WorldWideScience

Sample records for channel maintenance activities

  1. Forecasts of the Maintenance Activity

    Directory of Open Access Journals (Sweden)

    Marius Romocea

    2017-11-01

    Full Text Available Enhancing the machines’ technical and technological performance through constructive improvements that allow for the automation of orders, contributes to the increasing of the production capacity of the system and for maintaining an optimal level of product costs. In order to achieve these goals, it is necessary to organize the maintenance work well, in order to quickly diagnose the malfunctions and to shorten the repair time. Given the scale of the phenomena, this paper seeks to answer two questions, namely: how will be the maintenance in future and also how to prepare this activity.

  2. Activities of maintenance and outage

    International Nuclear Information System (INIS)

    Gracia-Orellan, J. M.; Gonzalez, P. L.; Verdu, M. F.; Fernandez, J. A.

    2004-01-01

    Iberinco Nuclear Generation Department have wanted to promote service activities in nuclear power plants for years besides its dedication to engineering activities. for it, in 1997 Nuclear Services Management was created to complement engineering activities and to be able to make an offer for products and turn key services. People involved in this Management have an extensive experience in Services Area, so that all type of maintenance works are promoted, as well other services like dismantling, fallout management and new equipment's for nuclear power plants services. Iberinco's experience in Support Services in Nuclear Power Plants allows to answer effectively to special workers during operation cycle and outages periods. These activities have been made in spanish nuclear power plants and Angra I and II plants, both of them in Brazil. In this area we provide Technical Consulting Management and Supervision to develop the following activities: - Improvement Maintenance Programs based in PSA: Implantation of Maintenance Rule in the plants. - Supervision and Assembly of design modifications in structures, systems and components. - Fulfilment of efficiency tests, inspection and turbo-group modification. - Noise and vibrations analysis. - Valves and rotative equipment calculations and diagnosis tests. Iberinco develop these outage activities itself or as contractors coordinator under its management. A lot of them have been working with Iberinco for many years and have a great experience in the Service Area they are developing. In this article, the main outage activities developed for Iberinco are detailed. (Author)

  3. Predictive techniques for river channel evolution and maintenance

    Science.gov (United States)

    Nelson, J.M.

    1996-01-01

    Predicting changes in alluvial channel morphology associated with anthropogenic and natural changes in flow and/or sediment supply is a critical part of the management of riverine systems. Over the past few years, advances in the understanding of the physics of sediment transport in conjunction with rapidly increasing capabilities in computational fluid dynamics have yielded now approaches to problems in river mechanics. Techniques appropriate for length scales ranging from reaches to bars and bedforms are described here. Examples of the use of these computational approaches are discussed for three cases: (1) the design of diversion scenarios that maintain channel morphology in steep cobble-bedded channels in Colorado, (2) determination of channel maintenance flows for the preservation of channel islands in the Snake River in Idaho, and (3) prediction of the temporal evolution of deposits in lateral separation zones for future assessment of the impacts of various dam release scenarios on lateral separation deposits in the Colorado River in Grand Canyon. With continued development of their scientific and technical components, the methodologies described here can provide powerful tools for the management of river environments in the future.

  4. A Dynamic Policy for Grouping Maintenance Activities

    NARCIS (Netherlands)

    R.E. Wildeman (Ralph); R. Dekker (Rommert); A.C.J.M. Smit

    1995-01-01

    textabstractA maintenance activity carried out on a technical system often involves a system-dependent set-up cost that is the same for all maintenance activities carried out on that system. Grouping activities thus saves costs since execution of a group of activities requires only one set-up. Many

  5. Inspection of licensee - Maintenance programme and activities

    International Nuclear Information System (INIS)

    2013-01-01

    An effective maintenance programme is critical to sustained safe and reliable operation of nuclear power plants. The Working Group on Inspection Practices (WGIP) concluded that when a licensee has an effective maintenance programme, the overall operating safety of the plant is improved and the protection of public health and safety enhanced. All Regulatory Bodies (RB) consider maintenance to be an important area for oversight. Although a variety of inspection practices are being used; RB are actively monitoring licensee performance. Specifically the following conclusions were reached and commendable practices identified: - Maintenance oversight by regulators appears to be in a stable continuous improvement state. Most regulators are executing inspection oversight based on an existing regulatory framework. - The performance of a licensee's maintenance programme is recognized as important part of maintaining nuclear safety. The result of the maintenance program assessment is included in the overall performance assessment of a license. - Maintenance inspection activities are recognized as an important part of the regulatory oversight process. Inspection activities are based on the safety significance and nature of work being performed by the licensee. - The effectiveness of the maintenance inspection activities is recognized to rely on properly qualified inspectors; who are adequately supported by specialists. Training and qualification of inspectors should be based on how the RB reviews and inspects licensee maintenance programmes. - Reporting requirements are identified to provide information on the licensees maintenance programme, and to help guide inspection activities. - Performance Indicators are recognized as a useful tool for helping focus regulatory activities. Basic PI are identified and tracked by the RB, and use of PI by the licensee is monitored. - Inspections are designed to confirm that the licensee is planning and scheduling maintenance with due

  6. New activities in nuclear power plants maintenance

    International Nuclear Information System (INIS)

    Mico, F.; Specht, J.

    2006-01-01

    This article addresses the consequences of the progressive deregulation of the electricity market of several key countries on the maintenance practices and techniques face to the growing pressures on O and M costs. The responses of the nuclear maintenance industry to make maintenance activities more efficient maintaining if not enhancing the safety of NPP's are briefly described. The article also analyses the potential impact of shortage of experience maintenance personnel to serve the NPP's in the future as some countries are already experiencing. (Author)

  7. Overview of maintenance principles and regulatory supervision of maintenance activities at nuclear power plants in Slovakia

    International Nuclear Information System (INIS)

    Rohar, S.; Cepcek, S.

    1997-01-01

    The maintenance represents one of the most important tools to ensure safe and reliable operation of nuclear power plants. The emphasis of Nuclear Regulatory Authority of the Slovak Republic to the maintenance issue is expressed by requirements in the regulations. The current practice of maintenance management in operated nuclear power plants in Slovak Republic is presented. Main aspects of maintenance, as maintenance programme, organization of maintenance, responsibilities for maintenance are described. Activities of nuclear regulatory authority in maintenance process are presented too. (author)

  8. Maintenance of an obstruction-forced pool in a gravel-bed channel: streamflow, channel morphology, and sediment transport.

    Science.gov (United States)

    Richard D. Woodsmith; Marwan A. Hassan

    2005-01-01

    Maintenance of pool morphology in a stream channel with a mobile bed requires hydraulic conditions at moderate to high flows that route bed load through the pool as it is delivered from upstream. Through field measurements of discharge, vertical velocity profiles, bed load transport, and streambed scour, fill, and grain-size distribution, we found that maintenance of a...

  9. Channel Bank Cohesion and the Maintenance of Suspension Rivers

    Science.gov (United States)

    Dunne, K. B. J.; Jerolmack, D. J.

    2017-12-01

    Gravel-bedded rivers organize their channel geometry and grain size such that transport is close to the threshold of motion at bankfull. Sand-bedded rivers, however, typically maintain bankfull fluid shear (or Shields) stresses far in excess of threshold; there is no widely accepted explanation for these "suspension rivers". We propose that all alluvial rivers are at the threshold of motion for their erosion-limiting material, i.e., the structural component of the river cross-section that is most difficult to mobilize. The entrainment threshold of gravel is large enough that bank cohesion has little influence on gravel-bed rivers. Sand, however, is the most easily entrained material; silt and clay can raise the entrainment threshold of sand by orders of magnitude. We examine a global dataset of river channel geometry and show that the shear stress range for sand-bedded channels is entirely within the range of entrainment thresholds for sand-mud mixtures - suggesting that rivers that suspend their sandy bed material are still threshold rivers in terms of bank material. We then present new findings from a New Jersey coastal-plain river examining if and how river-bank toe composition controls hydraulic geometry. We consider the toe because it is the foundation of the river bank, and its erosion leads to channel widening. Along a 20-km profile of the river we measure cross-section geometry, bed slope, and bed and bank composition, and we explore multiple methods of measuring the threshold shear stress of the the river-bank toe in-situ. As the composition of the river bed transitions from gravel to sand, we see preliminary evidence of a shift from bed-threshold to bank-threshold control on hydraulic geometry. We also observe that sub-bankfull flows are insufficient to erode (cohesive) bank materials, even though transport of sand is active at nearly all flows. Our findings highlight the importance of focusing on river-bank toe material, which in the studied stream is

  10. Laboratory simulation of maintenance activity

    International Nuclear Information System (INIS)

    Kantowitz, B.H.

    1988-01-01

    Laboratory research in highly controlled settings can augment, but not replace, studies in plant or training center locations. A laboratory simulation involves abstraction of the human information processing and social interactions required in prototypical maintenance tasks. A variety of independent variables can be studied quickly, efficiently, and at relatively low cost. Sources of human error can be identified in terms of models of human perception, cognition, action, attention, and social/organizational processes. This paper discusses research in progress at the Battelle Human Performance Laboratory. Both theoretical aspects and practical implications are considered. Directions for future human factors research are indicated

  11. Environmental favourable foundries through maintenance activities

    Directory of Open Access Journals (Sweden)

    A. Saniuk

    2015-10-01

    Full Text Available Maintenance today contributes to the aim of sustainable development in society, including environmental and energy saving, safety and economical aspects. The main purpose of the paper is to present the role of maintenance in the realization of sustainable developing practices in a foundry industry. Maintenance offers numerous opportunities to reduce the influence of foundry processes on the natural environment and utilize resources more efficiently. Maintenance activities can help to reduce losses and thereby improve the efficiency of the processes used in the production of resources (e.g. raw materials, energy, etc., especially in areas such as: the choice of an appropriate strategy for the maintenance of machinery and equipment (e.g. reactive, preventive, proactive; inventory management of spare parts; lubrication management and management of technology media.

  12. Fuel channel life limiting factors that dictate fuel channel maintenance requirements

    International Nuclear Information System (INIS)

    Richinson, P.J.; Wong, H.W.; Ellis, P.J.

    1995-01-01

    CANDU reactors have been operating for 33 years. The Nuclear Power Demonstration (NPD) Unit started up in 1962 and the prototype of CANDU, Douglas Point, started in 1967. The first commercial reactors, Pickering Units 1 and 2 both went into service in 1971 closely followed by Units 3 and 4 in 1972 and 1973 respectively. Operating commercial reactor experience represents over 10,000 pressure tubes, not including the replaced channels in all the Pickering A Units, and nearly 130,000 pressure tube operating years. No pressure tube has yet operated for its 30 year design lifetime of 210 KEFPH at 80% capacity factor. The longest operating time for pressure tubes to-date is about 120 KEFPH in Pickering Unit 4. Many lessons have been learned regarding pressure tube life limiting factors from the early CANDU units and these, together with the information obtained from an extensive pressure tube R and D program, have resulted in many design changes and improvements in material properties, mainly from manufacturing route changes. Reactors built recently are expected to achieve their 30 year design life. The development of Periodic and In-service Inspection programs and equipment, assessment methodologies and acceptance criteria, and the development of maintenance tooling and procedures are enabling the life limiting factors to be addressed in the currently operating units. The life limiting factors in currently operating Units are reviewed in relation to the experience gained from the early units, the R and D programs and the inspection and maintenance performed to date. (author)

  13. Management of maintenance activities at Cernavoda NPP

    International Nuclear Information System (INIS)

    Riedel, M.; Urjan, R.; Marinescu, S.; Corlan, L.

    2000-01-01

    After the Construction and Commissioning phases are over, the safety and economic performances of each nuclear unit are determined by the quality of its operation as well as by the combination of reliability and availability of the systems consisting the whole unit. Therefore, the value of a good maintenance program in making a safe and profitable operation is now widely recognized; all stations operating with excellent performance have well-defined policies and effectively administered programs to govern maintenance activities. The Cernavoda maintenance optimization initiative is derived from good practices suggested by such recognized bodies as the IAEA, INPO and EPRI. It should be noted that the program is evolving at this time and that this paper is an attempt to summarize the myriad of activities and plans required to finally implement the maintenance program. Although many of the elements of the program are in place, many are in various stages of completion. The main components of the program (policy's objectives, strategies used, specific subprograms, maintenance plans and their integration) are presented in brief along with the performance indicators used to assess the effectiveness of the program and maintenance activities. The new programs intended to be developed are also specified, as well as the main comments and recommendations received from IAEA experts. (author)

  14. Building Maintenance. Math Learning Activity Packet.

    Science.gov (United States)

    Grant, Shelia I.

    This collection of learning activities is intended for use in reinforcing mathematics instruction as it relates to building maintenance. Fifty activity sheets are provided. These are organized into units on the following topics: numeration, adding whole numbers, subtracting whole numbers, multiplying whole numbers, dividing whole numbers,…

  15. Regulatory oversight of maintenance activities at nuclear power plants

    International Nuclear Information System (INIS)

    Pape, M.

    1997-01-01

    Regulation of nuclear safety in the UK is based on monitoring of compliance with licence conditions. This paper discusses legislation aspects, license conditions, license requirements for maintenance and maintenance activities in the UK. It also addresses the regulator utility interaction, the regulatory inspection of maintenance and the trends in maintenance. (author)

  16. Regulatory control of maintenance activities in Argentine nuclear power plants

    International Nuclear Information System (INIS)

    Calvo, J.C.; Caruso, G.

    2000-01-01

    The main maintenance objective is to assure that the safety features of structures, components and systems of nuclear power plants are kept as designed. Therefore, there is a direct relationship between safety and maintenance. Owing to the above mentioned, maintenance activities are considered a relevant regulatory issue for the Argentine Nuclear Regulatory Authority (ARN). This paper describes the regulatory control to maintenance activities of Argentine nuclear power plants. It also addresses essential elements for maintenance control, routine inspections, special inspections during planned outages, audits and license conditions and requirements. (author)

  17. Activation of TRPV1 channels inhibits mechanosensitive Piezo channel activity by depleting membrane phosphoinositides

    Science.gov (United States)

    Borbiro, Istvan; Badheka, Doreen; Rohacs, Tibor

    2015-01-01

    Capsaicin is an activator of the heat-sensitive TRPV1 (transient receptor potential vanilloid 1) ion channels and has been used as a local analgesic. We found that activation of TRPV1 channels with capsaicin either in dorsal root ganglion neurons or in a heterologous expression system inhibited the mechanosensitive Piezo1 and Piezo2 channels by depleting phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] and its precursor PI(4)P from the plasma membrane through Ca2+-induced phospholipase Cδ (PLCδ) activation. Experiments with chemically inducible phosphoinositide phosphatases and receptor-induced activation of PLCβ indicated that inhibition of Piezo channels required depletion of both PI(4)P and PI(4,5)P2. The mechanically activated current amplitudes decreased substantially in the excised inside-out configuration, where the membrane patch containing Piezo1 channels is removed from the cell. PI(4,5)P2 and PI(4)P applied to these excised patches inhibited this decrease. Thus, we concluded that Piezo channel activity requires the presence of phosphoinositides, and the combined depletion of PI(4,5)P2 or PI(4)P reduces channel activity. In addition to revealing a role for distinct membrane lipids in mechanosensitive ion channel regulation, these data suggest that inhibition of Piezo2 channels may contribute to the analgesic effect of capsaicin. PMID:25670203

  18. Study on application of safety checklist in preventive maintenance activities

    International Nuclear Information System (INIS)

    Shi Jin; Chen Song; Liu Jingquan

    2013-01-01

    The paper describes the principles and the characteristics of safety checklist as a risk evaluation method. Examples of application of safety checklists to preventive maintenance activities such as criteria comparison and checkup items in place in nuclear power plants are illustrated in details with issues appeared in the checklist establishment. Checklist has a good application in the RCM analysis or in the actual preventive maintenance program for Chashma Nuclear Power Plant indicated by concrete instances. In the light of safety checklist which is used to sustain preventive maintenance as a simple and applicable risk analysis approach, we can get deep knowledge of risks of nuclear power plant to perfect preventive maintenance activities. (authors)

  19. Performing maintenance activities on-line a risk perspective

    International Nuclear Information System (INIS)

    Darling, S.S.; Mairs, T.P.

    1993-01-01

    The US nuclear power industry is attempting to deal with the issue of optimizing the performance of maintenance activities with the advent of the maintenance rule. The impact of any maintenance activity on the safe operation of a nuclear power facility can be quantified on the basis of risk. Therefore, it is incumbent upon the industry to accommodate the regulator and present a maintenance optimization program in terms that explicitly address the potential risk inherent in its concept and implementation. The motivation for utilities to perform maintenance while the plant is operating (i.e., on-line) is readily perceived to offer the following advantages: 1. greater system availability; 2. higher plant capacity factor; 3. more evenly distributed workload for the maintenance department; and 4. overall safer plant operation. However, risks inherent in undertaking maintenance activities during power operation could amount to the realization that reapportionment of these tasks is not cost-beneficial. Therefore, it is important that any utility considering implementing such a paradigm in maintenance philosophy establish a rigorous process to select, perform, and monitor maintenance tasks performed while operating

  20. Building Maintenance. Reading and Language Activities.

    Science.gov (United States)

    Kessman, William A.

    The student manual has been designed primarily for beginning students in a high school level vocational program in building maintenance. A workbook for special needs students, the document does not cover all areas of custodial training but has been written for the purpose of refining the basic skills of reading comprehension, vocabulary building,…

  1. Autonomous and professional maintenance in metallurgical enterprise as activities within total productive maintenance

    Directory of Open Access Journals (Sweden)

    B. Gajdzik

    2014-04-01

    Full Text Available The content of this publication consists of notions connected with Total Productive Maintenance (TPM in metallurgical enterprise. The basic areas of devices condition management through Autonomous and Professional Maintenance are described here. Mentioned areas of activities are performed in metallurgical enterprise ArcelorMittal Poland within pillars of World Class Manufacturing (WCM. The aims of UR programs are to maintain the basic functionality of the devices and decrease the number of failures in order to reach improvement of production efficiency.

  2. 76 FR 50489 - Agency Information Collection Activities: Harbor Maintenance Fee

    Science.gov (United States)

    2011-08-15

    ... Activities: Harbor Maintenance Fee AGENCY: U.S. Customs and Border Protection, Department of Homeland... Fee (CBP Forms 349 and 350). This is a proposed extension of an information collection that was... Fee. OMB Number: 1651-0055. Form Number: CBP Forms 349 and 350. Abstract: The Harbor Maintenance Fee...

  3. Heat sink considerations during SLAR defuelling activities of the 1995 maintenance outage at the Point Lepreau G.S

    International Nuclear Information System (INIS)

    Richard, R.A.; Edgar, D.J.; Gibb, R.A.

    1996-01-01

    In order to conduct the Spacer Locate and Reposition (SLAR) activities, individual fuel channels were de-fuelled using the SLAR tool itself. The presence of the tool in the channel containing fuel caused concerns over channel flow interruption. A methodology was devised to assess the limited fuel heat up during de-fuelling and during possible prolonged periods of complete flow interruption. Once established, the approach could be used to assess the effect of varying parameters which could effect channel heat-up. Costly delays in SLAR maintenance activities were avoided by ensuring the sequence of channel visits was such that the individual channel powers decayed sufficiently to prevent channel heat up beyond acceptable temperature limits. (author)

  4. Risk mitigation strategies for operations and maintenance activities.

    Science.gov (United States)

    2012-04-01

    The objective of this research was to investigate the application of integrated risk modeling to operations and maintenance activities, specifically moving operations, such as pavement testing, pavement marking, painting, snow removal, shoulder work,...

  5. Human Error Probability Assessment During Maintenance Activities of Marine Systems

    OpenAIRE

    Rabiul Islam; Faisal Khan; Rouzbeh Abbassi; Vikram Garaniya

    2018-01-01

    Background: Maintenance operations on-board ships are highly demanding. Maintenance operations are intensive activities requiring high man–machine interactions in challenging and evolving conditions. The evolving conditions are weather conditions, workplace temperature, ship motion, noise and vibration, and workload and stress. For example, extreme weather condition affects seafarers' performance, increasing the chances of error, and, consequently, can cause injuries or fatalities to personne...

  6. KCNQ4 channel activation by BMS-204352 and retigabine

    DEFF Research Database (Denmark)

    Schrøder, Rikke Louise K.; Jespersen, Thomas; Christophersen, P

    2001-01-01

    Activation of potassium channels generally reduces cellular excitability, making potassium channel openers potential drug candidates for the treatment of diseases related to hyperexcitabilty such as epilepsy, neuropathic pain, and neurodegeneration. Two compounds, BMS-204352 and retigabine, prese...

