Tests were performed to determine the statistical difference between the paired items. The results of Spearman's correlation testing indicated a statistically significant (p ethical, (e) managerial decisions, and (f) operations. Wilcoxon Signed Ranks testing indicated a significant difference for three paired items: strategic management; marketing and; legal and ethical. The results suggest that relationships exist for the majority of business items (6 of 8) however a statistically difference was demonstrated in only three of the paired business items. The implications of this study for social change include the potential to improve chiropractors' business knowledge and skills, enable practice success, enhance health services delivery and positively influence the profession as a viable career.

  13. RETURN ON CURRENT ASSETS, WORKING CAPITAL AND REQUIRED RATE OF RETURN ON EQUITY

    Directory of Open Access Journals (Sweden)

    Monika Bolek

    2014-08-01

    Full Text Available The problem of return on current assets and return on working capital related to the cost of equity invested in a company is analyzed in this paper. Risk – return and liquidity – profitability trade-offs influence the company’s equilibrium and management decisions. Liquidity is measured by the cash conversion cycle and it is related to the working capital strategy, measured by current ratio. Rate of return on current assets should be related to the rate of return on working capital that is linked to the cost of capital and the required rate of return. The results indicate that there is a positive relationship between the return on current assets and cash conversion cycle, a positive relationship between cost of equity and return on working capital meaning that the working capital, cash conversion cycle and current assets management are related to profitability, and cost of equity is determined by the required rate of return calculated based on the Capital Asset Pricing Model.

  14. Current control loop design and analysis based on resonant regulators for microgrid applications

    DEFF Research Database (Denmark)

    Federico, de Bosio; Pastorelli, Michelle; de Sousa Ribeiro, Luiz Antonio

    2015-01-01

    Voltage and current control loops play an important role in the performance of microgrids employing power electronics voltage source inverters. Correct design of feedback loops is essential for the proper operation of these systems. This paper analyzes the influence of state feedback cross......-coupling in the design of resonant regulators for inner current loops in power converters operating in standalone microgrids. It is also demonstrated that the effect of state feedback cross-coupling degrades the performance of the control loops by increasing the steady-state error. Different resonant regulators...... structures are analyzed and compared, performing experimental tests to validate the results of the theoretical analysis....

  15. 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.

  16. Differential regulation of potassium currents by FGF-1 and FGF-2 in embryonic Xenopus laevis myocytes.

    Science.gov (United States)

    Chauhan-Patel, R; Spruce, A E

    1998-10-01

    1. Fibroblast growth factors (FGFs) are involved in the regulation of many aspects of muscle development. This study investigated their role in regulating voltage-dependent K+ currents in differentiating Xenopus laevis myocytes. Both FGF-1 and FGF-2 are expressed by developing muscle cells, so their actions were compared. Experiments were performed on cultured myocytes isolated from stage 15 embryos. 2. Long-term exposure of the embryonic myocytes to FGF-1 downregulated inward rectifier K+ current (IK(IR)) density as well as both sustained and inactivating voltage-dependent outward K+ currents (IK,S and IK,I, respectively) and their densities. In contrast, FGF-2 upregulated these currents, although, because of an increase in capacitance caused by FGF-2, current density did not change with this factor. 3. The regulation of IK(IR) by FGF-1 was prevented by the cytoplasmic tyrosine kinase inhibitor herbimycin A, but that of IK,S and IK,I was unaffected, indicating that FGF-1 achieves its regulatory effects on electrical development via separate signalling pathways. The receptor tyrosine kinase inhibitor genistein in isolation suppressed K+ currents, but this may have occurred through a channel-blocking mechanism. 4. In many cells, IK, S was found to be composed of two components with differing voltage dependencies of activation. The FGFs brought about an alteration in the amount of total IK,S by equal effects on each component. Conversely, herbimycin A increased the proportion of low voltage-activated current without affecting total current amplitude. Therefore, we suggest that a single species of channel whose voltage dependence is shifted by tyrosine phosphorylation generates IK,S. 5. In summary, FGF-1 and FGF-2 exert opposite effects on voltage-dependent K+ currents in embryonic myocytes and, furthermore, FGF-1 achieves its effects on different K+ currents via separate second messenger pathways.

  17. Review of the neutral-beam current requirements for the Mirror Fusion Test Facility (MFTF)

    Energy Technology Data Exchange (ETDEWEB)

    Porter, G.D.; Coensgen, F.H.; Devoto, R.S.; Molvik, A.W.; Osher, J.F.; Shearer, J.W.; Thomassen, K.I.

    1978-08-04

    The techniques used to evaluate the beam-current requirements for MFTF are similar to those in previous studies but incorporate some improvements. In particular, we have enlarged the radial code BUILDUP to allow for a smaller radial grid and to improve the numerical accuracy; we have also improved the model of the atomic physics processes and the particle-equilibrium calculations. Also, a model of plasma containment that conserves energy as well as particles but does not include any effect of the expected drift-cyclotron loss-cone (DCLC) turbulence has been incorporated into the previous studies. We show that the DCLC turbulence increases the average ion energy and decreases the particle confinement time. Because these effects cancel to first order, the beam-current requirements are independent of the turbulence. We find that 24 of the present LBL source modules would sustain a plasma with an average beta of 0.46. This figure is within 10% of the MFTF goal and is within our calculational accuracies. We further show that the beam-current requirements for the large-diameter plasma are consistent with those of experiments to study the buildup of a field-reversed plasma.

  18. Down-regulation of endogenous KLHL1 decreases voltage-gated calcium current density.

    Science.gov (United States)

    Perissinotti, Paula P; Ethington, Elizabeth G; Cribbs, Leanne; Koob, Michael D; Martin, Jody; Piedras-Rentería, Erika S

    2014-05-01

    The actin-binding protein Kelch-like 1 (KLHL1) can modulate voltage-gated calcium channels in vitro. KLHL1 interacts with actin and with the pore-forming subunits of Cav2.1 and CaV3.2 calcium channels, resulting in up-regulation of P/Q and T-type current density. Here we tested whether endogenous KLHL1 modulates voltage gated calcium currents in cultured hippocampal neurons by down-regulating the expression of KLHL1 via adenoviral delivery of shRNA targeted against KLHL1 (shKLHL1). Control adenoviruses did not affect any of the neuronal properties measured, yet down-regulation of KLHL1 resulted in HVA current densities ~68% smaller and LVA current densities 44% smaller than uninfected controls, with a concomitant reduction in α(1A) and α(1H) protein levels. Biophysical analysis and western blot experiments suggest Ca(V)3.1 and 3.3 currents are also present in shKLHL1-infected neurons. Synapsin I levels, miniature postsynaptic current frequency, and excitatory and inhibitory synapse number were reduced in KLHL1 knockdown. This study corroborates the physiological role of KLHL1 as a calcium channel modulator and demonstrates a novel, presynaptic role.

  19. Exploring Faraday's Law of Electrolysis Using Zinc-Air Batteries with Current Regulative Diodes

    Science.gov (United States)

    Kamata, Masahiro; Paku, Miei

    2007-01-01

    Current regulative diodes (CRDs) are applied to develop new educational experiments on Faraday's law by using a zinc-air battery (PR2330) and a resistor to discharge it. The results concluded that the combination of zinc-air batteries and the CRD array is simpler, less expensive, and quantitative and gives accurate data.

  20. A NOVEL CONTROL STRATEGY FOR SINUSIODAL WAVE INVERTER WITH PI REGULATORS AND CAPACITOR CURRENT FEEDBACK

    Institute of Scientific and Technical Information of China (English)

    姜桂宾; 裴云庆; 王兆安

    2003-01-01

    Objective For the inverters used in UPS, it is important to maintain the pure sinusoidal AC output voltage waveform over all loading conditions and transients. Methods A novel sinusoidal output voltage control strategy is pregented in this paper. The output voltage is controlled by introducing filtering eapacitor current feedback. Two simple PI regulators are used for the current and voltage control loops. Results With the new control strategy, the inverter achieves very low output voltage distortion, good output voltage regulation and strong perturbation rejection, fast dynamic response, and good performance under nonlinear loads. The THD under capacitance rectifying load is better than 0.2%, the output voltage regulation within 0 to full load is less than 0.1%. The resting time under load transient is within 200μs. Conclusion The merits of the new control strategy include rapid response and good steady state stiffness.

  1. Characterization of Mg2+-regulated TRPM7-like current in human atrial myocytes

    Directory of Open Access Journals (Sweden)

    Macianskiene Regina

    2012-08-01

    Full Text Available Abstract Background TRPM7 (Transient Receptor Potential of the Melastatin subfamily proteins are highly expressed in the heart, however, electrophysiological studies, demonstrating and characterizing these channels in human cardiomyocytes, are missing. Methods We have used the patch clamp technique to characterize the biophysical properties of TRPM7 channel in human myocytes isolated from right atria small chunks obtained from 116 patients in sinus rhythm during coronary artery and valvular surgery. Under whole-cell voltage-clamp, with Ca2+ and K+ channels blocked, currents were generated by symmetrical voltage ramp commands to potentials between -120 and +80 mV, from a holding potential of -80 mV. Results We demonstrate that activated native current has dual control by intracellular Mg2+ (free-Mg2+ or ATP-bound form, and shows up- or down-regulation by its low or high levels, respectively, displaying outward rectification in physiological extracellular medium. High extracellular Mg2+ and Ca2+ block the outward current, while Gd3+, SpM4+, 2-APB, and carvacrol inhibit both (inward and outward currents. Besides, divalents also permeate the channel, and the efficacy sequence, at 20 mM, was Mg2+>Ni2+>Ca2+>Ba2+>Cd2+ for decreasing outward and Ni2+>Mg2+>Ba2+≥Ca2+>Cd2+ for increasing inward currents. The defined current bears many characteristics of heterologously expressed or native TRPM7 current, and allowed us to propose that current under study is TRPM7-like. However, the time of beginning and time to peak as well steady state magnitude (range from 1.21 to 11.63 pA/pF, ncells/patients = 136/77 of induced TRPM7-like current in atrial myocytes from different patients showed a large variability, while from the same sample of human atria all these parameters were very homogenous. We present new information that TRPM7-like current in human myocytes is less sensitive to Mg2+. In addition, in some myocytes (from 24 out of 77 patients that current

  2. Power lateral pnp transistor operating with high current density in irradiated voltage regulator

    Directory of Open Access Journals (Sweden)

    Vukić Vladimir Đ.

    2013-01-01

    Full Text Available The operation of power lateral pnp transistors in gamma radiation field was examined by detection of the minimum dropout voltage on heavily loaded low-dropout voltage regulators LM2940CT5, clearly demonstrating their low radiation hardness, with unacceptably low values of output voltage and collector-emitter voltage volatility. In conjunction with previous results on base current and forward emitter current gain of serial transistors, it was possible to determine the positive influence of high load current on a slight improvement of voltage regulator LM2940CT5 radiation hardness. The high-current flow through the wide emitter aluminum contact of the serial transistor above the isolation oxide caused intensive annealing of the positive oxide-trapped charge, leading to decrease of the lateral pnp transistor's current gain, but also a more intensive recovery of the small-signal npn transistors in the control circuit. The high current density in the base area of the lateral pnp transistor immediately below the isolation oxide decreased the concentration of negative interface traps. Consequently, the positive influence of the reduced concentration of the oxide-trapped charge on the negative feedback reaction circuit, together with the favourable effect of reduced interface traps concentration, exceeded negative influence of the annealed oxide-trapped charge on the serial pnp transistor's forward emitter current gain.

  3. Pacemaker neuron and network oscillations depend on a neuromodulator-regulated linear current

    Directory of Open Access Journals (Sweden)

    Shunbing Zhao

    2010-05-01

    Full Text Available Linear leak currents have been implicated in the regulation of neuronal excitability, generation of neuronal and network oscillations, and network state transitions. Yet, few studies have directly tested the dependence of network oscillations on leak currents or explored the role of leak currents on network activity. In the oscillatory pyloric network of decapod crustaceans neuromodulatory inputs are necessary for pacemaker activity. A large subset of neuromodulators is known to activate a single voltage-gated inward current IMI, which has been shown to regulate the rhythmic activity of the network and its pacemaker neurons. Using the dynamic clamp technique, we show that the crucial component of IMI for the generation of oscillatory activity is only a close-to-linear portion of the current-voltage relationship. The nature of this conductance is such that the presence or the absence of neuromodulators effectively regulates the amount of leak current and the input resistance in the pacemaker neurons. When deprived of neuromodulatory inputs, pyloric oscillations are disrupted; yet, a linear reduction of the total conductance in a single neuron within the pacemaker group recovers not only the pacemaker activity in that neuron, but also leads to a recovery of oscillations in the entire pyloric network. The recovered activity produces proper frequency and phasing that is similar to that induced by neuromodulators. These results show that the passive properties of pacemaker neurons can significantly affect their capacity to generate and regulate the oscillatory activity of an entire network, and that this feature is exploited by neuromodulatory inputs.

  4. The Investigators' Brochure: a comparison of the draft international conference on harmonisation guideline with current Food and Drug Administration requirements.

    Science.gov (United States)

    Cocchetto, D M

    1995-12-01

    For several years, the United States Food and Drug Administration (FDA) has participated in a collaborative effort to harmonize the technical procedures for development and regulatory approval of human pharmaceuticals in multiple countries. This harmonization effort is the work of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). At this point, ICH focuses on achieving harmonization of technical requirements in three major regions of the world, i.e., the United States, European Union, and Japan. One area for which harmonization is being sought is the format and content of the Investigators' Brochure. On October 27, 1993, the ICH Steering Committee agreed that the ICH draft guideline on the Investigators' Brochure should be made available for public comment. On August 9, 1994, the FDA published the draft guideline for the format and content of the Investigators' Brochure. This draft guideline was prepared by the Efficacy Expert Working Group of the ICH. FDA solicited written comments on this draft guideline until October 11, 1994. Ultimately, FDA intends to adopt the ICH Steering Committee's final guidelines on Investigators' Brochures as part of a larger document on good clinical practices. Therefore, the content of this draft guideline warrants careful attention. In this paper, ICH's proposed information for inclusion in the Investigators' Brochure is reviewed and compared with current regulatory requirements of FDA. The expanded contents of the Investigators' Brochure proposed by ICH, which represent potential new requirements beyond current FDA regulations, are highlighted. The major controversial elements of this draft guideline are summarized.

  5. [Current requirements for polymeric biomaterials in ear, nose and throat medicine].

    Science.gov (United States)

    Sternberg, K

    2009-05-01

    In recent years the ear, nose and throat medicine (ENT medicine) has been stimulated by numerous innovations in the field of implants which are based on new biomaterials and modern implant technologies. In this context, biomaterials integrated in living organisms have to allow for the technical requirements and the biological interactions between the implant and the tissue. With regard to their suitability, functional capability of the implant, which is complementary to the mechanical implant properties, sufficient stability against physiological media, as well as high biocompatibility are to be demanded. Another purpose of the use of biomaterials is the maintenance and the enhancement of biofunctionality over a long time period. These general requirements for biomaterials also have their validity in ENT medicine. Different materials are applied as biomaterials. Metals belong to the oldest biomaterials. In addition, alloys, ceramics, inorganic glasses and composites were tested. Furthermore, natural and synthetic polymers, which are primarily presented in this article regarding their properties and their applications as materials for cochlear implants, osteosynthesis implants, stents and novel scaffolds for tissue engineering, are increasingly applied. According to their use in permanent and temporary implants, polymers are to be differentiated between biostable and biodegradable polymers. The presented general and current requirements for biomaterials and biomaterial applications in ENT medicine demonstrate key aspects of the current biomaterial research in this field. They do as well document the high impact of the interdisciplinary collaboration of natural and medical scientists and engineers.

  6. 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...

  7. Sensory deprivation regulates the development of the hyperpolarization-activated current in auditory brainstem neurons.

    Science.gov (United States)

    Hassfurth, Benjamin; Magnusson, Anna K; Grothe, Benedikt; Koch, Ursula

    2009-10-01

    Hyperpolarization-activated and cyclic nucleotide-gated (HCN) channels are highly expressed in the superior olivary complex, the primary locus for binaural information processing. This hyperpolarization-activated current (I(h)) regulates the excitability of neurons and enhances the temporally precise analysis of the binaural acoustic cues. By using the whole-cell patch-clamp technique, we examined the properties of I(h) current in neurons of the lateral superior olive (LSO) and the medial nucleus of the trapezoid body (MNTB) before and after hearing onset. Moreover, we tested the hypothesis that I(h) currents are actively regulated by sensory input activity by performing bilateral and unilateral cochlear ablations before hearing onset, resulting in a chronic auditory deprivation. The results show that after hearing onset, I(h) currents are rapidly upregulated in LSO neurons, but change only marginally in neurons of the MNTB. We also found a striking difference in maximal current density, voltage dependence and activation time constant between the LSO and the MNTB in mature-like animals. Following bilateral cochlear ablations before hearing onset, the I(h) currents were scaled up in the LSO and scaled down in the MNTB. Consequently, in the LSO this resulted in a depolarized resting membrane potential and a lower input resistance of these neurons. This type of activity-dependent homeostatic change could thus result in an augmented response to the remaining inputs.

  8. Bench Test Modeling for Current and Future PCMO Fuel Economy Requirements

    Institute of Scientific and Technical Information of China (English)

    Mark T.Devlin; Cheng C.Kuo; John M.Pietras; Zhang Yun

    2008-01-01

    The automotive and lubricant industries have been placing increased emphasis on the fuel economy benefits of today's modern passenger car motor oils.The current ILSAC GF-4 specification has more stringent requirements than previous ILSAC specifications,and the proposed future ILSAC GF-5 specification is looking at further improvements in the lubricant's fuel economy performance.To address these needs,Afton Chemical has developed correlations between the rheological and frictional properties of oils and fuel economy measured in engine and field tests.In this paper we will present correlations between lubricauts' physical properties and fuel economy measured in vehicles and enginetests.These tools have been used to develop current commercial oils which are being used extensively to meet today's OEM needs.

  9. LED Current Balance Using a Variable Voltage Regulator with Low Dropout vDS Control

    Directory of Open Access Journals (Sweden)

    Hung-I Hsieh

    2017-02-01

    Full Text Available A cost-effective light-emitting diode (LED current balance strategy using a variable voltage regulator (VVR with low dropout vDS control is proposed. This can regulate the multiple metal-oxide-semiconductor field-effect transistors (MOSFETs of the linear current regulators (LCR, maintaining low dropout vDS on the flat vGS-characteristic curves and making all drain currents almost the same. Simple group LCRs respectively loaded with a string LED are employed to implement the theme. The voltage VVdc from a VVR is synthesized by a string LED voltage NvD, source voltage vR, and a specified low dropout vDS = VQ. The VVdc updates instantly, through the control loop of the master LCR, which means that all slave MOSFETs have almost the same biases on their flat vGS-characteristic curves. This leads to all of the string LED currents being equal to each other, producing an almost even luminance. An experimental setup with microchip control is built to verify the estimations. Experimental results show that the luminance of all of the string LEDs are almost equal to one another, with a maximum deviation below 1% during a wide dimming range, while keeping all vDS of the MOSFETs at a low dropout voltage, as expected.

  10. Modulation of inhibitory strength and kinetics facilitates regulation of persistent inward currents and motoneuron excitability following spinal cord injury.

    Science.gov (United States)

    Venugopal, Sharmila; Hamm, Thomas M; Crook, Sharon M; Jung, Ranu

    2011-11-01

    Spasticity is commonly observed after chronic spinal cord injury (SCI) and many other central nervous system disorders (e.g., multiple sclerosis, stroke). SCI-induced spasticity has been associated with motoneuron hyperexcitability partly due to enhanced activation of intrinsic persistent inward currents (PICs). Disrupted spinal inhibitory mechanisms also have been implicated. Altered inhibition can result from complex changes in the strength, kinetics, and reversal potential (E(Cl(-))) of γ-aminobutyric acid A (GABA(A)) and glycine receptor currents. Development of optimal therapeutic strategies requires an understanding of the impact of these interacting factors on motoneuron excitability. We employed computational methods to study the effects of conductance, kinetics, and E(Cl(-)) of a dendritic inhibition on PIC activation and motoneuron discharge. A two-compartment motoneuron with enhanced PICs characteristic of SCI and receiving recurrent inhibition from Renshaw cells was utilized in these simulations. This dendritic inhibition regulated PIC onset and offset and exerted its strongest effects at motoneuron recruitment and in the secondary range of the current-frequency relationship during PIC activation. Increasing inhibitory conductance compensated for moderate depolarizing shifts in E(Cl(-)) by limiting PIC activation and self-sustained firing. Furthermore, GABA(A) currents exerted greater control on PIC activation than glycinergic currents, an effect attributable to their slower kinetics. These results suggest that modulation of the strength and kinetics of GABA(A) currents could provide treatment strategies for uncontrollable spasms.

  11. p38 Activation Is Required Upstream of Potassium Current Enhancement and Caspase Cleavage in Thiol Oxidant-Induced Neuronal Apoptosis

    Science.gov (United States)

    McLaughlin, BethAnn; Pal, Sumon; Tran, Minhnga P.; Parsons, Andrew A.; Barone, Frank C.; Erhardt, Joseph A.; Aizenman, Elias

    2013-01-01

    Oxidant-induced neuronal apoptosis has been shown to involve potassium and zinc dysregulation, energetic dysfunction, activation of stress-related kinases, and caspase cleavage. The temporal ordering and interdependence of these events was investigated in primary neuronal cultures exposed to the sulfhydryl oxidizing agent 2,2′-dithiodipyridine (DTDP), a compound that induces the intracellular release of zinc. We previously observed that tetraethylammonium (TEA), high extracellular potassium, or cysteine protease inhibitors block apoptosis induced by DTDP. We now report that both p38 and extracellular signal-regulated kinase phosphorylation are evident in neuronal cultures within 2 hr of a brief exposure to 100 μm DTDP. However, only p38 inhibition is capable of blocking oxidant-induced toxicity. Cyclohexamide or actinomycin D does not attenuate DTDP-induced cell death, suggesting that posttranslational modification of existing targets, rather than transcriptional activation, is responsible for the deleterious effects of p38. Indeed, an early robust increase in TEA-sensitive potassium channel currents induced by DTDP is attenuated by p38 inhibition but not by caspase inhibition. Moreover, we found that activation of p38 is required for caspase 3 and 9 cleavage, suggesting that potassium currents enhancement is required for caspase activation. Finally, we observed that DTDP toxicity could be blocked with niacinamide or benzamide, inhibitors of poly (ADP-ribose) synthetase. Based on these findings, we conclude that oxidation of sulfhydryl groups on intracellular targets results in intracellular zinc release, p38 phosphorylation, enhancement of potassium currents, caspase cleavage, energetic dysfunction, and translationally independent apoptotic cell death. PMID:11331359

  12. Regulation of persistent sodium currents by glycogen synthase kinase 3 encodes daily rhythms of neuronal excitability

    Science.gov (United States)

    Paul, Jodi R.; Dewoskin, Daniel; McMeekin, Laura J.; Cowell, Rita M.; Forger, Daniel B.; Gamble, Karen L.

    2016-11-01

    How neurons encode intracellular biochemical signalling cascades into electrical signals is not fully understood. Neurons in the central circadian clock in mammals provide a model system to investigate electrical encoding of biochemical timing signals. Here, using experimental and modelling approaches, we show how the activation of glycogen synthase kinase 3 (GSK3) contributes to neuronal excitability through regulation of the persistent sodium current (INaP). INaP exhibits a day/night difference in peak magnitude and is regulated by GSK3. Using mathematical modelling, we predict and confirm that GSK3 activation of INaP affects the action potential afterhyperpolarization, which increases the spontaneous firing rate without affecting the resting membrane potential. Together, these results demonstrate a crucial link between the molecular circadian clock and electrical activity, providing examples of kinase regulation of electrical activity and the propagation of intracellular signals in neuronal networks.

  13. 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....

  14. 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.

  15. SOME CONSIDERATIONS OVER THE INTERNAL CONTROL IN THE CONTEXT OF CURRENT ACCOUNTANCY REGULATIONS

    Directory of Open Access Journals (Sweden)

    Lenghel Dorin Radu

    2012-12-01

    Full Text Available Through the hereby study we set ourselves on an analysis over the internal control and the ways of implementation of it. The final purpose of our research is represented by the characteristics of the internal control system in the context of current accountancy regulations. The accomplishment of such goal has forced us to research the current literature in the area and also the legal regulations over internal control. Of course, the accomplishment of such a research was possible only after a pertinent analysis over the opinions expressed in specialty literature regarding this area. Our research wants to be with a theoretical and applicative character. It is based on analysing the internal control system, the methodology of internal control and the steps of internal control. We express our belief that the implementation of the internal control in a very exact manner comes as an aid to the company management and lead to the accomplishment of managerial objectives and policies. In what it regards the result of our research there can be drawn the following conclusions: from the legal regulations (OMFP nr. 3055/2009, there is no clear conclusion over the procedures and policies of implementation of internal control applicable to an economical entity; also there are not presented in a detailed way the types (forms of internal control, being presented in the legal regulations only a classification of them after the time in which they are exercised (before, during or after the finishing of the operations; also in the legal regulations there is developed a certain way of internal control, represented by the financial and accounting internal control, as if the other forms of internal control do not exist; in what regards the components of internal control, these are presented in another way, but similar to already known models of internal control (Coso and Coco; also in the legal regulations there are presented the general objectives of internal control

  16. 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

  17. 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

  18. 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

  19. 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...

  20. 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....

  1. 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...

  2. 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

  3. 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...

  4. 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...

  5. 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.

  6. 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

  7. Shared Solar. Current Landscape, Market Potential, and the Impact of Federal Securities Regulation

    Energy Technology Data Exchange (ETDEWEB)

    Feldman, David [National Renewable Energy Lab. (NREL), Golden, CO (United States); Brockway, Anna M. [Office of Energy Efficiency and Renewable Energy (EERE), Washington, DC (United States); Ulrich, Elaine [Office of Energy Efficiency and Renewable Energy (EERE), Washington, DC (United States); Margolis, Robert [National Renewable Energy Lab. (NREL), Golden, CO (United States)

    2015-04-01

    This report provides a high-level overview of the current U.S. shared solar landscape and the impact that a given shared solar program’s structure has on requiring federal securities oversight, as well as an estimate of market potential for U.S. shared solar deployment.

  8. Shared Solar. Current Landscape, Market Potential, and the Impact of Federal Securities Regulation

    Energy Technology Data Exchange (ETDEWEB)

    Feldman, David [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Brockway, Anna M. [U.S. Department of Energy, Washington, DC (United States); Ulrich, Elaine [U.S. Department of Energy, Washington, DC (United States); Margolis, Robert [National Renewable Energy Laboratory (NREL), Golden, CO (United States)

    2015-04-07

    This report provides a high-level overview of the current U.S. shared solar landscape, the impact that a given shared solar program’s structure has on requiring federal securities oversight, as well as an estimate of market potential for U.S. shared solar deployment.

  9. 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

  10. [Requirements for quality indicators. The relevance of current developments in outcomes research for quality management].

    Science.gov (United States)

    Schmitt, Jochen; Petzold, Thomas; Eberlein-Gonska, Maria; Neugebauer, Edmund A M

    2013-01-01

    The assessment of the health state in patients and changes in their health state for the purpose of diagnosis, prognosis and monitoring of treatment response plays a central role in clinical practice. Quality criteria for measurements in medicine include validity, reliability, responsiveness, interpretability, and feasibility. High-quality measurement instruments are a prerequisite for evidence-based medicine. Therefore, international outcomes research groups have developed methods for quality assurance and for the standardisation of measurement instruments. Quality indicators are instruments to measure the quality of care. Due to the increasing relevance of quality assessment for all stakeholders in healthcare and due to the political intention to draw relevant conclusions from the assessment of the quality of care, quality indicators must at least meet the same high standards that are required for clinical trial end points. However, independent researchers and clinicians do not engage in the validation and standardisation of quality indicators in Germany; currently, only the AQUA institute (as assigned by the German GBA) deals with this important issue. Current activities concerning the validation of quality indicators do not meet the requirements of evidence-based healthcare. This is a critical barrier to achieving the political goals of quality medicine. Therefore, the authors propose a multi-step, multi-professional, evidence-driven and evidence-generating consensus process on the basis of established methods of outcomes research for the advancement of quality assessment with quality indicators in Germany. All relevant stakeholders should participate in this process. Copyright © 2013. Published by Elsevier GmbH.

  11. Current status and requirements for position-sensitive detectors in medicine

    CERN Document Server

    Speller, R

    2002-01-01

    This review considers the current status of detector developments for medical imaging using ionising radiation. This field is divided into two major areas; the use of X-rays for transmission imaging and the use of radioactive tracers in emission imaging (nuclear medicine). Until recently, most detector developments were for applications in nuclear medicine. However, in the past 5 years new developments in large area, X-ray-sensitive detectors have meant that both application domains are equally served. In X-ray imaging, work in CT and mammography are chosen as examples of sensor developments. Photodiode arrays in multi-slice spiral CT acquisitions are described and for mammography the use of amorphous silicon flat panel arrays is considered. The latter is an excellent example where new detector developments have required a re-think of traditional imaging methods. In gamma-ray imaging the recent developments in small area, task-specific cameras are described. Their limitations and current proposals to overcome...