  7. Physical activity maintenance in patients with rheumatoid arthritis

    DEFF Research Database (Denmark)

    Loeppenthin, K; Esbensen, Bente Appel; Østergaard, Mikkel

    2014-01-01

    OBJECTIVE: To describe the experience of physical activity maintenance in patients with rheumatoid arthritis. DESIGN: A qualitative salutogenic-oriented interview study. SETTING: A rheumatology outpatient clinic. SUBJECTS: A purposive sample of 16 physically active patients (mean age 50, range 37...... with non-arthritis populations. CONCLUSION: This study demonstrates that physical activity in patients with rheumatoid arthritis may be understood as a resource to resist disability and to feel and stay healthy while creating and sustaining meaningfulness in life.......-67) diagnosed with rheumatoid arthritis on average 21 years previously (range 4-46 years). METHODS: In-depth interviews were conducted using a semi-structured interview guide to illuminate how the phenomenon 'physical activity maintenance' was experienced by patients with rheumatoid arthritis. The interviews...

  8. Human Error Probability Assessment During Maintenance Activities of Marine Systems

    Directory of Open Access Journals (Sweden)

    Rabiul Islam

    2018-03-01

    Full Text Available Background: Maintenance operations on-board ships are highly demanding. Maintenance operations are intensive activities requiring high man–machine interactions in challenging and evolving conditions. The evolving conditions are weather conditions, workplace temperature, ship motion, noise and vibration, and workload and stress. For example, extreme weather condition affects seafarers' performance, increasing the chances of error, and, consequently, can cause injuries or fatalities to personnel. An effective human error probability model is required to better manage maintenance on-board ships. The developed model would assist in developing and maintaining effective risk management protocols. Thus, the objective of this study is to develop a human error probability model considering various internal and external factors affecting seafarers' performance. Methods: The human error probability model is developed using probability theory applied to Bayesian network. The model is tested using the data received through the developed questionnaire survey of >200 experienced seafarers with >5 years of experience. The model developed in this study is used to find out the reliability of human performance on particular maintenance activities. Results: The developed methodology is tested on the maintenance of marine engine's cooling water pump for engine department and anchor windlass for deck department. In the considered case studies, human error probabilities are estimated in various scenarios and the results are compared between the scenarios and the different seafarer categories. The results of the case studies for both departments are also compared. Conclusion: The developed model is effective in assessing human error probabilities. These probabilities would get dynamically updated as and when new information is available on changes in either internal (i.e., training, experience, and fatigue or external (i.e., environmental and operational conditions

  9. A quantized mechanism for activation of pannexin channels

    Science.gov (United States)

    Chiu, Yu-Hsin; Jin, Xueyao; Medina, Christopher B.; Leonhardt, Susan A.; Kiessling, Volker; Bennett, Brad C.; Shu, Shaofang; Tamm, Lukas K.; Yeager, Mark; Ravichandran, Kodi S.; Bayliss, Douglas A.

    2017-01-01

    Pannexin 1 (PANX1) subunits form oligomeric plasma membrane channels that mediate nucleotide release for purinergic signalling, which is involved in diverse physiological processes such as apoptosis, inflammation, blood pressure regulation, and cancer progression and metastasis. Here we explore the mechanistic basis for PANX1 activation by using wild type and engineered concatemeric channels. We find that PANX1 activation involves sequential stepwise sojourns through multiple discrete open states, each with unique channel gating and conductance properties that reflect contributions of the individual subunits of the hexamer. Progressive PANX1 channel opening is directly linked to permeation of ions and large molecules (ATP and fluorescent dyes) and occurs during both irreversible (caspase cleavage-mediated) and reversible (α1 adrenoceptor-mediated) forms of channel activation. This unique, quantized activation process enables fine tuning of PANX1 channel activity and may be a generalized regulatory mechanism for other related multimeric channels. PMID:28134257

  10. Anticipated affective consequences of physical activity adoption and maintenance.

    Science.gov (United States)

    Dunton, Genevieve Fridlund; Vaughan, Elaine

    2008-11-01

    The expected emotional consequences of future actions are thought to play an important role in health behavior change. This research examined whether anticipated affective consequences of success and failure vary across stages of physical activity change and differentially predict physical activity adoption as compared to maintenance. Using a prospective design over a 3-month period, a community sample of 329 healthy, middle-aged adults were assessed at 2 time points. Anticipated positive and negative emotions, stage of behavior change (precontemplation [PC], contemplation [C], preparation [P], action [A], maintenance [M]), and level of physical activity. At baseline, anticipated positive emotions were greater in C versus PC, whereas anticipated negative emotions were greater in M versus A and in M versus P. Higher anticipated positive but not negative emotions predicted physical activity adoption and maintenance after 3 months. Although the expected affective consequences of future success and failure differentiated among individuals in the early and later stages of physical activity change, respectively; only the anticipated affective consequences of success predicted future behavior.

  11. Public Tourism Infrastructure: Challenges in the Development and Maintenance Activities

    Directory of Open Access Journals (Sweden)

    Abdullah Shardy

    2014-01-01

    Full Text Available In Malaysia, the tourism sector is a major contributor to the nation’s development and is spearheaded by the government’s efforts in investing heavily towards providing sufficient and well-functioning public tourism infrastructure. This infrastructure should be ideally developed with a clear and systematic maintenance plan in hand. The challenge herein is not merely providing the necessary infrastructure to sustain tourism activities but rather a pro-active approach towards establishing and subsequently maintaining this infrastructure at its optimal level. The aim of this paper therefore is to identify critical aspects that need to be in place to further enhance the Malaysian tourism industry. The paper discusses the issues and challenges that need to be addressed as a precursor towards an effectively developed and maintained tourism infrastructure system. Development issues that have been identified revolve around the dimensions of quality, quantity and ability of the public agencies involved, particularly issues of inadequate infrastructure, quality of infrastructure and the capability of the agencies in undertaking efficient maintenance activities. These issues were found to lead towards challenges of working with resource constraints, lack of an effective maintenance culture and system as well as the need for clear and effective policies and strategies.

  12. Study on how to evaluate the effectiveness of maintenance activities for giant complex plant system

    International Nuclear Information System (INIS)

    Aoki, Takayuki

    2010-01-01

    If we try to check the effectiveness of maintenance activities in nuclear power plant, it is necessary to evaluate plant performance from the viewpoint of nuclear safety and economy. So, in this paper, after the relation among maintenance optimization, maintenance performance targets, maintenance effectiveness indicator and maintenance key parameters important to nuclear safety and economy was made clear, a method to evaluate the effectiveness of maintenance activities was discussed. As a result of consideration, it was concluded that the maintenance effectiveness indicator proposed in this paper can evaluate maintenance effectiveness and can show the direction of improving existing maintenance program because the relation with maintenance performance target and maintenance key parameters is clear. (author)

  13. BK channel activators and their therapeutic perspectives

    DEFF Research Database (Denmark)

    Bentzen, Bo Hjorth; Olesen, Søren-Peter; Rønn, Lars C B

    2014-01-01

    in intracellular calcium to outward hyperpolarizing potassium currents. Consequently, the channel has many important physiological roles including regulation of smooth muscle tone, neurotransmitter release and neuronal excitability. Additionally, cardioprotective roles have been revealed in recent years. After...

  14. Active Brownian motion in a narrow channel

    Science.gov (United States)

    Ao, X.; Ghosh, P. K.; Li, Y.; Schmid, G.; Hänggi, P.; Marchesoni, F.

    2014-12-01

    We review recent advances in rectification control of artificial microswimmers, also known as Janus particles, diffusing along narrow, periodically corrugated channels. The swimmer self-propulsion mechanism is modeled so as to incorporate a nonzero torque (propulsion chirality). We first summarize the effects of chirality on the autonomous current of microswimmers freely diffusing in channels of different geometries. In particular, left-right and upside-down asymmetric channels are shown to exhibit different transport properties. We then report new results on the dependence of the diffusivity of chiral microswimmers on the channel geometry and their own self-propulsion mechanism. The self-propulsion torque turns out to play a key role as a transport control parameter.

  15. A Study on the Maintenance Effectiveness Assessment for Active Components

    International Nuclear Information System (INIS)

    Lim, Woo Sang; Oh, Seung Jong

    2006-01-01

    One of the key tasks in the periodic safety review (PSR) of nuclear power plant is to assess the aging management of structures, systems and components (SSC). The evaluation can be categorized by two parts, passive and active components. Unlike the passive components, active components are periodically maintained and replaced with new components, so the evaluation of aging mechanism of the passive components such as erosion, corrosion is not applicable to the evaluation of active components of nuclear power plant. For active components, they will maintain capability to fulfill its design function if preventive maintenance effectiveness is proper. In this paper, the assessment based on the reliability and availability of the active components of the domestic nuclear power plants is examined

  16. Slack sodium-activated potassium channel membrane expression requires p38 mitogen-activated protein kinase phosphorylation.

    Science.gov (United States)

    Gururaj, Sushmitha; Fleites, John; Bhattacharjee, Arin

    2016-04-01

    p38 MAPK has long been understood as an inducible kinase under conditions of cellular stress, but there is now increasing evidence to support its role in the regulation of neuronal function. Several phosphorylation targets have been identified, an appreciable number of which are ion channels, implicating the possible involvement of p38 MAPK in neuronal excitability. The KNa channel Slack is an important protein to be studied as it is highly and ubiquitously expressed in DRG neurons and is important in the maintenance of their firing accommodation. We sought to examine if the Slack channel could be a substrate of p38 MAPK activity. First, we found that the Slack C-terminus contains two putative p38 MAPK phosphorylation sites that are highly conserved across species. Second, we show via electrophysiology experiments that KNa currents and further, Slack currents, are subject to tonic modulation by p38 MAPK. Third, biochemical approaches revealed that Slack channel regulation by p38 MAPK occurs through direct phosphorylation at the two putative sites of interaction, and mutating both sites prevented surface expression of Slack channels. Based on these results, we conclude that p38 MAPK is an obligate regulator of Slack channel function via the trafficking of channels into the membrane. The present study identifies Slack KNa channels as p38 MAPK substrates. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Priority ranking for maintenance activities on embankment dams

    International Nuclear Information System (INIS)

    Chouinard, L.E.; Andersen, G.R.; Robichaud, J.G.; Blanchette, G.; Gervais, R.

    1998-01-01

    Operators of dams in Canada and the U.S. are steadily shifting from construction of new facilities to the maintenance and repairs of existing ones. This paper emphasized the importance of prioritizing maintenance activities on embankment dams whose maintenance needs vary from structure to structure. Two parallel procedures were developed, one for monitoring devices and the other for defense groups. Both procedures are intended to be used together to rate the condition of the embankment dams. The term 'defense groups' is used to refer to the collection of physical components on dams to prevent adverse conditions from occurring that would result in an uncontrolled release of the reservoir. The priority rankings and condition indices developed by means of this procedure reflects the judgment of a panel of engineers and geologists who implement them. They are not to be interpreted as an index of dam safety. While the process is sufficiently well developed to warrant wide-spread distribution, it is considered to be still in the developmental stage. Therefore, it should be used in conjunction with other parallel processes evaluating structural, mechanical and electrical features of the structure under consideration. 6 refs., 7 tabs

  18. Physical activity and healthy weight maintenance from childhood to adulthood.

    Science.gov (United States)

    Cleland, Verity J; Dwyer, Terence; Venn, Alison J

    2008-06-01

    The objective of this study was to determine whether change in physical activity was associated with maintaining a healthy weight from childhood to adulthood. This prospective cohort study examined 1,594 young Australian adults (48.9% female) aged 27-36 years who were first examined at age 9-15 years as part of a national health and fitness survey. BMI was calculated from measured height and weight, and physical activity was self-reported at both time points; pedometers were also used at follow-up. Change in physical activity was characterized by calculating the difference between baseline and follow-up z-scores. Change scores were categorized as decreasing (large, moderate), stable, or increasing (large, moderate). Healthy weight was defined in childhood as a BMI less than international overweight cutoff points, and in adulthood as BMIdecreases in physical activity, females in all other groups were 25-37% more likely to be healthy weight maintainers, although associations differed according to the physical activity measure used at follow-up and few reached statistical significance. Although younger males whose relative physical activity moderately or largely increased were 27-34% more likely to be healthy weight maintainers than those whose relative physical activity largely decreased, differences were not statistically significant. In conclusion, relatively increasing and stable physical activity from childhood to adulthood was only weakly associated with healthy weight maintenance. Examining personal, social, and environmental factors associated with healthy weight maintenance will be an important next step in understanding why some groups avoid becoming overweight.

  19. Human temporal cortical single neuron activity during working memory maintenance.

    Science.gov (United States)

    Zamora, Leona; Corina, David; Ojemann, George

    2016-06-01

    The Working Memory model of human memory, first introduced by Baddeley and Hitch (1974), has been one of the most influential psychological constructs in cognitive psychology and human neuroscience. However the neuronal correlates of core components of this model have yet to be fully elucidated. Here we present data from two studies where human temporal cortical single neuron activity was recorded during tasks differentially affecting the maintenance component of verbal working memory. In Study One we vary the presence or absence of distracting items for the entire period of memory storage. In Study Two we vary the duration of storage so that distractors filled all, or only one-third of the time the memory was stored. Extracellular single neuron recordings were obtained from 36 subjects undergoing awake temporal lobe resections for epilepsy, 25 in Study one, 11 in Study two. Recordings were obtained from a total of 166 lateral temporal cortex neurons during performance of one of these two tasks, 86 study one, 80 study two. Significant changes in activity with distractor manipulation were present in 74 of these neurons (45%), 38 Study one, 36 Study two. In 48 (65%) of those there was increased activity during the period when distracting items were absent, 26 Study One, 22 Study Two. The magnitude of this increase was greater for Study One, 47.6%, than Study Two, 8.1%, paralleling the reduction in memory errors in the absence of distracters, for Study One of 70.3%, Study Two 26.3% These findings establish that human lateral temporal cortex is part of the neural system for working memory, with activity during maintenance of that memory that parallels performance, suggesting it represents active rehearsal. In 31 of these neurons (65%) this activity was an extension of that during working memory encoding that differed significantly from the neural processes recorded during overt and silent language tasks without a recent memory component, 17 Study one, 14 Study two

  20. Relationship challenges and relationship maintenance activities following disclosure of transsexualism.

    Science.gov (United States)

    Alegría, C Aramburu

    2010-12-01

    • Transsexual persons are increasing their visibility in society, and health care providers and others (such as social workers) will be called upon to help with issues that transsexual persons face. Challenges that face transsexual persons often include issues involving relationships. Psychiatric and mental health nurses and other caregivers can increase their therapeutic skills in working with couples that include transsexual persons by becoming aware of these challenges and subsequent activities that can help with them. • This research study looks at couple relationships in which one partner reveals male-to-female transsexual identity. These are relationships that were established as man-woman and now will transition into relationships that include a male-to-female person and a female partner. • Common challenges for these couples include issues related to: (1) sexual identity and relationship uncertainty; (2) male-to-female transition decision making; and (3) presenting in public. • Relationship maintenance activities that helped the couples in the study maintain and strengthen their relationships through these challenges include: (1) communication; (2) self-talk (for example, putting the situation in perspective); (3) social networks; (4) positive interactions; (5) impression management (for example, managing displays of affection in public); and (6) social activism. This qualitative study describes the relational dynamics that help sustain relationships of couples that include male-to-female transsexual persons (MTF) and their natal female partners (NF) following disclosure of transsexualism. Relationship challenges and relationship maintenance activities are identified. Each partner in 17 MTF-NF couples participated in individual surveys and interviews. The data were coded for themes related to relationship challenges and activities. MTF-NF couples experience challenges within the contexts of their relationships and of society. These challenges

  1. Human Temporal Cortical Single Neuron Activity During Working Memory Maintenance

    Science.gov (United States)

    Zamora, Leona; Corina, David; Ojemann, George

    2016-01-01

    The Working Memory model of human memory, first introduced by Baddeley and Hitch (1974), has been one of the most influential psychological constructs in cognitive psychology and human neuroscience. However the neuronal correlates of core components of this model have yet to be fully elucidated. Here we present data from two studies where human temporal cortical single neuron activity was recorded during tasks differentially affecting the maintenance component of verbal working memory. In Study One we vary the presence or absence of distracting items for the entire period of memory storage. In Study Two we vary the duration of storage so that distractors filled all, or only one-third of the time the memory was stored. Extracellular single neuron recordings were obtained from 36 subjects undergoing awake temporal lobe resections for epilepsy, 25 in Study one, 11 in Study two. Recordings were obtained from a total of 166 lateral temporal cortex neurons during performance of one of these two tasks, 86 study one, 80 study two. Significant changes in activity with distractor manipulation were present in 74 of these neurons (45%), 38 Study one, 36 Study two. In 48 (65%) of those there was increased activity during the period when distracting items were absent, 26 Study One, 22 Study Two. The magnitude of this increase was greater for Study One, 47.6%, than Study Two, 8.1%, paralleling the reduction in memory errors in the absence of distracters, for Study One of 70.3%, Study Two 26.3% These findings establish that human lateral temporal cortex is part of the neural system for working memory, with activity during maintenance of that memory that parallels performance, suggesting it represents active rehearsal. In 31 of these neurons (65%) this activity was an extension of that during working memory encoding that differed significantly from the neural processes recorded during overt and silent language tasks without a recent memory component, 17 Study one, 14 Study two

  2. Repair, Evaluation, Maintenance, and Rehabilitation Research Program. Geotechnical Aspects of Rock Erosion in Emergency Spillway Channels

    Science.gov (United States)

    1986-08-01

    8217 flooring unlined spillway channels. 115. Rippability was proposed by Weaver (1975) as a rock mass classifi- cation system that enables the assessment...geological. features which govern the as- sessment of rippability are rock type, haidness, weathering, stcucture, and fabric. Saismic P-wave velocity...channel. Rippability should be combined with a factor 84 that describes lithostratigraphic continuity in order to derive the erosion potential of the

  3. Cell volume and membrane stretch independently control K+ channel activity

    DEFF Research Database (Denmark)

    Bomholtz, Sofia Hammami; Willumsen, Niels J; Olsen, Hervør L

    2009-01-01

    A number of potassium channels including members of the KCNQ family and the Ca(2+) activated IK and SK, but not BK, are strongly and reversibly regulated by small changes in cell volume. It has been argued that this general regulation is mediated through sensitivity to changes in membrane stretch...... was not affected by membrane stretch. The results indicate that (1) activation of BK channels by local membrane stretch is not mimicked by membrane stress induced by cell swelling, and (2) activation of KCNQ1 channels by cell volume increase is not mediated by local tension in the cell membrane. We conclude....... To test this hypothesis we have studied the regulation of KCNQ1 and BK channels after expression in Xenopus oocytes. Results from cell-attached patch clamp studies (approximately 50 microm(2) macropatches) in oocytes expressing BK channels demonstrate that the macroscopic volume-insensitive BK current...

  4. Slack, Slick, and Sodium-Activated Potassium Channels

    Science.gov (United States)

    Kaczmarek, Leonard K.

    2013-01-01

    The Slack and Slick genes encode potassium channels that are very widely expressed in the central nervous system. These channels are activated by elevations in intracellular sodium, such as those that occur during trains of one or more action potentials, or following activation of nonselective cationic neurotransmitter receptors such as AMPA receptors. This review covers the cellular and molecular properties of Slack and Slick channels and compares them with findings on the properties of sodium-activated potassium currents (termed KNa currents) in native neurons. Human mutations in Slack channels produce extremely severe defects in learning and development, suggesting that KNa channels play a central role in neuronal plasticity and intellectual function. PMID:24319675

  5. Maintenance of Physical Activity among Faculty and Staff in University Settings

    Science.gov (United States)

    Whipple, Kerry; Kinney, Judy; Kattenbraker, Mark

    2008-01-01

    Previous studies have placed little emphasis on maintenance of healthy behaviors longer than six months. This study examined factors that contribute to maintenance of physical activity among faculty and staff in university settings. A 55-item survey on physical activity maintenance was used to assess attitudes towards exercise, exercise…

  6. Channel sialic acids limit hERG channel activity during the ventricular action potential.

    Science.gov (United States)

    Norring, Sarah A; Ednie, Andrew R; Schwetz, Tara A; Du, Dongping; Yang, Hui; Bennett, Eric S

    2013-02-01

    Activity of human ether-a-go-go-related gene (hERG) 1 voltage-gated K(+) channels is responsible for portions of phase 2 and phase 3 repolarization of the human ventricular action potential. Here, we questioned whether and how physiologically and pathophysiologically relevant changes in surface N-glycosylation modified hERG channel function. Voltage-dependent hERG channel gating and activity were evaluated as expressed in a set of Chinese hamster ovary (CHO) cell lines under conditions of full glycosylation, no sialylation, no complex N-glycans, and following enzymatic deglycosylation of surface N-glycans. For each condition of reduced glycosylation, hERG channel steady-state activation and inactivation relationships were shifted linearly by significant depolarizing ∼9 and ∼18 mV, respectively. The hERG window current increased significantly by 50-150%, and the peak shifted by a depolarizing ∼10 mV. There was no significant change in maximum hERG current density. Deglycosylated channels were significantly more active (20-80%) than glycosylated controls during phases 2 and 3 of action potential clamp protocols. Simulations of hERG current and ventricular action potentials corroborated experimental data and predicted reduced sialylation leads to a 50-70-ms decrease in action potential duration. The data describe a novel mechanism by which hERG channel gating is modulated through physiologically and pathophysiologically relevant changes in N-glycosylation; reduced channel sialylation increases hERG channel activity during the action potential, thereby increasing the rate of action potential repolarization.

  7. Determinants and benefits of physical activity maintenance in hospital employees.

    Science.gov (United States)

    Lavoie-Tremblay, Mélanie; Sounan, Charles; Martin, Kara; Trudel, Julie G; Lavigne, Genevieve L; Grover, Steven A; Lowensteyn, Ilka

    2014-01-01

    This study investigated whether the positive behavioral and anthropometric outcomes of a pedometer-based physical activity 8-week challenge were maintained 6 months after the end of the program. It further investigated the motivational profile of those who maintained their physical activity levels in the months following the end of the program and of those who did not. Hospital employees from a university-affiliated multisite health care center in Canada participated using a questionnaire. Of the 235 participants who completed the 8-week challenge, 157 questionnaires were returned 6 months later. Paired-samples t tests were conducted between the baseline and follow-up scores as well as between the postprogram and follow-up scores to detect significant differences between the measurement points. This study shows that the pedometer-based physical activity helped hospital employees maintain a high level of physical activity as well as maintain a healthy body mass index after 6 months. The results demonstrated that during maintenance the high physical activity group obtained higher scores for identified regulation and intrinsic regulation compared with the other groups. The results of the study revealed that identified and intrinsic regulations are important contributors to maintaining physical activity among hospital employees.

  8. Creating Value by Integrating Logistic Trains Services and Maintenance Activities

    NARCIS (Netherlands)

    Busstra, Marten; van Dongen, Leonardus Adriana Maria

    2015-01-01

    NedTrain is the Netherlands Railway's subsidiary responsible for rolling stock maintenance. Train sets are brought in for short-term routine maintenance after set intervals of some 75 to 120 days. When a major defect occurs, train sets are allocated to one of the three maintenance depots and are

  9. Use of probabilistic risk assessment in maintenance activities at Palo Verde

    International Nuclear Information System (INIS)

    Lindquist, R.C.; Pobst, D.S.

    1993-01-01

    Probabilistic risk assessment (PRA) is an important tool in addressing various maintenance activities. At the Palo Verde nuclear generating station (PVNGS), the PRA has been used in a variety of ways to support a wide and diverse selection of maintenance-related activities. For on-line or at-power maintenance, the PRA was used to evaluate combinations of maintenance activities possible with the 12-week or floating maintenance schedule. The maintenance schedule was evaluated to identify any higher risk, undesirable combinations of equipment outages, such as the sole steam-driven auxiliary feedwater pump and the same train emergency diesel generator. Table I is a sampling of the results from the maintenance schedule evaluation in terms of increase in conditional core damage frequency (CDF) above the base- line value due to maintenance on some important key safety systems and combinations thereof. The baseline CDF is 7.4 x 10 -7 per 72 h

  10. Imaging large cohorts of single ion channels and their activity

    Directory of Open Access Journals (Sweden)

    Katia eHiersemenzel

    2013-09-01

    Full Text Available As calcium is the most important signaling molecule in neurons and secretory cells, amongst many other cell types, it follows that an understanding of calcium channels and their regulation of exocytosis is of vital importance. Calcium imaging using calcium dyes such as Fluo3, or FRET-based dyes that have been used widely has provided invaluable information, which combined with modeling has estimated the sub-types of channels responsible for triggering the exocytotic machinery as well as inferences about the relative distances away from vesicle fusion sites these molecules adopt. Importantly, new super-resolution microscopy techniques, combined with novel Ca2+ indicators and imaginative imaging approaches can now define directly the nanoscale locations of very large cohorts of single channel molecules in relation to single vesicles. With combinations of these techniques the activity of individual channels can be visualized and quantified using novel Ca2+ indicators. Fluorescently labeled specific channel toxins can also be used to localize endogenous assembled channel tetramers. Fluorescence lifetime imaging microscopy and other single-photon-resolution spectroscopic approaches offer the possibility to quantify protein-protein interactions between populations of channels and the SNARE protein machinery for the first time. Together with simultaneous electrophysiology, this battery of quantitative imaging techniques has the potential to provide unprecedented detail describing the locations, dynamic behaviours, interactions and conductance activities of many thousands of channel molecules and vesicles in living cells.

  11. Spectrum scanning and reserve channel methods for link maintenance in cognitive radio systems

    OpenAIRE

    Subramani, S; Armour, SMD; Kaleshi, D; Fan, Z

    2008-01-01

    Underutilization of the limited spectrum sparked the need for dynamic spectrum access and flexible spectrum policies. Accurate estimation of spectrum occupancy is an essential step for spectrum access in distributed networks. This paper analyzes a bidirectional and dual scanning method that scans multiple spectrum bands to find a suitable free-channel. Moreover, secondary users' access to available spectrum could be interrupted by the arrival of primary spectrum users, forcing the well-behave...

  12. BAD and KATP channels regulate neuron excitability and epileptiform activity.

    Science.gov (United States)

    Martínez-François, Juan Ramón; Fernández-Agüera, María Carmen; Nathwani, Nidhi; Lahmann, Carolina; Burnham, Veronica L; Danial, Nika N; Yellen, Gary

    2018-01-25

    Brain metabolism can profoundly influence neuronal excitability. Mice with genetic deletion or alteration of Bad ( B CL-2 a gonist of cell d eath) exhibit altered brain-cell fuel metabolism, accompanied by resistance to acutely induced epileptic seizures; this seizure protection is mediated by ATP-sensitive potassium (K ATP ) channels. Here we investigated the effect of BAD manipulation on K ATP channel activity and excitability in acute brain slices. We found that BAD's influence on neuronal K ATP channels was cell-autonomous and directly affected dentate granule neuron (DGN) excitability. To investigate the role of neuronal K ATP channels in the anticonvulsant effects of BAD, we imaged calcium during picrotoxin-induced epileptiform activity in entorhinal-hippocampal slices. BAD knockout reduced epileptiform activity, and this effect was lost upon knockout or pharmacological inhibition of K ATP channels. Targeted BAD knockout in DGNs alone was sufficient for the antiseizure effect in slices, consistent with a 'dentate gate' function that is reinforced by increased K ATP channel activity. © 2018, Martínez-François et al.

  13. MYC activation is a hallmark of cancer initiation and maintenance.

    Science.gov (United States)

    Gabay, Meital; Li, Yulin; Felsher, Dean W

    2014-06-02

    The MYC proto-oncogene has been implicated in the pathogenesis of most types of human tumors. MYC activation alone in many normal cells is restrained from causing tumorigenesis through multiple genetic and epigenetically controlled checkpoint mechanisms, including proliferative arrest, apoptosis, and cellular senescence. When pathologically activated in a permissive epigenetic and/or genetic context, MYC bypasses these mechanisms, enforcing many of the "hallmark" features of cancer, including relentless tumor growth associated with DNA replication and transcription, cellular proliferation and growth, protein synthesis, and altered cellular metabolism. MYC mandates tumor cell fate, by inducing stemness and blocking cellular senescence and differentiation. Additionally, MYC orchestrates changes in the tumor microenvironment, including the activation of angiogenesis and suppression of the host immune response. Provocatively, brief or even partial suppression of MYC back to its physiological levels of activation can result in the restoration of intrinsic checkpoint mechanisms, resulting in acute and sustained tumor regression, associated with tumor cells undergoing proliferative arrest, differentiation, senescence, and apoptosis, as well as remodeling of the tumor microenvironment, recruitment of an immune response, and shutdown of angiogenesis. Hence, tumors appear to be "addicted" to MYC because of both tumor cell-intrinsic, cell-autonomous and host-dependent, immune cell-dependent mechanisms. Both the trajectory and persistence of many human cancers require sustained MYC activation. Multiscale mathematical modeling may be useful to predict when tumors will be addicted to MYC. MYC is a hallmark molecular feature of both the initiation and maintenance of tumorigenesis. Copyright © 2014 Cold Spring Harbor Laboratory Press; all rights reserved.

  14. PLC based control system and maintenance activities at NCAR, Bilaspur

    International Nuclear Information System (INIS)

    Dewangan, Jaidev; Trivedi, T.; Patel, Shiv P.; Malik, C.; Kumar, Rakesh; Gupta, Santosh Kumar; Bajpai, P.K.

    2015-01-01

    A 3.0 MV high current low energy Pelletron Accelerator facility (Model 9SDH-4, NEC, USA) with TORUIS (ion source for H + and He 2+ beam current H + ion ∼ 50μA @ 6 MeV, He 2+ at ∼ 10μA) and SNICS-II ion source for heavy ions has been commissioned as 'National Centre for Accelerator Based Research' in the Department of Pure and Applied Physics, Guru Ghasidas Vishwavidyalaya. In this paper, we detail out the control system developed and implemented at NCAR. The basic idea of controlling the machine is by providing the output signal through PLC to ACPC of accelerator using user interface points provided by the manufacturer. The PLC based system generates output signal once it receives the feedback signals from search and secure switches, door lock switches and scram switches interlocked with PLC. The output is controlled by ladder logic and is activated only when all the radiation monitors are in healthy state and outside radiations monitor having low radiation level. The details of control system and maintenance activities will be discussed in the paper

  15. Activation of purified calcium channels by stoichiometric protein phosphorylation

    Energy Technology Data Exchange (ETDEWEB)

    Nunoki, K.; Florio, V.; Catterall, W.A. (Univ. of Washington, Seattle (USA))

    1989-09-01

    Purified dihydropyridine-sensitive calcium channels from rabbit skeletal muscle were reconstituted into phosphatidylcholine vesicles to evaluate the effect of phosphorylation by cyclic AMP-dependent protein kinase (PK-A) on their function. Both the rate and extent of {sup 45}Ca{sup 2+} uptake into vesicles containing reconstituted calcium channels were increased severalfold after incubation with ATP and PK-A. The degree of stimulation of {sup 45}Ca{sup 2+} uptake was linearly proportional to the extent of phosphorylation of the alpha 1 and beta subunits of the calcium channel up to a stoichiometry of approximately 1 mol of phosphate incorporated into each subunit. The calcium channels activated by phosphorylation were determined to be incorporated into the reconstituted vesicles in the inside-out orientation and were completely inhibited by low concentrations of dihydropyridines, phenylalkylamines, Cd{sup 2+}, Ni{sup 2+}, and Mg{sup 2+}. The results demonstrate a direct relationship between PK-A-catalyzed phosphorylation of the alpha 1 and beta subunits of the purified calcium channel and activation of the ion conductance activity of the dihydropyridine-sensitive calcium channels.

  16. Activation of purified calcium channels by stoichiometric protein phosphorylation

    International Nuclear Information System (INIS)

    Nunoki, K.; Florio, V.; Catterall, W.A.

    1989-01-01

    Purified dihydropyridine-sensitive calcium channels from rabbit skeletal muscle were reconstituted into phosphatidylcholine vesicles to evaluate the effect of phosphorylation by cyclic AMP-dependent protein kinase (PK-A) on their function. Both the rate and extent of 45 Ca 2+ uptake into vesicles containing reconstituted calcium channels were increased severalfold after incubation with ATP and PK-A. The degree of stimulation of 45 Ca 2+ uptake was linearly proportional to the extent of phosphorylation of the alpha 1 and beta subunits of the calcium channel up to a stoichiometry of approximately 1 mol of phosphate incorporated into each subunit. The calcium channels activated by phosphorylation were determined to be incorporated into the reconstituted vesicles in the inside-out orientation and were completely inhibited by low concentrations of dihydropyridines, phenylalkylamines, Cd 2+ , Ni 2+ , and Mg 2+ . The results demonstrate a direct relationship between PK-A-catalyzed phosphorylation of the alpha 1 and beta subunits of the purified calcium channel and activation of the ion conductance activity of the dihydropyridine-sensitive calcium channels

  17. Parallel Evolution of Sperm Hyper-Activation Ca2+ Channels.

    Science.gov (United States)

    Cooper, Jacob C; Phadnis, Nitin

    2017-07-01

    Sperm hyper-activation is a dramatic change in sperm behavior where mature sperm burst into a final sprint in the race to the egg. The mechanism of sperm hyper-activation in many metazoans, including humans, consists of a jolt of Ca2+ into the sperm flagellum via CatSper ion channels. Surprisingly, all nine CatSper genes have been independently lost in several animal lineages. In Drosophila, sperm hyper-activation is performed through the cooption of the polycystic kidney disease 2 (pkd2) Ca2+ channel. The parallels between CatSpers in primates and pkd2 in Drosophila provide a unique opportunity to examine the molecular evolution of the sperm hyper-activation machinery in two independent, nonhomologous calcium channels separated by > 500 million years of divergence. Here, we use a comprehensive phylogenomic approach to investigate the selective pressures on these sperm hyper-activation channels. First, we find that the entire CatSper complex evolves rapidly under recurrent positive selection in primates. Second, we find that pkd2 has parallel patterns of adaptive evolution in Drosophila. Third, we show that this adaptive evolution of pkd2 is driven by its role in sperm hyper-activation. These patterns of selection suggest that the evolution of the sperm hyper-activation machinery is driven by sexual conflict with antagonistic ligands that modulate channel activity. Together, our results add sperm hyper-activation channels to the class of fast evolving reproductive proteins and provide insights into the mechanisms used by the sexes to manipulate sperm behavior. © The Author 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  18. Stretch-activated cation channel from larval bullfrog skin

    DEFF Research Database (Denmark)

    Hillyard, Stanley D; Willumsen, Niels J; Marrero, Mario B

    2010-01-01

    Cell-attached patches from isolated epithelial cells from larval bullfrog skin revealed a cation channel that was activated by applying suction (-1 kPa to -4.5 kPa) to the pipette. Activation was characterized by an initial large current spike that rapidly attenuated to a stable value and showed ...

  19. State of technology, system and solution supporting on-line maintenance - company's activities and products

    International Nuclear Information System (INIS)

    Nishitani, Junichi; Shimizu, Shunichi; Higasa, Hisakazu

    2010-01-01

    The new inspection system based on operator's maintenance and monitoring program of nuclear power plants was introduced in Japan more than one year ago and recommended on-line maintenance (maintenance during operation) will be carried out to increase capacity factor with safe and reliable operation of the plant. In this feature article, nine experts described the state of technology, system and solution supporting on-line maintenance - company's activities and products. These were titled as 'MHI's technology supporting on-line maintenance'. 'Technology supporting on-line maintenance - Toshiba's activities to upgrade monitoring and diagnostic service and maintenance management', 'AsahiKASEI's activities of on-line maintenance', 'Importance of information sharing of on-line maintenance and its ideal method-function of impact plan of IBM Maximo Asset Management for Nuclear', 'US's on-line maintenance and information systems', 'SmartProcedures realizing safe operation of nuclear power plant - proposal of computerized procedures', 'Ultrasonic leak detection system SDT170', 'Application of infrared thermography for equipment maintenance in nuclear power plant' and 'On-line condition monitoring system - condition eye'. (T. Tanaka)

  20. The role of affects and emotions in physical activity maintenance

    Directory of Open Access Journals (Sweden)

    Benjamin Wienke

    2016-08-01

    Full Text Available Although previous research has shown that affective variables are consistently associated with physical activity behavior, the working mechanisms are not understood to the extent of creating an intervention. The aim of this study is to identify situations and aspects of physical activity, which lead to positive affective reactions in people taking part in regular and long-term exercise. In this study 24 adults (12 female, 12 male distributed over three age groups (young, middle, and late adulthood that took part in sport programs (individual or team sport for at least five years. Semi-structured in depth interviews with questions about physical activity, long term participation and affective response in a sporting environment were conducted in order to ascertain those situations and aspects of the exercise program triggering positive affective states. Interviews were transcribed verbatim and followed Grounded Theory principles. Emerging concepts were grouped and merged into different categories representing the key aspects of exercise. Four factors were identified which are associated with emergence of positive emotions in sport and exercise. Firstly, perceived competence is one of the major factors influencing affective states during physical activity representing individual and collective success and progress, competition and challenge. Secondly, perceived social interaction is another factor comprising of all sorts of peer-related aspects such as communication with others, being part of a group and creating close relationships or friendships. Thirdly, novelty experience in contrast to other none-sporting activities such as work, family or other leisure activities was another factor. The last factor found was the perceived physical exertion comprising the degree of exhaustion, a possibly delayed turnaround in the affective response and the aspect of sport being a physical compensation for everyday sedentary life. The results of this study

  1. Maintenance of the JET active gas handling system

    International Nuclear Information System (INIS)

    Brennan, P.D.; Bell, A.C.; Brown, K.; Cole, C.; Cooper, B.; Gibbons, C.; Harris, M.; Jones, G.; Knipe, S.; Lewis, J.; Manning, C.; Miller, A.; Perevezentsev, A.; Skinner, N.; Stagg, R.; Stead, M.; Thomas, R.; Yorkshades, J.

    2003-01-01

    The JET active gas handling system (AGHS) has been in operation in conjunction with the JET machine since Spring 1997. The tritium levels within the vessel have remained sufficiently high, 6.2 g at the end of the DTE1 experiment and currently 1.5 g, such that the AGHS has been required to operate continuously to detritiate gases liberated during D-D operations and to maintain discharges to the environment to ALARP. Maintaining the system to ensure continued operation has been a key factor in guaranteeing the continued availability of the essential sub-systems. The operational history of the JET AGHS has been previously documented in a number of papers [R. Laesser, et al. Proc. of the 19th SOFT Conf. 1 (1996) 227; R. Laesser, et al., Fusion Eng. Des. 46 (1999) 307; P.D. Brennan, et al., 18th Symp. on Fusion Eng., 1999]. Operational downtime is minimised through well-engineered sub-systems that use high integrity components. Outage, contamination and operator dosage are minimised through pre-planned and prepared maintenance operations. The reliability of sub-system critical condition fault detection is demonstrated through routine testing of hard-wired alarms and interlocks

  2. Pharmacological targeting of native CatSper channels reveals a required role in maintenance of sperm hyperactivation.

    Directory of Open Access Journals (Sweden)

    Anne E Carlson

    2009-08-01

    Full Text Available The four sperm-specific CatSper ion channel proteins are required for hyperactivated motility and male fertility, and for Ca(2+ entry evoked by alkaline depolarization. In the absence of external Ca(2+, Na(+ carries current through CatSper channels in voltage-clamped sperm. Here we show that CatSper channel activity can be monitored optically with the [Na(+](i-reporting probe SBFI in populations of intact sperm. Removal of external Ca(2+ increases SBFI signals in wild-type but not CatSper2-null sperm. The rate of the indicated rise of [Na(+](i is greater for sperm alkalinized with NH(4Cl than for sperm acidified with propionic acid, reflecting the alkaline-promoted signature property of CatSper currents. In contrast, the [Na(+](i rise is slowed by candidate CatSper blocker HC-056456 (IC(50 approximately 3 microM. HC-056456 similarly slows the rise of [Ca(2+](i that is evoked by alkaline depolarization and reported by fura-2. HC-056456 also selectively and reversibly decreased CatSper currents recorded from patch-clamped sperm. HC-056456 does not prevent activation of motility by HCO(3 (- but does prevent the development of hyperactivated motility by capacitating incubations, thus producing a phenocopy of the CatSper-null sperm. When applied to hyperactivated sperm, HC-056456 causes a rapid, reversible loss of flagellar waveform asymmetry, similar to the loss that occurs when Ca(2+ entry through the CatSper channel is terminated by removal of external Ca(2+. Thus, open CatSper channels and entry of external Ca(2+ through them sustains hyperactivated motility. These results indicate that pharmacological targeting of the CatSper channel may impose a selective late-stage block to fertility, and that high-throughput screening with an optical reporter of CatSper channel activity may identify additional selective blockers with potential for male-directed contraception.