  12. A Review of Engine Seal Performance and Requirements for Current and Future Army Engine Platforms

    Science.gov (United States)

    Delgado, Irebert R.; Proctor, Margaret P.

    2008-01-01

    Sand ingestion continues to impact combat ground and air vehicles in military operations in the Middle East. The T-700 engine used in Apache and Blackhawk helicopters has been subjected to increased overhauls due to sand and dust ingestion during desert operations. Engine component wear includes compressor and turbine blades/vanes resulting in decreased engine power and efficiency. Engine labyrinth seals have also been subjected to sand and dust erosion resulting in tooth tip wear, increased clearances, and loss in efficiency. For the current investigation, a brief overview is given of the history of the T-700 engine development with respect to sand and dust ingestion requirements. The operational condition of labyrinth seals taken out of service from 4 different locations of the T-700 engine during engine overhauls are examined. Collaborative efforts between the Army and NASA to improve turbine engine seal leakage and life capability are currently focused on noncontacting, low leakage, compliant designs. These new concepts should be evaluated for their tolerance to sand laden air. Future R&D efforts to improve seal erosion resistance and operation in desert environments are recommended

  13. pH-regulated ionic current rectification in conical nanopores functionalized with polyelectrolyte brushes.

    Science.gov (United States)

    Zeng, Zhenping; Ai, Ye; Qian, Shizhi

    2014-02-14

    Mimicking biological ion channels capable of pH-regulated ionic transport, synthetic nanopores functionalized with pH-tunable polyelectrolyte (PE) brushes have been considered as versatile tools for active transport control of ions, fluids, and bioparticles on the nanoscale. The ionic current rectification (ICR) phenomenon through a conical nanopore functionalized with PE brushes whose charge highly depends upon the local solution properties (i.e., pH and background salt concentration) is studied theoretically for the first time. The results show that the rectification magnitude, as well as the preferential rectification direction, is sensitive to the pH stimulus. The bulk concentration of the background salt can also significantly influence the charge of the PE brushes and accordingly affect the ICR phenomenon. The obtained results provide an insightful understanding of the pH-regulated ICR and guidelines for designing nanopores functionalized with PE brushes for pH-tunable applications.

  14. Current practices and reform proposals for the regulation of advanced medicinal products in Canada.

    Science.gov (United States)

    Viswanathan, Sowmya; Bubela, Tania

    2015-01-01

    We describe the Canadian regulatory framework for evaluating advanced medicinal products based on current policies, guidance documents and regulations and analyze proposed reforms. Our analysis is based on a documentary review supplemented by discussions with Health Canada officials. We present an overview of the Canadian regulatory framework for cell and gene therapy, medical devices and manufacturing facilities. We use the approval of Prochymal™ to highlight Canada's conditional marketing approval system. Finally, we discuss proposed changes to the regulatory framework in response to identified gaps, stakeholder consultations and international harmonization initiatives. Based on our analyses, we suggest that Canadian regulators have taken a reasonable approach in applying their regulatory framework without compromising on product safety.

  15. Current Insights to Regulation and Role of Telomerase in Human Diseases

    Science.gov (United States)

    Ozturk, Mert Burak; Li, Yinghui; Tergaonkar, Vinay

    2017-01-01

    The telomerase ribonucleoprotein complex has a pivotal role in regulating the proliferation and senescence of normal somatic cells as well as cancer cells. This complex is comprised mainly of telomerase reverse transcriptase (TERT), telomerase RNA component (TERC) and other associated proteins that function to elongate telomeres localized at the end of the chromosomes. While reactivation of telomerase is a major hallmark of most cancers, together with the synergistic activation of other oncogenic signals, deficiency in telomerase and telomeric proteins might lead to aging and senescence-associated disorders. Therefore, it is critically important to understand the canonical as well as non-canonical functions of telomerase through TERT to develop a therapeutic strategy against telomerase-related diseases. In this review, we shed light on the regulation and function of telomerase, and current therapeutic strategies against telomerase in cancer and age-related diseases. PMID:28264499

  16. Carbohydrate Metabolism in Archaea: Current Insights into Unusual Enzymes and Pathways and Their Regulation

    Science.gov (United States)

    Esser, Dominik; Rauch, Bernadette

    2014-01-01

    SUMMARY The metabolism of Archaea, the third domain of life, resembles in its complexity those of Bacteria and lower Eukarya. However, this metabolic complexity in Archaea is accompanied by the absence of many “classical” pathways, particularly in central carbohydrate metabolism. Instead, Archaea are characterized by the presence of unique, modified variants of classical pathways such as the Embden-Meyerhof-Parnas (EMP) pathway and the Entner-Doudoroff (ED) pathway. The pentose phosphate pathway is only partly present (if at all), and pentose degradation also significantly differs from that known for bacterial model organisms. These modifications are accompanied by the invention of “new,” unusual enzymes which cause fundamental consequences for the underlying regulatory principles, and classical allosteric regulation sites well established in Bacteria and Eukarya are lost. The aim of this review is to present the current understanding of central carbohydrate metabolic pathways and their regulation in Archaea. In order to give an overview of their complexity, pathway modifications are discussed with respect to unusual archaeal biocatalysts, their structural and mechanistic characteristics, and their regulatory properties in comparison to their classic counterparts from Bacteria and Eukarya. Furthermore, an overview focusing on hexose metabolic, i.e., glycolytic as well as gluconeogenic, pathways identified in archaeal model organisms is given. Their energy gain is discussed, and new insights into different levels of regulation that have been observed so far, including the transcript and protein levels (e.g., gene regulation, known transcription regulators, and posttranslational modification via reversible protein phosphorylation), are presented. PMID:24600042

  17. 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.

  18. 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.

  19. An energy-efficient, adiabatic electrode stimulator with inductive energy recycling and feedback current regulation.

    Science.gov (United States)

    Arfin, Scott K; Sarpeshkar, Rahul

    2012-02-01

    In this paper, we present a novel energy-efficient electrode stimulator. Our stimulator uses inductive storage and recycling of energy in a dynamic power supply. This supply drives an electrode in an adiabatic fashion such that energy consumption is minimized. It also utilizes a shunt current-sensor to monitor and regulate the current through the electrode via feedback, thus enabling flexible and safe stimulation. Since there are no explicit current sources or current limiters, wasteful energy dissipation across such elements is naturally avoided. The dynamic power supply allows efficient transfer of energy both to and from the electrode and is based on a DC-DC converter topology that we use in a bidirectional fashion in forward-buck or reverse-boost modes. In an exemplary electrode implementation intended for neural stimulation, we show how the stimulator combines the efficiency of voltage control and the safety and accuracy of current control in a single low-power integrated-circuit built in a standard .35 μm CMOS process. This stimulator achieves a 2x-3x reduction in energy consumption as compared to a conventional current-source-based stimulator operating from a fixed power supply. We perform a theoretical analysis of the energy efficiency that is in accord with experimental measurements. This theoretical analysis reveals that further improvements in energy efficiency may be achievable with better implementations in the future. Our electrode stimulator could be widely useful for neural, cardiac, retinal, cochlear, muscular and other biomedical implants where low power operation is important.

  20. International evaluation of current and future requirements for environmental engineering education.

    Science.gov (United States)

    Morgenroth, E; Daigger, G T; Ledin, A; Keller, J

    2004-01-01

    The field of environmental engineering is developing as a result of changing environmental requirements. In response, environmental engineering education (E3) needs to ensure that it provides students with the necessary tools to address these challenges. In this paper the current status and future development of E3 is evaluated based on a questionnaire sent to universities and potential employers of E3 graduates. With increasing demands on environmental quality, the complexity of environmental engineering problems to be solved can be expected to increase. To find solutions environmental engineers will need to work in interdisciplinary teams. Based on the questionnaire there was a broad agreement that the best way to prepare students for these future challenges is to provide them with a fundamental education in basic sciences and related engineering fields. Many exciting developments in the environmental engineering profession will be located at the interface between engineering, science, and society. Aspects of all three areas need to be included in E3 and the student needs to be exposed to the tensions associated with linking the three.

  1. Radiation and Reason Why radiation at modest dose rates is quite harmless and current radiation safety regulations are flawed

    CERN Document Server

    CERN. Geneva

    2013-01-01

    Data on the impact of ionising radiation on life are examined in the light of evolutionary biology. This comparison confirms that fear of nuclear radiation is not justified by science itself; rather it originates in a failure of public trust in nuclear science, a relic of the international politics of the Cold War era. Current ionisation safety regulations appease this fear but without scientific support and they need fundamental reformulation. This should change the reaction to accidents like Fukushima, the cost of nuclear energy and the application of nuclear technology to the supply of food and fresh water. Such a boost to the world economy would require that more citizens study and appreciate the science involved – and then tell others -- not as much fun as the Higgs, perhaps, but no less important! www.radiationandreason.com

  2. Distributed Nonlinear Control with Event-Triggered Communication to Achieve Current-Sharing and Voltage Regulation in DC Microgrids

    DEFF Research Database (Denmark)

    Han, Renke; Meng, Lexuan; Guerrero, Josep M.

    2017-01-01

    A distributed nonlinear controller is presented to achieve both accurate current-sharing and voltage regulation simultaneously in dc microgrids considering different line impedances’ effects among converters. Then, an improved event-triggered principle for the controller is introduced through...... combining the state-dependent tolerance with a nonnegative offset. In order to design the event-triggered principle and guarantee the global stability, a generalized dc microgrid model is proposed and proven to be positive definite, based on which Lyapunov-based approach is applied. Furthermore, considering...... the effects from constant power loads, the damping performance of proposed controller is further improved and compared with the traditional V-I droop controller. The proposed event-triggered-based communication strategy can considerably reduce the communication traffic and significantly relax the requirement...

  3. A Small Potassium Current in AgRP/NPY Neurons Regulates Feeding Behavior and Energy Metabolism

    Directory of Open Access Journals (Sweden)

    Yanlin He

    2016-11-01

    Full Text Available Neurons that co-express agouti-related peptide (AgRP and neuropeptide Y (NPY are indispensable for normal feeding behavior. Firing activities of AgRP/NPY neurons are dynamically regulated by energy status and coordinate appropriate feeding behavior to meet nutritional demands. However, intrinsic mechanisms that regulate AgRP/NPY neural activities during the fed-to-fasted transition are not fully understood. We found that AgRP/NPY neurons in satiated mice express high levels of the small-conductance calcium-activated potassium channel 3 (SK3 and are inhibited by SK3-mediated potassium currents; on the other hand, food deprivation suppresses SK3 expression in AgRP/NPY neurons, and the decreased SK3-mediated currents contribute to fasting-induced activation of these neurons. Genetic mutation of SK3 specifically in AgRP/NPY neurons leads to increased sensitivity to diet-induced obesity, associated with chronic hyperphagia and decreased energy expenditure. Our results identify SK3 as a key intrinsic mediator that coordinates nutritional status with AgRP/NPY neural activities and animals’ feeding behavior and energy metabolism.

  4. Voltage-gated K+ currents in mouse articular chondrocytes regulate membrane potential.

    Science.gov (United States)

    Clark, Robert B; Hatano, Noriyuki; Kondo, Colleen; Belke, Darrell D; Brown, Barry S; Kumar, Sanjay; Votta, Bartholomew J; Giles, Wayne R

    2010-01-01

    of the joint cartilage, or in response to hypertonic challenge is expected to result in depolarization and Ca(2+) entry. Potassium currents are required to maintain the resting membrane potential.

  5. 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.

  6. A fast transient response low dropout regulator with current control methodology

    Institute of Scientific and Technical Information of China (English)

    Ma Zhuo; Guo Yang; Duan Zhikui; Xie Lunguo; Chen Jihua; Yu Jinshan

    2011-01-01

    A transient performance optimized CCL-LDO regulator is proposed.In the CCL-LDO,the control method of the charge pump phase-locked loop is adopted.A current control loop has the feedback signal and reference current to be compared,and then a loop filter generates the gate voltage of the power MOSFET by integrating the error current.The CCL-LDO has the optimized damping coefficient and natural resonant frequency,while its output voltage can be sub-1-V and is not restricted by the reference voltage.With a 1μF decoupling capacitor,the experimental results based on a 0.13 μm CMOS process show that the output voltage is 1.0 V; when the workload changes from 100 μA to 100 mA transiently,the stable dropout is 4.25 mV,the settling time is 8.2 μs and the undershoot is 5.11 mV; when the workload changes from 1 00 mA to 100 μA transiently,the stable dropout is 4.25 mV,the settling time is 23.3 μs and the overshoot is 6.21 mV.The PSRR value is more than -95 dB.Most of the attributes of the CCL-LDO are improved rapidly with a FOM value of 0.0097.

  7. 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

  8. Current regulators for I/SUP 2/L circuits to be operated from low-voltage power supplies

    DEFF Research Database (Denmark)

    Bruun, Erik; Hansen, Ole

    1980-01-01

    A new bandgap current reference is described which can be used to control the injector current of I/SUP 2/L circuits for supply voltages down to about 1 V. For small currents the total injector current is obtained as a mirror of the reference current. For large injector currents the current control...... is performed by a series regulator which compares the injector current of one I/SUP 2/L gate to the reference current. The described reference current can be adjusted to give a variation with temperature of about 60 ppm/°C over the temperature range -10 to +70°C. However, in some applications a nonzero......, but well controlled temperature coefficient is desired. It is shown how a temperature stable ring oscillator with I/SUP 2/L gates can be constructed by tailoring the temperature dependence of the supply current appropriately....

  9. Design of Current-Controller with PR-regulator for LCL-Filter Based Grid-Connected Converter

    DEFF Research Database (Denmark)

    Zeng, Guohong; Rasmussen, Tonny Wederberg

    2010-01-01

    In the application of LCL-filter based converters, the structure and parameters of current-controller is very important for the system stability and output current quality. This paper presents a filter-capacitor current feedback control scheme for grid-connected converter. The controller...... is consisted of a proportional-resonance regulator and a proportional regulator. Unlike the existing control strategy with unit capacitor current feedback, the proposed method applies the proportional regulator to the feedback path, which can decouple these two regulators, and simplify the tuning process....... Controller parameters are tuned with the aid of root-locus of system close-loop transfer function, which can provide better grasp of the influence of individual parameters and make the design process visible and holistic without any simplification and assumption. The validity, reasonability and feasibility...

  10. Investigation of the current requirements engineering practices among software developers at the Universiti Utara Malaysia Information Technology (UUMIT) centre

    Science.gov (United States)

    Hussain, Azham; Mkpojiogu, Emmanuel O. C.; Abdullah, Inam

    2016-08-01

    Requirements Engineering (RE) is a systemic and integrated process of eliciting, elaborating, negotiating, validating and managing of the requirements of a system in a software development project. UUM has been supported by various systems developed and maintained by the UUM Information Technology (UUMIT) Centre. The aim of this study was to assess the current requirements engineering practices at UUMIT. The main problem that prompted this research is the lack of studies that support software development activities at the UUMIT. The study is geared at helping UUMIT produce quality but time and cost saving software products by implementing cutting edge and state of the art requirements engineering practices. Also, the study contributes to UUM by identifying the activities needed for software development so that the management will be able to allocate budget to provide adequate and precise training for the software developers. Three variables were investigated: Requirement Description, Requirements Development (comprising: Requirements Elicitation, Requirements Analysis and Negotiation, Requirements Validation), and Requirement Management. The results from the study showed that the current practice of requirement engineering in UUMIT is encouraging, but still need further development and improvement because a few RE practices were seldom practiced.

  11. Current advances in DNA repair: regulation of enzymes and pathways involved in maintaining genomic stability.

    Science.gov (United States)

    Neher, Tracy M; Turchi, John J

    2011-06-15

    Novel discoveries in the DNA repair field have lead to continuous and rapid advancement of our understanding of not only DNA repair but also DNA replication and recombination. Research in the field transcends numerous areas of biology, biochemistry, physiology, and medicine, making significant connections across these broad areas of study. From early studies conducted in bacterial systems to current analyses in eukaryotic systems and human disease, the innovative research into the mechanisms of repair machines and the consequences of ineffective DNA repair has impacted a wide scientific community. This Forum contains a select mix of primary research articles in addition to a number of timely reviews covering a subset of DNA repair pathways where recent advances and novel discoveries are improving our understanding of DNA repair, its regulation, and implications to human disease.

  12. A role of stretch-activated potassium currents in the regulation of uterine smooth muscle contraction

    Institute of Scientific and Technical Information of China (English)

    Iain L O BUXTON; Nathanael HEYMAN; Yi-ying WU; Scott BARNETT; Craig ULRICH

    2011-01-01

    Rates of premature birth are alarming and threaten societies and healthcare systems worldwide. Premature labor results in premature birth in over 50% of cases. Preterm birth accounts for three-quarters of infant morbidity and mortality. Children that survive birth before 34 weeks gestation often face life-long disability. Current treatments for preterm labor are wanting. No treatment has been found to be generally effective and none are systematically evaluated beyond 48 h. New approaches to the treatment of preterm labor are desperately needed. Recent studies from our laboratory suggest that the uterine muscle is a unique compartment with regulation of uterine relaxation unlike that of other smooth muscles. Here we discuss recent evidence that the mechanically activated 2-pore potassium channel, TREK-1, may contribute to contraction-relaxation signaling in uterine smooth muscle and that TREK-1 gene variants associated with human labor and preterm labor may lead to a better understanding of preterm labor and its possible prevention.

  13. The methodologies and instruments of vehicle particulate emission measurement for current and future legislative regulations

    Science.gov (United States)

    Otsuki, Yoshinori; Nakamura, Hiroshi; Arai, Masataka; Xu, Min

    2015-09-01

    Since the health risks associated with fine particles whose aerodynamic diameters are smaller than 2.5 μm was first proven, regulations restricting particulate matter (PM) mass emissions from internal combustion engines have become increasingly severe. Accordingly, the gravimetric method of PM mass measurement is facing its lower limit of detection as the emissions from vehicles are further reduced. For example, the variation in the adsorption of gaseous components such as hydrocarbons from unburned fuel and lubricant oil and the presence of agglomerated particles, which are not directly generated in engine combustion but re-entrainment particulates from walls of sampling pipes, can cause uncertainty in measurement. The PM mass measurement systems and methodologies have been continuously refined in order to improve measurement accuracy. As an alternative metric, the particle measurement programme (PMP) within the United Nations Economic Commission for Europe (UNECE) developed a solid particle number measurement method in order to improve the sensitivity of particulate emission measurement from vehicles. Consequently, particle number (PN) limits were implemented into the regulations in Europe from 2011. Recently, portable emission measurement systems (PEMS) for in-use vehicle emission measurements are also attracting attention, currently in North America and Europe, and real-time PM mass and PN instruments are under evaluation.

  14. 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.

  15. 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.

  16. Leptin regulation of inward membrane currents, electrical activity and LH release in isolated bovine gonadotropes.

    Science.gov (United States)

    Domínguez-Mancera, Belisario; Barrientos-Morales, Manuel; Cervantes-Acosta, Patricia; Hernández-Beltrán, Antonio; Rodríguez-Andrade, Araceli; González-Ramírez, Ricardo; Monjaraz, Eduardo; Felix, Ricardo

    2017-09-09

    Leptin, a peptide hormone produced by adipocytes, is recognized as one of the signals involved in the onset of reproductive activity. The leptin receptor has been found in hypothalamic neurons and pituitary gonadotropes, suggesting that the hormone may act at both sites to stimulate the secretion of GnRH and consequently, FSH and LH. In response to a stimulus such as a hypothalamic secretagogue, gonadotropes respond with changes in electrical activity, intracellular Ca(2+) and hormone release. The main aim of this report was to investigate whether leptin promotes a change in the electrical and secretory activities of bovine gonadotropes. After 48 h of treatment with leptin (10 nM) significant changes in the action potential properties were observed in gonadotropes, which included an increase in amplitude, time-to-pike and post-hyperpolarization, as well as a decrease in firing threshold. Likewise, leptin induced a significant (∼1.3-fold) up-regulation of voltage-gated Na(+) channel current density, and a selective increase (∼2.1-fold) in Ca(2+) current density through high voltage-activated channels. Consistent with this, leptin enhanced GnRH-induced secretion of LH measured by ELISA. We suggest that leptin enhances membrane expression of voltage-gated Na(+) and Ca(2+) channels, which results in a modulation of the action potential properties and an increase in hormone release from gonadotropes. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Return on current assets, working capital and required rate of return on equity

    OpenAIRE

    2014-01-01

    The problem of return on current assets and return on working capital related to the cost of equity invested in a company is analyzed in this paper. Risk – return and liquidity – profitability trade-offs influence the company’s equilibrium and management decisions. Liquidity is measured by the cash conversion cycle and it is related to the working capital strategy, measured by current ratio. Rate of return on current assets should be related to the rate of return on working capital that is li...

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. Flexible Power Regulation and Current-limited Control of Grid-connected Inverter under Unbalanced Grid Voltage Faults

    DEFF Research Database (Denmark)

    Guo, Xiaoqiang; Liu, Wenzhao; Lu, Zhigang

    2017-01-01

    The grid-connected inverters may experience excessive current stress in case of unbalanced grid voltage Fault Ride Through (FRT), which significantly affects the reliability of the power supply system. In order to solve the problem, the inherent mechanisms of the excessive current phenomenon...... with the conventional FRT solutions are discussed. The quantitative analysis of three phase current peak values are conducted and a novel current-limited control strategy is proposed to achieve the flexible active and reactive power regulation and successful FRT in a safe current operation area with the aim...

  4. 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

  5. Current knowledge on the photoneuroendocrine regulation of reproduction in temperate fish species.

    Science.gov (United States)

    Migaud, H; Davie, A; Taylor, J F

    2010-01-01

    Seasonality is an important adaptive trait in temperate fish species as it entrains or regulates most physiological events such as reproductive cycle, growth profile, locomotor activity and key life-stage transitions. Photoperiod is undoubtedly one of the most predictable environmental signals that can be used by most living organisms including fishes in temperate areas. This said, however, understanding of how such a simple signal can dictate the time of gonadal recruitment and spawning, for example, is a complex task. Over the past few decades, many scientists attempted to unravel the roots of photoperiodic signalling in teleosts by investigating the role of melatonin in reproduction, but without great success. In fact, the hormone melatonin is recognized as the biological time-keeping hormone in fishes mainly due to the fact that it reflects the seasonal variation in daylength across the whole animal kingdom rather than the existence of direct evidences of its role in the entrainment of reproduction in fishes. Recently, however, some new studies clearly suggested that melatonin interacts with the reproductive cascade at a number of key steps such as through the dopaminergic system in the brain or the synchronization of the final oocyte maturation in the gonad. Interestingly, in the past few years, additional pathways have become apparent in the search for a fish photoneuroendocrine system including the clock-gene network and kisspeptin signalling and although research on these topics are still in their infancy, it is moving at great pace. This review thus aims to bring together the current knowledge on the photic control of reproduction mainly focusing on seasonal temperate fish species and shape the current working hypotheses supported by recent findings obtained in teleosts or based on knowledge gathered in mammalian and avian species. Four of the main potential regulatory systems (light perception, melatonin, clock genes and kisspeptin) in fish reproduction

  6. 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.

  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. Educational Electronic Information Dissemination and Broadcast Services: History, Current Infrastructure and Public Broadcasting Requirements.

    Science.gov (United States)

    Singh, Jai P.; Morgan, Robert P.

    This memorandum describes the results of a study on electronic educational information dissemination and broadcast services in the United States. Included are detailed discussions of the historical development and current infrastructure (both in terms of organization and physical plant) of the following services: educational radio and television…

  9. Minimum Field Requirements for Spin-Polarized Current Assisted Switching of Magnetization in Nanostructures with Uniaxial Anisotropy

    Directory of Open Access Journals (Sweden)

    GINDULESCU, A.

    2009-03-01

    Full Text Available The present paradigm of magnetic data storage is approaching its fundamental limits for areal storage density, as well as for speed in data processing. As a result, several magnetic recording alternatives, such as spin polarized current assisted recording, precessional switching, toggle switching, heat assisted recording are currently under intense research efforts. This article is aimed at providing a pertinent theoretical analysis of the spin polarized current assisted recording, emphasizing its performance with respect to minimum requirements for switching field. The first analytical derivation of the critical field curve in the presence of spin polarized currents is presented and the results are compared to the classical Stoner-Wohlfarth astroid. The analysis is performed under the framework of the Landau-Lifshitz-Gilbert-Slonczewski equation for describing the magnetization dynamics driven by external magnetic fields and spin polarized currents.

  10. Regulating Current Rectification and Nanoparticle Transport Through a Salt Gradient in Bipolar Nanopores.

    Science.gov (United States)

    Lin, Chih-Yuan; Yeh, Li-Hsien; Hsu, Jyh-Ping; Tseng, Shiojenn

    2015-09-16

    Tuning of ion and nanoparticle transport is validated through applying a salt gradient in two types of nanopores: the inner wall of a nanopore has bipolar charges and its outer wall neutral (type I), and both the inner and outer walls of a nanopore have bipolar charges (type II). The ion current rectification (ICR) behavior of these nanopores can be regulated by an applied salt gradient: if it is small, the degree of ICR in type II nanopore is more significant than that in type I nanopore; a reversed trend is observed at a sufficiently large salt gradient. If the applied salt gradient and electric field have the same direction, type I nanopore exhibits two significant features that are not observed in type II nanopore: (i) a cation-rich concentration polarization field and an enhanced funneling electric field are present near the cathode side of the nanopore, and (ii) the magnitude of the axial electric field inside the nanopore is reduced. These features imply that applying a salt gradient to type I nanopore is capable of simultaneously enhancing the nanoparticle capture into the nanopore and reducing its translocation velocity inside, so that high sensing performance and resolution can be achieved.

  11. 78 FR 4307 - Current Good Manufacturing Practice Requirements for Combination Products

    Science.gov (United States)

    2013-01-22

    ... streamlined regulatory framework for firms to use when demonstrating compliance with CGMP requirements for... framework that takes account of the fact that combination products are made up of drug, device, and biological product constituent parts. At the same time, commenters wanted to ensure that the framework...

  12. Socio-legal regulation of family relations: current status and prospects for development

    Directory of Open Access Journals (Sweden)

    Ludmila Saenko

    2013-04-01

    Full Text Available The crisis of the traditional family becomes important research methods and techniques of social regulation of family relations in contemporary reality. Particular attention is paid to the legal regulation of having the author's opinion the fundamental nature of the general system of social regulation, its value in the preservation of traditional moral family values.

  13. Thoughts on How to Regulate Behaviours: An Overview of the Current Debate

    Science.gov (United States)

    Etienne, Julien

    2013-01-01

    The article intends to present the debate on behaviour modification in the regulation studies literature, at a time of renewed interest among regulators for new ideas and strategies. As the financial crisis has led to the most public critique yet of the rational choice view of individuals that has informed regulation in the last few decades, other…

  14. 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...

  15. 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.

  16. 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.

  17. 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.

  18. Current issues in determining dietary protein and amino-acid requirements

    DEFF Research Database (Denmark)

    Pencharz, P; Jahoor, F; Kurpad, A

    2014-01-01

    Pregnancy and the first two years of life are periods of rapid growth and yet the knowledge of requirements for protein and dietary indispensable amino acids is very limited. The development of carbon oxidation methods opens the way to studies that should fill these important gaps in knowledge.Eu.......European Journal of Clinical Nutrition advance online publication, 15 January 2014; doi:10.1038/ejcn.2013.297....

  19. Validating Vegetable Production Unit (VPU) Plants, Protocols, Procedures and Requirements (P3R) using Currently Existing Flight Resources

    Science.gov (United States)

    Bingham, Gail; Bates, Scott; Bugbee, Bruce; Garland, Jay; Podolski, Igor; Levinskikh, Rita; Sychev, Vladimir; Gushin, Vadim

    2009-01-01

    Validating Vegetable Production Unit (VPU) Plants, Protocols, Procedures and Requirements (P3R) Using Currently Existing Flight Resources (Lada-VPU-P3R) is a study to advance the technology required for plant growth in microgravity and to research related food safety issues. Lada-VPU-P3R also investigates the non-nutritional value to the flight crew of developing plants on-orbit. The Lada-VPU-P3R uses the Lada hardware on the ISS and falls under a cooperative agreement between National Aeronautics and Space Administration (NASA) and the Russian Federal Space Association (FSA). Research Summary: Validating Vegetable Production Unit (VPU) Plants, Protocols, Procedures and Requirements (P3R) Using Currently Existing Flight Resources (Lada-VPU-P3R) will optimize hardware and

  20. International development assistance for renewable technologies: current programs and institutional requirements

    Energy Technology Data Exchange (ETDEWEB)

    Ashworth, J. H.; Meunier, R. E.

    1979-05-01

    Within the last several years, foreign-assistance donor agencies have begun to provide significant aid for the search for renewable energy sources for developing nations. This paper reports preliminary results from a survey of development--assistance projects in renewable energy sources, indicating which areas are extensions of traditional assistance areas and which are new areas of involvement. The last two portions of the paper indicate certain shortcomings in the current effort, and linkages that must be emphasized in order to incease the effectiveness of the range of donor activities.