  3. Proton and non-proton activation of ASIC channels.

    Directory of Open Access Journals (Sweden)

    Ivan Gautschi

    Full Text Available The Acid-Sensing Ion Channels (ASIC exhibit a fast desensitizing current when activated by pH values below 7.0. By contrast, non-proton ligands are able to trigger sustained ASIC currents at physiological pHs. To analyze the functional basis of the ASIC desensitizing and sustained currents, we have used ASIC1a and ASIC2a mutants with a cysteine in the pore vestibule for covalent binding of different sulfhydryl reagents. We found that ASIC1a and ASIC2a exhibit two distinct currents, a proton-induced desensitizing current and a sustained current triggered by sulfhydryl reagents. These currents differ in their pH dependency, their sensitivity to the sulfhydryl reagents, their ionic selectivity and their relative magnitude. We propose a model for ASIC1 and ASIC2 activity where the channels can function in two distinct modes, a desensitizing mode and a sustained mode depending on the activating ligands. The pore vestibule of the channel represents a functional site for binding non-proton ligands to activate ASIC1 and ASIC2 at neutral pH and to prevent channel desensitization.

  4. Proton and non-proton activation of ASIC channels.

    Science.gov (United States)

    Gautschi, Ivan; van Bemmelen, Miguel Xavier; Schild, Laurent

    2017-01-01

    The Acid-Sensing Ion Channels (ASIC) exhibit a fast desensitizing current when activated by pH values below 7.0. By contrast, non-proton ligands are able to trigger sustained ASIC currents at physiological pHs. To analyze the functional basis of the ASIC desensitizing and sustained currents, we have used ASIC1a and ASIC2a mutants with a cysteine in the pore vestibule for covalent binding of different sulfhydryl reagents. We found that ASIC1a and ASIC2a exhibit two distinct currents, a proton-induced desensitizing current and a sustained current triggered by sulfhydryl reagents. These currents differ in their pH dependency, their sensitivity to the sulfhydryl reagents, their ionic selectivity and their relative magnitude. We propose a model for ASIC1 and ASIC2 activity where the channels can function in two distinct modes, a desensitizing mode and a sustained mode depending on the activating ligands. The pore vestibule of the channel represents a functional site for binding non-proton ligands to activate ASIC1 and ASIC2 at neutral pH and to prevent channel desensitization.

  5. Maintenance of physical activity in patients with rheumaoid arthritis

    DEFF Research Database (Denmark)

    Løppenthin, Katrine; Esbensen, Bente Appel; Østergaard, Mikkel

    2012-01-01

    , described experiences of bodily well-being and liberation from illness provided and secured through PA maintenance. The understanding of regular PA as a resistance resource is essential to development future health promoting interventions aimed to encourage PA maintenance in patients with RA........ The analysis was discussed within a multidisciplinary team of qualitative researchers. Results: The analysis revealed tree dimensions of PA maintenance: (1) A bodily dimension: physical sensations of vitality, sparkling energy and liberation in movement; (2) a mental component: referring to experiences of self...... the participants to adjust to the uncertainty of the illness and preserving hope for a future dominated as little as possible by illness. Conclusions: While RA usually forces a negative attention to joint pain, fatigue and limited mobility in parts of the body, the participants in this study, despite illness...

  6. Large conductance Ca2+-activated K+ (BK channel: Activation by Ca2+ and voltage

    Directory of Open Access Journals (Sweden)

    RAMÓN LATORRE

    2006-01-01

    Full Text Available Large conductance Ca2+-activated K+ (BK channels belong to the S4 superfamily of K+ channels that include voltage-dependent K+ (Kv channels characterized by having six (S1-S6 transmembrane domains and a positively charged S4 domain. As Kv channels, BK channels contain a S4 domain, but they have an extra (S0 transmembrane domain that leads to an external NH2-terminus. The BK channel is activated by internal Ca2+, and using chimeric channels and mutagenesis, three distinct Ca2+-dependent regulatory mechanisms with different divalent cation selectivity have been identified in its large COOH-terminus. Two of these putative Ca2+-binding domains activate the BK channel when cytoplasmic Ca2+ reaches micromolar concentrations, and a low Ca2+ affinity mechanism may be involved in the physiological regulation by Mg2+. The presence in the BK channel of multiple Ca2+-binding sites explains the huge Ca2+ concentration range (0.1 μM-100 μM in which the divalent cation influences channel gating. BK channels are also voltage-dependent, and all the experimental evidence points toward the S4 domain as the domain in charge of sensing the voltage. Calcium can open BK channels when all the voltage sensors are in their resting configuration, and voltage is able to activate channels in the complete absence of Ca2+. Therefore, Ca2+ and voltage act independently to enhance channel opening, and this behavior can be explained using a two-tiered allosteric gating mechanism.

  7. Lessons Learned from Outsourcing the Pearl Harbor MK-48 Intermediate Maintenance Activity

    National Research Council Canada - National Science Library

    Gansler, Jacques S; Lucyshyn, William; Alarcon-Deza, Benigno

    2008-01-01

    During the 1990s, active duty sailor resources were in large demand. In an effort to return sailors to the fleet, the operation of the Navy's Heavyweight Torpedo Intermediate Maintenance Activities (IMA...

  8. A Naval Aviation Maintenance Organizational Activity Strategic Information System (OASIS)

    Science.gov (United States)

    1990-03-01

    Management Plan ( CIMP ) [Ref. 49] lists all information systems currently being planned or developed to support NAVAIR missions. The four "functional...in the CIMP . They include local area networks (LANs) for specific commands, systems dedicated to supporting one level of maintenance, systems designed

  9. Cell swelling activates K+ and Cl- channels as well as nonselective, stretch-activated cation channels in ehrlich ascites tumor cells

    DEFF Research Database (Denmark)

    Christensen, Ove; Hoffmann, Else Kay

    1992-01-01

    Cell-attached patch-clamp recordings from Ehrlich ascites tumor cells reveal nonselective cation channels which are activated by mechanical deformation of the membrane. These channels are seen when suction is applied to the patch pipette or after osmotic cell swelling. The channel activation does...... system. In isolated insideout patches a Ca2+-dependent, inwardly rectifying K+ channel is demonstrated. The single-channel conductance recorded with symmetrical 150 mm K+ solutions is for inward current estimated at 40 pS and for outward current at 15 pS. Activation of the K+ channel takes place after...... by membrane stretch (suction). The time-averaged number of open K+ channels during regulatory volume decrease (RVD) can be estimated at 40 per cell. The number of open K+ channels following addition of Ca2+ plus ionophore A23187 was estimated at 250 per cell. Concurrent activation in cell-attached patches...

  10. Peripheral hyperpolarization-activated cyclic nucleotide-gated channels contribute to inflammation-induced hypersensitivity of the rat temporomandibular joint.

    Science.gov (United States)

    Hatch, R J; Jennings, E A; Ivanusic, J J

    2013-08-01

    Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels conduct an inward cation current (Ih ) that contributes to the maintenance of neuronal membrane potential and have been implicated in a number of animal models of neuropathic and inflammatory pain. In the current study, we investigated HCN channel involvement in inflammatory pain of the temporomandibular joint (TMJ). The contribution of HCN channels to inflammation (complete Freund's adjuvant; CFA)-induced mechanical hypersensitivity of the rat TMJ was tested with injections of the HCN channel blocker ZD7288. Retrograde labelling and immunohistochemistry was used to explore HCN channel expression in sensory neurons that innervate the TMJ. Injection of CFA into the TMJ (n = 7) resulted in a significantly increased mechanical sensitivity relative to vehicle injection (n = 7) (p blocked by co-injection of ZD7288 with the CFA (n = 7). Retrograde labelling and immunohistochemistry experiments revealed expression predominantly of HCN1 and HCN2 channel subunits in trigeminal ganglion neurons that innervate the TMJ (n = 3). No change in the proportion or intensity of HCN channel expression was found in inflamed (n = 6) versus control (n = 5) animals at the time point tested. Our findings suggest a role for peripheral HCN channels in inflammation-induced pain of the TMJ. Peripheral application of a HCN channel blocker could provide therapeutic benefit for inflammatory TMJ pain and avoid side effects associated with activation of HCN channels in the central nervous system. © 2012 European Federation of International Association for the Study of Pain Chapters.

  11. Nuclear power plant maintenance optimisation SENUF network activity

    International Nuclear Information System (INIS)

    Ahlstrand, R.; Bieth, M.; Pla, P.; Rieg, C.; Trampus, P.

    2004-01-01

    During providing scientific and technical support to TACIS and PHARE nuclear safety programs a large amount of knowledge related to Russian design reactor systems has accumulated and led to creation of a new Network concerning Nuclear Safety in Central and Eastern Europe called ''Safety of Eastern European type Nuclear Facilities'' (SENUF). SENUF contributes to bring together all stakeholders of TACIS and PHARE: beneficiaries, end users, Eastern und Western nuclear industries, and thus, to favour fruitful technical exchanges and feedback of experience. At present the main focus of SENUF is the nuclear power plant maintenance as substantial element of plant operational safety as well as life management. A Working Group has been established on plant maintenance. One of its major tasks in 2004 is to prepare a status report on advanced strategies to optimise maintenance. Optimisation projects have an interface with the plant's overall life management program. Today, almost all plants involved in SENUF network have an explicit policy to extend their service life, thus, component ageing management, modernization and refurbishment actions became much more important. A database is also under development, which intends to help sharing the available knowledge and specific equipment and tools. (orig.)

  12. Nuclear power plant maintenance optimisation SENUF network activity

    Energy Technology Data Exchange (ETDEWEB)

    Ahlstrand, R.; Bieth, M.; Pla, P.; Rieg, C.; Trampus, P. [Inst. for Energy, EC DG Joint Research Centre, Petten (Netherlands)

    2004-07-01

    During providing scientific and technical support to TACIS and PHARE nuclear safety programs a large amount of knowledge related to Russian design reactor systems has accumulated and led to creation of a new Network concerning Nuclear Safety in Central and Eastern Europe called ''Safety of Eastern European type Nuclear Facilities'' (SENUF). SENUF contributes to bring together all stakeholders of TACIS and PHARE: beneficiaries, end users, Eastern und Western nuclear industries, and thus, to favour fruitful technical exchanges and feedback of experience. At present the main focus of SENUF is the nuclear power plant maintenance as substantial element of plant operational safety as well as life management. A Working Group has been established on plant maintenance. One of its major tasks in 2004 is to prepare a status report on advanced strategies to optimise maintenance. Optimisation projects have an interface with the plant's overall life management program. Today, almost all plants involved in SENUF network have an explicit policy to extend their service life, thus, component ageing management, modernization and refurbishment actions became much more important. A database is also under development, which intends to help sharing the available knowledge and specific equipment and tools. (orig.)

  13. Swell activated chloride channel function in human neutrophils

    Energy Technology Data Exchange (ETDEWEB)

    Salmon, Michael D. [Leukocyte and Ion Channel Research Laboratory, School of Health and Biosciences, University of East London, Stratford Campus, London E15 4LZ (United Kingdom); Ahluwalia, Jatinder, E-mail: j.ahluwalia@uel.ac.uk [Leukocyte and Ion Channel Research Laboratory, School of Health and Biosciences, University of East London, Stratford Campus, London E15 4LZ (United Kingdom)

    2009-04-17

    Non-excitable cells such as neutrophil granulocytes are the archetypal inflammatory immune cell involved in critical functions of the innate immune system. The electron current generated (I{sub e}) by the neutrophil NADPH oxidase is electrogenic and rapidly depolarises the membrane potential. For continuous function of the NADPH oxidase, I{sub e} has to be balanced to preserve electroneutrality, if not; sufficient depolarisation would prevent electrons from leaving the cell and neutrophil function would be abrogated. Subsequently, the depolarisation generated by the neutrophil NADPH oxidase I{sub e} must be counteracted by ion transport. The finding that depolarisation required counter-ions to compensate electron transport was followed by the observation that chloride channels activated by swell can counteract the NADPH oxidase membrane depolarisation. In this mini review, we discuss the research findings that revealed the essential role of swell activated chloride channels in human neutrophil function.

  14. Design and evaluation of augmented reality for nuclear power plant maintenance based on Cognitive activity analysis

    International Nuclear Information System (INIS)

    Koo, Jwa Jin

    2008-02-01

    The goals of maintenance in nuclear power plants (NPPs) are to reduce the number of plant shutdowns, to decrease the shutdown time, to minimize the maintenance costs, and to improve nuclear safety. Furthermore, it is difficult for maintenance personnel to conduct work in NPPs due to the complexity of NPPs, safety issues and time constraints. Human errors should be prevented to achieve maintenance goals in NPPs. Therefore, it is necessary to improve working conditions by introducing support systems. Augmented Reality (AR) has the potential to assist with maintenance tasks and make maintenance personnel concentrate on the tasks, which will then the reduced human error and improve human performance. The working procedure for AR systems was designed based on the characteristics of cognitive activity, and an experiment was conducted to estimate the effectiveness of AR. The results indicated that a well-designed working procedure for AR systems was more effective than a conventional paper-based working procedure. In addition, participants made fewer errors using the designed working procedure. AR systems as maintenance support systems are beneficial technology that can improve maintenance performance by reducing cognitive loads and unnecessary movements. An AR application can be attractive and effective at supporting maintenance task

  15. Dynamic contracting : An asset management tool in controlling infrastructure maintenance activities

    NARCIS (Netherlands)

    Demirel, H.C.; de Ridder, H.A.J.; Hertogh, M.J.C.M.

    2013-01-01

    Infrastructure road network is a complex system in a fast changing complicated environment and therefore subject to change. The changes refer to demands, requirements, regulations and financial possibilities as well as advanced technologies. Therefore outsourcing maintenance activities are rather

  16. Mutational analysis of an archaeal minichromosome maintenance protein exterior hairpin reveals critical residues for helicase activity and DNA binding

    Directory of Open Access Journals (Sweden)

    Brewster Aaron S

    2010-08-01

    Full Text Available Abstract Background The mini-chromosome maintenance protein (MCM complex is an essential replicative helicase for DNA replication in Archaea and Eukaryotes. While the eukaryotic complex consists of six homologous proteins (MCM2-7, the archaeon Sulfolobus solfataricus has only one MCM protein (ssoMCM, six subunits of which form a homohexamer. We have recently reported a 4.35Å crystal structure of the near full-length ssoMCM. The structure reveals a total of four β-hairpins per subunit, three of which are located within the main channel or side channels of the ssoMCM hexamer model generated based on the symmetry of the N-terminal Methanothermobacter thermautotrophicus (mtMCM structure. The fourth β-hairpin, however, is located on the exterior of the hexamer, near the exit of the putative side channels and next to the ATP binding pocket. Results In order to better understand this hairpin's role in DNA binding and helicase activity, we performed a detailed mutational and biochemical analysis of nine residues on this exterior β-hairpin (EXT-hp. We examined the activities of the mutants related to their helicase function, including hexamerization, ATPase, DNA binding and helicase activities. The assays showed that some of the residues on this EXT-hp play a role for DNA binding as well as for helicase activity. Conclusions These results implicate several current theories regarding helicase activity by this critical hexameric enzyme. As the data suggest that EXT-hp is involved in DNA binding, the results reported here imply that the EXT-hp located near the exterior exit of the side channels may play a role in contacting DNA substrate in a manner that affects DNA unwinding.

  17. Program plan for future regulatory activity in nuclear-power-plant maintenance

    International Nuclear Information System (INIS)

    Badalamente, R.V.

    1982-10-01

    The intent of this paper is to describe the results of a study of nuclear power plant (NPP) maintenance conducted by Battelle's Pacific Northwest Laboratories (PNL) for the Nuclear Regulatory Commission (NRC). The purpose of the study for the NRC was to determine problems affecting human performance in NPP maintenance, pinpoint those which adversely affect public health and safety, review strategies for overcoming the problems, and suggest the direction that regulatory activities should take. Results of the study were presented to the NRC (Division of Human Factors Safety) in the form of a recommended program plan for future regulatory activity in NPP maintenance

  18. Experience with ActiveX control for simple channel access

    International Nuclear Information System (INIS)

    Timossi, C.; Nishimura, H.; McDonald, J.

    2003-01-01

    Accelerator control system applications at Berkeley Lab's Advanced Light Source (ALS) are typically deployed on operator consoles running Microsoft Windows 2000 and utilize EPICS[2]channel access for data access. In an effort to accommodate the wide variety of Windows based development tools and developers with little experience in network programming, ActiveX controls have been deployed on the operator stations. Use of ActiveX controls for use in the accelerator control environment has been presented previously[1]. Here we report on some of our experiences with the use and development of these controls

  19. Activation of stretch-activated channels and maxi-K+ channels by membrane stress of human lamina cribrosa cells.

    LENUS (Irish Health Repository)

    Irnaten, Mustapha

    2009-01-01

    The lamina cribrosa (LC) region of the optic nerve head is considered the primary site of damage in glaucomatous optic neuropathy. Resident LC cells have a profibrotic potential when exposed to cyclical stretch. However, the mechanosensitive mechanisms of these cells remain unknown. Here the authors investigated the effects of membrane stretch on cell volume change and ion channel activity and examined the associated changes in intracellular calcium ([Ca(2+)](i)).

  20. The impact of self-efficacy on physical activity maintenance in patients with hip osteoarthritis

    DEFF Research Database (Denmark)

    Hammer, Nanna Maria; Bieler, Theresa; Beyer, Nina Ann-Marie

    2016-01-01

    Purpose: Understanding motivational factors related to physical activity (PA) maintenance is essential in promoting long-term exercise benefits. This study explored the impact of self-efficacy (SE) on post-intervention PA maintenance in patients with hip osteoarthritis. Method: An SE-theory based...... mixed-methods sub-study of a trial investigating the effects of 4 months supervised exercise in patients with hip osteoarthritis. Questionnaire data (n = 52; baseline and 12 months) on PA and SE (Arthritis Self-Efficacy Scale, ASES, score-range 10–100) were analysed (Mann–Whitney test) for differences...... from incorporating the self-efficacy theory in the planning and execution of exercise interventions to promote post-intervention physical activity maintenance and long term health benefits. •Post-intervention physical activity maintenance may be increased by focussing on the patients’ exercise self-efficacy...

  1. Expression, purification and functional reconstitution of slack sodium-activated potassium channels.

    Science.gov (United States)

    Yan, Yangyang; Yang, Youshan; Bian, Shumin; Sigworth, Fred J

    2012-11-01

    The slack (slo2.2) gene codes for a potassium-channel α-subunit of the 6TM voltage-gated channel family. Expression of slack results in Na(+)-activated potassium channel activity in various cell types. We describe the purification and reconstitution of Slack protein and show that the Slack α-subunit alone is sufficient for potassium channel activity activated by sodium ions as assayed in planar bilayer membranes and in membrane vesicles.

  2. Curcumin inhibits activation of TRPM2 channels in rat hepatocytes

    Directory of Open Access Journals (Sweden)

    E. Kheradpezhouh

    2016-04-01

    Full Text Available Oxidative stress is a hallmark of many liver diseases including viral and drug-induced hepatitis, ischemia-reperfusion injury, and non-alcoholic steatohepatitis. One of the consequences of oxidative stress in the liver is deregulation of Ca2+ homeostasis, resulting in a sustained elevation of the free cytosolic Ca2+ concentration ([Ca2+]c in hepatocytes, which leads to irreversible cellular damage. Recently it has been shown that liver damage induced by paracetamol and subsequent oxidative stress is, in large part, mediated by Ca2+ entry through Transient Receptor Potential Melastatin 2 (TRPM2 channels. Involvement of TRPM2 channels in hepatocellular damage induced by oxidative stress makes TRPM2 a potential therapeutic target for treatment of a range of oxidative stress-related liver diseases. We report here the identification of curcumin ((1E,6E-1,7-bis(4-hydroxy-3-methoxyphenyl-1,6-heptadiene-3,5-dione, a natural plant-derived polyphenol in turmeric spice, as a novel inhibitor of TRPM2 channel. Presence of 5 µM curcumin in the incubation medium prevented the H2O2- and paracetamol-induced [Ca2+]c rise in rat hepatocytes. Furthermore, in patch clamping experiments incubation of hepatocytes with curcumin inhibited activation of TRPM2 current by intracellular ADPR with IC50 of approximately 50 nM. These findings enhance understanding of the actions of curcumin and suggest that the known hepatoprotective properties of curcumin are, at least in part, mediated through inhibition of TRPM2 channels.