  1. Focused transcranial direct current stimulation (tDCS) over the dorsolateral prefrontal cortex modulates specific domains of self-regulation.

    Science.gov (United States)

    Pripfl, Jürgen; Lamm, Claus

    2015-02-01

    Recent neuroscience theories suggest that different kinds of self-regulation may share a common psychobiological mechanism. However, empirical evidence for a domain general self-regulation mechanism is scarce. The aim of this study was to investigate whether focused anodal transcranial direct current stimulation (tDCS), facilitating the activity of the dorsolateral prefrontal cortex (dlPFC), acts on a domain general self-regulation mechanism and thus modulates both affective and appetitive self-regulation. Twenty smokers participated in this within-subject sham controlled study. Effects of anodal left, anodal right and sham tDCS over the dlPFC on affective picture appraisal and nicotine craving-cue appraisal were assessed. Anodal right tDCS over the dlPFC reduced negative affect in emotion appraisal, but neither modulated regulation of positive emotion appraisal nor of craving appraisal. Anodal left stimulation did not induce any significant effects. The results of our study show that domain specific self-regulation networks are at work in the prefrontal cortex. Focused tDCS modulation of this specific self-regulation network could probably be used during the first phase of nicotine abstinence, during which negative affect might easily result in relapse. These findings have implications for neuroscience models of self-regulation and are of relevance for the development of brain stimulation based treatment methods for neuropsychiatric disorders associated with self-regulation deficits. Copyright © 2014 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

  2. 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.

  3. 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.

  4. Industrial cooling for mechanical engineering - requirements and current developments; Maschinenkuehlungen Anforderungen und aktuelle Tendenzen

    Energy Technology Data Exchange (ETDEWEB)

    Schmidt, K.P. [Kulmbacher Klimageraete-Werke GmbH, Geschaeftsbereich Riedel Kaeltetechnik (Germany)

    2002-06-01

    A special field of refrigeration is the cooling of technological processes. Common refrigeration technology mostly operates at lower evaporation temperatures and with shorter operating times, however the cooling of tool machines often demands higher temperatures and long-term operating times. This requires a special design and calculation of the components. (orig.) [German] Ein besonderes Einsatzgebiet der Kaeltetechnik stellt die Kuehlung technologischer Prozesse dar. Waehrend in der ueblichen Kaeltetechnik niedrige Verdampfungstemperaturen und relativ geringe Nutzungszeiten dominieren, sind bei der Waermeabfuhr von Maschinen haeufig hoehere Temperaturen und lange Betriebszeiten zu realisieren. Dies erfordert eine spezielle Auslegung der Komponenten. (orig.)

  5. Inhibitors of arachidonate-regulated calcium channel signaling suppress triggered activity induced by the late sodium current.

    Science.gov (United States)

    Wolkowicz, Paul; Umeda, Patrick K; Sharifov, Oleg F; White, C Roger; Huang, Jian; Mahtani, Harry; Urthaler, Ferdinand

    2014-02-05

    Disturbances in myocyte calcium homeostasis are hypothesized to be one cause for cardiac arrhythmia. The full development of this hypothesis requires (i) the identification of all sources of arrhythmogenic calcium and (ii) an understanding of the mechanism(s) through which calcium initiates arrhythmia. To these ends we superfused rat left atria with the late sodium current activator type II Anemonia sulcata toxin (ATXII). This toxin prolonged atrial action potentials, induced early afterdepolarization, and provoked triggered activity. The calmodulin-dependent protein kinase II (CaMKII) inhibitor KN-93 (N-[2-[[[3-(4-chlorophenyl)-2-propenyl]methylamino]methyl]phenyl]-N-(2-hydroxyethyl)-4-methoxybenzenesulphon-amide) suppressed ATXII triggered activity but its inactive congener KN-92 (2-[N-(4-methoxy benzenesulfonyl)]amino-N-(4-chlorocinnamyl)-N-methylbenzylamine) did not. Neither drug affected normal atrial contractility. Calcium entry via L-type channels or calcium leakage from sarcoplasmic reticulum stores are not critical for this type of ectopy as neither verapamil ((RS)-2-(3,4-dimethoxyphenyl)-5-{[2-(3,4-dimethoxyphenyl)ethyl]-(methyl)amino}-2-prop-2-ylpentanenitrile) nor ryanodine affected ATXII triggered activity. By contrast, inhibitors of the voltage independent arachidonate-regulated calcium (ARC) channel and the store-operated calcium channel specifically suppressed ATXII triggered activity without normalizing action potentials or affecting atrial contractility. Inhibitors of cytosolic calcium-dependent phospholipase A2 also suppressed triggered activity suggesting that this lipase, which generates free arachidonate, plays a key role in ATXII ectopy. Thus, increased left atrial late sodium current appears to activate atrial Orai-linked ARC and store operated calcium channels, and these voltage-independent channels may be unexpected sources for the arrhythmogenic calcium that underlies triggered activity.

  6. 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.

  7. 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…

  8. THE DUAL CONTROL – A REQUIREMENT OF THE CURRENT BANK MANAGEMENT

    Directory of Open Access Journals (Sweden)

    ALINA MATIȘ

    2014-10-01

    Full Text Available Security is a major problem of the banking industry, which bases its whole system on the reputation the bank manages to establish and then maintain among its customers. With the manifestation of information security issues and in interdependence with them, the need for internal control also developed, both of them being associated with modern risk management practices. As the concept of internal control has been rising as a major concern of management and corporate governance, there are attempts to classify it and distinguish it in different categories, dual control being a result of this process. The authors of this study attempt to present the concept of dual control as it is currently applied in the banking system, trying to analyze contextually its capacities of fraud detection, its consequences in terms of risk and costs as well as its effects on bank performance.

  9. Space Weather - Current Capabilities, Future Requirements, and the Path to Improved Forecasting

    Science.gov (United States)

    Mann, Ian

    2016-07-01

    We present an overview of Space Weather activities and future opportunities including assessments of current status and capabilities, knowledge gaps, and future directions in relation to both observations and modeling. The review includes input from the scientific community including from SCOSTEP scientific discipline representatives (SDRs), COSPAR Main Scientific Organizers (MSOs), and SCOSTEP/VarSITI leaders. The presentation also draws on results from the recent activities related to the production of the COSPAR-ILWS Space Weather Roadmap "Understanding Space Weather to Shield Society" [Schrijver et al., Advances in Space Research 55, 2745 (2015) http://dx.doi.org/10.1016/j.asr.2015.03.023], from the activities related to the United Nations (UN) Committee on the Peaceful Uses of Outer Space (COPUOS) actions in relation to the Long-term Sustainability of Outer Space (LTS), and most recently from the newly formed and ongoing efforts of the UN COPUOS Expert Group on Space Weather.

  10. Chloride current in mammalian cardiac myocytes. Novel mechanism for autonomic regulation of action potential duration and resting membrane potential

    OpenAIRE

    1990-01-01

    The properties of the autonomically regulated chloride current (ICl) were studied in isolated guinea pig ventricular myocytes. This current was elicited upon exposure to isoproterenol (ISO) and reversed upon concurrent exposure to acetylcholine (ACh). ICl was time independent and exhibited outward rectification. The responses to ISO and ACh could be blocked by propranolol and atropine, respectively, and ICl was also elicited by forskolin, 8-bromoadenosine 3',5'-cyclic monophosphate, and 3-iso...

  11. 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.

  12. The neuroendocrine regulation of food intake in fish: a review of current knowledge

    Directory of Open Access Journals (Sweden)

    Helene Volkoff

    2016-11-01

    Full Text Available Fish are the most diversified group of vertebrates and, although progress has been made in the past years, only relatively few fish species have been examined to date, with regards to the endocrine regulation of feeding in fish. In fish, as in mammals, feeding behavior is ultimately regulated by central effectors within feeding centers of the brain, which receive and process information from endocrine signals from both brain and peripheral tissues. Although basic endocrine mechanisms regulating feeding appear to be conserved among vertebrates, major physiological differences between fish and mammals and the diversity of fish, in particular in regard to feeding habits, digestive tract anatomy and physiology, suggest the existence of fish- and species-specific regulating mechanisms. This review provides an overview of hormones known to regulate food intake in fish, emphasizing on major hormones and the main fish groups studied to date.

  13. The Neuroendocrine Regulation of Food Intake in Fish: A Review of Current Knowledge.

    Science.gov (United States)

    Volkoff, Helene

    2016-01-01

    Fish are the most diversified group of vertebrates and, although progress has been made in the past years, only relatively few fish species have been examined to date, with regards to the endocrine regulation of feeding in fish. In fish, as in mammals, feeding behavior is ultimately regulated by central effectors within feeding centers of the brain, which receive and process information from endocrine signals from both brain and peripheral tissues. Although basic endocrine mechanisms regulating feeding appear to be conserved among vertebrates, major physiological differences between fish and mammals and the diversity of fish, in particular in regard to feeding habits, digestive tract anatomy and physiology, suggest the existence of fish- and species-specific regulating mechanisms. This review provides an overview of hormones known to regulate food intake in fish, emphasizing on major hormones and the main fish groups studied to date.

  14. Balancing functional and nutritional quality of oils and fats: Current requirements and future trends

    Directory of Open Access Journals (Sweden)

    Van den Bremt Karen

    2012-03-01

    Full Text Available Oils and fats play an important role in the structure, aroma and stability of a wide variety of food products, as well as in their nutritional properties. For Puratos, a producer of ingredients for bakery, patisserie and chocolate sector, functionality and taste are of utmost importance, but the company also wants to contribute to the balanced diet of consumers. Vegetable oils and fats are used in margarines and releasing agents, vegetable creams, compound chocolate, fillings and emulsifiers. Each application requires an oil or fat with specific physicochemical properties in order to ensure the optimal structure, stability and taste of the end product. Traditionally, (partially hydrogenated vegetable oils deliver important functional characteristics concerning crystallization behaviour, directly linked with the workability, melting properties, stability and mouth feel of the food product. However, due to negative nutritional implications, trans fats are to be replaced by healthier alternatives, preferably not by saturated fats. Consumers – and in some regions, legal instances – demand transfree or hydro-free products while not compromising on taste. Alternative fats and oils will be discussed concerning their functional and nutritional properties.

  15. 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

  16. AKAP150 participates in calcineurin/NFAT activation during the down-regulation of voltage-gated K(+) currents in ventricular myocytes following myocardial infarction.

    Science.gov (United States)

    Nieves-Cintrón, Madeline; Hirenallur-Shanthappa, Dinesh; Nygren, Patrick J; Hinke, Simon A; Dell'Acqua, Mark L; Langeberg, Lorene K; Navedo, Manuel; Santana, Luis F; Scott, John D

    2016-07-01

    The Ca(2+)-responsive phosphatase calcineurin/protein phosphatase 2B dephosphorylates the transcription factor NFATc3. In the myocardium activation of NFATc3 down-regulates the expression of voltage-gated K(+) (Kv) channels after myocardial infarction (MI). This prolongs action potential duration and increases the probability of arrhythmias. Although recent studies infer that calcineurin is activated by local and transient Ca(2+) signals the molecular mechanism that underlies the process is unclear in ventricular myocytes. Here we test the hypothesis that sequestering of calcineurin to the sarcolemma of ventricular myocytes by the anchoring protein AKAP150 is required for acute activation of NFATc3 and the concomitant down-regulation of Kv channels following MI. Biochemical and cell based measurements resolve that approximately 0.2% of the total calcineurin activity in cardiomyocytes is associated with AKAP150. Electrophysiological analyses establish that formation of this AKAP150-calcineurin signaling dyad is essential for the activation of the phosphatase and the subsequent down-regulation of Kv channel currents following MI. Thus AKAP150-mediated targeting of calcineurin to sarcolemmal micro-domains in ventricular myocytes contributes to the local and acute gene remodeling events that lead to the down-regulation of Kv currents.

  17. Investigating Relationships among Pre-Service Science Teachers' Conceptual Knowledge of Electric Current, Motivational Beliefs and Self-Regulation

    Science.gov (United States)

    Inaltun, Hüseyin; Ates, Salih

    2015-01-01

    The purpose of this study is to examine relationships among pre-service science teachers' conceptual knowledge of electric current, motivational beliefs, and self-regulation. One hundred and twenty-seven students (female = 107, male = 20) enrolled in the science education program of a public university in Ankara participated the study. A concept…

  18. Investigating Relationships among Pre-Service Science Teachers' Conceptual Knowledge of Electric Current, Motivational Beliefs and Self-Regulation

    Science.gov (United States)

    Inaltun, Hüseyin; Ates, Salih

    2015-01-01

    The purpose of this study is to examine relationships among pre-service science teachers' conceptual knowledge of electric current, motivational beliefs, and self-regulation. One hundred and twenty-seven students (female = 107, male = 20) enrolled in the science education program of a public university in Ankara participated the study. A concept…

  19. Regulation of voltage-gated sodium current by endogenous Src family kinases in cochlear spiral ganglion neurons in culture.

    Science.gov (United States)

    Feng, Shuang; Pflueger, Melissa; Lin, Shuang-Xiu; Groveman, Bradley R; Su, Jiping; Yu, Xian-Min

    2012-04-01

    Voltage-gated sodium (Na+) and potassium (K+)channels have been found to be regulated by Src family kinases(SFKs).However, how these channels are regulated by SFKs in cochlear spiral ganglion neurons (SGNs) remains unknown.Here, we report that altering the activity of endogenous SFKs modulated voltage-gated Na+, but not K+, currents recorded in embryonic SGNs in culture. Voltage-gated Na+ current was suppressed by inhibition of endogenous SFKs or just Src and potentiated by the activation of these enzymes. Detailed investigations showed that under basal conditions, SFK inhibitor application did not significantly affect the voltage-dependent activation, but shifted the steady-state inactivation curves of Na+ currents and delayed the recovery of Na+ currents from inactivation. Application of Src specific inhibitor, Src40–58,not only shifted the inactivation curve but also delayed the recovery of Na+ currents and moved the voltage-dependent activation curve towards the left. The pre-inhibition of SFKs occluded all the effects induced by Src40–58 application, except the left shift of the activation curve. The activation of SFKs did not change either steady-state inactivation or recovery of Na+ currents, but caused the left shift of the activation curve.SFK inhibitor application effectively prevented all the effects induced by SFK activation, suggesting that both the voltage-dependent activation and steady-state inactivation of Na+ current are subjects of SFK regulation. The different effects induced by activation versus inhibition of SFKs implied that under basal conditions, endogenously active and inactive SFKs might be differentially involved in the regulation of voltage-gated Na+ channels in SGNs.

  20. 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.

  1. 21 CFR 210.2 - Applicability of current good manufacturing practice regulations.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 4 2010-04-01 2010-04-01 false Applicability of current good manufacturing... AND HUMAN SERVICES (CONTINUED) DRUGS: GENERAL CURRENT GOOD MANUFACTURING PRACTICE IN MANUFACTURING, PROCESSING, PACKING, OR HOLDING OF DRUGS; GENERAL § 210.2 Applicability of current good...

  2. 21 CFR 210.1 - Status of current good manufacturing practice regulations.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 4 2010-04-01 2010-04-01 false Status of current good manufacturing practice... SERVICES (CONTINUED) DRUGS: GENERAL CURRENT GOOD MANUFACTURING PRACTICE IN MANUFACTURING, PROCESSING, PACKING, OR HOLDING OF DRUGS; GENERAL § 210.1 Status of current good manufacturing practice...

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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...

  9. The interaction of approach-alcohol action tendencies, working memory capacity, and current task goals predicts the inability to regulate drinking behavior.

    Science.gov (United States)

    Sharbanee, Jason M; Stritzke, Werner G K; Wiers, Reinout W; Young, Paul; Rinck, Mike; MacLeod, Colin

    2013-09-01

    The inability to regulate alcohol consumption has been attributed to an imbalance between stimulus-driven behavioral biases, or action tendencies, and the ability to exert goal-directed control, or working memory capacity (WMC). Previous research assessing the interaction between these variables has not considered the effect of whether individuals' current goals or task demands require goal-directed control. Our aim was to examine the potential interaction of appetitive action tendencies and the ability to exert control over these action tendencies as a function of whether task demands require applying control for successful task completion. Two groups of social drinkers (n = 40 per group) who differed in their ability to regulate their alcohol consumption completed a novel variant of the Approach-Avoidance Task (AAT), which separately assessed approach and avoid trials. The approach and avoidance responses differentially require goal-directed control, depending on whether the task-relevant response is incongruent with the stimulus-driven action tendency. Results indicated that (a) group differences in AAT indices were only observed on trials that required an avoidance movement, which are trials where the task-relevant response would be incongruent with an approach action tendency, and (b) the extent of the group differences for these avoidance trials was moderated by individual differences in WMC, such that problem drinkers with lower WMC showed greater behavioral bias toward alcohol than those with higher WMC. These findings suggest that difficulties in regulating alcohol consumption arise from a complex interaction of action-tendencies, WMC, and current goals or task demands.

  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. Selective regulation of current densities underlies spontaneous changes in the activity of cultured neurons.

    Science.gov (United States)

    Turrigiano, G; LeMasson, G; Marder, E

    1995-05-01

    We study the electrical activity patterns and the expression of conductances in adult stomatogastric ganglion (STG) neurons as a function of time in primary cell culture. When first plated in culture, these neurons had few active properties. After 1 d in culture they produced small action potentials that rapidly inactivated during maintained depolarization. After 2 d in culture they fired large action potentials tonically when depolarized, and their properties resembled very closely the properties of STG neurons pharmacologically isolated in the ganglion. After 3-4 d in culture, however, their electrical properties changed and they fired in bursts when depolarized. We characterized the currents expressed by these neurons in culture. They included two TTX-sensitive sodium currents, a calcium current, a delayed-rectifier-like current, a calcium-dependent potassium current, and two A-type currents. The changes in firing properties with time in culture were accompanied by an increase in inward and decrease in outward current densities. A single-compartment conductance-based model of an STG neuron was constructed by fitting the currents measured in the biological neurons. When the current densities in the model neuron were matched to those measured for the biological neurons in each activity state, the model neuron closely reproduced each state, indicating that the changes in current densities are sufficient to account for the changes in intrinsic properties. These data indicate that STG neurons isolated in culture change their intrinsic electrical properties by selectively adjusting the magnitudes of their ionic conductances.

  12. Methods for growth regulation of greenhouse produced ornamental pot- and bedding plants – a current review

    Directory of Open Access Journals (Sweden)

    Bergstrand Karl-Johan I.

    2017-06-01

    Full Text Available Chemical plant growth regulators (PGRs are used in the production of ornamental potted and bedding plants. Growth control is needed for maximizing production per unit area, reducing transportation costs and to obtain a desired visual quality. However, the use of PGRs is associated with toxicity risks to humans and the environment. In many countries the availability of PGRs is restricted as few substances are registered for use. A number of alternative methods have been suggested. The methods include genetic methods (breeding and crop cultivation practices such as fertigation, temperature and light management. A lot of research into “alternative” growth regulation was performed during the 1980-1990s, revealing several possible ways of using different climatic factors to optimize plant growth with respect to plant height. In recent years, the interest in climatic growth regulation has been resurrected, not least due to the coming phase-out of the plant growth regulator chlormequat chloride (CCC. Today, authorities in many countries are aiming towards reducing the use of agrochemicals. At the same time, there is a strong demand from consumers for products produced without chemicals. This article provides a broad overview of available methods for non-chemical growth control. It is concluded that a combination of plant breeding and management of temperature, fertigation and light management has the potential of replacing chemical growth regulators in the commercial production of ornamental pot- and bedding plants.

  13. Marketing of research institutes in view of current organizational and legal regulations

    Directory of Open Access Journals (Sweden)

    Anna Slotorsz

    2013-03-01

    Full Text Available The paper aims to describe in a synthetic way some issues related to the marketing activity of R&D organizations in view of legal acts regulating the functioning of the state owned R&D organizations. The authors have analyzed some regulations related to research institutes and their activities, regulations specifying criteria and procedures of granting of a scientific category to a R&D unit as well as principals of financing of the science and regulations related to the operations of certifying units. Basing on this analysis the authors have elaborated a set of marketing goals of the R&D organizations, a list of their stakeholders and of specific marketing tools both accessible in view of the rules and regulations in force and effective from the point of view of parametric evaluation of the R&D organizations. Among the analyzed tools there are the following: issues related to publications, organization of conferences, seminars and popularizing events, organization of education and training courses, internal policies related to intellectual property protection, internal policies related to technology transfer, membership in international organizations and applied quality policies.

  14. Process stabilization by peak current regulation in reactive high-power impulse magnetron sputtering of hafnium nitride

    Science.gov (United States)

    Shimizu, T.; Villamayor, M.; Lundin, D.; Helmersson, U.

    2016-02-01

    A simple and cost effective approach to stabilize the sputtering process in the transition zone during reactive high-power impulse magnetron sputtering (HiPIMS) is proposed. The method is based on real-time monitoring and control of the discharge current waveforms. To stabilize the process conditions at a given set point, a feedback control system was implemented that automatically regulates the pulse frequency, and thereby the average sputtering power, to maintain a constant maximum discharge current. In the present study, the variation of the pulse current waveforms over a wide range of reactive gas flows and pulse frequencies during a reactive HiPIMS process of Hf-N in an Ar-N2 atmosphere illustrates that the discharge current waveform is a an excellent indicator of the process conditions. Activating the reactive HiPIMS peak current regulation, stable process conditions were maintained when varying the N2 flow from 2.1 to 3.5 sccm by an automatic adjustment of the pulse frequency from 600 Hz to 1150 Hz and consequently an increase of the average power from 110 to 270 W. Hf-N films deposited using peak current regulation exhibited a stable stoichiometry, a nearly constant power-normalized deposition rate, and a polycrystalline cubic phase Hf-N with (1 1 1)-preferred orientation over the entire reactive gas flow range investigated. The physical reasons for the change in the current pulse waveform for different process conditions are discussed in some detail.

  15. 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.

  16. The evolution, current status, and regulation of ostomy products in the United States.

    Science.gov (United States)

    Turnbull, G B

    2001-01-01

    In today's rapidly evolving health care environment in which quality, cost-effectiveness, and outcomes are among the most valued health care products, significant issues must be addressed regarding ostomy products and how persons with stomas use them. The lack of evidence-based data has a direct impact on the pocketbooks of many patients with ostomies in the United States and their quality of life. Regulators, accrediting agencies, and payers look to WOC nurses and other providers to provide a sound scientific base upon which they can develop standards and regulations that will improve the life of Americans with stomas.

  17. 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...

  18. A fresh look at electron cyclotron current drive power requirements for stabilization of tearing modes in ITER

    Energy Technology Data Exchange (ETDEWEB)

    La Haye, R. J., E-mail: lahaye@fusion.gat.com [General Atomics, P.O. Box 85608, San Diego, California 92186-5608 (United States)

    2015-12-10

    ITER is an international project to design and build an experimental fusion reactor based on the “tokamak” concept. ITER relies upon localized electron cyclotron current drive (ECCD) at the rational safety factor q=2 to suppress or stabilize the expected poloidal mode m=2, toroidal mode n=1 neoclassical tearing mode (NTM) islands. Such islands if unmitigated degrade energy confinement, lock to the resistive wall (stop rotating), cause loss of “H-mode” and induce disruption. The International Tokamak Physics Activity (ITPA) on MHD, Disruptions and Magnetic Control joint experiment group MDC-8 on Current Drive Prevention/Stabilization of Neoclassical Tearing Modes started in 2005, after which assessments were made for the requirements for ECCD needed in ITER, particularly that of rf power and alignment on q=2 [1]. Narrow well-aligned rf current parallel to and of order of one percent of the total plasma current is needed to replace the “missing” current in the island O-points and heal or preempt (avoid destabilization by applying ECCD on q=2 in absence of the mode) the island [2-4]. This paper updates the advances in ECCD stabilization on NTMs learned in DIII-D experiments and modeling during the last 5 to 10 years as applies to stabilization by localized ECCD of tearing modes in ITER. This includes the ECCD (inside the q=1 radius) stabilization of the NTM “seeding” instability known as sawteeth (m/n=1/1) [5]. Recent measurements in DIII-D show that the ITER-similar current profile is classically unstable, curvature stabilization must not be neglected, and the small island width stabilization effect from helical ion polarization currents is stronger than was previously thought [6]. The consequences of updated assumptions in ITER modeling of the minimum well-aligned ECCD power needed are all-in-all favorable (and well-within the ITER 24 gyrotron capability) when all effects are included. However, a “wild card” may be broadening of the localized

  19. A fresh look at electron cyclotron current drive power requirements for stabilization of tearing modes in ITER

    Science.gov (United States)

    La Haye, R. J.

    2015-12-01

    ITER is an international project to design and build an experimental fusion reactor based on the "tokamak" concept. ITER relies upon localized electron cyclotron current drive (ECCD) at the rational safety factor q=2 to suppress or stabilize the expected poloidal mode m=2, toroidal mode n=1 neoclassical tearing mode (NTM) islands. Such islands if unmitigated degrade energy confinement, lock to the resistive wall (stop rotating), cause loss of "H-mode" and induce disruption. The International Tokamak Physics Activity (ITPA) on MHD, Disruptions and Magnetic Control joint experiment group MDC-8 on Current Drive Prevention/Stabilization of Neoclassical Tearing Modes started in 2005, after which assessments were made for the requirements for ECCD needed in ITER, particularly that of rf power and alignment on q=2 [1]. Narrow well-aligned rf current parallel to and of order of one percent of the total plasma current is needed to replace the "missing" current in the island O-points and heal or preempt (avoid destabilization by applying ECCD on q=2 in absence of the mode) the island [2-4]. This paper updates the advances in ECCD stabilization on NTMs learned in DIII-D experiments and modeling during the last 5 to 10 years as applies to stabilization by localized ECCD of tearing modes in ITER. This includes the ECCD (inside the q=1 radius) stabilization of the NTM "seeding" instability known as sawteeth (m/n=1/1) [5]. Recent measurements in DIII-D show that the ITER-similar current profile is classically unstable, curvature stabilization must not be neglected, and the small island width stabilization effect from helical ion polarization currents is stronger than was previously thought [6]. The consequences of updated assumptions in ITER modeling of the minimum well-aligned ECCD power needed are all-in-all favorable (and well-within the ITER 24 gyrotron capability) when all effects are included. However, a "wild card" may be broadening of the localized ECCD by the presence of

  20. A compilation of current regulations, standards and guidelines in remote afterloading brachytherapy

    Energy Technology Data Exchange (ETDEWEB)

    Tortorelli, J.P.; Simion, G.P.; Kozlowski, S.D. [Idaho National Engineering Lab., Idaho Falls, ID (United States)

    1994-10-01

    Over a dozen government and professional organizations in the United States and Europe have issued regulations and guidance concerning quality management in the practice of remote afterloading brachytherapy. Information from the publications of these organizations was collected and collated for this report. This report provides the brachytherapy licensee access to a broad field of quality management information in a single, topically organized document.

  1. 76 FR 19282 - Direct Investment Surveys: Alignment of Regulations With Current Practices

    Science.gov (United States)

    2011-04-07

    ... eliminated from the regulations are: a survey of foreign direct investment in the U.S. seafood industry (BE... Foreign Person BE-21, Survey of Foreign Direct Investment in U.S. Business Enterprises Engaged in the... established or acquired entity. The data had been used to measure the amount of new foreign direct......

  2. A small potassium current in AgRP/NPY neurons regulates feeding behavior and enery metabolism

    Science.gov (United States)

    Neurons that co-express agouti-related peptide (AgRP) and neuropeptide Y (NPY) are indispensable for normal feeding behavior. Firing activities of AgRP/NPY neurons are dynamically regulated by energy status and coordinate appropriate feeding behavior to meet nutritional demands. However, intrinsic m...

  3. 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...

  4. 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...

  5. Dissociation of the store-operated calcium current ICRAC and the Mg-nucleotide-regulated metal ion current MagNuM

    Science.gov (United States)

    Hermosura, Meredith C; Monteilh-Zoller, Mahealani K; Scharenberg, Andrew M; Penner, Reinhold; Fleig, Andrea

    2002-01-01

    Rat basophilic leukaemia cells (RBL-2H3-M1) were used to study the characteristics of the store-operated Ca2+ release-activated Ca2+ current (ICRAC) and the magnesium-nucleotide-regulated metal cation current (MagNuM) (which is conducted by the LTRPC7 channel). Pipette solutions containing 10 mm BAPTA and no added ATP induced both currents in the same cell, but the time to half-maximal activation for MagNuM was about two to three times slower than that of ICRAC. Differential suppression of ICRAC was achieved by buffering free [Ca2+]i to 90 nm and selective inhibition of MagNuM was accomplished by intracellular solutions containing 6 mm Mg.ATP, 1.2 mm free [Mg2+]i or 100 μm GTP-γ-S, allowing investigations on these currents in relative isolation. Removal of extracellular Ca2+ and Mg2+ caused both currents to be carried significantly by monovalent ions. In the absence or presence of free [Mg2+]i, ICRAC carried by monovalent ions inactivated more rapidly and more completely than MagNuM carried by monovalent ions. Since several studies have used divalent-free solutions on either side of the membrane to study selectivity and single-channel behaviour of ICRAC, these experimental conditions would have favoured the contribution of MagNuM to monovalent conductance and call for caution in interpreting results where both ICRAC and MagNuM are activated. PMID:11882677

  6. Exploring end users' system requirements for a handheld computer supporting both sepsis test workflow and current IT solutions.

    Science.gov (United States)

    Samson, Lasse Lefevre; Pape-Haugaard, Louise Bilenberg; Søgaard, Mette; Schønheyder, Henrik Carl; Hejlesen, Ole K

    2013-01-01

    Sepsis is a systemic response associated with very high mortality. Early initiation of the correct antimicrobial therapy remains a cornerstone in the treatment of sepsis. Currently, a new microbiological test is under development, which aims to detect major, prevalent pathogens in positive blood cultures within an hour. Concurrently, a tablet-based data entry and reporting system will be developed to facilitate the workflow of the test. This study investigated the system requirements for the tablet-based data entry and reporting system in order to support the clinical workflow. By observing the workflow of the blood culture analysis and through interviews with medical laboratory technicians, four main system requirements were identified. The system requirements are; the ability to receive and send data to the laboratory information system, support for the use of barcodes, the ability to access a browser based instruction system, and communication of results between medical laboratory technicians and physicians. These system requirements will be used as a basis in the future development of the tablet-based data entry and reporting system.

  7. Visinin-like neuronal calcium sensor proteins regulate the slow calcium-activated afterhyperpolarizing current in the rat cerebral cortex

    Science.gov (United States)

    Villalobos, Claudio; Andrade, Rodrigo

    2010-01-01

    Many neurons in the nervous systems express afterhyperpolarizations that are mediated by a slow calcium-activated potassium current. This current shapes neuronal firing and is inhibited by neuromodulators, suggesting an important role in the regulation of neuronal function. Surprisingly, very little is currently known about the molecular basis for this current or how it is gated by calcium. Recently, the neuronal calcium sensor protein hippocalcin was identified as a calcium sensor for the slow afterhyperpolarizing current in the hippocampus. However, while hippocalcin is very strongly expressed in the hippocampus, this protein shows a relatively restricted distribution in the brain. Furthermore, the genetic deletion of this protein only partly reduces the slow hyperpolarizing current in hippocampus. These considerations question whether hippocalcin can be the sole calcium sensor for the slow afterhyperpolarizing current. Here we use loss of function and overexpression strategies to show that hippocalcin functions as a calcium sensor for the slow afterhyperpolarizing current in the cerebral cortex, an area where hippocalcin is expressed at much lower levels than in hippocampus. In addition we show that neurocalcin δ, but not VILIP-2, can also act as a calcium sensor for the slow afterhyperpolarizing current. Finally we show that hippocalcin and neurocalcin δ both increase the calcium sensitivity of the afterhyperpolarizing current but do not alter its sensitivity to inhibition by carbachol acting through the Gαq-11-PLCβ signaling cascade. These results point to a general role for a subgroup of visinin-like neuronal calcium sensor proteins in the activation of the slow calcium-activated afterhyperpolarizing current. PMID:20980592

  8. 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...

  9. A Low Input Current and Wide Conversion Ratio Buck Regulator with 75% Efficiency for High-Voltage Triboelectric Nanogenerators

    Science.gov (United States)

    Luo, Li-Chuan; Bao, De-Chun; Yu, Wu-Qi; Zhang, Zhao-Hua; Ren, Tian-Ling

    2016-01-01

    It is meaningful to research the Triboelectric Nanogenerators (TENG), which can create electricity anywhere and anytime. There are many researches on the structures and materials of TENG to explain the phenomenon that the maximum voltage is stable and the current is increasing. The output voltage of the TENG is high about 180-400 V, and the output current is small about 39 μA, which the electronic devices directly integration of TENG with Li-ion batteries will result in huge energy loss due to the ultrahigh TENG impedance. A novel interface circuit with the high-voltage buck regulator for TENG is introduced firstly in this paper. The interface circuit can transfer the output signal of the TENG into the signal fit to a lithium ion battery. Through the circuit of the buck regulator, the average output voltage is about 4.0 V and the average output current is about 1.12 mA. Further, the reliability and availability for the lithium ion battery and the circuit are discussed. The interface circuit is simulated using the Cadence software and verified through PCB experiment. The buck regulator can achieve 75% efficiency for the High-Voltage TENG. This will lead to a research hot and industrialization applications.

  10. Inward Rectifier K+ Currents Are Regulated by CaMKII in Endothelial Cells of Primarily Cultured Bovine Pulmonary Arteries.

    Directory of Open Access Journals (Sweden)

    Lihui Qu

    Full Text Available Endothelium lines the interior surface of vascular walls and regulates vascular tones. The endothelial cells sense and respond to chemical and mechanical stimuli in the circulation, and couple the stimulus signals to vascular smooth muscles, in which inward rectifier K+ currents (Kir play an important role. Here we applied several complementary strategies to determine the Kir subunit in primarily cultured pulmonary arterial endothelial cells (PAECs that was regulated by the Ca2+/calmodulin (CaM-dependent protein kinase II (CaMKII. In whole-cell voltage clamp, the Kir currents were sensitive to micromolar concentrations of extracellular Ba2+. In excised inside-out patches, an inward rectifier K+ current was observed with single-channel conductance 32.43 ± 0.45 pS and Popen 0.27 ± 0.04, which were consistent with known unitary conductance of Kir 2.1. RT-PCR and western blot results showed that expression of Kir 2.1 was significantly stronger than that of other subtypes in PAECs. Pharmacological analysis of the Kir currents demonstrated that insensitivity to intracellular ATP, pinacidil, glibenclamide, pH, GDP-β-S and choleratoxin suggested that currents weren't determined by KATP, Kir2.3, Kir2.4 and Kir3.x. The currents were strongly suppressed by exposure to CaMKII inhibitor W-7 and KN-62. The expression of Kir2.1 was inhibited by knocking down CaMKII. Consistently, vasodilation was suppressed by Ba2+, W-7 and KN-62 in isolated and perfused pulmonary arterial rings. These results suggest that the PAECs express an inward rectifier K+ current that is carried dominantly by Kir2.1, and this K+ channel appears to be targeted by CaMKII-dependent intracellular signaling systems.

  11. Drought Predictability and Prediction in a Changing Climate: Assessing Current Predictive Knowledge and Capabilities, User Requirements and Research Priorities

    Science.gov (United States)

    Schubert, Siegfried

    2011-01-01

    Drought is fundamentally the result of an extended period of reduced precipitation lasting anywhere from a few weeks to decades and even longer. As such, addressing drought predictability and prediction in a changing climate requires foremost that we make progress on the ability to predict precipitation anomalies on subseasonal and longer time scales. From the perspective of the users of drought forecasts and information, drought is however most directly viewed through its impacts (e.g., on soil moisture, streamflow, crop yields). As such, the question of the predictability of drought must extend to those quantities as well. In order to make progress on these issues, the WCRP drought information group (DIG), with the support of WCRP, the Catalan Institute of Climate Sciences, the La Caixa Foundation, the National Aeronautics and Space Administration, the National Oceanic and Atmospheric Administration, and the National Science Foundation, has organized a workshop to focus on: 1. User requirements for drought prediction information on sub-seasonal to centennial time scales 2. Current understanding of the mechanisms and predictability of drought on sub-seasonal to centennial time scales 3. Current drought prediction/projection capabilities on sub-seasonal to centennial time scales 4. Advancing regional drought prediction capabilities for variables and scales most relevant to user needs on sub-seasonal to centennial time scales. This introductory talk provides an overview of these goals, and outlines the occurrence and mechanisms of drought world-wide.

  12. Shared Solar: Current Landscape, Market Potential, and the Impact of Federal Securities Regulation; NREL (National Renewable Energy Laboratory)

    Energy Technology Data Exchange (ETDEWEB)

    None

    2015-05-27

    This presentation provides a high-level overview of the current U.S. shared solar landscape, the impact that a given shared solar program's structure has on requiring federal securities oversight, as well as an estimate of market potential for U.S. shared solar deployment.

  13. 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...

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  18. 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....

  19. 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.

  20. Implementation issues on the design of current loops based on resonant regulators for isolated microgrids

    DEFF Research Database (Denmark)

    Federico, de Bosio; Pastorelli, Michele; Antonio DeSouza Ribeiro, Luiz;

    2016-01-01

    This paper analyzes the influence of state feedback coupling between the capacitor voltage and inductor current in voltage source inverters (VSI) operating in stand-alone microgrids. A decoupling technique is proposed as an effective measure to enhance the dynamics. Further implementation issues ...

  1. Regulation of calcium currents and secretion by magnesium in crustacean peptidergic neurons.

    Science.gov (United States)

    Richmond, J E; Sher, E; Keller, R; Haylett, B; Reichwein, B; Cooke, I M

    1995-12-01

    The effect of varying the external Mg2+ concentration on Ca2+ currents through voltage-operated Ca2+ channels has been examined with the patch-clamp technique in acutely isolated neuronal somata from the X-organ-sinus gland (XOSG) of the crab, Cardisoma carnifex. Neurons from this neurosecretory system were selected for morphology associated with crustacean hyperglycemic hormone (CHH) content. In parallel, the effects of Mg2+ concentration on K(+)-evoked secretion of CHH from isolated, intact XOSGs have been assayed by ELISA. At physiological Ca2+ levels the high-voltage-activated Ca2+ currents were attenuated with increasing Mg2+ concentration, with 50% inhibition at approximately 75 mM. Mg2+ block was voltage-dependent, relief from block occurring with increasing depolarization. Thus, in 24 mM Mg2+ inhibition of the Ca2+ current was approximately 55% at -10 mV. Secretion of CHH varied almost linearly with the log of Mg2+ concentration; in 2.4 mM Mg2+ it was double that in 24 mM Mg2+ and almost completely inhibited in 100 mM. Thus, Mg2+ produces a parallel inhibition of Ca2+ currents and CHH secretion and may play a role as a physiological modulator of neuronal activity and secretion in the XOSG of these crabs.

  2. Soft regulations in pharmaceutical policy making : an overview of current approaches and their consequences

    NARCIS (Netherlands)

    Wettermark, B.; Godman, B.; Jacobsson, B.; Haaijer-Ruskamp, F.M.

    2009-01-01

    It is a challenge to improve public health within limited resources. Pharmaceutical policy making is a greater challenge due to conflicting interests between key stakeholder groups. This paper reviews current and future strategies to help improve the quality and efficiency of care, with special emph

  3. GABAA receptor β3 subunit expression regulates tonic current in developing striatopallidal medium spiny neurons

    Directory of Open Access Journals (Sweden)

    Megan eJanssen

    2011-07-01

    Full Text Available The striatum is a key structure for movement control, but the mechanisms that dictate the output of distinct subpopulations of medium spiny projection neurons (MSNs, striatonigral projecting and dopamine D1 receptor- (D1+ or striatopallidal projecting and dopamine D2 receptor- (D2+ expressing neurons, remains poorly understood. GABA-mediated tonic inhibition largely controls neuronal excitability and action potential firing rates, and we previously suggested with pharmacological analysis that the GABAA receptor β3 subunit plays a large role in the basal tonic current seen in D2+ MSNs from young mice (Ade et al, 2008; Janssen et al, 2009. In this study, we demonstrated the essential role of the β3 GABAA receptor subunit in mediating MSN tonic currents using conditional β3 subunit knock-out (β3f/fDrd2 mice. Cre-lox genetics were used to generate conditional knock-out animals where Cre recombinase was expressed under the D2 receptor (Drd2 promoter. We show that while the wild-type MSN tonic current pattern demonstrates a high degree of variability, tonic current patterns from β3f/fDrd2 mice are narrow, suggesting that the β3 subunit is essential to striatal MSN GABA-mediated tonic current. Our data also suggest that a distinct population of synaptic receptors upregulate due to β3 subunit removal. Further, deletion of this subunit significantly decreases the D2+ MSN excitability. These results offer insight for target mechanisms in Parkinson’s disease, where symptoms arise due to the imbalance in striatal D1+ and D2+ MSN excitability and output.

  4. CURRENT WAGE AND LABOR REGULATION PROBLEMS IN RUSSIAN RESTRAURANTS END HOTELS

    Directory of Open Access Journals (Sweden)

    E. Sh. Kachalova

    2012-01-01

    Full Text Available At present, considerable disproportions take place in Russia between organizational, regulatory, social and economic relations actually existing in restaurant and hotel business enterprises on one part and provisions of their legal law basic norms and regulations on the other part. Labor organization and normative practice improvement directions are proposed for enterprises of the branch. Economic essence of the “tipping” phenomenon is revealed. Measures aimed at elimination of overcoming wage rise rates in comparison with those of the labor productivity presently existing in the branch.

  5. Influence of electroosmotic flow on the ionic current rectification in a pH-regulated, conical nanopore.

    Science.gov (United States)

    Lin, Dong-Huei; Lin, Chih-Yuan; Tseng, Shiojenn; Hsu, Jyh-Ping

    2015-09-01

    The ionic current rectification (ICR) is studied theoretically by considering a pH-regulated, conical nanopore. In particular, the effect of electroosmotic flow (EOF), which was often neglected in previous studies, is investigated by solving a set of coupled Poisson, Nernst-Planck, and Navier-Stokes equations. The behaviors of ICR under various conditions are examined by varying solution pH, bulk ionic concentration, and applied electric potential bias. We show that the EOF effect is significant when the bulk ionic concentration is medium high, the pH is far away from the iso-electric point, and the electric potential bias is high. The percentage deviation in the current rectification ratio arising from neglecting the EOF effect can be on the order of 100%. In addition, the behavior of the current rectification ratio at a high pH taking account of EOF is different both qualitatively and quantitatively from that without taking account of EOF.

  6. 76 FR 39260 - Direct Investment Surveys: Alignment of Regulations With Current Practices

    Science.gov (United States)

    2011-07-06

    ... is eliminating the reporting requirements for two surveys of new foreign direct investment in the... several direct investment surveys. The surveys are: BE-13, Initial Report on a Foreign Person's Direct or... to measure the amount of new foreign direct investment in the United States and assess its impact...

  7. Current concepts of IgE regulation and impact of genetic determinants.

    Science.gov (United States)

    Potaczek, D P; Kabesch, M

    2012-06-01

    Immunoglobulin E (IgE) mediated immune responses seem to be directed against parasites and neoplasms, but are best known for their involvement in allergies. The IgE network is tightly controlled at different levels as outlined in this review. Genetic determinants were suspected to influence IgE regulation and IgE levels considerably for many years. Linkage and candidate gene studies suggested a number of loci and genes to correlate with total serum IgE levels, and recently genome-wide association studies (GWAS) provided the power to identify genetic determinants for total serum IgE levels: 1q23 (FCER1A), 5q31 (RAD50, IL13, IL4), 12q13 (STAT6), 6p21.3 (HLA-DRB1) and 16p12 (IL4R, IL21R). In this review, we analyse the potential role of these GWAS hits in the IgE network and suggest mechanisms of how genes and genetic variants in these loci may influence IgE regulation.

  8. Current view on regulation of voltage-gated sodium channels by calcium and auxiliary proteins.

    Science.gov (United States)

    Pitt, Geoffrey S; Lee, Seok-Yong

    2016-09-01

    In cardiac and skeletal myocytes, and in most neurons, the opening of voltage-gated Na(+) channels (NaV channels) triggers action potentials, a process that is regulated via the interactions of the channels' intercellular C-termini with auxiliary proteins and/or Ca(2+) . The molecular and structural details for how Ca(2+) and/or auxiliary proteins modulate NaV channel function, however, have eluded a concise mechanistic explanation and details have been shrouded for the last decade behind controversy about whether Ca(2+) acts directly upon the NaV channel or through interacting proteins, such as the Ca(2+) binding protein calmodulin (CaM). Here, we review recent advances in defining the structure of NaV intracellular C-termini and associated proteins such as CaM or fibroblast growth factor homologous factors (FHFs) to reveal new insights into how Ca(2+) affects NaV function, and how altered Ca(2+) -dependent or FHF-mediated regulation of NaV channels is perturbed in various disease states through mutations that disrupt CaM or FHF interaction.

  9. Contribution of presynaptic calcium-activated potassium currents to transmitter release regulation in cultured Xenopus nerve-muscle synapses.

    Science.gov (United States)

    Pattillo, J M; Yazejian, B; DiGregorio, D A; Vergara, J L; Grinnell, A D; Meriney, S D

    2001-01-01

    Using Xenopus nerve-muscle co-cultures, we have examined the contribution of calcium-activated potassium (K(Ca)) channels to the regulation of transmitter release evoked by single action potentials. The presynaptic varicosities that form on muscle cells in these cultures were studied directly using patch-clamp recording techniques. In these developing synapses, blockade of K(Ca) channels with iberiotoxin or charybdotoxin decreased transmitter release by an average of 35%. This effect would be expected to be caused by changes in the late phases of action potential repolarization. We hypothesize that these changes are due to a reduction in the driving force for calcium that is normally enhanced by the local hyperpolarization at the active zone caused by potassium current through the K(Ca) channels that co-localize with calcium channels. In support of this hypothesis, we have shown that when action potential waveforms were used as voltage-clamp commands to elicit calcium current in varicosities, peak calcium current was reduced only when these waveforms were broadened beginning when action potential repolarization was 20% complete. In contrast to peak calcium current, total calcium influx was consistently increased following action potential broadening. A model, based on previously reported properties of ion channels, faithfully reproduced predicted effects on action potential repolarization and calcium currents. From these data, we suggest that the large-conductance K(Ca) channels expressed at presynaptic varicosities regulate transmitter release magnitude during single action potentials by altering the rate of action potential repolarization, and thus the magnitude of peak calcium current.

  10. β-Secretase BACE1 regulates hippocampal and reconstituted M-currents in a β-subunit-like fashion.

    Science.gov (United States)

    Hessler, Sabine; Zheng, Fang; Hartmann, Stephanie; Rittger, Andrea; Lehnert, Sandra; Völkel, Meike; Nissen, Matthias; Edelmann, Elke; Saftig, Paul; Schwake, Michael; Huth, Tobias; Alzheimer, Christian

    2015-02-25

    The β-secretase BACE1 is widely known for its pivotal role in the amyloidogenic pathway leading to Alzheimer's disease, but how its action on transmembrane proteins other than the amyloid precursor protein affects the nervous system is only beginning to be understood. We report here that BACE1 regulates neuronal excitability through an unorthodox, nonenzymatic interaction with members of the KCNQ (Kv7) family that give rise to the M-current, a noninactivating potassium current with slow kinetics. In hippocampal neurons from BACE1(-/-) mice, loss of M-current enhanced neuronal excitability. We relate the diminished M-current to the previously reported epileptic phenotype of BACE1-deficient mice. In HEK293T cells, BACE1 amplified reconstituted M-currents, altered their voltage dependence, accelerated activation, and slowed deactivation. Biochemical evidence strongly suggested that BACE1 physically associates with channel proteins in a β-subunit-like fashion. Our results establish BACE1 as a physiologically essential constituent of regular M-current function and elucidate a striking new feature of how BACE1 impacts on neuronal activity in the intact and diseased brain. Copyright © 2015 the authors 0270-6474/15/353298-14$15.00/0.

  11. 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.

  12. ErbB receptors and PKC regulate PC12 neuronal-like differentiation and sodium current elicitation.

    Science.gov (United States)

    García, L; Castillo, C; Carballo, J; Rodríguez, Y; Forsyth, P; Medina, R; Martínez, J C; Longart, M

    2013-04-16

    Excitability, neurite outgrowth and their specification are very important features in the establishment of neuronal differentiation. We have studied a conditioned medium (CM) from sciatic nerve which is able to induce a neuronal-like differentiation of PC12 cells. Previously, we have demonstrated that supplementing this CM with a generic inhibitor (k252a), which mainly inhibits tropomyosin-related kinase receptors (Trk receptors) and protein kinase C (PKC), caused neurite elongation, sodium current induction and axon development. In the present work, we are showing that the enhancement of neurite length and induction of sodium currents induced by CM+k252a were prevented by ErbB receptor inhibition. Additionally, we demonstrated that specific inhibition of PKC produced a similar effect to that exerted by k252a in CM-treated cells, specifically by increasing the percentage of differentiated cells with long neurites and inducing sodium currents. Moreover, CM changed the mRNA levels for ErbB2 and ErbB3 increasing them 6- and 36-folds respectively compared to their control. The inclusion of k252a with CM changed the ErbB1, ErbB2 and ErbB3 mRNA proportions increasing those eight-, seven- and fivefolds respectively. From this point, it is clear that appropriate ErbB receptor levels and PKC inhibition are necessary to enhance the effect of the CM in inducing the neuronal-like differentiation of PC12 cells. In summary, we demonstrated the involvement of ErbB receptors in the regulation of neurite elongation and sodium current induction in PC12 cells and propose that these processes could be initiated by ErbB receptors followed by a fine regulation of PKC signaling. These findings might implicate a novel interplay between ErbB receptors and PKC in the regulation of these molecular mechanisms.

  13. 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.

  14. 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.

  15. Review of current and anticipated regulations on air protection in the Czech Republic

    Energy Technology Data Exchange (ETDEWEB)

    Jilek, P.; Novotny, V. [Ministry of the Environment of the Czech Republic, Prague (Czechoslovakia)

    1995-12-01

    Environmental issues, especially the solution of the air pollution problem, have taken on great significance in the Czech Republic (which was a part of the Czech and Slovak Federal Republic until the end of 1992) since the 1989 {open_quotes}Velvet{close_quotes} Revolution. The former CSFR Federal Committee for the Environment and both the Republic Ministries started immediately with creating new environmental legislation, which is the main governmental tool for protecting the environment in the newly developing democracy state system with a market oriented economy. The inspiration for that activity was found in legislation of developed countries - member states of the European Union, and in German environmental law in particular. This paper surveys the major laws and regulations that gradually came into force in the Czech Republic since 1990. The provisions of the primary significance are the Act No.309/1991 S.B., dated July 9, 1991, on the protection of the air against pollutants - The Clean Air Act, the Act No.218/1992 S.B., dated April 27, 1992, which changes and supplements the Act No.309 - The Clean Air Act, the Measure of the Federal Committee for the Environment of October 1, 1991 to the Clean Air Act, and its amended wordings of June 23, 1992, 84/1991 S.B., and 84/1992 S.B., the Act No.389/1991 S.B., dated September 10, 1991 on the state administration of air protection and charges for the pollution of air, and several regulations based on the Act No.389/1991 S.B., issued in the period 1992 -1993.

  16. 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.

  17. Aspartame: a safety evaluation based on current use levels, regulations, and toxicological and epidemiological studies.

    Science.gov (United States)

    Magnuson, B A; Burdock, G A; Doull, J; Kroes, R M; Marsh, G M; Pariza, M W; Spencer, P S; Waddell, W J; Walker, R; Williams, G M

    2007-01-01

    Aspartame is a methyl ester of a dipeptide used as a synthetic nonnutritive sweetener in over 90 countries worldwide in over 6000 products. The purpose of this investigation was to review the scientific literature on the absorption and metabolism, the current consumption levels worldwide, the toxicology, and recent epidemiological studies on aspartame. Current use levels of aspartame, even by high users in special subgroups, remains well below the U.S. Food and Drug Administration and European Food Safety Authority established acceptable daily intake levels of 50 and 40 mg/kg bw/day, respectively. Consumption of large doses of aspartame in a single bolus dose will have an effect on some biochemical parameters, including plasma amino acid levels and brain neurotransmitter levels. The rise in plasma levels of phenylalanine and aspartic acid following administration of aspartame at doses less than or equal to 50 mg/kg bw do not exceed those observed postprandially. Acute, subacute and chronic toxicity studies with aspartame, and its decomposition products, conducted in mice, rats, hamsters and dogs have consistently found no adverse effect of aspartame with doses up to at least 4000 mg/kg bw/day. Critical review of all carcinogenicity studies conducted on aspartame found no credible evidence that aspartame is carcinogenic. The data from the extensive investigations into the possibility of neurotoxic effects of aspartame, in general, do not support the hypothesis that aspartame in the human diet will affect nervous system function, learning or behavior. Epidemiological studies on aspartame include several case-control studies and one well-conducted prospective epidemiological study with a large cohort, in which the consumption of aspartame was measured. The studies provide no evidence to support an association between aspartame and cancer in any tissue. The weight of existing evidence is that aspartame is safe at current levels of consumption as a nonnutritive

  18. 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...

  19. 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...

  20. 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...

  1. 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...

  2. Inner Current Loop Analysis and Design Based on Resonant Regulators for Isolated Microgrids

    DEFF Research Database (Denmark)

    Federico, de Bosio; de Sousa Ribeiro, Luiz Antonio; Soares Lima, Marcel

    2015-01-01

    coupling in the design of proportional resonant controllers for these inner loops in voltage source inverters operating in islanded microgrids. It is also shown that the state feedback coupling has an important effect in the performance of the control loops by increasing the steady-state error......Inner current and voltage loops are fundamental in achieving good performance of microgrids based on power electronics voltage source inverters. The analysis and design of these loops are essential for the adequate operation of these systems. This paper investigates the effect of state feedback....... A comparison between different types of proportional+resonant controllers is done. Experimental results verify the theoretical assumptions done....

  3. 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.

  4. Recent experimental results of effects of perfluoroalkyl substances in laboratory animals - Relation to current regulations and guidance values.

    Science.gov (United States)

    Lilienthal, Hellmuth; Dieter, Hermann H; Hölzer, Jürgen; Wilhelm, Michael

    2017-03-02

    The detection of perfluoroalkyl substances (PFAS) in surface and drinking water from various countries raised the attention to the presence of these chemicals in environmental probes and led to several regulatory actions to limit exposure in human beings. There was particular concern about perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS), due to their former wide-spread use. Recently, several institutions published revisions of former regulatory or recommended maximum concentrations in drinking water and food, which are markedly lower than the former values. The present short overview describes the current regulations for PFAS and compares them with the outcome of several experimental studies in laboratory animals at low-level exposure to PFOA and PFOS. In addition, regulations for short-chain PFAS are presented which, due to lack of toxicological information, are evaluated according to the concepts of Threshold of Toxicological Concern (TTC) or the Health-related Indication Values (HRIV).

  5. 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...

  6. 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.

  7. 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...

  8. Cystic fibrosis transmembrane conductance regulator modulators in cystic fibrosis: current perspectives

    Directory of Open Access Journals (Sweden)

    Schmidt BZ

    2016-09-01

    Full Text Available Béla Z Schmidt,1 Jérémy B Haaf,2 Teresinha Leal,2 Sabrina Noel,2 1Stem Cell Biology and Embryology, Department of Development and Regeneration, Katholieke Universiteit Leuven, Leuven, 2Louvain Center for Toxicology and Applied Pharmacology, Université Catholique de Louvain, Brussels, Belgium Abstract: Mutations of the CFTR gene cause cystic fibrosis (CF, the most common recessive monogenic disease worldwide. These mutations alter the synthesis, processing, function, or half-life of CFTR, the main chloride channel expressed in the apical membrane of epithelial cells in the airway, intestine, pancreas, and reproductive tract. Lung disease is the most critical manifestation of CF. It is characterized by airway obstruction, infection, and inflammation that lead to fatal tissue destruction. In spite of great advances in early and multidisciplinary medical care, and in our understanding of the pathophysiology, CF is still considerably reducing the life expectancy of patients. This review highlights the current development in pharmacological modulators of CFTR, which aim at rescuing the expression and/or function of mutated CFTR. While only Kalydeco® and Orkambi® are currently available to patients, many other families of CFTR modulators are undergoing preclinical and clinical investigations. Drug repositioning and personalized medicine are particularly detailed in this review as they represent the most promising strategies for restoring CFTR function in CF. Keywords: high-throughput screening, drug repositioning, personalized medicine, precision medicine, potentiators, correctors

  9. 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.

  10. 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.

  11. Regulation of zebrafish sleep and arousal states: current and prospective approaches

    Directory of Open Access Journals (Sweden)

    Cindy N Chiu

    2013-04-01

    Full Text Available Every day, we shift among various states of sleep and arousal to meet the many demands of our bodies and environment. A central puzzle in neurobiology is how the brain controls these behavioral states, which are essential to an animal’s well-being and survival. Mammalian models have predominated sleep and arousal research, although in the past decade, invertebrate models have made significant contributions to our understanding of the genetic underpinnings of behavioral states. More recently, the zebrafish has emerged as a promising model system for sleep and arousal research. Here we review experimental evidence that the zebrafish, a diurnal vertebrate, exhibits fundamental behavioral and neurochemical characteristics of mammalian sleep and arousal. We also propose how specific advantages of the zebrafish can be harnessed to advance the field. These include tractable genetics to identify and manipulate molecular and cellular regulators of behavioral states, optical transparency to facilitate in vivo observation of neural structure and function, and amenability to high-throughput drug screens to discover novel therapies for neurological disorders.