  3. BAX channel activity mediates lysosomal disruption linked to Parkinson disease.

    Science.gov (United States)

    Bové, Jordi; Martínez-Vicente, Marta; Dehay, Benjamin; Perier, Celine; Recasens, Ariadna; Bombrun, Agnes; Antonsson, Bruno; Vila, Miquel

    2014-05-01

    Lysosomal disruption is increasingly regarded as a major pathogenic event in Parkinson disease (PD). A reduced number of intraneuronal lysosomes, decreased levels of lysosomal-associated proteins and accumulation of undegraded autophagosomes (AP) are observed in PD-derived samples, including fibroblasts, induced pluripotent stem cell-derived dopaminergic neurons, and post-mortem brain tissue. Mechanistic studies in toxic and genetic rodent PD models attribute PD-related lysosomal breakdown to abnormal lysosomal membrane permeabilization (LMP). However, the molecular mechanisms underlying PD-linked LMP and subsequent lysosomal defects remain virtually unknown, thereby precluding their potential therapeutic targeting. Here we show that the pro-apoptotic protein BAX (BCL2-associated X protein), which permeabilizes mitochondrial membranes in PD models and is activated in PD patients, translocates and internalizes into lysosomal membranes early following treatment with the parkinsonian neurotoxin MPTP, both in vitro and in vivo, within a time-frame correlating with LMP, lysosomal disruption, and autophagosome accumulation and preceding mitochondrial permeabilization and dopaminergic neurodegeneration. Supporting a direct permeabilizing effect of BAX on lysosomal membranes, recombinant BAX is able to induce LMP in purified mouse brain lysosomes and the latter can be prevented by pharmacological blockade of BAX channel activity. Furthermore, pharmacological BAX channel inhibition is able to prevent LMP, restore lysosomal levels, reverse AP accumulation, and attenuate mitochondrial permeabilization and overall nigrostriatal degeneration caused by MPTP, both in vitro and in vivo. Overall, our results reveal that PD-linked lysosomal impairment relies on BAX-induced LMP, and point to small molecules able to block BAX channel activity as potentially beneficial to attenuate both lysosomal defects and neurodegeneration occurring in PD.

  4. Assessment of complex microwaves occupational exposure in radar maintenance activity

    International Nuclear Information System (INIS)

    Danulescu, R.

    1996-01-01

    The modern of the society teas determined the increase of thousand times greater than the natural fond of the humankind exposure to a complex combination of electromagnetic man-made fields and radiations of extremely various strength and frequencies. A special contribution to this environmental change has had in the last decade the appearance and the explosive development of the microwaves generating appliances such as radars used in a great variety of military and civilian applications and which essentially contributes to the electromagnetic pollution. In the above mentioned content which firstly interests the occupational environment, it is necessary to improve the exposure limits, as well as, the emission standards, in order to better protect the human health and well-being. From this point of view, the estimation of the microwave occupational exposure risk constitutes, alongside the health status assessment, one of the priorities of the Occupational Health because the theoretical and practical problems related to the bioeffects of this kind of radiations are far to be clarified. Our study has been carried out in a factory where one performs research, production and especially maintenance of microwaves generating devices. (author)

  5. Formation and maintenance of single-thread tie channels entering floodplain lakes: Observations from three diverse river systems

    Science.gov (United States)

    Rowland, J. C.; Dietrich, W. E.; Day, G.; Parker, G.

    2009-06-01

    Tie channels connect rivers to floodplain lakes on many lowland rivers and thereby play a central role in floodplain sedimentology and ecology; yet they are generally unrecognized and little studied. Here we report the results of field studies focused on tie channel origin and morphodynamics in the following three contrasting systems: the Middle Fly River (Papua New Guinea), the Lower Mississippi River, and Birch Creek in Alaska. Across these river systems, tie channels vary by an order of magnitude in size but exhibit the same characteristic morphology and appear to develop and evolve by a similar set of processes. In all three systems, the channels are characterized by a narrow, leveed, single-thread morphology with maximum width approximately one tenth the width of the mainstem river. The channels typically have a V-shaped cross section, unlike most fluvial channels. These channels develop as lakes become isolated from the river by sedimentation. Narrowing of the connection between river and lake causes a sediment-laden jet to develop. Levees develop along the margins of the jet leading to channel emergence and eventual levee aggradation to the height of the mainstem levees. Bidirectional flow in these channels is common. Outflows from the lake scour sediment and prevent channel blockage. We propose that channel geometry and size are then controlled by a dynamic balance between channel narrowing by suspended sediment deposition and incision and widening by mass failure of banks during outflows. Tie channels are laterally stable and may convey flow for hundreds to a few thousand of years.

  6. The formation and maintenance of single-thread tie channels entering floodplain lakes: observations from three diverse river systems

    Energy Technology Data Exchange (ETDEWEB)

    Rowland, Joel C [Los Alamos National Laboratory; Dietrich, William E [UC BERKELEY; Day, Geoff [NEWCREST MINING; Parker, Gary [UNIV OF ILLINOIS

    2009-01-01

    Tie channels connect rivers to floodplain lakes on many lowland rivers and thereby play a central role in floodplain sedimentology and ecology, yet they are generally unrecognized and little studied. here we report the results of field studies focused on tie channel origin and morphodynamics in three contrasting systems: the Middle Fly River, Papua New Guinea, the Lower Mississippi River, and Birch Creek in Alaska. Across these river systems, tie channels vary by an order of magnitude in size but exhibit the same characteristic morphology and appear to develop and evolve by a similar set of processes. In all three systems, the channels are characterized by a narrow, leveed single-thread morphology with maximum width approximately one tenth the width of the mainstem river. The channels typically have a V shaped cross-section, unlike most fluvial channels. These channels develop as lakes become isolated from the river by sedimentation. Narrowing of the connection between river and lake causes a sediment-laden jet to develop. Levees develop along the margins of the jet leading to channel emergence and eventual levee aggradation to the height of the mainstem levees. Bi-directional flow in these channels is common. Outflows from the lake scour sediment and prevent channel blockage. We propose that channel geometry and size are then controlled by a dynamic balance between channel narrowing by suspended sediment deposition and incision and widening by mass failure of banks during outflows. Tie channels are laterally stable and may convey flow for hundreds to a few thousand of years.

  7. Summary of typical routine maintenance activities at Tokai Reprocessing Plant. Supplement (March, 2002)

    International Nuclear Information System (INIS)

    2002-03-01

    Typical maintenance activities, such as replacement of worn out parts and cleaning of filter elements, routinely performed during steady operation are summarized. [The Summary of Typical Routine Maintenance Activities at Tokai Reprocessing Plant] (JNC TN 8450 2001-006) was already prepared in September, 2001. The purpose of this summary is to give elementary understanding on these activities to people who are responsible for explanation them to the public. At this time, the same kind of summary is prepared as a supplement of the previous one. (author)

  8. Maintenance and decline of physical activity during adolescence: insights from a qualitative study

    Directory of Open Access Journals (Sweden)

    Filion Annie

    2011-10-01

    Full Text Available Abstract Purpose Better knowledge on why some individuals succeed in maintaining participation in physical activity throughout adolescence is needed to guide the development of effective interventions to increase and then maintain physical activity levels. Despite allowing an in-depth understanding, qualitative designs have infrequently been used to study physical activity maintenance. We explored factors contributing to the maintenance and the decline of physical activity during adolescence. Methods Questionnaires were administered to 515 grade 10-12 students. The Physical Activity Questionnaire for Adolescents was used to determine physical activity level at the end of adolescence. An adapted version of this questionnaire was used to estimate physical activity in early adolescence. Among both genders, we identified participants who maintained a high level of physical activity since grade 7 and some whose activity level declined. For each category, groups of 10 students were randomly selected to take part in focus group discussions. Results Seven focus groups with 5 to 8 participants in each were held. Both maintainers and decliners associated physical activity with positive health outcomes. Maintenance of physical activity was associated with supportive social environments and heightened feelings of competence and attractiveness. A decline in physical activity was associated with negative social validation, poor social support and barriers related to access. Conclusions Although maintainers and decliners associate physical activity with similar themes, the experiences of both groups differ substantially with regards to those themes. Taking both perspectives in consideration could help improve interventions to increase and maintain physical activity levels of adolescents.

  9. Milrinone-Induced Postconditioning Requires Activation of Mitochondrial Ca2+-sensitive Potassium (mBKCa) Channels

    NARCIS (Netherlands)

    Behmenburg, Friederike; Trefz, Lara; Dorsch, Marianne; Ströthoff, Martin; Mathes, Alexander; Raupach, Annika; Heinen, André; Hollmann, Markus W.; Berger, Marc M.; Huhn, Ragnar

    2017-01-01

    Cardioprotection by postconditioning requires activation of mitochondrial large-conductance Ca2+-sensitive potassium (mBKCa) channels. The involvement of these channels in milrinone-induced postconditioning is unknown. The authors determined whether cardioprotection by milrinone-induced

  10. Integrated fuzzy analytic hierarchy process and VIKOR method in the prioritization of pavement maintenance activities

    Directory of Open Access Journals (Sweden)

    Peyman Babashamsi

    2016-03-01

    Full Text Available Maintenance activities and pavement rehabilitation require the allocation of massive finances. Yet due to budget shortfalls, stakeholders and decision-makers must prioritize projects in maintenance and rehabilitation. This article addresses the prioritization of pavement maintenance alternatives by integrating the fuzzy analytic hierarchy process (AHP with the VIKOR method (which stands for ‘VlseKriterijumska Optimizacija I Kompromisno Resenje,’ meaning multi-criteria optimization and compromise solution for the process of multi-criteria decision analysis (MCDA by considering various pavement network indices. The indices selected include the pavement condition index (PCI, traffic congestion, pavement width, improvement and maintenance costs, and the time required to operate. In order to determine the weights of the indices, the fuzzy AHP is used. Subsequently, the alternatives’ priorities are ranked according to the indices weighted with the VIKOR model. The choice of these two independent methods was motivated by the fact that integrating fuzzy AHP with the VIKOR model can assist decision makers with solving MCDA problems. The case study was conducted on a pavement network within the same particular region in Tehran; three main streets were chosen that have an empirically higher maintenance demand. The most significant factors were evaluated and the project with the highest priority was selected for urgent maintenance. By comparing the index values of the alternative priorities, Delavaran Blvd. was revealed to have higher priority over the other streets in terms of maintenance and rehabilitation activities. Keywords: Maintenance and rehabilitation prioritization, Fuzzy analysis hierarchy process, VIKOR model, Pavement condition index, Multi-criteria decision analysis

  11. Quantification and distribution of big conductance Ca2+-activated K+ channels in kidney epithelia

    DEFF Research Database (Denmark)

    Grunnet, Morten; Hay-Schmidt, Anders; Klaerke, Dan A

    2005-01-01

    and immunohistochemical studies. In cortical collecting ducts, BK channels were exclusively located in principal cells while no channels could be found in intercalated cells. The abundant and distinct distribution in kidney epithelia talks in favor for BK channels being important contributors in maintaining salt......Big conductance Ca2+ activated K+ channels (BK channels) is an abundant channel present in almost all kind of tissue. The accurate quantity and especially the precise distribution of this channel in kidney epithelia are, however, still debated. The aim of the present study has therefore been...... to examine the presence of BK channels in kidney epithelia and determine the actual number and distribution of these channels. For this purpose, a selective peptidyl ligand for BK channels called iberiotoxin or the radiolabeled double mutant analog 125I-IbTX-D19Y/Y36F has been employed. The presence of BK...

  12. Persistent discharges in dentate gyrus perisoma-inhibiting interneurons require hyperpolarization-activated cyclic nucleotide-gated channel activation.

    Science.gov (United States)

    Elgueta, Claudio; Köhler, Johannes; Bartos, Marlene

    2015-03-11

    Parvalbumin (PV)-expressing perisoma-inhibiting interneurons (PIIs) of the dentate gyrus integrate rapidly correlated synaptic inputs and generate short-duration action potentials that propagate along the axon to their output synapses, supporting fast inhibitory signaling onto their target cells. Here we show that PV-PIIs in rat and mouse dentate gyrus (DG) integrate their intrinsic activity over time and can turn into a persistent firing mode characterized by the ability to generate long-lasting trains of action potentials at ∼50 Hz in the absence of additional inputs. Persistent firing emerges in the axons remote from the axon initial segment and markedly depends on hyperpolarization-activated cyclic nucleotide-gated channel (HCNC) activation. Persistent firing properties are modulated by intracellular Ca(2+) levels and somatic membrane potential. Detailed computational single-cell PIIs models reveal that HCNC-mediated conductances can contribute to persistent firing during conditions of a shift in their voltage activation curve to more depolarized potentials. Paired recordings from PIIs and their target granule cells show that persistent firing supports strong inhibitory output signaling. Thus, persistent firing may emerge during conditions of intense activation of the network, thereby providing silencing to the circuitry and the maintenance of sparse activity in the dentate gyrus. Copyright © 2015 the authors 0270-6474/15/354131-09$15.00/0.

  13. Remote maintenance design activities and research and development accomplishments for the Compact Ignition Tokamak

    International Nuclear Information System (INIS)

    Spampinato, P.T.

    1988-01-01

    The use of deuterium-tritium (D-T) fuel for the Compact Ignition Tokamak (CIT) requires the use of remote handling technology to carry out maintenance operations. The remote operations consist of removing and replacing such components as first wall armor protection tiles, radio-frequency (rf) heating modules, and diagnostic modules. The major pieces of equipment being developed for maintenance activities internal to the vacuum vessel include an articulated boom manipulator (ABM), an inspection manipulator, and special tooling. For activities external to the vessel, the equipment includes a bridge-mounted manipulator system, decontamination equipment, hot cell equipment, and solid radiation-waste (rad-waste) handling and packaging equipment. The CIT Project is completing the conceptual design phase; research and development (R and D) activities, which include demonstrations of remote maintenance operations on full-size partial mock-ups are under way. 5 figs

  14. The Function of the Novel Mechanical Activated Ion Channel Piezo1 in the Human Osteosarcoma Cells

    OpenAIRE

    Jiang, Long; Zhao, Yi-ding; Chen, Wei-xiang

    2017-01-01

    Background The Piezo1 protein ion channel is a novel mechanical activated ion channel which is related to mechanical signal transduction. However, the function of the mechanically activated ion channel Piezo1 had not been explored. In this study, we explored the function of the Piezo1 ion channel in human osteosarcoma (OS) cells related to apoptosis, invasion, and the cell proliferation. Material/Methods Reverse transcription polymerase chain reaction (RT-PCR) and western-blotting were used t...

  15. 25 CFR Appendix A to Subpart G - List of Activities Eligible for Funding Under BIA Transportation Facility Maintenance Program

    Science.gov (United States)

    2010-04-01

    ... Transportation Facility Maintenance Program A Appendix A to Subpart G Indians BUREAU OF INDIAN AFFAIRS, DEPARTMENT OF THE INTERIOR LAND AND WATER INDIAN RESERVATION ROADS PROGRAM BIA Road Maintenance Pt. 170... Transportation Facility Maintenance Program The following activities are eligible for BIA Transportation Facility...

  16. Homogenization of independent inspection criteria of Quality Assurance in the maintenance activities

    International Nuclear Information System (INIS)

    Gomez Cardinanos, R.

    2012-01-01

    Among the official documents governing the operation of Spanish nuclear power plants is the Quality Manual and Quality Assurance. This manual gives a formal answer to the requirements of the regulations and the Nuclear Safety Council. To monitor the compliance of its requirements, the plants are controlled by independent organizations. Since 2007, in nuclear Sta. Maria de Garona is developed a project to improve the performance of the QA inspectors during maintenance activities. Results of the project have been edited data sheets on specific activities of maintenance. The project aims to incorporate sheets that help QA inspectors in their daily inspection.

  17. Opportunities and supporting activities to promote preventive maintenance of NPPs in Japan

    International Nuclear Information System (INIS)

    Sanada, A.; Shinkawa, T.; Sakurada, M.

    1998-01-01

    With increase of the number of NPPs and operation history, enhancement of the safety regulation is becoming important for such long-operated NPPs. Centering on the comprehensive preventive maintenance, periodic safety reviews by utilities and its review and evaluation by MITI are in progress. The first reviews have not revealed essential and critical indications to be newly implemented. This means that the most of activities to secure reliability and safety have been done steadily. The present paper addresses the mechanism of on-going preventive maintenance and its essential elements: opportunities to identify issues and problems, and supporting activities to promote decision-makings on feedback, upgrading and modernization of NPPs. (author)

  18. Integrity Checking and Maintenance with Active Rules in XML Databases

    DEFF Research Database (Denmark)

    Christiansen, Henning; Rekouts, Maria

    2007-01-01

    While specification languages for integrity constraints for XML data have been considered in the literature, actual technologies and methodologies for checking and maintaining integrity are still in their infancy. Triggers, or active rules, which are widely used in previous technologies for the p...... updates, the method indicates trigger conditions and correctness criteria to be met by the trigger code supplied by a developer or possibly automatic methods. We show examples developed in the Sedna XML database system which provides a running implementation of XML triggers....

  19. Basic Characteristics of Human Erroneous Actions during Test and Maintenance Activities Leading to Unplanned Reactor Trips

    International Nuclear Information System (INIS)

    Kim, Jae Whan; Park, Jin Kyun

    2010-01-01

    Test and maintenance (T and M) activities of nuclear power plants are essential for sustaining the safety of a power plant and maintaining the reliability of plant systems and components. However, the potential of human errors during T and M activities has also the potential to induce unplanned reactor trips or power derate or making safety-related systems unavailable. According to the major incident/accident reports of nuclear power plants in Korea, contribution of human errors takes up about 20% of the total events. The previous study presents that most of human-related unplanned reactor trip events during normal power operation are associated with T and M activities (63%), which are comprised of plant maintenance activities such as a 'periodic preventive maintenance (PPM)', a 'planned maintenance (PM)' and a 'corrective maintenance (CM)'. This means that T and M activities should be a major subject for reducing the frequency of human-related unplanned reactor trips. This paper aims to introduce basic characteristics of human erroneous actions involved in the test and maintenance-induced unplanned reactor trip events that have occurred between 1986 and 2006 in Korean nuclear power plants. The basic characteristics are described by dividing human erroneous actions into planning-based errors and execution-based errors. For the events associated with planning failures, they are, firstly, classified according to existence of the work procedure and then described for what aspects of the procedure or work plan have deficiency or problem. On the other hand, for the events associated with execution failures, they are described from the aspect of external error modes

  20. Localization of Ca2+ -activated big-conductance K+ channels in rabbit distal colon

    DEFF Research Database (Denmark)

    Hay-Schmidt, Anders; Grunnet, Morten; Abrahamse, Salomon L

    2003-01-01

    Big-conductance Ca(2+)-activated K(+) channels (BK channels) may play an important role in the regulation of epithelial salt and water transport, but little is known about the expression level and the precise localization of BK channels in epithelia. The aim of the present study was to quantify a...

  1. The Sodium-Activated Potassium Channel Slack Is Required for Optimal Cognitive Flexibility in Mice

    Science.gov (United States)

    Bausch, Anne E.; Dieter, Rebekka; Nann, Yvette; Hausmann, Mario; Meyerdierks, Nora; Kaczmarek, Leonard K.; Ruth, Peter; Lukowski, Robert

    2015-01-01

    "Kcnt1" encoded sodium-activated potassium channels (Slack channels) are highly expressed throughout the brain where they modulate the firing patterns and general excitability of many types of neurons. Increasing evidence suggests that Slack channels may be important for higher brain functions such as cognition and normal intellectual…

  2. Transfer of technology in maintenance tools

    International Nuclear Information System (INIS)

    Delorme, H.; Cartry, J.P.

    1985-10-01

    For maximum reliability of maintenance operations, FRAMATOME has aimed to improve the accessibility of highly irradiated and contaminated reactor coolant system equipment by the active promotion of an automation and robotization strategy. A number of robot designs have been developed providing replacement of operators inside the steam generator channel head, smooth handling of heavy or cumbersome objects in this zone, easy marking of steam generator tubes and increased speed of repetitive channel head operations: maintenance spider, theta-theta prime robotic service arm, R-theta robotic service arm, remote manipulator arm. Two operations are described: channel head folding cover plate and, operation of the torquing machine for head cover plate

  3. Human area MT+ shows load-dependent activation during working memory maintenance with continuously morphing stimulation.

    Science.gov (United States)

    Galashan, Daniela; Fehr, Thorsten; Kreiter, Andreas K; Herrmann, Manfred

    2014-07-11

    Initially, human area MT+ was considered a visual area solely processing motion information but further research has shown that it is also involved in various different cognitive operations, such as working memory tasks requiring motion-related information to be maintained or cognitive tasks with implied or expected motion.In the present fMRI study in humans, we focused on MT+ modulation during working memory maintenance using a dynamic shape-tracking working memory task with no motion-related working memory content. Working memory load was systematically varied using complex and simple stimulus material and parametrically increasing retention periods. Activation patterns for the difference between retention of complex and simple memorized stimuli were examined in order to preclude that the reported effects are caused by differences in retrieval. Conjunction analysis over all delay durations for the maintenance of complex versus simple stimuli demonstrated a wide-spread activation pattern. Percent signal change (PSC) in area MT+ revealed a pattern with higher values for the maintenance of complex shapes compared to the retention of a simple circle and with higher values for increasing delay durations. The present data extend previous knowledge by demonstrating that visual area MT+ presents a brain activity pattern usually found in brain regions that are actively involved in working memory maintenance.