  12. 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.

  13. Speaker-Oriented Classroom Acoustics Design Guidelines in the Context of Current Regulations in European Countries

    DEFF Research Database (Denmark)

    Pelegrin Garcia, David; Brunskog, Jonas; Rasmussen, Birgit

    2014-01-01

    experienced at work. With the aim of improving working conditions for teachers, this article presents guidelines for classroom acoustics design that meet simultaneously criteria of vocal comfort and speech intelligibility, which may be of use in future discussions for updating regulatory requirements...... in classroom acoustics. Two room acoustic parameters are shown relevant for a speaker: the voice support, linked to vocal effort, and the decay time derived from an oral-binaural impulse response, linked to vocal comfort. Theoretical prediction models for room-averaged values of these parameters are combined...... with a model of speech intelligibility based on the useful-to-detrimental ratio and empirical models of signal-to-noise ratio in classrooms in order to derive classroom acoustic guidelines, taking into account physical volume restrictions linked to the number of students present in a classroom. The recommended...

  14. 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.

  15. 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....

  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. A 3 A sink/source current fast transient response low-dropout Gm driven linear regulator*

    Institute of Scientific and Technical Information of China (English)

    Chu Xiuqin; Li Qingwei; Lai Xinquan; Yuan Bing; LiYanming; Zhao Yongrui

    2011-01-01

    A 3 A sink/source Gm-driven CMOS low-dropout regulator (LDO), specially designed for low input voltage and low cost, is presented by utilizing the structure of a current mirror Gm (transconductance) driving technique, which provides high stability as well as a fast load transient response. The proposed LDO was fabricated by a 0.5μm standard CMOS process, and the die size is as small as 1.0 mm2. The proposed LDO dissipates 220 μA of quiescent current in no-load conditions and is able to deliver up to 3 A of load current. The measured results show that the output voltage can be resumed within 2 μs with a less than l mV overshoot and undershoot in the output current step from -1.8 to 1.8 A with a 0.1μs rising and falling time at three 10μF ceramic capacitors.

  19. CURRENT SHEET REGULATION OF SOLAR NEAR-RELATIVISTIC ELECTRON INJECTION HISTORIES

    Energy Technology Data Exchange (ETDEWEB)

    Agueda, N.; Sanahuja, B. [Departament d' Astronomia i Meteorologia, Institut de Ciencies del Cosmos, Universitat de Barcelona (Spain); Vainio, R. [Department of Physics, University of Helsinki (Finland); Dalla, S. [Jeremiah Horrocks Institute, University of Central Lancashire (United Kingdom); Lario, D. [Applied Physics Laboratory, Johns Hopkins University (United States)

    2013-03-10

    We present a sample of three large near-relativistic (>50 keV) electron events observed in 2001 by both the ACE and the Ulysses spacecraft, when Ulysses was at high-northern latitudes (>60 Degree-Sign ) and close to 2 AU. Despite the large latitudinal distance between the two spacecraft, electrons injected near the Sun reached both heliospheric locations. All three events were associated with large solar flares, strong decametric type II radio bursts and accompanied by wide (>212 Degree-Sign ) and fast (>1400 km s{sup -1}) coronal mass ejections (CMEs). We use advanced interplanetary transport simulations and make use of the directional intensities observed in situ by the spacecraft to infer the electron injection profile close to the Sun and the interplanetary transport conditions at both low and high latitudes. For the three selected events, we find similar interplanetary transport conditions at different heliolatitudes for a given event, with values of the mean free path ranging from 0.04 AU to 0.27 AU. We find differences in the injection profiles inferred for each spacecraft. We investigate the role that sector boundaries of the heliospheric current sheet (HCS) have on determining the characteristics of the electron injection profiles. Extended injection profiles, associated with coronal shocks, are found if the magnetic footpoints of the spacecraft lay in the same magnetic sector as the associated flare, while intermittent sparse injection episodes appear when the spacecraft footpoints are in the opposite sector or a wrap in the HCS bounded the CME structure.

  20. [Role of calcineurin in down-regulation of left ventricular transmural voltage- dependent K(+) currents in mice with heart failure].

    Science.gov (United States)

    Shi, Chen-Xia; Dong, Fang; Chang, Yan-Chao; Wang, Xiao-Feng; Xu, Yan-Fang

    2015-08-25

    The aim of the present study was to investigate the role of calcineurin in the down-regulation of left ventricular transmural voltage-dependent K(+) currents in heart failure. Transverse aorta was banded by using microsurgical techniques to create mouse heart failure model. Sham-operated (Sham) or aorta banded (Band) mice were randomized to receive calcineurin inhibitor cyclosporine A (CsA) or vehicle. The densities and kinetic properties of voltage-dependent K(+) currents, as well as action potential (AP), of left ventricular subendocardial (Endo) and subepicardial (Epi) myocytes were determined by using whole-cell patch-clamp technique. The results showed that calcineurin activity was significant higher in Endo myocytes than that in Epi ones in all the groups. Compared with Sham group, Band mice showed significantly increased calcineurin activity both in Endo and Epi myocytes. CsA significantly reduced calcineurin activity in Band mice. CsA treatment in Band mice partially reversed the down-regulation of Ito density, completely reversed the down-regulation of IK,slow density both in Endo and Epi myocytes, and Iss density in Endo myocytes. In addition, CsA treatment in Band mice partially antagonized the prolongation of action potential duration (APD), and APD at 50% (APD50) and 90% repolarization (APD90) were significantly reduced. Because of non-parallel shortening of APD in Endo and Epi myocytes, the ratio of Endo/Epi APD90 was reduced from 4.8:1 in Band mice to 2.6:1 in CsA-treated mice, which was close to that in Sham mice. The results suggest that non-parallel activation of calcineurin in Endo and Epi myocytes contributes to the down-regulation of transmural voltage-dependent K(+) currents and the amplification of transmural dispersion of repolarization (TDR) in left ventricular failure hearts. Inhibition of calcineurin may be a potential new therapeutic strategy to prevent and cure arrhythmias and sudden death in heart failure.

  1. 77 FR 34927 - National Pollutant Discharge Elimination System-Proposed Regulations To Establish Requirements...

    Science.gov (United States)

    2012-06-12

    ... policy options and current conditions. EPA solicits public comment on all aspects of the study, including the methodology used, the strengths and weaknesses of stated preferences methods generally, and the... options differ in the levels of environmental improvements and ] impose different costs on the...

  2. 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.

  3. 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

  4. 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...

  5. 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...

  6. 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

  7. 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...

  8. 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...

  9. 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...

  10. 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...

  11. 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.

  12. Ultralow-quiescent-current and wide-load-range low-dropout linear regulator with self-biasing technique for micropower battery management

    Science.gov (United States)

    Ozaki, Toshihiro; Hirose, Tetsuya; Asano, Hiroki; Kuroki, Nobutaka; Numa, Masahiro

    2017-04-01

    In this paper, we present a 151 nA quiescent and 6.8 mA maximum-output-current low-dropout (LDO) linear regulator for micropower battery management. The LDO regulator employs self-biasing and multiple-stacked cascode techniques to achieve efficient, accurate, and high-voltage-input-tolerant operation. Measurement results demonstrated that the proposed LDO regulator operates with an ultralow quiescent current of 151 nA. The maximum output currents with a 4.16 V output were 1.0 and 6.8 mA when the input voltages were 4.25 and 5.0 V, respectively.

  13. Role of calcium mobilization in the regulation of spontaneous transient outward currents in porcine coronary artery myocytes

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The purpose of the present study was to further study the characteristics and regulation of spontaneous transient outward currents (STOCs) in freshly isolated porcine coronary artery smooth muscle cells (ASMCs). STOCs were recorded using the perforated whole-cell patch-clamp configuration. STOCs were voltage-dependent and superimposed stochastically onto whole-cell Ca2+-activated-K+ (BKCa) currents. Charybdotoxin (ChTX, 200 nmol/L), a selective blocker of BKCa channels, completely inhibited STOCs within 10 min. STOCs activity was greatly suppressed when extracellular Ca2+ concentration decreased from 1.8 mmol/L to 200 nmol/L, further removal of Ca2+ abolished STOCs activity. Ca2+ ionophore A23187 (10 μmol/L) increased STOCs activity significantly. Verapamil (20 μmol/L) and CdCl2 (200 μmol/L), two kinds of organic L-type voltage-dependent Ca2+ channels (L-VDCCs) antagonists, had little effect on STOCs. In addition, the ryanodine receptors (RyRs) agonist caffeine (5 mmol/L) significantly activated STOCs. Application of ryanodine (50 μmol/L) to block RyRs abolished STOCs, subsequent washout of ryanodine or application of caffeine failed to reproduce STOCs activity. Inhibition of inositol 1,4,5-trisphosphate receptors (IP3Rs) by 2APB (40 μmol/L) greatly suppressed the activity of STOCs, application of caffeine (5 mmol/L) in the presence of 2APB caused a burst of outward currents followed by inhibition of STOCs. These results suggest that STOCs in porcine coronary ASMCs are mediated by BKCa channels. Extracellular Ca2+ is essential for STOCs activity, while Ca2+ entry through L-VDCCs has little effect on STOCs. Intracellular Ca2+ release induced by RyRs is responsible for the regulation of STOCs, whereas IP3Rs might also be involved.

  14. Determining K+ channel activation curves from K+ channel currents often requires the Goldman-Hodgkin-Katz equation

    Directory of Open Access Journals (Sweden)

    john r Clay

    2009-12-01

    Full Text Available Potassium ion current in nerve membrane, IK, has traditionally been described by IK = gK(V-EK, where gK is the K ion conductance, V is membrane potential, and EK is the K+ Nernst potential. This description has been unchallenged by most investigators in neuroscience since its introduction almost sixty years ago. The problem with the IK ~ (V-EK proportionality is that it is inconsistent with the unequal distribution of K ions in the intra- and extracellular bathing media. Under physiological conditions the intracellular K+ concentraion is significantly higher than the extracellular concentration. Consequently, the slope conductance at potentials positive to EK cannot be the same as that for potentials negative to EK, as the linear proportionality requires. Instead IK has a non-linear dependence on (V-EK which is well described by the Goldman-Hodgkin-Katz equation. The implications of this result for K+ channel gating and membrane excitability are reviewed in this report.

  15. 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.

  16. 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

  17. 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...

  18. 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.

  19. 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.

  20. Single-channel and whole-cell recordings of mitogen-regulated inward currents in human cloned helper T lymphocytes.

    Science.gov (United States)

    Kuno, M; Goronzy, J; Weyand, C M; Gardner, P

    Cytoplasmic free Ca2+ [( Ca2+]i) appears to be an important signal for DNA synthesis in early stages of lymphocyte activation. In spite of many experimental studies which employ fluorescent Ca2+ indicator dye to demonstrate an early increase of [Ca2+]i in T-lymphocytes after stimulation with lectins, specific antigens, and monoclonal antibodies to T-lymphocyte receptors, the mechanism responsible for the rise of [Ca2+]i is unknown. We have used the extracellular patch clamp technique to investigate this mechanism. Unitary inward currents, mediated by Ca2+ or Ba2+, were recorded in the membrane of T-lymphocytes. The inward current channel was characterized by a conductance of 7 pS and extrapolated reversal potential (Erev) 110 mV positive to resting potential (Vr). While gating kinetic parameters were not affected by membrane potential changes, the probability of channel opening markedly increased upon activation of the T-lymphocyte by the mitogenic lectin, phytohaemagglutinin (PHA). PHA also evoked a cadmium-sensitive, inward Ba2+ current on whole-cell clamp. We suggest that this mitogen-regulated channel introduces Ca2+ into the cytoplasm upon activation and represents a new class of voltage-independent Ca2+ channels.

  1. Control of the light-regulated current in rod photoreceptors by cyclic GMP, calcium, and l-cis-diltiazem.

    Science.gov (United States)

    Stern, J H; Kaupp, U B; MacLeish, P R

    1986-02-01

    The effect of calcium ions on the cGMP-activated current of outer segment membrane was examined by the excised-patch technique. Changes in the extracellular calcium concentration had marked effects on the cGMP-activated current, while changes in intracellular calcium concentration were ineffective. Changes in calcium concentration in the absence of cGMP had little, if any, effect on membrane conductance. These results suggest that both intracellular cGMP and extracellular calcium can directly affect the conductance underlying the light response in rod cells. The pharmacological agent l-cis-diltiazem reversibly inhibited the cGMP-activated current when applied to the intracellular side of an excised patch. When superfused over intact rod cells, l-cis-diltiazem reversibly blocked much of the normal light response. The isomer, d-cis-diltiazem, did not significantly affect either patches or intact rod cells. Thus, the light-regulated conductance has binding sites for both calcium and cGMP that may interact during the normal light response in rod cells and a site specific for l-cis-diltiazem that can be used to identify and further study the conductance mechanism.

  2. 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.

  3. 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.

  4. 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

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. Mechanism for Muscarinic Inhibitory Regulation of the L-type Ca2 + Current in Cardiac Ventricular Myocytes

    Institute of Scientific and Technical Information of China (English)

    蒋彬; 杨向军; 惠杰; 蒋廷波; 宋建平; 刘志华

    2004-01-01

    @@ Objective The autonomic nervous system plays a key role in regulating cardiac function by modifying heart rate, contractility and impulse. The parasympathetic neurotransmitter acetyl-choline and muscarinic agonist carbachol (Cch) inhibit excitation-contraction coupling in cardiac ventricular myocytes. Muscarinic agonists suppress adenylyl cyclase (AC) acitivity and,by reducing activation of the cAMP/protein kinase A (PKA)cascade, inhibit the L-type Ca2+ current (ICa(L) ). They also increase the content of cGMP by stimulating guanylyl cyclase (GC) activity. The role of nitric oxide (NO)/cGMP in muscarinic inhibition has undergone considerable scrutiny. The role of the NO/cGMP pathway in the inhibition of ICa(L) by Cch was examined in guinea-pig ventricular myocytes.

  14. [National system of protection against electromagnetic fields 0 Hz-300 GHz in the light of current legal regulations].

    Science.gov (United States)

    Aniołczyk, Halina

    2006-01-01

    Exposure to electromagnetic fields (EMF) occurs when man is exposed to the effect of electric, magnetic and electromagnetic fields and contact currents different from those resulting from physiological processes in the organism or other natural phenomena. In Poland, the system of protection against EMF has been functioning for over 35 years. In 2001, when the Minister of Labor and Social Policy issued the regulation introducing the maximum admissible intensities (MAI) for electromagnetic fields and radiation within the range of 0 Hz-300 GHz, the system was directed mainly towards evaluation of exposure to EMF occurring in the occupational environment. The system is linked via MAI values with human protection in the natural environment. In this paper, the background, principles and the range of the national system of protection against EMF and its monitoring are presented. The project of implementation of EU directives, following Poland's accession to the European Union is also discussed.

  15. 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...

  16. 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.

  17. Derivation of linearized transfer functions for switching-mode regulations. Phase A: Current step-up and voltage step-up converters

    Science.gov (United States)

    Wong, R. C.; Owen, H. A., Jr.; Wilson, T. G.

    1981-01-01

    Small-signal models are derived for the power stage of the voltage step-up (boost) and the current step-up (buck) converters. The modeling covers operation in both the continuous-mmf mode and the discontinuous-mmf mode. The power stage in the regulated current step-up converter on board the Dynamics Explorer Satellite is used as an example to illustrate the procedures in obtaining the small-signal functions characterizing a regulated converter.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. The persistent sodium current generates pacemaker activities in the central pattern generator for locomotion and regulates the locomotor rhythm.

    Science.gov (United States)

    Tazerart, Sabrina; Vinay, Laurent; Brocard, Frédéric

    2008-08-20

    Rhythm generation in neuronal networks relies on synaptic interactions and pacemaker properties. Little is known about the contribution of the latter mechanisms to the integrated network activity underlying locomotion in mammals. We tested the hypothesis that the persistent sodium current (I(NaP)) is critical in generating locomotion in neonatal rodents using both slice and isolated spinal cord preparations. After removing extracellular calcium, 75% of interneurons in the area of the central pattern generator (CPG) for locomotion exhibited bursting properties and I(NaP) was concomitantly upregulated. Putative CPG interneurons such as commissural and Hb9 interneurons also expressed I(NaP)-dependent (riluzole-sensitive) bursting properties. Most bursting cells exhibited a pacemaker-like behavior (i.e., burst frequency increased with depolarizing currents). Veratridine upregulated I(NaP), induced riluzole-sensitive bursting properties, and slowed down the locomotor rhythm. This study provides evidence that I(NaP) generates pacemaker activities in CPG interneurons and contributes to the regulation of the locomotor activity.

  7. 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.

  8. Volume regulated anion channel currents of rat hippocampal neurons and their contribution to oxygen-and-glucose deprivation induced neuronal death.

    Directory of Open Access Journals (Sweden)

    Huaqiu Zhang

    Full Text Available Volume-regulated anion channels (VRAC are widely expressed chloride channels that are critical for the cell volume regulation. In the mammalian central nervous system, the physiological expression of neuronal VRAC and its role in cerebral ischemia are issues largely unknown. We show that hypoosmotic medium induce an outwardly rectifying chloride conductance in CA1 pyramidal neurons in rat hippocampal slices. The induced chloride conductance was sensitive to some of the VRAC inhibitors, namely, IAA-94 (300 µM and NPPB (100 µM, but not to tamoxifen (10 µM. Using oxygen-and-glucose deprivation (OGD to simulate ischemic conditions in slices, VRAC activation appeared after OGD induced anoxic depolarization (AD that showed a progressive increase in current amplitude over the period of post-OGD reperfusion. The OGD induced VRAC currents were significantly inhibited by inhibitors for glutamate AMPA (30 µM NBQX and NMDA (40 µM AP-5 receptors in the OGD solution, supporting the view that induction of AD requires an excessive Na(+-loading via these receptors that in turn to activate neuronal VRAC. In the presence of NPPB and DCPIB in the post-OGD reperfusion solution, the OGD induced CA1 pyramidal neuron death, as measured by TO-PRO-3-I staining, was significantly reduced, although DCPIB did not appear to be an effective neuronal VRAC blocker. Altogether, we show that rat hippocampal pyramidal neurons express functional VRAC, and ischemic conditions can initial neuronal VRAC activation that may contribute to ischemic neuronal damage.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. Pesticide regulation, utilization, and retailers' selling practices in Trinidad and Tobago, West Indies: current situation and needed changes.

    Science.gov (United States)

    Pinto Pereira, Lexley M; Boysielal, Kim; Siung-Chang, Avril

    2007-08-01

    To explore pesticide regulation in Trinidad and Tobago, and to ascertain pesticide utilization and retailers' selling practices on Trinidad, which is the larger of twin islands that constitute the republic of Trinidad and Tobago. Between February and June 2005, agrochemical retailers in Trinidad were surveyed about the most frequently sold pesticides and their knowledge and practices of pesticide sale. The Poisons and Toxic Chemicals Control Board of the Ministry of Health informed on legislature. Of 107 actively trading licensed pesticide outlets, 97 participated (91% response rate) in the survey. Currently only 2.9% (21) of 720 registered products from four chemical classes are frequently utilized. Paraquat, methomyl, and alpha-cypermethrin (respective trade names are Gramoxone, Lannate, and Fastac) from World Health Organization (WHO) Hazard Classes I and II, and glyphosate isopropylamine (Swiper, Class U) are the most frequently purchased pesticides. Pet shops constitute 39.2% (38) of retail shops selling pesticides. No regulations guide pesticide sale to agriculturists, and children may purchase them. Inadequate human and technical resources render legislative controls ineffective and disciplinary action against offenders is weak. Extensive governmental resources are employed in legislative procedures and product approval for the very low, 2.9% utilization rate, negatively impacting on monitoring pesticide sales. The Poisons Information Centre (PIC) does not liaise with the Poisons and Toxic Chemicals Control Board or provide educational interventions for the community. As a result of this survey, it was possible to develop the first database to include the chemical, brand, and colloquial names of pesticides used in Trinidad and Tobago; WHO classification of approved pesticides; manufacturers; packaging; and antidotes and their availability for use by the Board and health professionals in Trinidad. Urgent critical evaluation of legislation regarding pesticide

  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. Learned EEG-based regulation of motor-related brain oscillations during application of transcranial electric currents: feasibility and limitations

    Directory of Open Access Journals (Sweden)

    Surjo R Soekadar

    2014-03-01

    Full Text Available Objective: Transcranial direct current stimulation (tDCS improves motor learning and can influence emotional processing or attention. However, it remained unclear whether learned electroencephalography (EEG-based brain-machine interface (BMI control during tDCS is feasible and how application of transcranial electric currents during BMI control would interfere with feature-extraction of physiological brain signals. Here we tested this combination and evaluated stimulation-dependent artifacts across different EEG frequencies and stability of motor imagery-based BMI control. Approach: Ten healthy volunteers were invited to two BMI-sessions, each comprising two 60-trial blocks. During the trials, modulation of mu-rhythms (8-15Hz associated with motor imagery recorded over C4 was translated into online cursor movements on a computer screen. During block 2, either sham (session A or anodal tDCS (session B was applied at 1mA with the stimulation electrode placed 1cm anterior of C4. Main results: tDCS was associated with a significant signal power increase in the lower frequencies most evident in the signal spectrum of the EEG channel closest to the stimulation electrode. Stimulation-dependent signal power increase exhibited a decay of 12dB per decade, leaving frequencies above 9Hz unaffected. Analysis of BMI control performance did not indicate a difference between blocks and tDCS conditions. Conclusion: Application of tDCS during learned EEG-based self-regulation of brain oscillations above 9Hz is feasible and safe, and might improve applicability of BMI systems in patient populations.

  16. 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.

  17. Historical perspective and current status of the physical education graduation requirement at American 4-year colleges and universities.

    Science.gov (United States)

    Cardinal, Bradley J; Sorensen, Spencer D; Cardinal, Marita K

    2012-12-01

    This study gives an overview of the history of required physical education in America's 4 year colleges and universities and provides an update on the requirement status. After randomly identifying 354 institutions, we searched their respective websites to determine whether physical education was a requirement to earn a baccalaureate degree. The major finding was that the physical education requirement declined from an all-time high of 97% in the 1920s and 1930s to an all-time low of 39.55% in 2010. Given society's ongoing health challenges and the important role of physical activity in maintaining health, this seems counterintuitive. It is also inconsistent with the National Physical Activity Plan (2010).

  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. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. Fragile X mental retardation protein is required for rapid experience-dependent regulation of the potassium channel Kv3.1b.

    Science.gov (United States)

    Strumbos, John G; Brown, Maile R; Kronengold, Jack; Polley, Daniel B; Kaczmarek, Leonard K

    2010-08-04

    Fragile X mental retardation protein (FMRP) is an RNA-binding protein that regulates synaptic plasticity by repressing translation of specific mRNAs. We found that FMRP binds mRNA encoding the voltage-gated potassium channel Kv3.1b in brainstem synaptosomes. To explore the regulation of Kv3.1b by FMRP, we investigated Kv3.1b immunoreactivity and potassium currents in the auditory brainstem sound localization circuit of male mice. The unique features of this circuit allowed us to control neuronal activity in vivo by exposing animals to high-frequency, amplitude-modulated stimuli, which elicit predictable and stereotyped patterns of input to the anterior ventral cochlear nucleus (AVCN) and medial nucleus of the trapezoid body (MNTB). In wild-type (WT) animals, Kv3.1b is expressed along a tonotopic gradient in the MNTB, with highest levels in neurons at the medial, high-frequency end. At baseline, Fmr1(-/-) mice, which lack FMRP, displayed dramatically flattened tonotopicity in Kv3.1b immunoreactivity and K(+) currents relative to WT controls. Moreover, after 30 min of acoustic stimulation, levels of Kv3.1b immunoreactivity were significantly elevated in both the MNTB and AVCN of WT, but not Fmr1(-/-), mice. These results suggest that FMRP is necessary for maintenance of the gradient in Kv3.1b protein levels across the tonotopic axis of the MNTB, and are consistent with a role for FMRP as a repressor of protein translation. Using numerical simulations, we demonstrate that Kv3.1b tonotopicity may be required for accurate encoding of stimulus features such as modulation rate, and that disruption of this gradient, as occurs in Fmr1(-/-) animals, degrades processing of this information.

  13. Corrosion protection of Arctic offshore structures: Final report. [Effects of temperature and salinity on required cathodic protection current

    Energy Technology Data Exchange (ETDEWEB)

    Sackinger, W.M.; Rogers, J.C.; Feyk, C.; Theuveny, B.

    1985-10-01

    Results are presented for a research program on corrosion prevention for Arctic offshore structures which are in contact with sea ice for a significant portion of the year. The electrical method most adaptable for structure protection involves the injection of impressed current from several remote anodes buried just beneath the sea floor. The electrical resistivity of annual sea ice as a function of temperature and salinity is presented. Details of the interface layers formed between sea ice and steel in the presence of current injection are shown. A computer program was developed to enable the calculation of protective current density into the structure, in the presence of ice rubble and ridges around the structure. The program and the results of an example calculation are given for a caisson- retained island structure. 81 refs., 103 figs., 3 tabs.

  14. Heating and current drive requirements for ideal MHD stability and ITB sustainment in ITER steady state scenarios

    Science.gov (United States)

    Poli, Francesca

    2012-10-01

    Steady state scenarios envisaged for ITER aim at optimizing the bootstrap current, while maintaining sufficient confinement and stability to provide the necessary fusion yield. Non-inductive scenarios will need to operate with Internal Transport Barriers (ITBs) in order to reach adequate fusion gain at typical currents of 9 MA. However, the large pressure gradients associated with ITBs in regions of weak or negative magnetic shear can be conducive to ideal MHD instabilities in a wide range of βN, reducing the no-wall limit. Scenarios are established as relaxed flattop states with time-dependent transport simulations with TSC [1]. Fully non-inductive configurations with current in the range of 7-10 MA and various heating mixes (NB, EC, IC and LH) have been studied against variations of the pressure profile peaking and of the Greenwald fraction. It is found that stable equilibria have qmin> 2 and moderate ITBs at 2/3 of the minor radius [2]. The ExB flow shear from toroidal plasma rotation is expected to be low in ITER, with a major role in the ITB dynamics being played by magnetic geometry. Combinations of H&CD sources that maintain reverse or weak magnetic shear profiles throughout the discharge and ρ(qmin)>=0.5 are the focus of this work. The ITER EC upper launcher, designed for NTM control, can provide enough current drive off-axis to sustain moderate ITBs at mid-radius and maintain a non-inductive current of 8-9MA and H98>=1.5 with the day one heating mix. LH heating and current drive is effective in modifying the current profile off-axis, facilitating the formation of stronger ITBs in the rampup phase, their sustainment at larger radii and larger bootstrap fraction. The implications for steady state operation and fusion performance are discussed.[4pt] [1] Jardin S.C. et al, J. Comput. Phys. 66 (1986) 481[0pt] [2] Poli F.M. et al, Nucl. Fusion 52 (2012) 063027.

  15. SEMICONDUCTOR INTEGRATED CIRCUITS: A current-mode voltage regulator with an embedded sub-threshold reference for a passive UHF RFID transponder

    Science.gov (United States)

    Zhongqi, Liu; Chun, Zhang; Yongming, Li; Zhihua, Wang

    2010-06-01

    This paper presents a current-mode voltage regulator for a passive UHF RFID transponder. The passive tag power is extracted from RF energy through the RF-to-DC rectifier. Due to huge variations of the incoming RF power, the rectifier output voltage should be regulated to achieve a stable power supply. By accurately controlling the current flowing into the load with an embedded sub-threshold reference, the regulated voltage varies in a range of 1-1.3 V from -20 to 80 °C, and a bandwidth of about 100 kHz is achieved for a fast power recovery. The circuit is fabricated in UMC 0.18 μm mixed-mode CMOS technology, and the current consumption is only 1 μA.

  16. 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.

  17. 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.

  18. 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

  19. 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

  20. 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.

  1. Regulating the private security industry

    CERN Document Server

    Percy, Sarah

    2002-01-01

    The under-regulation of the private security industry has increasingly become a topic of media and academic interest. This Adelphi Paper enters the debate by explaining why the industry requires further regulation, and what is wrong with the current system. It begins by briefly defining the industry and explaining the need for more effective regulation, before analysing three types of regulation: domestic, international and informal (including self-regulation).

  2. Two cystic fibrosis transmembrane conductance regulator mutations have different effects on both pulmonary phenotype and regulation of outwardly rectified chloride currents.

    OpenAIRE

    Fulmer, S B; Schwiebert, E M; M.M. Morales; Guggino, W B; Cutting, G R

    1995-01-01

    Cystic fibrosis (CF), a disorder of electrolyte transport manifest in the lungs, pancreas, sweat duct, and vas deferens, is caused by mutations in the CF transmembrane conductance regulator (CFTR). The CFTR protein has been shown to function as a cAMP-activated chloride channel and also regulates a separate protein, the outwardly rectifying chloride channel (ORCC). To determine the consequence of disease-producing mutations upon these functions, mutant CFTR was transiently expressed in Xenopu...