  4. Immunolocalization and expression of small-conductance calcium-activated potassium channels in human myometrium

    DEFF Research Database (Denmark)

    Rosenbaum, Sofia T; Svalø, Julie; Nielsen, Karsten

    2012-01-01

    Small-conductance calcium-activated potassium (SK3) channels have been detected in human myometrium and we have previously shown a functional role of SK channels in human myometrium in vitro. The aims of this study were to identify the precise localization of SK3 channels and to quantify SK3 m....... This is the first report to provide evidence for a possible role of SK3 channels in human uterine telocytes....

  5. Chloride Channel 3 Channels in the Activation and Migration of Human Blood Eosinophils in Allergic Asthma.

    Science.gov (United States)

    Gaurav, Rohit; Bewtra, Againdra K; Agrawal, Devendra K

    2015-08-01

    Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase is responsible for respiratory burst in immune cells. Chloride channel 3 (CLC3) has been linked to the respiratory burst in eosinophils and neutrophils. The effect of cytokines and the involvement of CLC3 in the regulation of NADPH-dependent oxidative stress and on cytokine-mediated migration of eosinophils are not known. Human peripheral blood eosinophils were isolated from healthy individuals and from individuals with asthma by negative selection. Real-time PCR was used to detect the expression of NADPH oxidases in eosinophils. Intracellular reactive oxygen species (ROS) measurement was done with flow cytometry. Superoxide generation was measured with transforming growth factor (TGF)-β, eotaxin, and CLC3 blockers. CLC3 dependence of eosinophils in TGF-β- and eotaxin-induced migration was also examined. The messenger RNA (mRNA) transcripts of NADPH oxidase (NOX) 2, dual oxidase (DUOX) 1, and DUOX2 were detected in blood eosinophils, with very low expression of NOX1, NOX3, and NOX5 and no NOX4 mRNA. The level of NOX2 mRNA transcripts increased with disease severity in the eosinophils of subjects with asthma compared with healthy nonatopic volunteers. Change in granularity and size in eosinophils, but no change in intracellular ROS, was observed with phorbol myristate acetate (PMA). PMA, TGF-β, and eotaxin used the CLC3-dependent pathway to increase superoxide radicals. TGF-β and eotaxin induced CLC3-dependent chemotaxis of eosinophils. These findings support the requirement of CLC3 in the activation and migration of human blood eosinophils and may provide a potential novel therapeutic target to regulate eosinophil hyperactivity in allergic airway inflammation in asthma.

  6. Regulation of cloned, Ca2+-activated K+ channels by cell volume changes

    DEFF Research Database (Denmark)

    Grunnet, Morten; MacAulay, Nanna; Jorgensen, Nanna K

    2002-01-01

    Ca2+-activated K+ channels of big (hBK), intermediate (hIK) or small (rSK3) conductance were co-expressed with aquaporin 1 (AQP1) in Xenopus laevis oocytes. hBK channels were activated by depolarization, whereas hIK and rSK3 channels were activated by direct injection of Ca2+ or Cd2+ into the ooc...

  7. Recent activities within Nuclear Electric related to the qualification of digital systems and the extension of maintenance intervals

    International Nuclear Information System (INIS)

    Holt, G.P.

    1993-01-01

    Nuclear Electric is pursuing a program of modernization and enhancement of existing control and safety (C ampersand I) systems to improve power plant availability and increase power output. This paper describes recent experience in qualifying digital systems for safety applications. Also, described is the work currently in hand to reduce the frequency of maintenance periods for safety C ampersand I equipment. The qualification of digital systems is shown to be a more rigorous and much more extensive (and thus costly) exercise than the qualification of conventional equipment. This is illustrated by brief references to two software-based safety systems, the Single Channel Trip System at the Dungeness open-quotes Bclose quotesAGR and the Primary Protection System for the new PWR at Sizewell open-quotes Bclose quotes. The emphasis of the paper is on the use of Application Specific Integrated Circuits (ASICs) in safety systems, to reduce, or eliminate dependence on software for the fulfillment of requirements which need implementations that are complex or of a high functional density. Recent experience in the development of a prototype safety system based on ASICs, The Gaseous Activity Monitor and Trip Equipment (GAM), is used to illustrate their possible benefits and potential problems. When using complex integrated circuits like microprocessors or ASICs in digital systems a difficulty arises in analyzing the failure modes of the system. Efforts to overcome this difficulty led to the development of a method for performing failure analyses based on functional blocks. The techniques and benefits of such an approach are outlined. Finally, efforts to reduce maintenance costs, yet at the same time enhance safety, are leading towards an extension of maintenance intervals, for some safety equipment, from 3 months to 4 months and also towards lengthening the period between AGR outages from 2 years to 3 years

  8. Relationship between volition, physical activity and weight loss maintenance: Study rationale, design, methods and baseline characteristics.

    Science.gov (United States)

    Dandanell, Sune; Elbe, Anne-Marie; Pfister, Gertrud; Elsborg, Peter; W Helge, Jørn

    2017-05-01

    To investigate the relationship between volition, physical activity and weight loss maintenance. We recruited 84 sedentary (maximal oxygen uptake: 25 ± 5 ml/min), overweight and obese (Body mass index (BMI) 38 ± 7 m/h 2 , fat 44 ± 7 %) women ( n = 55) and men ( n = 29) for an interdisciplinary prospective study with follow-up. The change in lifestyle and weight loss is promoted via a 3-month intensive lifestyle intervention at a private health school. The intervention consists of supervised training (1-3 hours/day), a healthy hypo-caloric diet (-500 to -700 kCal/day) and education in healthy lifestyle in classes/groups. The participants' body weight and composition (Dual Energy X-ray absorptiometry), volitional skills (questionnaire), physical activity level (heart rate accelerometer/questionnaire) and maximal oxygen uptake (indirect calorimetry) are to be monitored before, after, and 3 and 12 months after the intervention. At the 12-month follow-up, three different groups will be established: Clinical weight loss maintenance (> 10% weight loss from baseline), moderate weight loss maintenance (1-10% weight loss) and no weight loss (or weight regain). A linear mixed model analysis will be used to compare levels of volitional skills, physical activity and maximal oxygen uptake over time, between the three groups. Correlational analyses will be used to investigate possible associations between volition, maximal oxygen uptake, physical activity level and weight loss maintenance. If specific volitional skills are identified as predictors of adherence to physical activity and success in clinical weight loss maintenance, these can be trained in future intensive lifestyle interventions in order to optimize the success rate.

  9. Remote maintenance design activities and research and development accomplishments for the compact ignition tokamak

    International Nuclear Information System (INIS)

    Spampinato, P.T.

    1989-01-01

    The use of deuterium-tritium (D-T) fuel for the Compact Ignition Tokamak (CIT) requires the use of remote handling technology in order to carry out maintenance operations. The remote operations consist of removing and replacing such components as first wall armor protection tiles, radio-frequency (RF) heating modules, and diagnostic modules. The major pieces of equipment being developed for maintenance operations internal to the vacuum vessel include an articulated boom manipulator (ABM), an inspection manipulator, and special tooling. For operations external to the vessel, the equipment includes a bridge-mounted manipulator system, decontamination equipment, hot cell equipment, and solid radioactive waste (rad-waste) handling and packaging equipment. The CIT Project is completing the conceptual design phase; research and development (R and D) activities, which include demonstrations of remote maintenance operations on full-size partial mock-ups are under way. (orig.)

  10. Coassembly of big conductance Ca2+-activated K+ channels and L-type voltage-gated Ca2+ channels in rat brain

    DEFF Research Database (Denmark)

    Grunnet, Morten; Kaufmann, Walter A

    2004-01-01

    Based on electrophysiological studies, Ca(2+)-activated K(+) channels and voltage-gated Ca(2+) channels appear to be located in close proximity in neurons. Such colocalization would ensure selective and rapid activation of K(+) channels by local increases in the cytosolic calcium concentration...

  11. Activation of ERG2 potassium channels by the diphenylurea NS1643

    DEFF Research Database (Denmark)

    Elmedyb, Pernille; Olesen, Søren-Peter; Grunnet, Morten

    2007-01-01

    Three members of the ERG potassium channel family have been described (ERG1-3 or Kv 11.1-3). ERG1 is by far the best characterized subtype and it constitutes the molecular component of the cardiac I(Kr) current. All three channel subtypes are expressed in neurons but their function remains unclear....... The lack of functional information is at least partly due to the lack of specific pharmacological tools. The compound NS1643 has earlier been reported as an ERG1 channel activator. We found that NS1643 also activates the ERG2 channel; however, the molecular mechanism of the activation differs between...... the ERG1 and ERG2 channels. This is surprising since ERG1 and ERG2 channels have very similar biophysical and structural characteristics. For ERG2, NS1643 causes a left-ward shift of the activation curve, a faster time-constant of activation and a slower time-constant of inactivation as well...

  12. Atomic basis for therapeutic activation of neuronal potassium channels

    DEFF Research Database (Denmark)

    Kim, Robin Y; Yau, Michael C; Galpin, Jason D

    2015-01-01

    Retigabine is a recently approved anticonvulsant that acts by potentiating neuronal M-current generated by KCNQ2-5 channels, interacting with a conserved Trp residue in the channel pore domain. Using unnatural amino-acid mutagenesis, we subtly altered the properties of this Trp to reveal specific...

  13. Repair, Evaluation, Maintenance, and Rehabilitation Research Program: Geotechnical Aspects of Rock Erosion in Emergency Spillway Channels. Report 3. Remediation

    Science.gov (United States)

    1988-09-01

    exposed, hard sandstone ledge which floors the upper portion of the unlined channel at the Saylorville (Iowa) spillway still bears the elongate scars ...this alluvium was placed 60-cm (2-ft) armour of angular cobble and boulder-size basalt riprap (US Soil Conservation Services 1932). All of the riprap

  14. Two Salt Bridges Differentially Contribute to the Maintenance of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Channel Function*

    Science.gov (United States)

    Cui, Guiying; Freeman, Cody S.; Knotts, Taylor; Prince, Chengyu Z.; Kuang, Christopher; McCarty, Nael A.

    2013-01-01

    Previous studies have identified two salt bridges in human CFTR chloride ion channels, Arg352-Asp993 and Arg347-Asp924, that are required for normal channel function. In the present study, we determined how the two salt bridges cooperate to maintain the open pore architecture of CFTR. Our data suggest that Arg347 not only interacts with Asp924 but also interacts with Asp993. The tripartite interaction Arg347-Asp924-Asp993 mainly contributes to maintaining a stable s2 open subconductance state. The Arg352-Asp993 salt bridge, in contrast, is involved in stabilizing both the s2 and full (f) open conductance states, with the main contribution being to the f state. The s1 subconductance state does not require either salt bridge. In confirmation of the role of Arg352 and Asp993, channels bearing cysteines at these sites could be latched into a full open state using the bifunctional cross-linker 1,2-ethanediyl bismethanethiosulfonate, but only when applied in the open state. Channels remained latched open even after washout of ATP. The results suggest that these interacting residues contribute differently to stabilizing the open pore in different phases of the gating cycle. PMID:23709221

  15. Bi-objective analysis of water-sediment regulation for channel scouring and delta maintenance: A study of the lower Yellow River

    Science.gov (United States)

    Kong, D.; Miao, C.; Duan, Q.

    2016-12-01

    Long-term hydrological data and remotely-sensed satellite images were used to analyze the effects of the water-sediment regulation scheme (WSRS) implemented in the lower Yellow River (LYR), China, between 1983 and 2013. The WSRS aimed to control channel scouring in the LYR and maintain the Yellow River Delta (YRD). Channel erosion in the LYR has primarily depended on the incoming sediment concentration at Xiaolangdi, where the concentration must be lower than approximately 9.17 × 10-3 t m-3 to avoid rising of the riverbed. In 1996, an artificial diversion altered the evolution of the YRD. To maintain delta equilibrium, an average sediment load of about 441 × 106 t year-1 was required before 1996, after which this value decreased to 167 × 106 t year-1. We provide a preliminary estimate of the incoming water and sediment conditions required at the Xiaolangdi station to guarantee both LYR channel scouring and maintenance of the YRD. Our results show that it is feasible to transport sediment originally deposited in the LYR to the river mouth to maintain the delta, which is of great significance for the future management and environmental protection of the LYR.

  16. Brain Activation during Associative Short-Term Memory Maintenance is Not Predictive for Subsequent Retrieval

    Directory of Open Access Journals (Sweden)

    Heiko eBergmann

    2015-09-01

    Full Text Available Performance on working memory (WM tasks may partially be supported by long-term memory (LTM processing. Hence, brain activation recently being implicated in WM may actually have been driven by (incidental LTM formation. We examined which brain regions actually support successful WM processing, rather than being confounded by LTM processes, during the maintenance and probe phase of a WM task. We administered a four-pair (faces and houses associative delayed-match-to-sample (WM task using event-related fMRI and a subsequent associative recognition LTM task, using the same stimuli. This enabled us to analyze subsequent memory effects for both the WM and the LTM test by contrasting correctly recognized pairs with incorrect pairs for either task. Critically, with respect to the subsequent WM effect, we computed this analysis exclusively for trials that were forgotten in the subsequent LTM recognition task. Hence, brain activity associated with successful WM processing was less likely to be confounded by incidental LTM formation. The subsequent LTM effect, in contrast, was analyzed exclusively for pairs that previously had been correctly recognized in the WM task, disclosing brain regions involved in successful LTM formation after successful WM processing. Results for the subsequent WM effect showed no significantly activated brain areas for WM maintenance, possibly due to an insensitivity of fMRI to mechanisms underlying active WM maintenance. In contrast, a correct decision at WM probe was linked to activation in the retrieval success network (anterior and posterior midline brain structures. The subsequent LTM analyses revealed greater activation in left dorsolateral prefrontal cortex and posterior parietal cortex in the early phase of the maintenance stage. No supra-threshold activation was found during the WM probe. Together, we obtained clearer insights in which brain regions support successful WM and LTM without the potential confound of the

  17. Brain activation during associative short-term memory maintenance is not predictive for subsequent retrieval.

    Science.gov (United States)

    Bergmann, Heiko C; Daselaar, Sander M; Beul, Sarah F; Rijpkema, Mark; Fernández, Guillén; Kessels, Roy P C

    2015-01-01

    Performance on working memory (WM) tasks may partially be supported by long-term memory (LTM) processing. Hence, brain activation recently being implicated in WM may actually have been driven by (incidental) LTM formation. We examined which brain regions actually support successful WM processing, rather than being confounded by LTM processes, during the maintenance and probe phase of a WM task. We administered a four-pair (faces and houses) associative delayed-match-to-sample (WM) task using event-related functional MRI (fMRI) and a subsequent associative recognition LTM task, using the same stimuli. This enabled us to analyze subsequent memory effects for both the WM and the LTM test by contrasting correctly recognized pairs with incorrect pairs for either task. Critically, with respect to the subsequent WM effect, we computed this analysis exclusively for trials that were forgotten in the subsequent LTM recognition task. Hence, brain activity associated with successful WM processing was less likely to be confounded by incidental LTM formation. The subsequent LTM effect, in contrast, was analyzed exclusively for pairs that previously had been correctly recognized in the WM task, disclosing brain regions involved in successful LTM formation after successful WM processing. Results for the subsequent WM effect showed no significantly activated brain areas for WM maintenance, possibly due to an insensitivity of fMRI to mechanisms underlying active WM maintenance. In contrast, a correct decision at WM probe was linked to activation in the "retrieval success network" (anterior and posterior midline brain structures). The subsequent LTM analyses revealed greater activation in left dorsolateral prefrontal cortex and posterior parietal cortex in the early phase of the maintenance stage. No supra-threshold activation was found during the WM probe. Together, we obtained clearer insights in which brain regions support successful WM and LTM without the potential confound of

  18. The temperature dependence of the BK channel activity - kinetics, thermodynamics, and long-range correlations.

    Science.gov (United States)

    Wawrzkiewicz-Jałowiecka, Agata; Dworakowska, Beata; Grzywna, Zbigniew J

    2017-10-01

    Large-conductance, voltage dependent, Ca 2+ -activated potassium channels (BK) are transmembrane proteins that regulate many biological processes by controlling potassium flow across cell membranes. Here, we investigate to what extent temperature (in the range of 17-37°C with ΔT=5°C step) is a regulating parameter of kinetic properties of the channel gating and memory effect in the series of dwell-time series of subsequent channel's states, at membrane depolarization and hyperpolarization. The obtained results indicate that temperature affects strongly the BK channels' gating, but, counterintuitively, it exerts no effect on the long-range correlations, as measured by the Hurst coefficient. Quantitative differences between dependencies of appropriate channel's characteristics on temperature are evident for different regimes of voltage. Examining the characteristics of BK channel activity as a function of temperature allows to estimate the net activation energy (E act ) and changes of thermodynamic parameters (ΔH, ΔS, ΔG) by channel opening. Larger E act corresponds to the channel activity at membrane hyperpolarization. The analysis of entropy and enthalpy changes of closed to open channel's transition suggest the entropy-driven nature of the increase of open state probability during voltage activation and supports the hypothesis about the voltage-dependent geometry of the channel vestibule. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Intracellular zinc activates KCNQ channels by reducing their dependence on phosphatidylinositol 4,5-bisphosphate.

    Science.gov (United States)

    Gao, Haixia; Boillat, Aurélien; Huang, Dongyang; Liang, Ce; Peers, Chris; Gamper, Nikita

    2017-08-01

    M-type (Kv7, KCNQ) potassium channels are proteins that control the excitability of neurons and muscle cells. Many physiological and pathological mechanisms of excitation operate via the suppression of M channel activity or expression. Conversely, pharmacological augmentation of M channel activity is a recognized strategy for the treatment of hyperexcitability disorders such as pain and epilepsy. However, physiological mechanisms resulting in M channel potentiation are rare. Here we report that intracellular free zinc directly and reversibly augments the activity of recombinant and native M channels. This effect is mechanistically distinct from the known redox-dependent KCNQ channel potentiation. Interestingly, the effect of zinc cannot be attributed to a single histidine- or cysteine-containing zinc-binding site within KCNQ channels. Instead, zinc dramatically reduces KCNQ channel dependence on its obligatory physiological activator, phosphatidylinositol 4,5-bisphosphate (PIP 2 ). We hypothesize that zinc facilitates interactions of the lipid-facing interface of a KCNQ protein with the inner leaflet of the plasma membrane in a way similar to that promoted by PIP 2 Because zinc is increasingly recognized as a ubiquitous intracellular second messenger, this discovery might represent a hitherto unknown native pathway of M channel modulation and provide a fresh strategy for the design of M channel activators for therapeutic purposes.

  20. The inhibitor of volume-regulated anion channels DCPIB activates TREK potassium channels in cultured astrocytes

    Czech Academy of Sciences Publication Activity Database

    Minieri, L.; Pivoňková, Helena; Caprini, M.; Harantová, Lenka; Anděrová, Miroslava; Ferroni, S.

    2013-01-01

    Roč. 168, č. 5 (2013), s. 1240-1254 ISSN 0007-1188 R&D Projects: GA ČR GAP303/10/1338 Institutional support: RVO:68378041 Keywords : two-pore-domain potassium channels * patch clamp * neuroprotection Subject RIV: FH - Neurology Impact factor: 4.990, year: 2013

  1. Financial Motivation Undermines Maintenance in an Intensive Diet and Activity Intervention

    Science.gov (United States)

    Moller, Arlen C.; McFadden, H. Gene; Hedeker, Donald; Spring, Bonnie

    2012-01-01

    Financial incentives are widely used in health behavior interventions. However, self-determination theory posits that emphasizing financial incentives can have negative consequences if experienced as controlling. Feeling controlled into performing a behavior tends to reduce enjoyment and undermine maintenance after financial contingencies are removed (the undermining effect). We assessed participants' context-specific financial motivation to participate in the Make Better Choices trial—a trial testing four different strategies for improving four health risk behaviors: low fruit and vegetable intake, high saturated fat intake, low physical activity, and high sedentary screen time. The primary outcome was overall healthy lifestyle change; weight loss was a secondary outcome. Financial incentives were contingent upon meeting behavior goals for 3 weeks and became contingent upon merely providing data during the 4.5-month maintenance period. Financial motivation for participation was assessed at baseline using a 7-item scale (α = .97). Across conditions, a main effect of financial motivation predicted a steeper rate of weight regained during the maintenance period, t(165) = 2.15, P = .04. Furthermore, financial motivation and gender interacted significantly in predicting maintenance of healthy diet and activity changes, t(160) = 2.42, P = .016, such that financial motivation had a more deleterious influence among men. Implications for practice and future research on incentivized lifestyle and weight interventions are discussed. PMID:22548152

  2. Review on Japanese activities in the field of maintenance and repair of LMFBR steam generators

    International Nuclear Information System (INIS)

    Tsuchiya, T.; Fukuda, T.; Sato, M.; Okabayashi, K.; Takahashi, T.

    2002-01-01

    Summary of Japanese activities on maintenance and repair of LMFBR steam generators (SG) is described in this paper. The concept (adoption of helical coil tube etc.) of MONJU SG was established in conceptional design started from 1968, and research and development (R and D) program was prepared. Parallel with basic studies such as material, welding, sodium water reaction and etc., overall verification tests using mock up SGs were conducted. As the first step, 1 Mw SG with two active helical tubes (and eight dummy tubes) was fabricated and operated, and many maintenance and repair experiences were accumulated through two small water leak troubles. Two 50Mw SGs, 1/5 scale of MONJU SG, were constructed and operated for long time. Post test examinations were carried out for No.1 50 Mw SG and feasibility of this type of SG was confirmed. In regard to maintenance and repair techniques, explosive and welding method for tube plugging and UT and ECT techniques for inspection of tube integrity are under development. Overall verification test for on-site and in-factory maintenance and repair techniques was conducted using No.2 50Mw SG evaporator and applicability of those techniques to real plant was evaluated. Many experiences were accumulated for removal and cleaning of sodium water reaction products after sodium water reaction in the cooling system and pressure relief system, using the Large Sodium Water Reaction Test Facility (SWAT-1 and 3). (author)

  3. Financial Motivation Undermines Maintenance in an Intensive Diet and Activity Intervention

    Directory of Open Access Journals (Sweden)

    Arlen C. Moller

    2012-01-01

    Full Text Available Financial incentives are widely used in health behavior interventions. However, self-determination theory posits that emphasizing financial incentives can have negative consequences if experienced as controlling. Feeling controlled into performing a behavior tends to reduce enjoyment and undermine maintenance after financial contingencies are removed (the undermining effect. We assessed participants’ context-specific financial motivation to participate in the Make Better Choices trial—a trial testing four different strategies for improving four health risk behaviors: low fruit and vegetable intake, high saturated fat intake, low physical activity, and high sedentary screen time. The primary outcome was overall healthy lifestyle change; weight loss was a secondary outcome. Financial incentives were contingent upon meeting behavior goals for 3 weeks and became contingent upon merely providing data during the 4.5-month maintenance period. Financial motivation for participation was assessed at baseline using a 7-item scale (=.97. Across conditions, a main effect of financial motivation predicted a steeper rate of weight regained during the maintenance period, (165=2.15, =.04. Furthermore, financial motivation and gender interacted significantly in predicting maintenance of healthy diet and activity changes, (160=2.42, =.016, such that financial motivation had a more deleterious influence among men. Implications for practice and future research on incentivized lifestyle and weight interventions are discussed.

  4. Organizational-economic maintenance of innovation activity in the region: comparative assessment

    Directory of Open Access Journals (Sweden)

    Nadezhda Igorevna Antipina

    2014-05-01

    Full Text Available The article proposes the approach to evaluate the organizational-economic maintenance of innovation activity in the regions in quantitative and qualitative indicators, as well as the method to calculate the assessment of regulatory support of this activity. It justifies the author’s approach of comparative efficiency evaluation of innovation legislation and regions’ innovation development level. The article gives the qualitative estimation of regulatory support of innovation development in the regions that are innovation leaders. It singles out key directions to develop regulatory support of innovation activity, which encourage RF subjects’ innovation activity

  5. Operation and Maintenance, 9-Foot Navigation Channel, Upper Mississippi River, Head of Navigation to Guttenberg, Iowa. Volume 2. Exhibits.

    Science.gov (United States)

    1974-08-01

    Walleye and Sauger 18,000 19,000 R c. Northern Pike 3,000 6,000 p d. Other 27000 25.000 S TOTAL (Fish) 196,000 134 ,OO0 0 F Catch Rates (Fish Caught per...0 0 0 R 00000000 𔃺 0,~. 1.IpC 0 o t 0 Fi. C Z C Z 2 4 .tt 04 ’ 𔃻. IsO 00 cC cO ~~It N Ea Et OPRTO AND0MAINTENANCE OFC NINElOOTl NAIATO

  6. Quality Control Activities Related to Mechanical Maintenance of Safety Related Components at Krsko NPP

    International Nuclear Information System (INIS)

    Djakovic, D.

    2016-01-01

    For successful, safe and reliable operation of nuclear power plant, maintenance processes have to be systematically controlled and procedures for quality control of maintenance activities shall be established. This is requested by the quality assurance program, which shall provide control over activities affecting the quality of structures, systems, and components, considering their importance to safety. As a part of Quality and Nuclear Oversight Division (QNOD; SKV), the Quality Control Department (QC) provides quality control activities, which are deeply involved in maintenance processes at Krsko NPP, both on safety related and non-safety related (non-nuclear safety) components. QC activities on safety related components have to fulfil all requirements, which will enable the components to perform their intended safety functions. This paper describes quality control activities related to mechanical maintenance of safety related components at Krsko NPP and significant role of the Krsko plant QC Department in three particular maintenance cases connected with safety related components. In these three specific cases, the QC has confirmed its importance in compliance with quality assurance program and presented its significant added value in providing safe and reliable operation of the plant. The first maintenance activity was installation of nozzle check valves in the scope of a modification for improving regulation of spent fuel pit pumps. The QC Department performed receipt inspection of the valves. Using non-destructive examination methods and X-ray spectrometry, it was found out that the valve diffuser was made of improper material, which could cause progressive corrosion of the valve diffuser in borated water and consequently a loss of safety function of the valves followed by long-term consequences. The second one was the receipt inspection of containment ventilation fan coolers. The coolers were claimed and sent back to the supplier because the QC Department

  7. Terrestrial gamma ray flash production by active lightning leader channels

    OpenAIRE

    İnan, Umran Savaş; Carlson, B. E.; Lehtinen, N. G.

    2017-01-01

    The production of terrestrial gamma ray flashes (TGFs) requires a seed energetic electron source and a strong electric field. Lightning leaders naturally provide seed electrons by cold runaway and strong electric fields by charge accumulation on the channel. We model possible TGF production in such fields by simulating the charges and currents on the channel. The resulting electric fields then drive simulations of runaway relativistic electron avalanche and photon emission. Photon spectra and...

  8. Novel Biomarkers of Physical Activity Maintenance in Midlife Women: Preliminary Investigation

    Directory of Open Access Journals (Sweden)

    Kelly A. Bosak

    2018-04-01

    Full Text Available The precision health initiative is leading the discovery of novel biomarkers as important indicators of biological processes or responses to behavior, such as physical activity. Neural biomarkers identified by magnetic resonance imaging (MRI hold promise to inform future research, and ultimately, for transfer to the clinical setting to optimize health outcomes. This study investigated resting-state and functional brain biomarkers between midlife women who were maintaining physical activity in accordance with the current national guidelines and previously acquired age-matched sedentary controls. Approval was obtained from the Human Subjects Committee. Participants included nondiabetic, healthy weight to overweight (body mass index 19–29.9 kg/m2 women (n = 12 aged 40–64 years. Control group data were used from participants enrolled in our previous functional MRI study and baseline resting-state MRI data from a subset of sedentary (<500 kcal of physical activity per week midlife women who were enrolled in a 9-month exercise intervention conducted in our imaging center. Differential activation of the inferior frontal gyrus (IFG and greater connectivity with the dorsolateral prefrontal cortex (dlPFC was identified between physically active women and sedentary controls. After correcting for multiple comparisons, these differences in biomarkers of physical activity maintenance did not reach statistical significance. Preliminary evidence in this small sample suggests that neural biomarkers of physical activity maintenance involve activations in the brain region associated with areas involved in implementing goal-directed behavior. Specifically, activation of the IFG and connectivity with the dlPFC is identified as a neural biomarker to explain and predict long-term physical activity maintenance for healthy aging. Future studies should evaluate these biomarker links with relevant clinical correlations.

  9. 5-HT1A receptors modulate small-conductance Ca2+-activated K+ channels

    DEFF Research Database (Denmark)

    Grunnet, Morten; Jespersen, Thomas; Perrier, Jean-François

    2004-01-01

    Small-conductance calcium-activated potassium channels (SK) are responsible for the medium afterhyperpolarisation (mAHP) following action potentials in neurons. Here we tested the ability of serotonin (5-HT) to modulate the activity of SK channels by coexpressing 5-HT1A receptors with different...

  10. Present activity in ASME Section XI regarding risk-informed maintenance

    International Nuclear Information System (INIS)

    Hedden, Owen; Chockie, Alan

    2005-01-01

    Since 1996 Section XI of the ASME Boiler and Pressure Vessel Code has actively incorporated risk-informed concepts. The risk-informed process provides a framework for allocating inspection resources in a cost-effective manner and helps focus inspections where most critical for plant safety. Based on the success of the risk-informed ISI piping applications at US and non-US plants, Section XI has refined existing Code Cases and expanded the use of the risk-informed process to a variety of high-risk components and systems. The risk informed approach started in the area of inspection and is now being expanded to other plant maintenance activities. This article summarizes the Section XI actions and the continued development of the risk-informed process to improve nuclear plant maintenance. (author)

  11. Discovery Channel Telescope active optics system early integration and test

    Science.gov (United States)

    Venetiou, Alexander J.; Bida, Thomas A.

    2012-09-01

    The Discovery Channel Telescope (DCT) is a 4.3-meter telescope with a thin meniscus primary mirror (M1) and a honeycomb secondary mirror (M2). The optical design is an f/6.1 Ritchey-Chrétien (RC) with an unvignetted 0.5° Field of View (FoV) at the Cassegrain focus. We describe the design, implementation and performance of the DCT active optics system (AOS). The DCT AOS maintains collimation and controls the figure of the mirror to provide seeing-limited images across the focal plane. To minimize observing overhead, rapid settling times are achieved using a combination of feed-forward and low-bandwidth feedback control using a wavefront sensing system. In 2011, we mounted a Shack-Hartmann wavefront sensor at the prime focus of M1, the Prime Focus Test Assembly (PFTA), to test the AOS with the wavefront sensor, and the feedback loop. The incoming wavefront is decomposed using Zernike polynomials, and the mirror figure is corrected with a set of bending modes. Components of the system that we tested and tuned included the Zernike to Bending Mode transformations. We also started open-loop feed-forward coefficients determination. In early 2012, the PFTA was replaced by M2, and the wavefront sensor moved to its normal location on the Cassegrain instrument assembly. We present early open loop wavefront test results with the full optical system and instrument cube, along with refinements to the overall control loop operating at RC Cassegrain focus.

  12. Selective activation of heteromeric SK channels contributes to action potential repolarization in mouse atrial myocytes.

    Science.gov (United States)

    Hancock, Jane M; Weatherall, Kate L; Choisy, Stéphanie C; James, Andrew F; Hancox, Jules C; Marrion, Neil V

    2015-05-01

    Activation of small conductance calcium-activated potassium (SK) channels is proposed to contribute to repolarization of the action potential in atrial myocytes. This role is controversial, as these cardiac SK channels appear to exhibit an uncharacteristic pharmacology. The objectives of this study were to resolve whether activation of SK channels contributes to atrial action potential repolarization and to determine the likely subunit composition of the channel. The effect of 2 SK channel inhibitors was assessed on outward current evoked in voltage clamp and on action potential duration in perforated patch and whole-cell current clamp recording from acutely isolated mouse atrial myocytes. The presence of SK channel subunits was assessed using immunocytochemistry. A significant component of outward current was reduced by the SK channel blockers apamin and UCL1684. Block by apamin displayed a sensitivity indicating that this current was carried by homomeric SK2 channels. Action potential duration was significantly prolonged by UCL1684, but not by apamin. This effect was accompanied by an increase in beat-to-beat variability and action potential triangulation. This pharmacology was matched by that of expressed heteromeric SK2-SK3 channels in HEK293 cells. Immunocytochemistry showed that atrial myocytes express both SK2 and SK3 channels with an overlapping expression pattern. Only proposed heteromeric SK2-SK3 channels are physiologically activated to contribute to action potential repolarization, which is indicated by the difference in pharmacology of evoked outward current and prolongation of atrial action potential duration. The effect of blocking this channel on the action potential suggests that SK channel inhibition during cardiac function has the potential to be proarrhythmic. Copyright © 2015 Heart Rhythm Society. Published by Elsevier Inc. All rights reserved.

  13. Downregulation of Kv7.4 channel activity in primary and secondary hypertension

    DEFF Research Database (Denmark)

    Jepps, Thomas Andrew; Chadha, Preet S; Davis, Alison J

    2011-01-01

    Voltage-gated potassium (K(+)) channels encoded by KCNQ genes (Kv7 channels) have been identified in various rodent and human blood vessels as key regulators of vascular tone; however, nothing is known about the functional impact of these channels in vascular disease. We ascertained the effect of...... structurally different activators of Kv7.2 through Kv7.5 channels (BMS-204352, S-1, and retigabine) on blood vessels from normotensive and hypertensive animals.......Voltage-gated potassium (K(+)) channels encoded by KCNQ genes (Kv7 channels) have been identified in various rodent and human blood vessels as key regulators of vascular tone; however, nothing is known about the functional impact of these channels in vascular disease. We ascertained the effect of 3...

  14. Neutron induced activation in the EVEDA accelerator materials: Implications for the accelerator maintenance

    International Nuclear Information System (INIS)

    Sanz, J.; Garcia, M.; Sauvan, P.; Lopez, D.; Moreno, C.; Ibarra, A.; Sedano, L.

    2009-01-01

    The Engineering Validation and Engineering Design Activities (EVEDA) phase of the International Fusion Materials Irradiation Facility project should result in an accelerator prototype for which the analysis of the dose rates evolution during the beam-off phase is a necessary task for radioprotection and maintenance feasibility purposes. Important aspects of the computational methodology to address this problem are discussed, and dose rates for workers inside the accelerator vault are assessed and found to be not negligible.

  15. Neutron induced activation in the EVEDA accelerator materials: Implications for the accelerator maintenance

    Energy Technology Data Exchange (ETDEWEB)

    Sanz, J. [Department of Power Engineering, Universidad Nacional de Educacion a Distancia (UNED), C/Juan del Rosal 12, 28040 Madrid (Spain); Institute of Nuclear Fusion, UPM, 28006 Madrid (Spain)], E-mail: jsanz@ind.uned.es; Garcia, M.; Sauvan, P.; Lopez, D. [Department of Power Engineering, Universidad Nacional de Educacion a Distancia (UNED), C/Juan del Rosal 12, 28040 Madrid (Spain); Institute of Nuclear Fusion, UPM, 28006 Madrid (Spain); Moreno, C.; Ibarra, A.; Sedano, L. [CIEMAT, 28040 Madrid (Spain)

    2009-04-30

    The Engineering Validation and Engineering Design Activities (EVEDA) phase of the International Fusion Materials Irradiation Facility project should result in an accelerator prototype for which the analysis of the dose rates evolution during the beam-off phase is a necessary task for radioprotection and maintenance feasibility purposes. Important aspects of the computational methodology to address this problem are discussed, and dose rates for workers inside the accelerator vault are assessed and found to be not negligible.

  16. A structural view of ligand-dependent activation in thermoTRP channels

    Directory of Open Access Journals (Sweden)

    Ximena eSteinberg

    2014-05-01

    Full Text Available Transient Receptor Potential (TRP proteins are a large family of ion channels, grouped intoseven sub-families. Although great advances have been made regarding the activation andmodulation of TRP channel activity, detailed molecular mechanisms governing TRPchannel gating are still needed. Sensitive to electric, chemical, mechanical, and thermalcues, TRP channels are tightly associated with the detection and integration of sensoryinput, emerging as a model to study the polymodal activation of ion channel proteins.Among TRP channels, the temperature-activated kind constitute a subgroup by itself,formed by Vanilloid receptors 1-4, Melastatin receptors 2, 4, 5 and 8, TRPC5, and TRPA1.Some of the so-called thermoTRP channels participate in the detection of noxious stimulimaking them an interesting pharmacological target for the treatment of pain. However, thepoor specificity of the compounds available in the market represents an important obstacleto overcome. Understanding the molecular mechanics underlying ligand-dependentmodulation of TRP channels may help with the rational design of novel syntheticanalgesics. The present review focuses on the structural basis of ligand-dependentactivation of TRPV1 and TRPM8 channels. Special attention is drawn to the dissection ofligand-binding sites within TRPV1, PIP 2 -dependent modulation of TRP channels, and thestructure of natural and synthetic ligands.

  17. Organization of industrial maintenance in oil pipeline and terminals activities: a project for the implantation of a corporative management system

    Energy Technology Data Exchange (ETDEWEB)

    Correia, Paulo de Tarso Arruda [TRANSPETRO - PETROBRAS Transporte S.A., Rio de Janeiro, RJ (Brazil). Diretoria de Terminais e Oleodutos; Arruda, Daniela Mendonca; Oliveira, Italo Luiz [TRANSPETRO - PETROBRAS Transporte S.A., Rio de Janeiro, RJ (Brazil)

    2009-07-01

    This paper discusses the definition and implementation of a corporate management model for industrial maintenance within TRANSPETRO's Terminals and Oil Pipelines business segment. A project is under way to answer the company's needs for better coordination of its maintenance activities as well as to face the challenge of increasing performance with higher systems and equipment' availability, at lower costs. To achieve this target, the project has been employing updating management tools for industrial maintenance planning and control. Its scope comprises the definition of: industrial maintenance process guidelines; industrial maintenance normative requirements and operational procedures; industrial maintenance performance indicators; local organizational charts; training of maintenance personnel; qualification and certification policies; conceptualization of maintenance engineering and maintenance based on risk; updating technologies such as asset management; warehousing and logistics for maintenance support in the supply of spare parts and materials. The project will also consider the results of a previous strategic sourcing study, concerning the definition of the most suitable out sourcing strategy for each operational unit, considering its characteristics as well as the characteristics of the region where it is located. (author)

  18. Organization of industrial maintenance in oil pipeline and terminals activities: a project for the implantation of a corporative management system

    Energy Technology Data Exchange (ETDEWEB)

    Correia, Paulo de Tarso Arruda [TRANSPETRO - PETROBRAS Transporte S.A., Rio de Janeiro, RJ (Brazil). Diretoria de Terminais e Oleodutos; Arruda, Daniela Mendonca; Oliveira, Italo Luiz [TRANSPETRO - PETROBRAS Transporte S.A., Rio de Janeiro, RJ (Brazil)

    2009-07-01

    This paper discusses the definition and implementation of a corporate management model for industrial maintenance within TRANSPETRO's Terminals and Oil Pipelines business segment. A project is under way to answer the company's needs for better coordination of its maintenance activities as well as to face the challenge of increasing performance with higher systems and equipment' availability, at lower costs. To achieve this target, the project has been employing updating management tools for industrial maintenance planning and control. Its scope comprises the definition of: industrial maintenance process guidelines; industrial maintenance normative requirements and operational procedures; industrial maintenance performance indicators; local organizational charts; training of maintenance personnel; qualification and certification policies; conceptualization of maintenance engineering and maintenance based on risk; updating technologies such as asset management; warehousing and logistics for maintenance support in the supply of spare parts and materials. The project will also consider the results of a previous strategic sourcing study, concerning the definition of the most suitable out sourcing strategy for each operational unit, considering its characteristics as well as the characteristics of the region where it is located. (author)

  19. Cell swelling activates separate taurine and chloride channels in Ehrlich mouse ascites tumor cells

    DEFF Research Database (Denmark)

    Lambert, Ian Henry; Hoffmann, Else Kay

    1994-01-01

    The taurine efflux from Ehrlich ascites tumor cells is stimulated by hypotonic cell swelling. The swelling-activated taurine efflux is unaffected by substitution of gluconate for extracellular Cl– but inhibited by addition of MK196 (anion channel blocker) and 4,4 -diisothiocyanostilbene-2......,2 -disulfonic acid (DIDS; anion channel and anion exchange blocker) and by depolarization of the cell membrane. This is taken to indicate that taurine does not leave the osmotically swollen Ehrlich cells in exchange for extracellular Cl–, i.e., via the anion exchanger but via a MK196- and DIDS-sensitive channel...... that is potential dependent. An additional stimulation of the swelling-activated taurine efflux is seen after addition of arachidonic acid and oleic acid. Cell swelling also activates a Mini Cl– channel. The Cl– efflux via this Cl– channel, in contrast to the swelling-activated taurine efflux, is unaffected by DIDS...