  3. Current Requirements of the Society to the Professional Training of Specialists in Information Technology Industry in Japan

    Directory of Open Access Journals (Sweden)

    Pododimenko Inna

    2014-06-01

    Full Text Available The problem of professional training of skilled human personnel in the industry of information communication technology, the urgency of which is recognized at the state level of Ukraine and the world, has been considered. It has been traced that constantly growing requirements of the labour market, swift scientific progress require the use of innovative approaches to the training of future ІТ specialists with the aim to increase their professional level. The content of standards of professional training and development of information technologies specialists in foreign countries, particularly in Japan, has been analyzed and generalized. On the basis of analysis of educational and professional standards of Japan, basic requirements to the engineer in industry of information communication technology in the conditions of competitive environment at the labour market have been comprehensively characterized. The competencies that graduate students of educational qualification level of bachelor in the conditions of new state policy concerning upgrading the quality of higher education have been considered. The constituents of professional competence in the structure of an engineer-programmer’s personality, necessary on different levels of professional improvement of a specialist for the development of community of highly skilled ІТ specialists, have been summarized. Positive features of foreign experience and the possibility of their implementation into the native educational space have been distinguished. Directions for modernization and upgrading of the quality of higher education in Ukraine and the prospects for further scientific research concerning the practice of specialists in information technologies training have been suggested

  4. 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...

  5. 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.

  6. 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.

  7. 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

  8. 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.

  9. 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®

  10. 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

  11. 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.

  12. 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.

  13. Current practices of diagnostic techniques requir- ing the use of ophthalmic drugs among KwaZulu- Natal optometrists*

    Directory of Open Access Journals (Sweden)

    K. P. Mashige

    2009-12-01

    Full Text Available In anendeavour to improve the quality of optometric eye care services in South Africa, the scope of practice was expanded to include the use of ocular diagnostic procedures such as goniosco-py that require the use of ophthalmic drugs. The purpose of this study was to assess the practices of specific diagnostic techniques (contact tonometry, 78 D/90 D lens fundus examination, binocular indirect ophthalmoscopy and gonioscopy requiring the use of ophthalmic drugs among optometrists in KwaZulu-Natal (KZN province. These specific techniques are referred to as diagnostic procedures in this article. A questionnaire containing information on demography and practice of these specific techniques was sent to all 213 KwaZulu-Natal registered optometrists who owned private practices. One hundred and thirty two completed questionnaires were received, a response rate of 62%. One hundred and seventeen (55% of the questionnaires were included in the analysis of which 55% of the respondents were females and 45% were males. Sixty two optometrists (53% were certified in di-agnostic procedures but many procedures were not being practiced. These procedures and the percentage respondents were: Contact tonometry (60%, 78 D/90 D lens fundus examination (60%, binocular indirect ophthalmoscopy (84% and gonioscopy (78%. Also, among these certified respondents (62 optometrists, a significant proportion (60% disagreed when asked if they were confident and proficient in performing the relevant diagnostic procedures. Many, (61% agreed that lack of incentives discouraged them from routinely performing the procedures. More than half (58%, agreed that chair time was an important factor in deciding whether or not to perform these diagnostic procedures. Of the total respondents (117, 86% agreed that they were confident about the accuracy of their referrals and less than half (45% disagreed that diagnostic procedures should be the sole responsibility of ophthalmologists. Less than

  14. Current state of nuclear fuel cycles in nuclear engineering and trends in their development according to the environmental safety requirements

    Science.gov (United States)

    Vislov, I. S.; Pischulin, V. P.; Kladiev, S. N.; Slobodyan, S. M.

    2016-08-01

    The state and trends in the development of nuclear fuel cycles in nuclear engineering, taking into account the ecological aspects of using nuclear power plants, are considered. An analysis of advantages and disadvantages of nuclear engineering, compared with thermal engineering based on organic fuel types, was carried out. Spent nuclear fuel (SNF) reprocessing is an important task in the nuclear industry, since fuel unloaded from modern reactors of any type contains a large amount of radioactive elements that are harmful to the environment. On the other hand, the newly generated isotopes of uranium and plutonium should be reused to fabricate new nuclear fuel. The spent nuclear fuel also includes other types of fission products. Conditions for SNF handling are determined by ecological and economic factors. When choosing a certain handling method, one should assess these factors at all stages of its implementation. There are two main methods of SNF handling: open nuclear fuel cycle, with spent nuclear fuel assemblies (NFAs) that are held in storage facilities with their consequent disposal, and closed nuclear fuel cycle, with separation of uranium and plutonium, their purification from fission products, and use for producing new fuel batches. The development of effective closed fuel cycles using mixed uranium-plutonium fuel can provide a successful development of the nuclear industry only under the conditions of implementation of novel effective technological treatment processes that meet strict requirements of environmental safety and reliability of process equipment being applied. The diversity of technological processes is determined by different types of NFA devices and construction materials being used, as well as by the composition that depends on nuclear fuel components and operational conditions for assemblies in the nuclear power reactor. This work provides an overview of technological processes of SNF treatment and methods of handling of nuclear fuel

  15. Current practices of diagnostic techniques requiring the use of ophthalmic drugs among KwaZulu- Natal optometrists*

    Directory of Open Access Journals (Sweden)

    K. P. Mashige

    2009-12-01

    Full Text Available In anendeavour to improve the quality of optometric eye care services in South Africa, the scope of practice was expanded to include the use of ocular diagnostic procedures such as gonioscopy that require the use of ophthalmic drugs. The purpose of this study was to assess the practices of specific diagnostic techniques (contact tonometry, 78 D/90 D lens fundus examination, binocular indirect ophthalmoscopy and gonioscopy requiring the use of ophthalmic drugs among optometrists in KwaZulu-Natal (KZN province. These specific techniques are referred to as diagnostic procedures in this article. A questionnaire containing information on demography and practice of these specific techniques was sent to all 213 KwaZulu-Natal registered optometrists who owned private practices. One hundred and thirty two completed questionnaires were received, a response rate of 62%. One hundred and seventeen (55% of the questionnaires were included in the analysis of which 55% of the respondents were females and 45% were males. Sixty two optometrists (53% were certified in di-agnostic procedures but many procedures were not being practiced. These procedures and the percentage respondents were: Contact tonometry (60%, 78 D/90 D lens fundus examination (60%, binocular indirect ophthalmoscopy (84% and gonioscopy (78%. Also, among these certified respondents (62 optometrists, a significant proportion (60% disagreed when asked if they were confident and proficient in performing the relevant diagnostic procedures. Many, (61% agreed that lack of incentives discouraged them from routinely performing the procedures. More than half (58%, agreed that chair time was an important factor in deciding whether or not to perform these diagnostic procedures. Of the total respondents (117, 86% agreed that they were confident about the accuracy of their referrals and less than half (45% disagreed that diagnostic procedures should be the sole responsibility of ophthalmologists. Less than

  16. Power requirements for electron cyclotron current drive and ion cyclotron resonance heating for sawtooth control in ITER

    CERN Document Server

    Chapman, I T; Sauter, O; Zucca, C; Asunta, O; Buttery, R J; Coda, S; Goodman, T; Igochine, V; Johnson, T; Jucker, M; La Haye, R J; Lennholm, M; Contributors, JET-EFDA

    2013-01-01

    13MW of electron cyclotron current drive (ECCD) power deposited inside the q = 1 surface is likely to reduce the sawtooth period in ITER baseline scenario below the level empirically predicted to trigger neo-classical tearing modes (NTMs). However, since the ECCD control scheme is solely predicated upon changing the local magnetic shear, it is prudent to plan to use a complementary scheme which directly decreases the potential energy of the kink mode in order to reduce the sawtooth period. In the event that the natural sawtooth period is longer than expected, due to enhanced alpha particle stabilisation for instance, this ancillary sawtooth control can be provided from > 10MW of ion cyclotron resonance heating (ICRH) power with a resonance just inside the q = 1 surface. Both ECCD and ICRH control schemes would benefit greatly from active feedback of the deposition with respect to the rational surface. If the q = 1 surface can be maintained closer to the magnetic axis, the efficacy of ECCD and ICRH schemes sig...

  17. Cholesterol modulates the volume-regulated anion current in Ehrlich-Lettre ascites cells via effects on Rho and F-actin

    DEFF Research Database (Denmark)

    Klausen, Thomas Kjaer; Hougaard, Charlotte; Hoffmann, Else K

    2006-01-01

    The mechanisms controlling the volume-regulated anion current (VRAC) are incompletely elucidated. Here, we investigate the modulation of VRAC by cellular cholesterol and the potential involvement of F-actin, Rho, Rho kinase, and phosphatidylinositol-(4,5)-bisphosphate [PtdIns(4,5)P(2)] in this pr......The mechanisms controlling the volume-regulated anion current (VRAC) are incompletely elucidated. Here, we investigate the modulation of VRAC by cellular cholesterol and the potential involvement of F-actin, Rho, Rho kinase, and phosphatidylinositol-(4,5)-bisphosphate [PtdIns(4,5)P(2...... in cellular cholesterol content increased cortical and stress fiber-associated F-actin content in swollen cells. Cholesterol depletion increased VRAC activation rate and maximal current after a modest (15%), but not after a severe (36%) reduction in extracellular osmolarity. The cholesterol depletion...

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. Multi-step constant-current charging method for electric vehicle, valve-regulated, lead/acid batteries during night time for load-levelling

    Energy Technology Data Exchange (ETDEWEB)

    Ikeya, Tomohiko; Mita, Yuichi; Ishihara, Kaoru [Central Research Inst. of Electric Power Industry, Tokyo (Japan); Sawada, Nobuyuki [Hokkaido Electric Power, Sapporo (Japan); Takagi, Sakae; Murakami, Jun-ichi [Tohoku Electric Power, Sendai (Japan); Kobayashi, Kazuyuki [Tokyo Electric Power, Yokohama (Japan); Sakabe, Tetsuya [Chubu Electric Power, Nagoya (Japan); Kousaka, Eiichi [Hokuriku Electric Power, Toyama (Japan); Yoshioka, Haruki [The Kansai Electric Power, Osaka (Japan); Kato, Satoru [The Chugoku Electric Power, Hiroshima (Japan); Yamashita, Masanori [Shikoku Research Inst., Takamatsu (Japan); Narisoko, Hayato [The Okinawa Electric Power, Naha (Japan); Nishiyama, Kazuo [The Central Electric Power Council, Tokyo (Japan); Adachi, Kazuyuki [Kyushu Electric Power, Fukuoka (Japan)

    1998-09-01

    For the popularization of electric vehicles (EVs), the conditions for charging EV batteries with available current patterns should allow complete charging in a short time, i.e., less than 5 to 8 h. Therefore, in this study, a new charging condition is investigated for the EV valve-regulated lead/acid battery system, which should allow complete charging of EV battery systems with multi-step constant currents in a much shorter time with longer cycle life and higher energy efficiency compared with two-step constant-current charging. Although a high magnitude of the first current in the two-step constant-current method prolongs cycle life by suppressing the softening of positive active material, too large a charging current magnitude degrades cells due to excess internal evolution of heat. A charging current magnitude of approximately 0.5 C is expected to prolong cycle life further. Three-step charging could also increase the magnitude of charging current in the first step without shortening cycle life. Four-or six-step constant-current methods could shorten the charging time to less than 5 h, as well as yield higher energy efficiency and enhanced cycle life of over 400 cycles compared with two-step charging with the first step current of 0.5 C. Investigation of the degradation mechanism of the batteries revealed that the conditions of multi-step constant-current charging suppressed softening of positive active material and sulfation of negative active material, but, unfortunately, advanced the corrosion of the grids in the positive plates. By adopting improved grids and cooling of the battery system, the multistep constant-current method may enhance the cycle life. (orig.)

  4. 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

  5. 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...

  6. 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

  7. 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...

  8. 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.

  9. Surface profile gradient in amorphous Ta2O5 semi conductive layers regulates nanoscale electric current stability

    Science.gov (United States)

    Cefalas, A. C.; Kollia, Z.; Spyropoulos-Antonakakis, N.; Gavriil, V.; Christofilos, D.; Kourouklis, G.; Semashko, V. V.; Pavlov, V.; Sarantopoulou, E.

    2017-02-01

    A link between the morphological characteristics and the electric properties of amorphous layers is established by means of atomic, conductive, electrostatic force and thermal scanning microscopy. Using amorphous Ta2O5 (a-Ta2O5) semiconductive layer, it is found that surface profile gradients (morphological gradient), are highly correlated to both the electron energy gradient of trapped electrons in interactive Coulombic sites and the thermal gradient along conductive paths and thus thermal and electric properties are correlated with surface morphology at the nanoscale. Furthermore, morphological and electron energy gradients along opposite conductive paths of electrons intrinsically impose a current stability anisotropy. For either long conductive paths (L > 1 μm) or along symmetric nanodomains, current stability for both positive and negative currents i is demonstrated. On the contrary, for short conductive paths along non-symmetric nanodomains, the set of independent variables (L, i) is spanned by two current stability/intability loci. One locus specifies a stable state for negative currents, while the other locus also describes a stable state for positive currents.

  10. 24 CFR 280.25 - Other Federal requirements.

    Science.gov (United States)

    2010-04-01

    ... housing marketing requirements at 24 CFR part 200, subpart M, and the implementing regulations at 24 CFR part 108. (2) Racial and ethnic data collection requirements. Recipients must maintain current data...

  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. The effect of current military maintenance practices and regulations on the implementation of Integrated Vehicle Health Management technology

    OpenAIRE

    Esperon Miguez, Manuel; John, Philip; Ian K. Jennions

    2012-01-01

    Health monitoring tools can be used to diagnose failures and estimate the remaining useful life of certain components, generating information that can be used to improve the management of logistics and maintenance activities in what is known as Integrated Vehicle Health Management (IVHM). The work presented here analyzes the effect of military practices and regulations on the benefits that can be expected from installing health monitoring tools on military aircraft. The findings on the impact...

  13. 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.

  14. 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.

  15. 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...

  16. 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

  17. 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

  18. Serotonin Regulates the Firing of Principal Cells of the Subiculum by Inhibiting a T-type Ca(2+) Current

    DEFF Research Database (Denmark)

    Petersen, Anders V; Jensen, Camilla S; Crépel, Valérie

    2017-01-01

    and calcium imaging, we demonstrate that 5-HT2C receptors reduce bursting activity by inhibiting a low-threshold calcium current mediated by T-type Ca(2+) channels in principal cells of the subiculum. In addition, we show that the activation of this novel pathway decreases bursting activity and the occurrence...

  19. Region-specific regulation of voltage-gated intrinsic currents in the developing optic tectum of the Xenopus tadpole.

    Science.gov (United States)

    Hamodi, Ali S; Pratt, Kara G

    2014-10-01

    Across the rostrocaudal (RC) axis of the Xenopus tadpole optic tectum exists a developmental gradient. This gradient has served as a useful model to study many aspects of synapse and dendrite maturation. To compliment these studies, we characterized how the intrinsic excitability, the ease in which a neuron can fire action potentials, might also be changing across the same axis. Whole-cell recordings from tectal neurons at different points along the RC axis revealed a graded increase in intrinsic excitability: compared with neurons at the caudal end of the tectum, neurons at the rostral end fired more action potentials in response to current injection and expressed greater peak Na⁺ and K⁺ currents, the major intrinsic currents in these neurons that underlie the action potential. We also observed, along the same axis and in the same direction, a previously described increase in the amount of synaptic drive received by individual neurons (Wu GY, Malinow R, Cline HT. Science 274: 972-976, 1996). Thus as synaptic activity ramps up across the RC axis, so does intrinsic excitability. The reduction of overall circuit activity induced a compensatory scaling up of peak Na⁺ and K⁺ currents only in the caudal portion of the tectum, suggesting a region-specific, compensatory form of plasticity.

  20. 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.

  1. Guar gum and similar soluble fibers in the regulation of cholesterol metabolism: Current understandings and future research priorities

    Directory of Open Access Journals (Sweden)

    Todd C Rideout

    2008-10-01

    Full Text Available Todd C Rideout1, Scott V Harding1, Peter JH Jones1, Ming Z Fan21Richardson Centre for Functional Foods and Nutraceuticals, University of Manitoba, Winnipeg, Manitoba, Canada; 2Centre for Nutrition Modeling, Department of Animal and Poultry Science, University of Guelph, Guelph, Ontario, CanadaAbstract: The hypocholesterolemic effects associated with soluble fiber consumption are clear from animal model and human clinical investigations. Moreover, the modulation of whole-body cholesterol metabolism in response to dietary fiber consumption, including intestinal cholesterol absorption and fecal sterol and bile acid loss, has been the subject of many published reports. However, our understanding of how dietary fibers regulate molecular events at the gene/protein level and alter cellular cholesterol metabolism is limited. The modern emphasis on molecular nutrition and rapid progress in ‘high-dimensional’ biological techniques will permit further explorations of the role of genetic polymorphisms in determining the variable interindividual responses to soluble fibers. Furthermore, with traditional molecular biology tools and the application of ‘omic’ technology, specific insight into how fibers modulate the expression of genes and proteins that regulate intestinal cholesterol absorption and alter hepatic sterol balance will be gained. Detailed knowledge of the molecular mechanisms by which soluble fibers reduce plasma cholesterol concentrations is paramount to developing novel fiber-based “cocktails” that target specific metabolic pathways to gain maximal cholesterol reductions.Keywords: dietary fiber, cholesterol, bile acids, gene, protein

  2. 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...

  3. 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

  4. 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

  5. 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

  6. Improving self-regulation in adolescents: current evidence for the role of mindfulness-based cognitive therapy

    Science.gov (United States)

    Perry-Parrish, Carisa; Copeland-Linder, Nikeea; Webb, Lindsey; Shields, Ashley H; Sibinga, Erica MS

    2016-01-01

    Mindfulness-based cognitive therapy (MBCT) was introduced in 1995 to address the problem of recurrent depression. MBCT is based on the notion that meditation helps individuals effectively deploy and regulate attention to effectively manage and treat a range of psychological symptoms, including emotional responses to stress, anxiety, and depression. Several studies demonstrate that mindfulness approaches can effectively reduce negative emotional reactions that result from and/or exacerbate psychiatric difficulties and exposure to stressors among children, adolescents, and their parents. Mindfulness may be particularly relevant for youth with maladaptive cognitive processes such as rumination. Clinical experience regarding the utility of mindfulness-based approaches, including MBCT, is being increasingly supported by empirical studies to optimize the effective treatment of youth with a range of challenging symptoms. This paper provides a description of MBCT, including mindfulness practices, theoretical mechanisms of action, and targeted review of studies in adolescents. PMID:27695378

  7. The impact of Egypt's current challenges in adopting a conservative approach in its nuclear regulations regarding population considerations

    Directory of Open Access Journals (Sweden)

    Samia Wafik Morsy

    2015-10-01

    The paper outlines chronic problems the Egyptian population is suffering from in normal country state operation. It also examines the three major nuclear accidents and looks at the roles their governments played in protecting the population by defining the urgent actions such as sheltering, evacuation, distribution of iodine tablets and relocation. It argues that because developing countries are less able to deal with emergency crises as developed countries, a conservatory factor should increase distances away from the plant in nuclear regulations and reduce population numbers near the plant designated for developing countries. It is a simple way of protecting the population in developing countries where promptness for human rights of the citizens is not very much observed because of socio economic reasons.

  8. 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.

  9. 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...

  10. 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...

  11. Regulation of voltage-gated Ca(2+) currents by Ca(2+)/calmodulin-dependent protein kinase II in resting sensory neurons.

    Science.gov (United States)

    Kostic, Sandra; Pan, Bin; Guo, Yuan; Yu, Hongwei; Sapunar, Damir; Kwok, Wai-Meng; Hudmon, Andy; Wu, Hsiang-En; Hogan, Quinn H

    2014-09-01

    Calcium/calmodulin-dependent protein kinase II (CaMKII) is recognized as a key element in encoding depolarization activity of excitable cells into facilitated voltage-gated Ca(2+) channel (VGCC) function. Less is known about the participation of CaMKII in regulating VGCCs in resting cells. We examined constitutive CaMKII control of Ca(2+) currents in peripheral sensory neurons acutely isolated from dorsal root ganglia (DRGs) of adult rats. The small molecule CaMKII inhibitor KN-93 (1.0μM) reduced depolarization-induced ICa by 16-30% in excess of the effects produced by the inactive homolog KN-92. The specificity of CaMKII inhibition on VGCC function was shown by the efficacy of the selective CaMKII blocking peptide autocamtide-2-related inhibitory peptide in a membrane-permeable myristoylated form, which also reduced VGCC current in resting neurons. Loss of VGCC currents is primarily due to reduced N-type current, as application of mAIP selectively reduced N-type current by approximately 30%, and prior N-type current inhibition eliminated the effect of mAIP on VGCCs, while prior block of L-type channels did not reduce the effect of mAIP on total ICa. T-type currents were not affected by mAIP in resting DRG neurons. Transduction of sensory neurons in vivo by DRG injection of an adeno-associated virus expressing AIP also resulted in a loss of N-type currents. Together, these findings reveal a novel molecular adaptation whereby sensory neurons retain CaMKII support of VGCCs despite remaining quiescent.

  12. Electronic regulation of the SPS extraction quadrupole current pulse shape for improved stability of the extracted beam

    CERN Document Server

    Carlier, E; Vossenberg, Eugène B; CERN. Geneva. SPS and LEP Division

    1996-01-01

    In order to minimise the event pile-up and therefore optimise the detection efficiency, Chorus and Nomad experiments ask for a long and rectangular spill profile. At present the fast-slow extractio n is generated by driving the beam into a quadrupolar-octopolar resonance by exciting a quadrupole magnet with a semi-trapezoidal current [1]. The trapezoidal pulse shape is obtained by dischargin g a capacitor into the magnet coils. After a few milliseconds of undamped discharge a fixed resistor is switched into the circuit. The attenuation is then higher and the sine wave continues with a lower gradient. The two gradients can be adjusted by varying the initial capacitor voltage and the time at which the resistor is switched into the circuit. A further degree of freedom in determini ng the spill shape has been added by allowing the possibility of changing the second slope value independently of the initial conditions. This task is achieved by means of a variable current sour ce added in parallel to the fixed resis...

  13. 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...

  14. 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.

  15. Improving self-regulation in adolescents: current evidence for the role of mindfulness-based cognitive therapy

    Directory of Open Access Journals (Sweden)

    Perry-Parrish C

    2016-09-01

    Full Text Available Carisa Perry-Parrish,1 Nikeea Copeland-Linder,2 Lindsey Webb,2 Ashley Shields,1 Erica MS Sibinga2 1Department of Psychiatry and Behavioral Sciences, Division of Child and Adolescent Psychiatry, Pediatric Medical Psychology Program, 2Department of Pediatrics and Adolescent Medicine, Johns Hopkins School of Medicine, Baltimore, MD, USA Abstract: Mindfulness-based cognitive therapy (MBCT was introduced in 1995 to address the problem of recurrent depression. MBCT is based on the notion that meditation helps individuals effectively deploy and regulate attention to effectively manage and treat a range of psychological symptoms, including emotional responses to stress, anxiety, and depression. Several studies demonstrate that mindfulness approaches can effectively reduce negative emotional reactions that result from and/or exacerbate psychiatric difficulties and exposure to stressors among children, adolescents, and their parents. Mindfulness may be particularly relevant for youth with maladaptive cognitive processes such as rumination. Clinical experience regarding the utility of mindfulness-based approaches, including MBCT, is being increasingly supported by empirical studies to optimize the effective treatment of youth with a range of challenging symptoms. This paper provides a description of MBCT, including mindfulness practices, theoretical mechanisms of action, and targeted review of studies in adolescents. Keywords: youth, mindfulness-based cognitive therapy, adolescents

  16. Analysis on Current Laws and Regulations of Medical Device Clinical Trial in China%对我国现阶段医疗器械临床试验法规热点的解读

    Institute of Scientific and Technical Information of China (English)

    王悦; 李天萍; 梁宁霞

    2014-01-01

    A series of laws and regulations are the essential legal requirement in the field of clinical trial of medical device currently in China, especial y the Provision for Clinical Trial of Medical Device. On the basis of current situation of medical device clinical trial, systemic analysis on the hot spot topics in the regulations was conducted to explore the way of improving the control system of clinical trial of medical device in China, which wil provide the reference for medical device industry and the investigators of the clinical trial of medical device.%《医疗器械临床试验规定》等一系列法规是目前我国进行医疗器械临床试验的主要法律依据。该文基于我国医疗器械临床研究的现状,对这些法规中若干热点问题加以解读和剖析,旨在探索完善我国医疗器械临床试验监管体系的途径,为医疗器械业内人士及医疗器械临床试验的研究者和管理者提供参考。

  17. 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.

  18. 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.

  19. A review of the current geographic distribution of and debate surrounding electronic cigarette clean air regulations in the United States.

    Science.gov (United States)

    Kadowaki, Joy; Vuolo, Mike; Kelly, Brian C

    2015-01-01

    In this article, we present the results of a systematic review of state, county, and municipal restrictions on the use of electronic cigarettes (e-cigarettes) in public spaces within the United States, alongside an overview of the current legal landscape. The lack of federal guidance leaves lower-level jurisdictions to debate the merits of restrictions on use in public spaces without sufficient scientific research. As we show through a geographic assessment of restrictions, this has resulted in an inconsistent patchwork of e-cigarette use bans across the United States of varying degrees of coverage. Bans have emerged over time in a manner that suggests a "bottom up" diffusion of e-cigarette clean air policies. Ultimately, the lack of clinical and scientific knowledge on the risks and potential harm reduction benefits has led to precautionary policymaking, which often lacks grounding in empirical evidence and results in spatially uneven diffusion of policy.

  20. 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.

  1. 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.

  2. A Power Regulation and Droop Mode Control Method for a Stand-Alone Load Fed from a PV-Current Source Inverter

    Science.gov (United States)

    Khayamy, Mehdy; Ojo, Olorunfemi

    2015-04-01

    A current source inverter fed from photovoltaic cells is proposed to power an autonomous load when operating under either power regulation or voltage and frequency drooping modes. Input-output linearization technique is applied to the overall nonlinear system to achieve a globally stable system under feasible operating conditions. After obtaining the steady-state model that demarcates the modes of operation, computer Simulation results for variations in irradiance and the load power of the controlled system are generated in which an acceptable dynamic response of the power generator system under the two modes of operation is observed.

  3. Changes in neuronal excitability by activated microglia: Differential Na+ current up-regulation in pyramid-shaped and bipolar neurons by TNF-α and IL-18

    Directory of Open Access Journals (Sweden)

    Lars eKlapal

    2016-03-01

    Full Text Available Microglia are activated during pathological events in the brain and are capable of releasing various types of inflammatory cytokines. Here we demonstrate that the addition of 5% microglia activated by 1 µg/ml lipopolysaccharides (LPS to hippocampal cultures up-regulates Na+ current densities (INavD of bipolar as well as pyramid-shaped neurons, thereby increasing their excitability. Deactivation of microglia by the addition of 10 ng/ml transforming growth factor-β (TGF-β decreases INavD below control levels suggesting that the residual activated microglial cells influence neuronal excitability in control cultures. Preincubation of hippocampal cultures with 10 ng/ml tumor necrosis factor-α (TNF-α, a major cytokine released by activated microglia, up-regulated INavD significantly by ~30% in bipolar cells, whereas in pyramid-shaped cells the up-regulation only reached an increase of ~14%. Incubation of the cultures with antibodies against either TNF-receptor 1 or 2 blocked the up-regulation of INavD in bipolar cells, whereas in pyramid-shaped cells increases in INavD were exclusively blocked by antibodies against TNF-receptor 2, suggesting that both cell types respond differently to TNF-α exposure. Since additional cytokines, such as interleukin-18 (IL-18, are released from activated microglia we tested potential effects of IL-18 on INavD in both cell types. Exposure to 5-10 ng/ml IL-18 for 4 days increased INavD in both pyramid-shaped as well as bipolar neurons, albeit the dose-response curves were shifted to lower concentrations in bipolar cells. Our results suggest that by secretion of cytokines microglial cells up-regulate Na+ current densities in bipolar and pyramid-shaped neurons to some extent differentially. Depending on the exact cytokine composition and concentration released this could change the balance between the activity of inhibitory bipolar and excitatory pyramid-shaped cells. Since bipolar cells show a larger up-regulation of

  4. 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.

  5. 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.

  6. 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.

  7. 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...

  8. 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.

  9. Study of quality assurance regulations for linear accelerators in Korea: A comparison study between the current status in Korea and the international guidelines

    CERN Document Server

    Lee, Hyunho; Jo, Yunhui; Yoon, Myonggeun

    2015-01-01

    Quality assurance (QA) for medical linear accelerators is indispensable for appropriate cancer treatment. Some international organizations and western advanced countries provide QA guidelines for linear accelerators. Currently, QA regulations for linear accelerators in Korean hospitals specify a system in which each hospital stipulates its independent hospital-based protocols for QA procedures (HP_QAPs) and conducts QA based on these HP_QAPs while regulatory authorities verify whether items under these HP_QAPs have been performed. However, because this regulatory method cannot guarantee the quality of universal treatment, and QA items with tolerance criteria are different in many hospitals, the presentation of standardized QA items and tolerance criteria is essential. In this study, QA items in HP_QAPs from various hospitals and those presented by international organizations. Concordance rates between QA items for linear accelerators that were presented by the aforementioned organizations and those currently ...

  10. [Requirements for the prevention of nosocomial infections. German Guideline 2009 and reality. Current data from hospitals in Frankfurt am Main, Germany].

    Science.gov (United States)

    Heudorf, U; Exner, M

    2011-03-01

    In 2009, the new directive of the German Commission for Hospital Hygiene and Infection Prevention (Kommission für Krankenhaushygiene und Infektionsprävention, KRINKO) entitled Human and Organizational Requirements for the Prevention of Nosocomial Infections was published, including detailed information on the needs of hygiene professionals in hospital settings. Compared to the needs calculated according to the above policy, the current staff hygiene health professionals (HHPs) in the hospitals of Frankfurt am Main (Frankfurt/M), Germany, was on average 27.6%: 36% in the large hospitals (>600 beds), 21.6% in medium hospitals (300-600 beds), and 19.8% in small hospitals (hospitals had a full-time hygienist. The demands of the KRINKO policy have not been met by any of the hospitals. Hospitals with lower percentages of hospital hygiene staff not only had a lower rating of hygienic quality, they also showed a lower usage of hand disinfection per patient-day. In Germany, there is currently a lack of trained HHPs and hygienists to meet the needs of the KRINKO policy. Therefore, the reactions of the hospitals in Frankfurt/M ranged from the establishment of additional jobs for HHPs to changes in structures and organization of hospital hygiene. Thus, the new KRINKO guideline in Frankfurt/M did not result in a wave of recruitment of health professionals, but at least resulted in organizational and structural improvements in hygiene.

  11. Postnatal development of temporal integration, spike timing and spike threshold regulation by a dendrotoxin-sensitive K⁺ current in rat CA1 hippocampal cells.

    Science.gov (United States)

    Giglio, Anna M; Storm, Johan F

    2014-01-01

    Spike timing and network synchronization are important for plasticity, development and maturation of brain circuits. Spike delays and timing can be strongly modulated by a low-threshold, slowly inactivating, voltage-gated potassium current called D-current (ID ). ID can delay the onset of spiking, cause temporal integration of multiple inputs, and regulate spike threshold and network synchrony. Recent data indicate that ID can also undergo activity-dependent, homeostatic regulation. Therefore, we have studied the postnatal development of ID -dependent mechanisms in CA1 pyramidal cells in hippocampal slices from young rats (P7-27), using somatic whole-cell recordings. At P21-27, these neurons showed long spike delays and pronounced temporal integration in response to a series of brief depolarizing current pulses or a single long pulse, whereas younger cells (P7-20) showed shorter discharge delays and weak temporal integration, although the spike threshold became increasingly negative with maturation. Application of α-dendrotoxin (α-DTX), which blocks ID , reduced the spiking latency and temporal integration most strongly in mature cells, while shifting the spike threshold most strongly in a depolarizing direction in these cells. Voltage-clamp analysis revealed an α-DTX-sensitive outward current (ID ) that increased in amplitude during development. In contrast to P21-23, ID in the youngest group (P7-9) showed smaller peri-threshold amplitude. This may explain why long discharge delays and robust temporal integration only appear later, 3 weeks postnatally. We conclude that ID properties and ID -dependent functions develop postnatally in rat CA1 pyramidal cells, and ID may modulate network activity and plasticity through its effects on synaptic integration, spike threshold, timing and synchrony.

  12. Voltage-gated potassium currents are targets of diurnal changes in estradiol feedback regulation and kisspeptin action on gonadotropin-releasing hormone neurons in mice.

    Science.gov (United States)

    Pielecka-Fortuna, Justyna; DeFazio, R Anthony; Moenter, Suzanne M

    2011-11-01

    Estradiol has both negative and positive feedback actions upon gonadotropin-releasing hormone (GnRH) release; the latter actions trigger the preovulatory GnRH surge. Although neurobiological mechanisms of the transitions between feedback modes are becoming better understood, the roles of voltage-gated potassium currents, major contributors to neuronal excitability, are unknown. Estradiol alters two components of potassium currents in these cells: a transient current, I(A), and a sustained current, I(K). Kisspeptin is a potential mediator between estradiol and GnRH neurons and can act directly on GnRH neurons. We examined how estradiol, time of day, and kisspeptin interact to regulate these conductances in a mouse model exhibiting daily switches between estradiol negative (morning) and positive feedback (evening). Whole-cell voltage clamp recordings were made from GnRH neurons in brain slices from ovariectomized (OVX) mice and from OVX mice treated with estradiol (OVX+E). There were no diurnal changes in either I(A) or I(K) in GnRH neurons from OVX mice. In contrast, in GnRH neurons from OVX+E mice, I(A) and I(K) were greater during the morning when GnRH neuron activity is low and smaller in the evening when GnRH neuron activity is high. Estradiol increased I(A) in the morning and decreased it in the evening, relative to that in cells from OVX mice. Exogenously applied kisspeptin reduced I(A) regardless of time of day or estradiol status. Estradiol, interacting with time of day, and kisspeptin both depolarized I(A) activation. These findings extend our understanding of both the neurobiological mechanisms of estradiol negative vs. positive regulation of GnRH neurons and of kisspeptin action on these cells.

  13. User requirements and user acceptance of current and next-generation satellite mission and sensor complement, oriented toward the monitoring of water resources

    Science.gov (United States)

    Castruccio, P. A.; Loats, H. L., Jr.; Fowler, T. R.; Robinson, P.

    1975-01-01

    Principal water resources users were surveyed to determine the applicability of remotely sensed data to their present and future requirements. Analysis of responses was used to assess the levels of adequacy of LANDSAT 1 and 2 in fulfilling hydrological functions, and to derive systems specifications for future water resources-oriented remote sensing satellite systems. The analysis indicates that water resources applications for all but the very large users require: (1) resolutions on the order of 15 meters, (2) a number of radiometric levels of the same order as currently used in LANDSAT 1 (64), (3) a number of spectral bands not in excess of those used in LANDSAT 1, and (4) a repetition frequency on the order of 2 weeks. The users had little feel for the value of new sensors (thermal IR, passive and active microwaves). What is needed in this area is to achieve specific demonstrations of the utility of these sensors and submit the results to the users to evince their judgement.

  14. Current and proposed revisions, changes, and modifications to American codes and standards to address packaging, handling, and transportation of radioactive materials and how they relate to comparable international regulations

    Energy Technology Data Exchange (ETDEWEB)

    Borter, W.H.; Froehlich, C.H. [BNFL Fuel Solutions, Campbell, CA (United States)

    2004-07-01

    This paper addresses current and proposed revisions, additions, and modifications to American Society of Mechanical Engineers (ASME) Boiler and Pressure Vessel Code (BPVC) (i.e., ''ASMEthe Code'') Section III, Division 3 and American National Standards Institute (ANSI)/ASME N14.6. It provides insight into the ongoing processes of the associated committees and highlights important revisions, changes, and modifications to this Code and Standard. The ASME Code has developed and issued Division 3 to address items associated with the transportation and storage of radioactive materials. It currently only addresses ''General Requirements'' in Subsections WA and ''Class TP (Type B) Containments'' (Transportation Packages) in Subsection WB, but is in the process of adding a new Subsection WC to address ''Class SC'' (Storage Containments). ANSI/ASME Standard N14.6 which interacts with components constructed to Division 3 by addressinges special lifting devices for radioactive material shipping containers. This Standard is in the process of a complete re-write. This Code and Standard can be classified as ''dynamic'' in that their committees meet at least four times a year to evaluate proposed modifications and additions that reflect current safety practices in the nuclear industry. These evaluations include the possible addition of new materials, fabrication processes, examination methods, and testing requirements. An overview of this ongoing process is presented in this paper along with highlights of the more important proposed revisions, changes, and modifications and how they relate to United States (US) and international regulations and guidance like International Atomic Energy Agency (IAEA) Requirement No. TS-R-1.

  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. 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...

  17. 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...

  18. 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....

  19. 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...

  20. 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.

  1. 交流电机离散时间电流调节器的设计%Discrete-time Current Regulator Design for AC Machine

    Institute of Scientific and Technical Information of China (English)

    杨立永; 田安民

    2011-01-01

    分析了运行在较高基频与采样频率的比值下异步电机的离散时间电流调节器的特点,该情况普遍应用在高速列车驱动和大型牵引驱动上.若电流调节器的设计不能很好地满足控制系统离散特性,在较高的基频与采样频率比值下,系统会发生严重的振荡或者不稳定响应.通过一个含SVPWM延迟的离散时域交流电机模型,设计离散时域复矢量电流调节器.仿真和实验结果证明该调节器具有非常优越的稳定性和鲁棒性,充分满足了控制系统的离散特性.%The behaviors of discrete-time current regulators for AC machines operating at high ratios of fundamental-tosampling frequencies are analyzed, a situation common for high speed automotive drives and large traction drives. When the current regulator design does not properly incorporate the effects of the discrete nature of the controller, the highly oscillatory,or unstable response can occur,at high ratios of fundamental-to-sampling frequencies. As a part of this investigation,a discrete-time domain AC machine model is developed that includes the delays associated with SVPWM. This model is used to design a discrete-time domain version of the complex vector current regulator that demonstrates improved response compared with the other regulators studied. Simulation and the experimental results prove that the complex vector peoformance indice has very superior stability and robustness, fully meets the discrete control system.

  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. HSP90 Regulation of P2X7 Receptor Function Requires an Intact Cytoplasmic C-Terminus.

    Science.gov (United States)

    Migita, Keisuke; Ozaki, Taku; Shimoyama, Shuji; Yamada, Junko; Nikaido, Yoshikazu; Furukawa, Tomonori; Shiba, Yuko; Egan, Terrance M; Ueno, Shinya

    2016-08-01

    P2X7 receptors (P2X7Rs) are ATP-gated ion channels that display the unusual property of current facilitation during long applications of agonists. Here we show that facilitation disappears in chimeric P2X7Rs containing the C-terminus of the P2X2 receptor (P2X2R), and in a truncated P2X7R missing the cysteine-rich domain of the C-terminus. The chimeric and truncated receptors also show an apparent decreased permeability to N-methyl-d-glucamine(+) (NMDG(+)). The effects of genetic modification of the C-terminus on NMDG(+) permeability were mimicked by preapplication of the HSP90 antagonist geldanamycin to the wild-type receptor. Further, the geldanamycin decreased the shift in the reversal potential of the ATP-gated current measured under bi-ionic NMDG(+)/Na(+) condition without affecting the ability of the long application of agonist to facilitate current amplitude. Taken together, the results suggest that HSP90 may be essential for stabilization and function of P2X7Rs through an action on the cysteine-rich domain of the cytoplasmic the C-terminus.

  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. The up-regulation of voltage-gated sodium channels subtypes coincides with an increased sodium current in hippocampal neuronal culture model.

    Science.gov (United States)

    Guo, Feng; Xu, Xiaoxue; Cai, Jiqun; Hu, Huiyuan; Sun, Wei; He, Guilin; Shao, Dongxue; Wang, Lei; Chen, Tianbao; Shaw, Chris; Zhu, Tong; Hao, Liying

    2013-02-01

    Voltage-gated sodium channels (VGSC) have been linked to inherited forms of epilepsy. The expression and biophysical properties of VGSC in the hippocampal neuronal culture model have not been clarified. In order to evaluate mechanisms of epileptogenesis that are related to VGSC, we examined the expression and function of VGSC in the hippocampal neuronal culture model in vitro and spontaneously epileptic rats (SER) in vivo. Our data showed that the peak amplitude of transient, rapidly-inactivating Na(+) current (I(Na,T)) in model neurons was significantly increased compared with control neurons, and the activation curve was shifted to the negative potentials in model neurons in whole cell recording by patch-clamp. In addition, channel activity of persistent, non-inactivating Na(+) current (I(Na,P)) was obviously increased in the hippocampal neuronal culture model as judged by single-channel patch-clamp recording. Furthermore, VGSC subtypes Na(V)1.1, Na(V)1.2 and Na(V)1.3 were up-regulated at the protein expression level in model neurons and SER as assessed by Western blotting. Four subtypes of VGSC proteins in SER were clearly present throughout the hippocampus, including CA1, CA3 and dentate gyrus regions, and neurons expressing VGSC immunoreactivity were also detected in hippocampal neuronal culture model by immunofluorescence. These findings suggested that the up-regulation of voltage-gated sodium channels subtypes in neurons coincided with an increased sodium current in the hippocampal neuronal culture model, providing a possible explanation for the observed seizure discharge and enhanced excitability in epilepsy.

  12. Up-regulation of the transient A-type K+ current (IA) in the differentiation of neural stem cells of the early postnatal rat hippocampus

    Institute of Scientific and Technical Information of China (English)

    GUO Hong-bo; HUANG Lian-yan; ZOU Yu-xi; ZOU Fei

    2010-01-01

    Background Neural stem cells (NSCs) not only are essential to cell replacement therapy and transplantation in clinical settings, but also provide a unique model for the research into neurogenesis and epigenesis. However, little attention has been paid to the electrophysiological characterization of NSC development. This work aimed to identify whether the morphological neuronal differentiation process in NSCs included changes in the electrophysiological properties of transient A-type K+ currents (IA).Methods NSCs were isolated from early postnatal rat hippocampus and were multiplied in basic serum-free medium containing basic fibroblast growth factor. Potassium currents were investigated and compared using whole-cell patch-clamp techniques and one-way analysis of variance (ANOVA), respectively.Results Compared with NSC-derived neurons, cloned NSCs (cNSCs) had a more positive resting membrane potential, a higher input resistance, and a lower membrane capacitance. Part of cNSCs and NSC-derived neurons possessed both delayed-rectifier K+ currents (IDR) and IA, steady-state activation of IA in cNSCs (half-maximal activation at (21.34±4.37) mV) occurred at a more positive voltage than in NSC-derived neurons at 1-6 days in vitro (half-maximal activation at (12.85±4.19) mV).Conclusions Our research revealed a developmental up-regulation of the IA component during differentiation of postnatal NSCs. Together with the marked developmental up-regulation of IDR in vitro neuronal differentiation we have previously found, the voltage-gated potassium channels may participate in neuronal maturation process.

  13. 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.

  14. An APC:WNT counter-current-like mechanism regulates cell division along the colonic crypt axis: a mechanism that explains how APC mutations induce proliferative abnormalities that drive colon cancer development.

    Directory of Open Access Journals (Sweden)

    Bruce M Boman

    2013-11-01

    Full Text Available APC normally down-regulates WNT signaling in human colon, and APC mutations cause proliferative abnormalities in premalignant crypts leading to colon cancer, but the mechanisms are unclear at the level of spatial and functional organization of the crypt. Accordingly, we postulated a counter-current-like mechanism based on gradients of factors (APC;WNT that regulate colonocyte proliferation along the crypt axis. During crypt renewal, stem cells (SCs at the crypt bottom generate non-SC daughter cells that proliferate and differentiate while migrating upwards. The APC concentration is low at the crypt bottom and high at the top (where differentiated cells reside. WNT signaling, in contrast, is high at the bottom (where SCs reside and low at the top. Given that WNT and APC gradients are counter to one another, we hypothesized that a counter-current-like mechanism exists. Since both APC and WNT signaling components (e.g. survivin are required for mitosis, this mechanism establishes a zone in the lower crypt where conditions are optimal for maximal cell division and mitosis orientation (symmetric versus asymmetric. APC haploinsufficiency diminishes the APC gradient, shifts the proliferative zone upwards, and increases symmetric division, which causes SC overpopulation. In homozygote mutant crypts, these changes are exacerbated. Thus, APC-mutation-induced changes in the counter-current-like mechanism cause expansion of proliferative populations (SCs, rapidly-proliferating cells during tumorigenesis. We propose this mechanism also drives crypt fission, functions in the crypt cycle, and underlies adenoma development. Novel chemoprevention approaches designed to normalize the two gradients and readjust the proliferative zone downwards, might thwart progression of these premalignant changes.

  15. Digital constant current source based on the adjustable voltage regulator%基于可调稳压芯片的数控恒流源

    Institute of Scientific and Technical Information of China (English)

    苗新法

    2011-01-01

    Briefly discusses the design of DC constant current source, and discusses the existing devices, analyses their disadvantages,mainly introduces an efficient digital controlled constant current source which is based on adjustable voltage regulator. The schematic diagram of the circuit is given. The author introduces the principle of the system and the relation parameter in detail. Kernel model and feedback model are introduced from the point of view of system composition. The principle of the system is using the kernel chip's feedback pin, changing the parallel feedback to serial feedback. This system has function of current preset and low current ripple etc. An instrument developed by the author employs the constant current circuit designed in this paper and has got popular applications.%针对数控直流恒流源的设计进行了探讨,对比已有的恒流源实现方式,分析了其缺点,重点介绍1种高效率的基于可调稳压芯片的数控恒流源.给出了电路原理图,就整机工作原理、相关参数计算做了详细介绍.从系统组成角度分别介绍了核心模块、反馈模块及其实现方法.其基本原理是利用核心芯片的反馈引脚,把并联反馈改为串联反馈,从而实现恒流性能.系统具有输出电流可预置,输出纹波电流低等特性.采用设计的恒流源电路在自行研制的某仪器已推广应用.

  16. Cholesterol modulates the volume-regulated anion current in Ehrlich-Lettre ascites cells via effects on Rho and F-actin.

    Science.gov (United States)

    Klausen, Thomas Kjaer; Hougaard, Charlotte; Hoffmann, Else K; Pedersen, Stine F

    2006-10-01

    The mechanisms controlling the volume-regulated anion current (VRAC) are incompletely elucidated. Here, we investigate the modulation of VRAC by cellular cholesterol and the potential involvement of F-actin, Rho, Rho kinase, and phosphatidylinositol-(4,5)-bisphosphate [PtdIns(4,5)P(2)] in this process. In Ehrlich-Lettre ascites (ELA) cells, a current with biophysical and pharmacological properties characteristic of VRAC was activated by hypotonic swelling. A 44% increase in cellular cholesterol content had no detectable effects on F-actin organization or VRAC activity. A 47% reduction in cellular cholesterol content increased cortical and stress fiber-associated F-actin content in swollen cells. Cholesterol depletion increased VRAC activation rate and maximal current after a modest (15%), but not after a severe (36%) reduction in extracellular osmolarity. The cholesterol depletion-induced increase in maximal VRAC current was prevented by F-actin disruption using latrunculin B (LB), while the current activation rate was unaffected by LB, but dependent on Rho kinase. Rho activity was decreased by approximately 20% in modestly, and approximately 50% in severely swollen cells. In modestly swollen cells, this reduction was prevented by cholesterol depletion, which also increased isotonic Rho activity. Thrombin, which stimulates Rho and causes actin polymerization, potentiated VRAC in modestly swollen cells. VRAC activity was unaffected by inclusion of a water-soluble PtdIns(4,5)P(2) analogue or a PtdIns(4,5)P(2)-blocking antibody in the pipette, or neomycin treatment to sequester PtdIns(4,5)P(2). It is suggested that in ELA cells, F-actin and Rho-Rho kinase modulate VRAC magnitude and activation rate, respectively, and that cholesterol depletion potentiates VRAC at least in part by preventing the hypotonicity-induced decrease in Rho activity and eliciting actin polymerization.

  17. 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.

  18. Differential regulation of voltage-gated Ca2+ currents and metabotropic glutamate receptor activity by measles virus infection in rat cortical neurons.

    Science.gov (United States)

    Günther, Christine; Laube, Mandy; Liebert, Uwe-Gerd; Kraft, Robert

    2012-01-06

    Measles virus (MV) infection may lead to severe chronic CNS disease processes, including MV-induced encephalitis. Because the intracellular Ca(2+) concentration ([Ca(2+)](i)) is a major determinant of the (patho-)physiological state in all cells we asked whether important Ca(2+) conducting pathways are affected by MV infection in cultured cortical rat neurons. Patch-clamp measurements revealed a decrease in voltage-gated Ca(2+) currents during MV-infection, while voltage-gated K(+) currents and NMDA-evoked currents were unaffected. Calcium-imaging experiments using 50mM extracellular KCl showed reduced [Ca(2+)](i) increases in MV-infected neurons, confirming a decreased activity of voltage-gated Ca(2+) channels. In contrast, the group-I metabotropic glutamate receptor (mGluR) agonist DHPG evoked changes in [Ca(2+)](i) that were increased in MV-infected cells. Our results show that MV infection conversely regulates Ca(2+) signals induced by group-I mGluRs and by voltage-gated Ca(2+) channels, suggesting that these physiological impairments may contribute to an altered function of cortical neurons during MV-induced encephalitis.

  19. 75 FR 58387 - Federal Acquisition Regulation; Information Collection; American Recovery and Reinvestment Act...

    Science.gov (United States)

    2010-09-24

    ... Regulation; Information Collection; American Recovery and Reinvestment Act-Reporting Requirements--One-Time... an extension of a currently approved information collection requirement concerning the American Recovery and Reinvestment Act-Reporting Requirements--One Time- Reporting, Compensation...

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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

  7. The amplitude and inactivation properties of the delayed potassium currents are regulated by protein kinase activity in hair cells of the frog semicircular canals.

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    Marta Martini

    Full Text Available In hair cells dissected from the frog crista ampullaris, the combination of a calcium-dependent (IKCa and a purely voltage-dependent component (IKV gives rise to the delayed potassium current complex (IKD. These currents have been recently reported to display slow depolarization-induced inactivation and biphasic inactivation removal by hyperpolarization. The amplitude and inactivation kinetics of both IKCa and IKV are drastically modulated by a previously unrecognized mechanism of protein phosphorylation (sensitive to kinase inhibitors H89 and KT5823, which does not interfere with the transient potassium current (IA or the calcium current (ICa. IKD amplitude was stable in cells patched with pipettes containing 8 mM ATP or under perforated-patch; under these conditions, a 10 min treatment with 10 µM H89 or 1-10 µM KT5823 reduced IKD amplitude by a mean of 67% at +40 mV. Similarly affected was the isolated IKV component (ICa blocked with Cd(2+. Thus, a large potassium conductance can be activated by depolarization, but it is made available to the cell to a variable extent that depends on membrane potential and protein kinase activity. The total gKD ranged 4.6-44.0 nS in control cells, according to the level of steady-state inactivation, and was reduced to 1.4-2.7 nS after protein kinase inhibition. When sinusoidal membrane potential changes in the -70/-10 mV range were applied, to mimic receptor response to hair bundle deflection, IKD proved the main current dynamically activated and the only one regulated by PK: H89 decreased the total outward charge during each cycle by 60%. Phosphorylation appears to control both the amount of IKCa and IKV conductance activated by depolarization and the fraction thereof which can be rescued by removal of inactivation. The balance between the depolarizing transduction current and the repolarizing potassium current, and eventually the transmitter release at the cytoneural junction, are therefore modulated by a

  8. Choral Conducting Studies and Tools Required for the Development of Children´s Choirs. Current Situation in Costa Rica and Spain

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    Carlos Fernando Chaves-Cordero

    2016-12-01

    Full Text Available This qualitative research seeks to understand the current situation of choral conducting studies in Costa Rica and Spain, emphasizing on the tools required to work with children´s choirs. The curriculum of choral conducting careers at the Universidad Nacional de Costa Rica and the Real Conservatorio Superior de Música de Madrid was studied by means of an analysis of documents and an analytic comparison with the SWOT method. The study of plans primarily focused on subjects related to pedagogy and singing as essential foundations in the specific area of children’s choirs. Additionally, an analysis of interviews to choral directors from both countries visualized the experiences of directors in their performance with children´s choirs, what skills were acquired in their undergraduate education, as well their perception of what should change or adjust in the curriculum. Significant shortcomings were perceived in teacher’s training in relation with the subjects of the career. A strengthening with more contents in teaching methodologies and pedagogical resources was suggested.

  9. 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

  10. Conforming with current regulation in Turkey regarding the freezing of oocytes: A case report of the first pregnancy in Turkey achieved through oocyte vitrification

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    Semra Kahraman MD

    2017-01-01

    Full Text Available Objectives: To present the first pregnancy achieved in Turkey with frozen–warmed oocytes in a case with previous nine unsuccessful assisted reproductive technology (ART attempts. Methods: The clinical follow-up of a 33-year-old female applying to our ART centre after a long and complicated history of infertility is described. Results: In April 2013, the woman attempted our centre for her 10th ART trial. She informed us on oocyte pick-up (OPU day that her husband had been hospitalized following a car crush in Albania and was unable to travel to our clinic to give a sperm sample. We were therefore placed in the position of having to make an emergency decision. OPU was done and seven oocytes were retrieved. Six metaphase II (MII oocytes out of seven Cumulus Oocyte Complexes (COCs were vitrified using the Kitazato Vitrification Cryotop Kit. Six months later, in November 2013, the patient applied for transfer. Two blastocysts were transferred and the ART trial resulted with a singleton pregnancy and the birth of a healthy new-born at term via cesarean section. Conclusion: Regulation Codes on Assisted Reproductive Procedures and Assisted Reproductive Technology Centres, published in the Official Gazette of the Republic of Turkey, on 6 March 2010 forbade the freezing of gonad cells and tissues except when essential for medical reasons and stated that this would be specified later. However, the Regulation Codes published in the Official Gazette of the Republic of Turkey, on 30 September 2014 provided no further clarification. Unfortunately, the wording of the regulations did not specifically address this unexpected emergency situation. However, we saw our decision to cryopreserve the oocytes as a valid interpretation of the regulations, bearing in mind also the requirement that sperm and oocyte in the IVF process must be those of a married couple. Turkish medicolegal regulations should be revised to increase the chances of more women taking advantage

  11. Conforming with current regulation in Turkey regarding the freezing of oocytes: A case report of the first pregnancy in Turkey achieved through oocyte vitrification

    Science.gov (United States)

    Kahraman, Semra; Ulker, Kahraman; Pirkevi Çetinkaya, Caroline; Kumtepe Colakoglu, Yesim; Yelke, Hakan K.; Yilanlioglu, Cihangir

    2017-01-01

    Objectives: To present the first pregnancy achieved in Turkey with frozen–warmed oocytes in a case with previous nine unsuccessful assisted reproductive technology (ART) attempts. Methods: The clinical follow-up of a 33-year-old female applying to our ART centre after a long and complicated history of infertility is described. Results: In April 2013, the woman attempted our centre for her 10th ART trial. She informed us on oocyte pick-up (OPU) day that her husband had been hospitalized following a car crush in Albania and was unable to travel to our clinic to give a sperm sample. We were therefore placed in the position of having to make an emergency decision. OPU was done and seven oocytes were retrieved. Six metaphase II (MII) oocytes out of seven Cumulus Oocyte Complexes (COCs) were vitrified using the Kitazato Vitrification Cryotop Kit. Six months later, in November 2013, the patient applied for transfer. Two blastocysts were transferred and the ART trial resulted with a singleton pregnancy and the birth of a healthy new-born at term via cesarean section. Conclusion: Regulation Codes on Assisted Reproductive Procedures and Assisted Reproductive Technology Centres, published in the Official Gazette of the Republic of Turkey, on 6 March 2010 forbade the freezing of gonad cells and tissues except when essential for medical reasons and stated that this would be specified later. However, the Regulation Codes published in the Official Gazette of the Republic of Turkey, on 30 September 2014 provided no further clarification. Unfortunately, the wording of the regulations did not specifically address this unexpected emergency situation. However, we saw our decision to cryopreserve the oocytes as a valid interpretation of the regulations, bearing in mind also the requirement that sperm and oocyte in the IVF process must be those of a married couple. Turkish medicolegal regulations should be revised to increase the chances of more women taking advantage of the procedure

  12. Regulated expression of HCN channels and cAMP levels shape the properties of the h current in developing rat hippocampus.

    Science.gov (United States)

    Surges, Rainer; Brewster, Amy L; Bender, Roland A; Beck, Heinz; Feuerstein, Thomas J; Baram, Tallie Z

    2006-07-01

    The hyperpolarization-activated current (I(h)) contributes to intrinsic properties and network responses of neurons. Its biophysical properties depend on the expression profiles of the underlying hyperpolarization-activated, cyclic nucleotide-gated (HCN) channels and the presence of cyclic AMP (cAMP) that potently and differentially modulates I(h) conducted by HCN1, HCN2 and/or HCN4. Here, we studied the properties of I(h) in hippocampal CA1 pyramidal cells, the developmental evolution of the HCN-subunit isoforms that contribute to this current, and their interplay with age-dependent free cAMP concentrations, using electrophysiological, molecular and biochemical methods. I(h) amplitude increased progressively during the first four postnatal weeks, consistent with the observed overall increased expression of HCN channels. Activation kinetics of the current accelerated during this period, consonant with the quantitative reduction of mRNA and protein expression of the slow-kinetics HCN4 isoform and increased levels of HCN1. The sensitivity of I(h) to cAMP, and the contribution of the slow component to the overall I(h), decreased with age. These are likely a result of the developmentally regulated transition of the complement of HCN channel isoforms from cAMP sensitive to relatively cAMP insensitive. Thus, although hippocampal cAMP concentrations increased over twofold during the developmental period studied, the coordinated changes in expression of three HCN channel isoforms resulted in reduced effects of this signalling molecule on neuronal h currents.

  13. 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

  14. 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.

  15. 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.

  16. 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.

  17. A dual action of saturated fatty acids on electrical activity in rat pancreatic β-cells. Role of volume-regulated anion channel and KATP channel currents.

    Science.gov (United States)

    Best, L; Jarman, E; Brown, P D

    2011-03-15

    Free fatty acids (FFAs) exert complex actions on pancreatic β-cells. Typically, an initial potentiation of insulin release is followed by a gradual impairment of β-cell function, the latter effect being of possible relevance to hyperlipidaemia in type 2 diabetes mellitus. The molecular actions of FFAs are poorly understood. The present study investigated the acute effects of saturated FFAs on electrophysiological responses of rat pancreatic β-cells. Membrane potential and KATP channel activity were recorded using the perforated patch technique. Volume-regulated anion channel (VRAC) activity was assessed from conventional whole-cell recordings. Cell volume regulation was measured using a video-imaging technique. Addition of octanoate caused a transient potentiation of glucose-induced electrical activity, followed by a gradual hyper-polarisation and a prolonged inhibition of electrical activity. Octanoate caused an initial increase in VRAC activity followed by a secondary inhibition coinciding with increased KATP channel activity. Similar effects were observed with palmitate and 2-bromopalmitate whereas butyrate was virtually ineffective. Octanoate and palmitate also exerted a dual effect on electrical activity evoked by tolbutamide. Octanoate significantly attenuated cell volume regulation in hypotonic solutions, consistent with VRAC inhibition. It is concluded that medium and long chain FFAs have a dual action on glucose-induced electrical activity in rat pancreatic β-cells: an initial stimulatory effect followed by a secondary inhibition. These effects appear to be the result of reciprocal actions on VRAC and KATP channel currents, and could contribute towards the stimulatory and inhibitory actions of FFAs on pancreatic β-cell function.

  18. The Carboxy-terminus of BAK1 regulates kinase activity and is required for normal growth of Arabidopsis

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    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.

  19. 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.

  20. A Myb transcription factor of Phytophthora sojae, regulated by MAP kinase PsSAK1, is required for zoospore development.

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    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.

  1. 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

  2. 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.

  3. 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.

  4. 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.

  5. 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

  6. Review of current severe accident management approaches in Europe and identification of related modelling requirements for the computer code ASTEC V2.1

    Energy Technology Data Exchange (ETDEWEB)

    Hermsmeyer, S. [European Commission JRC, Petten (Netherlands). Inst. for Energy and Transport; Herranz, L.E.; Iglesias, R. [CIEMAT, Madrid (Spain); and others

    2015-07-15

    The severe accident at the Fukushima-Daiichi nuclear power plant (NPP) has led to a worldwide review of nuclear safety approaches and is bringing a refocussing of R and D in the field. To support these efforts several new Euratom FP7 projects have been launched. The CESAM project focuses on the improvement of the ASTEC computer code. ASTEC is jointly developed by IRSN and GRS and is considered as the European reference code for Severe Accident Analyses since it capitalizes knowledge from the extensive Euro-pean R and D in the field. The project aims at the code's enhancement and extension for use in Severe Accident Management (SAM) analysis of the NPPs of Generation II-III presently under operation or foreseen in the near future in Europe, spent fuel pools included. The work reported here is concerned with the importance, for the further development of the code, of SAM strategies to be simulated. To this end, SAM strategies applied in the EU have been compiled. This compilation is mainly based on the public information made available in the frame of the EU ''stress tests'' for NPPs and has been complemented by information pro-vided by the different CESAM partners. The context of SAM is explained and the strategies are presented. The modelling capabilities for the simulation of these strategies in the current production version 2.0 of ASTEC are discussed. Furthermore, the requirements for the next version of ASTEC V2.1 that is supported in the CESAM project are highlighted. They are a necessary complement to the list of code improvements that is drawn from consolidating new fields of application, like SFP and BWR model enhancements, and from new experimental results on severe accident phenomena.

  7. Role of tumor necrosis factor-αin the regulation of T-type calcium channel current in HL-1 cells

    Institute of Scientific and Technical Information of China (English)

    RAO Fang; SHAN Zhi-xin; ZHU Jie-ning; XIE Zhi; WU Shu-lin; DENG Chun-yu; XUE Yu-mei; YU Xi-yong; WEI Wei; LIU Fang-zhou; YANG Hui; KUANG Su-juan; CHEN Shao-xian; XIAO Ding-zhang

    2016-01-01

    AIM:Increasing evidence indicates that inflammation contributes to the initiation and perpetuation of atrial fibrillation ( AF) .Al-though tumor necrosis factor ( TNF)-αlevels are increased in patients with AF , the role of TNF-αin the pathogenesis of AF remains unclear.Recent research has revealed that T-type Ca2+currents ( ICa,T ) play an important role in the pathogenesis of AF .METH-ODS:In this study , we used the whole-cell voltage-clamp technique and biochemical assays to explore the role of TNF-αin the regula-tion of ICa,T in atrial myocytes.RESULTS:We found that compared with sinus rhythm (SR) controls, T-type calcium channel (TCC) subunit mRNA levels were decreased , while TNF-αexpression levels were increased , in human atrial tissue from patients with AF .In murine atrial myocyte HL-1 cells, after cultured for 24 h, 12.5, 25 and 50 μg/L TNF-αsignificantly reduced the protein expression levels of the TCC α1G subunit in a concentration-dependent manner .The peak current was reduced by the application of 12.5 or 25μg/L TNF-αin a concentration-dependent manner [from ( -15.08 ±1.11) pA/pF in controls to ( -11.89 ±0.83) pA/pF and (-8.54 ±1.55) pA/pF in 12.5 and 25 μg/L TNF-αgroups, respectively].TNF-αapplication also inhibited voltage-dependent inactivation of ICa,T shifted the inactivation curve to the left .CONCLUSION:These results suggest that TNF-αis involved in the path-ogenesis of AF, probably via decreasing ICa,T function in atrium-derived myocytes through impaired channel function and down -regula-tion of channel protein expression .This pathway thus represents a potential pathogenic mechanism in AF .

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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

  16. 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.

  17. 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.

  18. 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

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. TtsI, a key regulator of Rhizobium species NGR234 is required for type III-dependent protein secretion and synthesis of rhamnose-rich polysaccharides.

    Science.gov (United States)

    Marie, Corinne; Deakin, William J; Ojanen-Reuhs, Tuula; Diallo, Ericka; Reuhs, Brad; Broughton, William J; Perret, Xavier

    2004-09-01

    Formation of nitrogen-fixing nodules on legume roots by Rhizobium sp. NGR234 requires an array of bacterial factors, including nodulation outer proteins (Nops) secreted through a type III secretion system (TTSS). Secretion of Nops is abolished upon inactivation of ttsI (formerly y4xI), a protein with characteristics of two-component response regulators that was predicted to activate transcription of TTSS-related genes. During the symbiotic interaction, the phenotype of NGR omega ttsI differs from that of a mutant with a nonfunctional secretion machine, however. This indicated that TtsI regulates the synthesis of other symbiotic factors as well. Conserved sequences, called tts boxes, proposed to act as binding sites for TtsI, were identified not only within the TTSS cluster but also in the promoter regions of i) genes predicted to encode homologs of virulence factors secreted by pathogenic bacteria, ii) loci involved in the synthesis of a rhamnose-rich component (rhamnan) of the lipopolysaccharides (LPS), and iii) open reading frames that play roles in plasmid partitioning. Transcription studies showed that TtsI and tts boxes are required for the activation of TTSS-related genes and those involved in rhamnose synthesis. Furthermore, extraction of polysaccharides revealed that inactivation of ttsI abolishes the synthesis of the rhamnan component of the LPS. The phenotypes of mutants impaired in TTSS-dependent protein secretion, rhamnan synthesis, or in both functions were compared to assess the roles of some of the TtsI-controlled factors during symbiosis.

  5. Regulation of TRP-like muscarinic cation current in gastrointestinal smooth muscle with special reference to PLC/InsP3/Ca2+ system

    Institute of Scientific and Technical Information of China (English)

    Alexander V ZHOLOS

    2006-01-01

    Acetylcholine,the main enteric excitatory neuromuscular transmitter,evokes membrane depolarization and contraction of gastrointestinal smooth muscle cells by activating G protein-coupled muscarinic receptors.Although the cholinergic excitation is generally underlined by the multiplicity of ion channel effects,the primary event appears to be the opening of cation-selective channels;among them the 60 pS channel has been recently identified as the main target for the acetylcholine action in gastrointestinal myocytes.The evoked cation current,termed mICAT,causes either an oscillatory or a more sustained membrane depolarization response,which in turn leads to increases of the open probability of voltage-gated Ca2+ channels.thus providing Ca2+ entry in parallel with Ca2+ release for intracellular Ca2+ concentration rise and contraction.In recent years there have been several significant developments in our understanding of the signaling processes underlying mIcAT generation.They have revealed important synergistic interactions between M2 and M3 receptor subtypes,single channel mechanisms,and the involvement of TRPC-encoded proteins as essential components of native muscarinic cation channels.This review summarizes these recent findings and in particular discusses the roles of the phospholipase C/InsP3/intracellular Ca2+ release system in the mICAT physiological regulation.

  6. Activation of H2O2-induced VSOR Cl- currents in HTC cells require phospholipase Cgamma1 phosphorylation and Ca2+ mobilisation

    DEFF Research Database (Denmark)

    Varela, Diego; Simon, Felipe; Olivero, Pablo

    2007-01-01

    Volume-sensitive outwardly rectifying (VSOR) Cl(-) channels participate in several physiological processes such as regulatory volume decrease, cell cycle regulation, proliferation and apoptosis. Recent evidence points to a significant role of hydrogen peroxide (H(2)O(2)) in VSOR Cl(-) channel act...

  7. The Regulation of Technology-Assisted Distance Counseling and Supervision in the United States: An Analysis of Current Extent, Trends, and Implications

    Science.gov (United States)

    McAdams, Charles R.; Wyatt, Kristi Lee

    2010-01-01

    Counseling licensure boards report emerging needs to regulate technology-assisted distance counseling and supervision. An analysis of published regulations and telephone interviews with board administrators nationwide suggests that boards agree generally on 7 aspects of technology-assisted distance practice that need to be regulated. Nevertheless,…

  8. Optimal screening of children with acute malnutrition requires a change in current WHO guidelines as MUAC and WHZ identify different patient groups

    DEFF Research Database (Denmark)

    Laillou, Arnaud; Prak, Sophonneary; de Groot, Richard;

    2014-01-01

    BACKGROUND: Timely treatment of acute malnutrition in children 500,000 deaths annually. Screening at community level is essential to identify children with malnutrition. Current WHO guidelines for community screening for malnutrition recommend a Mid Upper Arm...

  9. Tonic 5nM DA stabilizes neuronal output by enabling bidirectional activity-dependent regulation of the hyperpolarization activated current via PKA and calcineurin.

    Directory of Open Access Journals (Sweden)

    Wulf-Dieter C Krenz

    Full Text Available Volume transmission results in phasic and tonic modulatory signals. The actions of tonic dopamine (DA at type 1 DA receptors (D1Rs are largely undefined. Here we show that tonic 5nM DA acts at D1Rs to stabilize neuronal output over minutes by enabling activity-dependent regulation of the hyperpolarization activated current (I h. In the presence but not absence of 5nM DA, I h maximal conductance (G max was adjusted according to changes in slow wave activity in order to maintain spike timing. Our study on the lateral pyloric neuron (LP, which undergoes rhythmic oscillations in membrane potential with depolarized plateaus, demonstrated that incremental, bi-directional changes in plateau duration produced corresponding alterations in LP I hG max when preparations were superfused with saline containing 5nM DA. However, when preparations were superfused with saline alone there was no linear correlation between LP I hGmax and duty cycle. Thus, tonic nM DA modulated the capacity for activity to modulate LP I h G max; this exemplifies metamodulation (modulation of modulation. Pretreatment with the Ca2+-chelator, BAPTA, or the specific PKA inhibitor, PKI, prevented all changes in LP I h in 5nM DA. Calcineurin inhibitors blocked activity-dependent changes enabled by DA and revealed a PKA-mediated, activity-independent enhancement of LP I hG max. These data suggested that tonic 5nM DA produced two simultaneous, PKA-dependent effects: a direct increase in LP I h G max and a priming event that permitted calcineurin regulation of LP I h. The latter produced graded reductions in LP I hG max with increasing duty cycles. We also demonstrated that this metamodulation preserved the timing of LP's first spike when network output was perturbed with bath-applied 4AP. In sum, 5nM DA permits slow wave activity to provide feedback that maintains spike timing, suggesting that one function of low-level, tonic modulation is to stabilize specific features of a dynamic output.

  10. Tonic 5nM DA stabilizes neuronal output by enabling bidirectional activity-dependent regulation of the hyperpolarization activated current via PKA and calcineurin.

    Science.gov (United States)

    Krenz, Wulf-Dieter C; Rodgers, Edmund W; Baro, Deborah J

    2015-01-01

    Volume transmission results in phasic and tonic modulatory signals. The actions of tonic dopamine (DA) at type 1 DA receptors (D1Rs) are largely undefined. Here we show that tonic 5nM DA acts at D1Rs to stabilize neuronal output over minutes by enabling activity-dependent regulation of the hyperpolarization activated current (I h). In the presence but not absence of 5nM DA, I h maximal conductance (G max) was adjusted according to changes in slow wave activity in order to maintain spike timing. Our study on the lateral pyloric neuron (LP), which undergoes rhythmic oscillations in membrane potential with depolarized plateaus, demonstrated that incremental, bi-directional changes in plateau duration produced corresponding alterations in LP I hG max when preparations were superfused with saline containing 5nM DA. However, when preparations were superfused with saline alone there was no linear correlation between LP I hGmax and duty cycle. Thus, tonic nM DA modulated the capacity for activity to modulate LP I h G max; this exemplifies metamodulation (modulation of modulation). Pretreatment with the Ca2+-chelator, BAPTA, or the specific PKA inhibitor, PKI, prevented all changes in LP I h in 5nM DA. Calcineurin inhibitors blocked activity-dependent changes enabled by DA and revealed a PKA-mediated, activity-independent enhancement of LP I hG max. These data suggested that tonic 5nM DA produced two simultaneous, PKA-dependent effects: a direct increase in LP I h G max and a priming event that permitted calcineurin regulation of LP I h. The latter produced graded reductions in LP I hG max with increasing duty cycles. We also demonstrated that this metamodulation preserved the timing of LP's first spike when network output was perturbed with bath-applied 4AP. In sum, 5nM DA permits slow wave activity to provide feedback that maintains spike timing, suggesting that one function of low-level, tonic modulation is to stabilize specific features of a dynamic output.

  11. Optimal screening of children with acute malnutrition requires a change in current WHO guidelines as MUAC and WHZ identify different patient groups

    DEFF Research Database (Denmark)

    Laillou, Arnaud; Prak, Sophonneary; de Groot, Richard

    2014-01-01

    BACKGROUND: Timely treatment of acute malnutrition in children 500,000 deaths annually. Screening at community level is essential to identify children with malnutrition. Current WHO guidelines for community screening for malnutrition recommend a Mid Upper Arm...... Circumference (MUAC) of malnutrition (SAM). However, it is currently unclear how MUAC relates to the other indicator used to define acute malnutrition: weight-for-height Z-score (WHZ). METHODS: Secondary data from >11,000 Cambodian children, obtained by different surveys between...... 2010 and 2012, was used to calculate sensitivity and ROC curves for MUAC and WHZ. FINDINGS: The secondary analysis showed that using the current WHO cut-off of 115 mm for screening for severe acute malnutrition over 90% of children with a weight-for-height z-score (WHZ)

  12. Requirements of Multiple Domains of SLI-1, a Caenorhabditis elegans Homologue of c-Cbl, and an Inhibitory Tyrosine in LET-23 in Regulating Vulval Differentiation

    Science.gov (United States)

    Yoon, Charles H.; Chang, Chieh; Hopper, Neil A.; Lesa, Giovanni M.; Sternberg, Paul W.

    2000-01-01

    SLI-1, a Caenorhabditis elegans homologue of the proto-oncogene product c-Cbl, is a negative regulator of LET-23-mediated vulval differentiation. Lack of SLI-1 activity can compensate for decreased function of the LET-23 epidermal growth factor receptor, the SEM-5 adaptor, but not the LET-60 RAS, suggesting that SLI-1 acts before RAS activation. SLI-1 and c-Cbl comprise an N-terminal region (termed SLI-1:N/Cbl-N, containing a four-helix bundle, an EF hand calcium-binding domain, and a divergent SH2 domain) followed by a RING finger domain and a proline-rich C-terminus. In a transgenic functional assay, the proline-rich C-terminal domain is not essential for sli-1(+) function. A protein lacking the SH2 and RING finger domains has no activity, but a chimeric protein with the SH2 and RING finger domains of SLI-1 replaced by the equivalent domains of c-Cbl has activity. The RING finger domain of c-Cbl has been shown recently to enhance ubiquitination of active RTKs by acting as an E3 ubiquitin–protein ligase. We find that the RING finger domain of SLI-1 is partially dispensable. Further, we identify an inhibitory tyrosine of LET-23 requiring sli-1(+) for its effects: removal of this tyrosine closely mimics the loss of sli-1 but not of another negative regulator, ark-1. Thus, we suggest that this inhibitory tyrosine mediates its effects through SLI-1, which in turn inhibits signaling upstream of LET-60 RAS in a manner not wholly dependent on the ubiquitin–ligase domain. PMID:11071924

  13. A proline-rich sequence unique to MEK1 and MEK2 is required for raf binding and regulates MEK function.

    Science.gov (United States)

    Catling, A D; Schaeffer, H J; Reuter, C W; Reddy, G R; Weber, M J

    1995-10-01

    Mammalian MEK1 and MEK2 contain a proline-rich (PR) sequence that is absent both from the yeast homologs Ste7 and Byr1 and from a recently cloned activator of the JNK/stress-activated protein kinases, SEK1/MKK4. Since this PR sequence occurs in MEKs that are regulated by Raf family enzymes but is missing from MEKs and SEKs activated independently of Raf, we sought to investigate the role of this sequence in MEK1 and MEK2 regulation and function. Deletion of the PR sequence from MEK1 blocked the ability of MEK1 to associate with members of the Raf family and markedly attenuated activation of the protein in vivo following growth factor stimulation. In addition, this sequence was necessary for efficient activation of MEK1 in vitro by B-Raf but dispensable for activation by a novel MEK1 activator which we have previously detected in fractionated fibroblast extracts. Furthermore, we found that a phosphorylation site within the PR sequence of MEK1 was required for sustained MEK1 activity in response to serum stimulation of quiescent fibroblasts. Consistent with this observation, we observed that MEK2, which lacks a phosphorylation site at the corresponding position, was activated only transiently following serum stimulation. Finally, we found that deletion of the PR sequence from a constitutively activated MEK1 mutant rendered the protein nontransforming in Rat1 fibroblasts. These observations indicate a critical role for the PR sequence in directing specific protein-protein interactions important for the activation, inactivation, and downstream functioning of the MEKs.

  14. The ChrSA and HrrSA Two-Component Systems Are Required for Transcriptional Regulation of the hemA Promoter in Corynebacterium diphtheriae

    Science.gov (United States)

    Burgos, Jonathan M.

    2016-01-01

    ABSTRACT Corynebacterium diphtheriae utilizes heme and hemoglobin (Hb) as iron sources for growth in low-iron environments. In C. diphtheriae, the two-component signal transduction systems (TCSs) ChrSA and HrrSA are responsive to Hb levels and regulate the transcription of promoters for hmuO, hrtAB, and hemA. ChrSA and HrrSA activate transcription at the hmuO promoter and repress transcription at hemA in an Hb-dependent manner. In this study, we show that HrrSA is the predominant repressor at hemA and that its activity results in transcriptional repression in the presence and absence of Hb, whereas repression of hemA by ChrSA is primarily responsive to Hb. DNA binding studies showed that both ChrA and HrrA bind to the hemA promoter region at virtually identical sequences. ChrA binding was enhanced by phosphorylation, while binding to DNA by HrrA was independent of its phosphorylation state. ChrA and HrrA are phosphorylated in vitro by the sensor kinase ChrS, whereas no kinase activity was observed with HrrS in vitro. Phosphorylated ChrA was not observed in vivo, even in the presence of Hb, which is likely due to the instability of the phosphate moiety on ChrA. However, phosphorylation of HrrA was observed in vivo regardless of the presence of the Hb inducer, and genetic analysis indicates that ChrS is responsible for most of the phosphorylation of HrrA in vivo. Phosphorylation studies strongly suggest that HrrS functions primarily as a phosphatase and has only minimal kinase activity. These findings collectively show a complex mechanism of regulation at the hemA promoter, where both two-component systems act in concert to optimize expression of heme biosynthetic enzymes. IMPORTANCE Understanding the mechanism by which two-component signal transduction systems function to respond to environmental stimuli is critical to the study of bacterial pathogenesis. The current study expands on the previous analyses of the ChrSA and HrrSA TCSs in the human pathogen C

  15. CYB5D2 requires heme-binding to regulate HeLa cell growth and confer survival from chemotherapeutic agents.

    Directory of Open Access Journals (Sweden)

    Anthony Bruce

    Full Text Available The cytochrome b5 domain containing 2 (CYB5D2; Neuferricin protein has been reported to bind heme, however, the critical residues responsible for heme-binding are undefined. Furthermore, the relationship between heme-binding and CYB5D2-mediated intracellular functions remains unknown. Previous studies examining heme-binding in two cytochrome b5 heme-binding domain-containing proteins, damage-associated protein 1 (Dap1; Saccharomyces cerevisiae and human progesterone receptor membrane component 1 (PGRMC1, have revealed that conserved tyrosine (Y 73, Y79, aspartic acid (D 86, and Y127 residues present in human CYB5D2 may be involved in heme-binding. CYB5D2 binds to type b heme, however, only the substitution of glycine (G at D86 (D86G within its cytochrome b5 heme-binding (cyt-b5 domain abolished its heme-binding ability. Both CYB5D2 and CYB5D2(D86G localize to the endoplasmic reticulum. Ectopic CYB5D2 expression inhibited cell proliferation and anchorage-independent colony growth of HeLa cells. Conversely, CYB5D2 knockdown and ectopic CYB5D2(D86G expression increased cell proliferation and colony growth. As PGRMC1 has been reported to regulate the expression and activities of cytochrome P450 proteins (CYPs, we examined the role of CYB5D2 in regulating the activities of CYPs involved in sterol synthesis (CYP51A1 and drug metabolism (CYP3A4. CYB5D2 co-localizes with cytochrome P450 reductase (CYPOR, while CYB5D2 knockdown reduced lanosterol demethylase (CYP51A1 levels and rendered HeLa cells sensitive to mevalonate. Additionally, knockdown of CYB5D2 reduced CYP3A4 activity. Lastly, CYB5D2 expression conferred HeLa cell survival from chemotherapeutic agents (paclitaxel, cisplatin and doxorubicin, with its ability to promote survival being dependent on its heme-binding ability. Taken together, this study provides evidence that heme-binding is critical for CYB5D2 in regulating HeLa cell growth and survival, with endogenous CYB5D2 being required to

  16. 75 FR 58388 - Federal Acquisition Regulation; Information Collection; American Recovery and Reinvestment Act...

    Science.gov (United States)

    2010-09-24

    ... Regulation; Information Collection; American Recovery and Reinvestment Act-Reporting Requirements--Quarterly... an extension of a currently approved information collection requirement concerning the American Recovery and Reinvestment Act--Quarterly Reporting for Prime Contractors. Public comments are...

  17. Iron-regulated metabolites produced by Pseudomonas fluorescens WCS374r are not required for eliciting induced systemic resistance against Pseudomonas syringae pv. tomato in Arabidopsis.

    Science.gov (United States)

    Djavaheri, Mohammad; Mercado-Blanco, Jesús; Versluis, C; Meyer, J-M; Loon, L C; Bakker, Peter A H M

    2012-09-01

    The plant growth-promoting rhizobacterium Pseudomonas fluorescens WCS374r produces several iron-regulated metabolites, including the fluorescent siderophore pseudobactin (Psb374), salicylic acid (SA), and pseudomonine (Psm), a siderophore that contains a SA moiety. After purification of Psb374 from culture supernatant of WCS374r, its structure was determined following isoelectrofocusing and tandem mass spectrometry, and found to be identical to the fluorescent siderophore produced by P. fluorescens ATCC 13525. To study the role of SA and Psm production in colonization of Arabidopsis thaliana roots and in induced systemic resistance (ISR) against Pseudomonas syringae pv. tomato (Pst) by strain WCS374r, mutants disrupted in the production of these metabolites were obtained by homologous recombination. These mutants were further subjected to transposon Tn5 mutagenesis to generate mutants also deficient in Psb374 production. The mutants behaved similar to the wild type in both their Arabidopsis rhizosphere-colonizing capacity and their ability to elicit ISR against Pst. We conclude that Psb374, SA, and Psm production by P. fluorescens WCS374r are not required for eliciting ISR in Arabidopsis.

  18. Efficient design and simulation of an expandable hybrid (wind-photovoltaic) power system with MPPT and inverter input voltage regulation features in compliance with electric grid requirements

    Energy Technology Data Exchange (ETDEWEB)

    Skretas, Sotirios B.; Papadopoulos, Demetrios P. [Electrical Machines Laboratory, Department of Electrical and Computer Engineering, Democritos University of Thrace (DUTH), 12 V. Sofias, 67100 Xanthi (Greece)

    2009-09-15

    In this paper an efficient design along with modeling and simulation of a transformer-less small-scale centralized DC - bus Grid Connected Hybrid (Wind-PV) power system for supplying electric power to a single phase of a three phase low voltage (LV) strong distribution grid are proposed and presented. The main components of the hybrid system are: a PV generator (PVG); and an array of horizontal-axis, fixed-pitch, small-size, variable-speed wind turbines (WTs) with direct-driven permanent magnet synchronous generator (PMSG) having an embedded uncontrolled bridge rectifier. An overview of the basic theory of such systems along with their modeling and simulation via Simulink/MATLAB software package are presented. An intelligent control method is applied to the proposed configuration to simultaneously achieve three desired goals: to extract maximum power from each hybrid power system component (PVG and WTs); to guarantee DC voltage regulation/stabilization at the input of the inverter; to transfer the total produced electric power to the electric grid, while fulfilling all necessary interconnection requirements. Finally, a practical case study is conducted for the purpose of fully evaluating a possible installation in a city site of Xanthi/Greece, and the practical results of the simulations are presented. (author)

  19. Human regulator of telomere elongation helicase 1 (RTEL1) is required for the nuclear and cytoplasmic trafficking of pre-U2 RNA.

    Science.gov (United States)

    Schertzer, Michael; Jouravleva, Karina; Perderiset, Mylene; Dingli, Florent; Loew, Damarys; Le Guen, Tangui; Bardoni, Barbara; de Villartay, Jean-Pierre; Revy, Patrick; Londoño-Vallejo, Arturo

    2015-02-18

    Hoyeraal-Hreidarsson syndrome (HHS) is a severe form of Dyskeratosis congenita characterized by developmental defects, bone marrow failure and immunodeficiency and has been associated with telomere dysfunction. Recently, mutations in Regulator of Telomere ELongation helicase 1 (RTEL1), a helicase first identified in Mus musculus as being responsible for the maintenance of long telomeres, have been identified in several HHS patients. Here we show that RTEL1 is required for the export and the correct cytoplasmic trafficking of the small nuclear (sn) RNA pre-U2, a component of the major spliceosome complex. RTEL1-HHS cells show abnormal subcellular partitioning of pre-U2, defects in the recycling of ribonucleotide proteins (RNP) in the cytoplasm and splicing defects. While most of these phenotypes can be suppressed by re-expressing the wild-type protein in RTEL1-HHS cells, expression of RTEL1 mutated variants in immortalized cells provokes cytoplasmic mislocalizations of pre-U2 and other RNP components, as well as splicing defects, thus phenocopying RTEL1-HHS cellular defects. Strikingly, expression of a cytoplasmic form of RTEL1 is sufficient to correct RNP mislocalizations both in RTEL1-HHS cells and in cells expressing nuclear mutated forms of RTEL1. This work unravels completely unanticipated roles for RTEL1 in RNP trafficking and strongly suggests that defects in RNP biogenesis pathways contribute to the pathology of HHS. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

  20. Enterohemorrhagic Escherichia coli O157:H7 requires quorum sensing transcriptional regulators QseA and SdiA for colonization and persistence in the bovine intestinal tract

    Science.gov (United States)

    QseA and SdiA are two of several transcriptional regulators that regulate virulence gene expression of enterohemorrhagic Escherichia coli (EHEC) O157:H7 via quorum sensing (QS). QseA regulates the expression of the locus of enterocyte effacement (LEE). LEE encodes for a type III secretion (T3S) sys...