  20. Optimal Medical Equipment Maintenance Service Proposal Decision Support System combining Activity Based Costing (ABC) and the Analytic Hierarchy Process (AHP).

    Science.gov (United States)

    da Rocha, Leticia; Sloane, Elliot; M Bassani, Jose

    2005-01-01

    This study describes a framework to support the choice of the maintenance service (in-house or third party contract) for each category of medical equipment based on: a) the real medical equipment maintenance management system currently used by the biomedical engineering group of the public health system of the Universidade Estadual de Campinas located in Brazil to control the medical equipment maintenance service, b) the Activity Based Costing (ABC) method, and c) the Analytic Hierarchy Process (AHP) method. Results show the cost and performance related to each type of maintenance service. Decision-makers can use these results to evaluate possible strategies for the categories of equipment.

  1. Flexibility within working memory and the focus of attention for sequential verbal information does not depend on active maintenance.

    Science.gov (United States)

    Sandry, Joshua; Schwark, Jeremy D; MacDonald, Justin

    2014-10-01

    The focus of attention seems to be a static element within working memory when verbal information is serially presented, unless additional time is available for processing or active maintenance. Experiment 1 manipulated the reward associated with early and medial list positions in a probe recognition paradigm and found evidence that these nonterminal list positions could be retrieved faster and more accurately if participants were appropriately motivated-without additional time for processing or active maintenance. Experiment 2 used articulatory suppression and demonstrated that the underlying maintenance mechanism cannot be attributed to rehearsal, leaving attentional refreshing as the more likely mechanism. These findings suggest that the focus of attention within working memory can flexibly maintain nonterminal early and medial list representations at the expense of other list representations even when there is not additional time for processing or active maintenance. Maintenance seems to be accomplished through an attentional refreshing mechanism.

  2. Structural mechanism underlying capsaicin binding and activation of TRPV1 ion channel

    OpenAIRE

    Yang, Fan; Xiao, Xian; Cheng, Wei; Yang, Wei; Yu, Peilin; Song, Zhenzhen; Yarov-Yarovoy, Vladimir; Zheng, Jie

    2015-01-01

    Capsaicin bestows spiciness by activating TRPV1 channel with exquisite potency and selectivity. Capsaicin-bound channel structure was previously resolved by cryo-EM at 4.2-to-4.5 ? resolution, however important details required for mechanistic understandings are unavailable: capsaicin was registered as a small electron density, reflecting neither its chemical structure nor specific ligand-channel interactions. We obtained the missing atomic-level details by iterative computation, which were c...

  3. 77 FR 70846 - Regulatory Guide 1.182, “Assessing and Managing Risk Before Maintenance Activities at Nuclear...

    Science.gov (United States)

    2012-11-27

    ... NUCLEAR REGULATORY COMMISSION [NRC-2012-0285] Regulatory Guide 1.182, ``Assessing and Managing Risk Before Maintenance Activities at Nuclear Power Plants'' AGENCY: Nuclear Regulatory Commission... withdrawing Regulatory Guide (RG)1.182, Revision (Rev.) 0, ``Assessing and Managing Risk Before Maintenance...

  4. Business process modeling of industrial maintenance at TRANSPETRO: integrating oil pipeline and marine terminals activities

    Energy Technology Data Exchange (ETDEWEB)

    Arruda, Daniela Mendonca; Oliveira, Italo Luiz [TRANSPETRO - PETROBRAS Transporte S.A., Rio de Janeiro, RJ (Brazil). Diretoria de Terminais e Oleodutos; Almeida, Maria Fatima Ludovico de [Pontificia Universidade Catolica do Rio de Janeiro (PUC-Rio), Rio de Janeiro, RJ (Brazil). Programa de Pos-Graduacao em Metrologia para Qualidade e Inovacao

    2009-07-01

    This paper describes the experience of TRANSPETRO in remodeling industrial maintenance activities focusing on: preparing for business process modeling (BPM); mapping and analyzing 'As-Is' process; designing 'To-Be' process; implementing remodeled process; improving process continuously. The conceptual model and results achieved will contribute to several areas within the company as: reliability engineering; human resources, including employees' selective processes, training and development, and certifications; standardization process encompassing standard and operational procedures adoption according to up-dating external normative references and legal requirements; health, safety and environment (HSE) performance improvement. These are some of potential benefits from BPM focusing on TRANSPETRO's industrial maintenance area in the search of operational excellence. (author)

  5. Activation of protein kinase C alters the intracellular distribution and mobility of cardiac Na+ channels.

    Science.gov (United States)

    Hallaq, Haifa; Wang, Dao W; Kunic, Jennifer D; George, Alfred L; Wells, K Sam; Murray, Katherine T

    2012-02-01

    Na(+) current derived from expression of the cardiac isoform SCN5A is reduced by receptor-mediated or direct activation of protein kinase C (PKC). Previous work has suggested a possible role for loss of Na(+) channels at the plasma membrane in this effect, but the results are controversial. In this study, we tested the hypothesis that PKC activation acutely modulates the intracellular distribution of SCN5A channels and that this effect can be visualized in living cells. In human embryonic kidney cells that stably expressed SCN5A with green fluorescent protein (GFP) fused to the channel COOH-terminus (SCN5A-GFP), Na(+) currents were suppressed by an exposure to PKC activation. Using confocal microscopy, colocalization of SCN5A-GFP channels with the plasma membrane under control and stimulated conditions was quantified. A separate population of SCN5A channels containing an extracellular epitope was immunolabeled to permit temporally stable labeling of the plasma membrane. Our results demonstrated that Na(+) channels were preferentially trafficked away from the plasma membrane by PKC activation, with a major contribution by Ca(2+)-sensitive or conventional PKC isoforms, whereas stimulation of protein kinase A (PKA) had the opposite effect. Removal of the conserved PKC site Ser(1503) or exposure to the NADPH oxidase inhibitor apocynin eliminated the PKC-mediated effect to alter channel trafficking, indicating that both channel phosphorylation and ROS were required. Experiments using fluorescence recovery after photobleaching demonstrated that both PKC and PKA also modified channel mobility in a manner consistent with the dynamics of channel distribution. These results demonstrate that the activation of protein kinases can acutely regulate the intracellular distribution and molecular mobility of cardiac Na(+) channels in living cells.

  6. Cell swelling activates cloned Ca(2+)-activated K(+) channels: a role for the F-actin cytoskeleton

    DEFF Research Database (Denmark)

    Jorgensen, Nanna K; Pedersen, Stine F; Rasmussen, Hanne B

    2003-01-01

    Cloned Ca(2+)-activated K(+) channels of intermediate (hIK) or small (rSK3) conductance were expressed in HEK 293 cells, and channel activity was monitored using whole-cell patch clamp. hIK and rSK3 currents already activated by intracellular calcium were further increased by 95% and 125......%, respectively, upon exposure of the cells to a 33% decrease in extracellular osmolarity. hIK and rSK3 currents were inhibited by 46% and 32%, respectively, by a 50% increase in extracellular osmolarity. Cell swelling and channel activation were not associated with detectable increases in [Ca(2+)](i), evidenced...... by population and single-cell measurements. In addition, inhibitors of IK and SK channels significantly reduced the rate of regulatory volume decrease (RVD) in cells expressing these channels. Cell swelling induced a decrease, and cell shrinkage an increase, in net cellular F-actin content. The swelling...

  7. Modulation of KCNQ4 channel activity by changes in cell volume

    DEFF Research Database (Denmark)

    Hougaard, Charlotte; Klaerke, Dan A; Hoffmann, Else K

    2004-01-01

    KCNQ4 channels expressed in HEK 293 cells are sensitive to cell volume changes, being activated by swelling and inhibited by shrinkage, respectively. The KCNQ4 channels contribute significantly to the regulatory volume decrease (RVD) process following cell swelling. Under isoosmotic conditions...

  8. Inhibition of small-conductance Ca2+-activated K+ channels terminates and protects against atrial fibrillation

    DEFF Research Database (Denmark)

    Diness, Jonas Goldin; Sørensen, Ulrik S; Nissen, Jakob Dahl

    2010-01-01

    Recently, evidence has emerged that small-conductance Ca(2+)-activated K(+) (SK) channels are predominantly expressed in the atria in a number of species including human. In rat, guinea pig, and rabbit ex vivo and in vivo models of atrial fibrillation (AF), we used 3 different SK channel inhibito...

  9. The Importance of Providing Multiple-Channel Sections in Dredging Activities to Improve Fish Habitat Environments

    Directory of Open Access Journals (Sweden)

    Hung-Pin Chiu

    2016-01-01

    Full Text Available After Typhoon Morakot, dredging engineering was conducted while taking the safety of humans and structures into consideration, but partial stream reaches were formed in the multiple-channel sections in Cishan Stream because of anthropogenic and natural influences. This study mainly explores the distribution of each fish species in both the multiple- and single-channel sections in the Cishan Stream. Parts of the environments did not exhibit significant differences according to a one-way ANOVA comparing the multiple- and single-channel sections, but certain areas of the multiple-channel sections had more diverse habitats. Each fish species was widely distributed by non-metric multidimensional scaling in the multiple-channel sections as compared to those in the single-channel sections. In addition, according to the principal component analysis, each fish species has a preferred environment, and all of them have a wide choice of habitat environments in the multiple-channel sections. Finally, the existence of multiple-channel sections could significantly affect the existence of the fish species under consideration in this study. However, no environmental factors were found to have an influence on fish species in the single-channel sections, with the exception of Rhinogobius nantaiensis. The results show that providing multiple-channel sections in dredging activities could improve fish habitat environments.

  10. Fragile X mental retardation protein controls ion channel expression and activity.

    Science.gov (United States)

    Ferron, Laurent

    2016-10-15

    Fragile X-associated disorders are a family of genetic conditions resulting from the partial or complete loss of fragile X mental retardation protein (FMRP). Among these disorders is fragile X syndrome, the most common cause of inherited intellectual disability and autism. FMRP is an RNA-binding protein involved in the control of local translation, which has pleiotropic effects, in particular on synaptic function. Analysis of the brain FMRP transcriptome has revealed hundreds of potential mRNA targets encoding postsynaptic and presynaptic proteins, including a number of ion channels. FMRP has been confirmed to bind voltage-gated potassium channels (K v 3.1 and K v 4.2) mRNAs and regulates their expression in somatodendritic compartments of neurons. Recent studies have uncovered a number of additional roles for FMRP besides RNA regulation. FMRP was shown to directly interact with, and modulate, a number of ion channel complexes. The sodium-activated potassium (Slack) channel was the first ion channel shown to directly interact with FMRP; this interaction alters the single-channel properties of the Slack channel. FMRP was also shown to interact with the auxiliary β4 subunit of the calcium-activated potassium (BK) channel; this interaction increases calcium-dependent activation of the BK channel. More recently, FMRP was shown to directly interact with the voltage-gated calcium channel, Ca v 2.2, and reduce its trafficking to the plasma membrane. Studies performed on animal models of fragile X syndrome have revealed links between modifications of ion channel activity and changes in neuronal excitability, suggesting that these modifications could contribute to the phenotypes observed in patients with fragile X-associated disorders. © 2016 The Authors. The Journal of Physiology © 2016 The Physiological Society.

  11. Activation of TRPM7 channels by small molecules under physiological conditions.

    Science.gov (United States)

    Hofmann, T; Schäfer, S; Linseisen, M; Sytik, L; Gudermann, T; Chubanov, V

    2014-12-01

    Transient receptor potential cation channel, subfamily M, member 7 (TRPM7) is a cation channel covalently linked to a protein kinase domain. TRPM7 is ubiquitously expressed and regulates key cellular processes such as Mg(2+) homeostasis, motility, and proliferation. TRPM7 is involved in anoxic neuronal death, cardiac fibrosis, and tumor growth. The goal of this work was to identify small molecule activators of the TRPM7 channel and investigate their mechanism of action. We used an aequorin bioluminescence-based assay to screen for activators of the TRPM7 channel. Valid candidates were further characterized using patch clamp electrophysiology. We identified 20 drug-like compounds with various structural backbones that can activate the TRPM7 channel. Among them, the δ opioid antagonist naltriben was studied in greater detail. Naltriben's action was selective among the TRP channels tested. Naltriben activates TRPM7 currents without prior depletion of intracellular Mg(2+) even under conditions of low PIP2. Moreover, naltriben interfered with the effect of the TRPM7 inhibitor NS8593. Finally, our experiments with TRPM7 variants carrying mutations in the pore, TRP, and kinase domains indicate that the site of TRPM7 activation by this small-molecule ligand is most likely located in or near the TRP domain. In conclusion, we identified the first organic small-molecule activators of TRPM7 channels, thus providing new experimental tools to study TRPM7 function in native cellular environments.

  12. PMS2 endonuclease activity has distinct biological functions and is essential for genome maintenance.

    Science.gov (United States)

    van Oers, Johanna M M; Roa, Sergio; Werling, Uwe; Liu, Yiyong; Genschel, Jochen; Hou, Harry; Sellers, Rani S; Modrich, Paul; Scharff, Matthew D; Edelmann, Winfried

    2010-07-27

    The DNA mismatch repair protein PMS2 was recently found to encode a novel endonuclease activity. To determine the biological functions of this activity in mammals, we generated endonuclease-deficient Pms2E702K knock-in mice. Pms2EK/EK mice displayed increased genomic mutation rates and a strong cancer predisposition. In addition, class switch recombination, but not somatic hypermutation, was impaired in Pms2EK/EK B cells, indicating a specific role in Ig diversity. In contrast to Pms2-/- mice, Pms2EK/EK male mice were fertile, indicating that this activity is dispensable in spermatogenesis. Therefore, the PMS2 endonuclease activity has distinct biological functions and is essential for genome maintenance and tumor suppression.

  13. Two-photon activation of endogenous store-operated calcium channels without optogenetics

    Science.gov (United States)

    Cheng, Pan; Tang, Wanyi; He, Hao

    2018-02-01

    Store-operated calcium (SOC) channels, regulated by intracellular Ca2+ store, are the essential pathway of calcium signaling and participate in a wide variety of cellular activities such as gene expression, secretion and immune response1. However, our understanding and regulation of SOC channels are mainly based on pharmacological methods. Considering the unique advantages of optical control, optogenetic control of SOC channels has been developed2. However, the process of genetic engineering to express exogenous light-sensitive protein is complicated, which arouses concerns about ethic difficulties in some research of animal and applications in human. In this report, we demonstrate rapid, robust and reproducible two-photon activation of endogenous SOC channels by femtosecond laser without optogenetics. We present that the short-duration two-photon scanning on subcellular microregion induces slow Ca2+ influx from extracellular medium, which can be eliminated by removing extracellular Ca2+. Block of SOC channels using various pharmacological inhibitors or knockdown of SOC channels by RNA interference reduce the probability of two-photon activated Ca2+ influx. On the contrary, overexpression of SOC channels can increase the probability of Ca2+ influx by two-photon scanning. These results collectively indicate Ca2+ influx through two-photon activated SOC channels. Different from classical pathway of SOC entry activated by Ca2+ store depletion, STIM1, the sensor protein of Ca2+ level in endoplasmic reticulum, does not show any aggregation or migration in this two-photon activated Ca2+ influx, which rules out the possibility of intracellular Ca2+ store depletion. Thereby, we propose this all-optical method of two-photon activation of SOC channels is of great potential to be widely applied in the research of cell calcium signaling and related biological research.

  14. Regulation of KV channel voltage-dependent activation by transmembrane β subunits

    Directory of Open Access Journals (Sweden)

    Xiaohui eSun

    2012-04-01

    Full Text Available Voltage-activated K+ (KV channels are important for shaping action potentials and maintaining resting membrane potential in excitable cells. KV channels contain a central pore-gate domain (PGD surrounded by four voltage-sensing domains (VSD. The VSDs will change conformation in response to alterations of the membrane potential thereby inducing the opening of the PGD. Many KV channels are heteromeric protein complexes containing auxiliary β subunits. These β subunits modulate channel expression and activity to increase functional diversity and render tissue specific phenotypes. This review focuses on the KV β subunits that contain transmembrane (TM segments including the KCNE family and the β subunits of large conductance, Ca2+- and voltage-activated K+ (BK channels. These TM β subunits affect the voltage-dependent activation of KV α subunits. Experimental and computational studies have described the structural location of these β subunits in the channel complexes and the biophysical effects on VSD activation, PGD opening and VSD-PGD coupling. These results reveal some common characteristics and mechanistic insights into KV channel modulation by TM β subunits.

  15. Numerical study of mixed convection heat transfer enhancement in a channel with active flow modulation

    Science.gov (United States)

    Billah, Md. Mamun; Khan, Md Imran; Rahman, Mohammed Mizanur; Alam, Muntasir; Saha, Sumon; Hasan, Mohammad Nasim

    2017-06-01

    A numerical study of steady two dimensional mixed convention heat transfer phenomena in a rectangular channel with active flow modulation is carried out in this investigation. The flow in the channel is modulated via a rotating cylinder placed at the center of the channel. In this study the top wall of the channel is subjected to an isothermal low temperature while a discrete isoflux heater is positioned on the lower wall. The fluid flow under investigation is assumed to have a Prandtl number of 0.71 while the Reynolds No. and the Grashof No. are varied in wide range for four different situations such as: i) plain channel with no cylinder, ii) channel with stationary cylinder, iii) channel with clockwise rotating cylinder and iv) channel with counter clockwise rotating cylinder. The results obtained in this study are presented in terms of the distribution of streamlines, isotherms in the channel while the heat transfer process from the heat source is evaluated in terms of the local Nusselt number, average Nusselt number. The outcomes of this study also indicate that the results are strongly dependent on the type of configuration and direction of rotation of the cylinder and that the average Nusselt number value rises with an increase in Reynolds and Grashof numbers but the correlation between these parameters at higher values of Reynolds and Grashof numbers becomes weak.

  16. Differential distribution of the sodium‐activated potassium channels slick and slack in mouse brain

    Science.gov (United States)

    Knaus, Hans‐Günther; Schwarzer, Christoph

    2015-01-01

    ABSTRACT The sodium‐activated potassium channels Slick (Slo2.1, KCNT2) and Slack (Slo2.2, KCNT1) are high‐conductance potassium channels of the Slo family. In neurons, Slick and Slack channels are involved in the generation of slow afterhyperpolarization, in the regulation of firing patterns, and in setting and stabilizing the resting membrane potential. The distribution and subcellular localization of Slick and Slack channels in the mouse brain have not yet been established in detail. The present study addresses this issue through in situ hybridization and immunohistochemistry. Both channels were widely distributed and exhibited distinct distribution patterns. However, in some brain regions, their expression overlapped. Intense Slick channel immunoreactivity was observed in processes, varicosities, and neuronal cell bodies of the olfactory bulb, granular zones of cortical regions, hippocampus, amygdala, lateral septal nuclei, certain hypothalamic and midbrain nuclei, and several regions of the brainstem. The Slack channel showed primarily a diffuse immunostaining pattern, and labeling of cell somata and processes was observed only occasionally. The highest Slack channel expression was detected in the olfactory bulb, lateral septal nuclei, basal ganglia, and distinct areas of the midbrain, brainstem, and cerebellar cortex. In addition, comparing our data obtained from mouse brain with a previously published study on rat brain revealed some differences in the expression and distribution of Slick and Slack channels in these species. J. Comp. Neurol. 524:2093–2116, 2016. © 2015 The Authors The Journal of Comparative Neurology Published by Wiley Periodicals, Inc. PMID:26587966

  17. Food activities and identity maintenance in old age: a systematic review and meta-synthesis.

    Science.gov (United States)

    Plastow, Nicola Ann; Atwal, Anita; Gilhooly, Mary

    2015-01-01

    Services provided to older people should be developed based on active ageing policies. Nutrition is one aspect of active ageing, but little is known about how food activities contribute to psychological well-being in later life. This is a systematic review of qualitative and quantitative research that answers the question 'What is known about the relationship between food activities and the maintenance of identities in old age?'. We followed the preferred reporting items for systematic reviews and meta-analyses guidelines and used quality assessment parameters to complete a systematic review and narrative synthesis. Academic Search Premier, MEDLINE, CINAHL Plus, and PsycINFO databases were searched. We initially identified 8016 articles, of which 167 full-text articles were screened for inclusion. Twenty-two articles were included in the review. There was moderate evidence from nine qualitative and two quantitative studies, of variable quality, that food activities contribute to the maintenance of women's gendered identities, the ethnic identities of men and women, and community identities. There was moderate evidence from 10 qualitative studies, of variable quality, that a change in food choice and deteriorating health changed food activity participation. These changes threatened identities. Most studies included both younger adults and older adults. In later life, there are many life experiences leading to change. Further research is needed to develop understanding of how identity and mental well-being are maintained, despite changes in everyday activities like cooking and eating. This may enable health care professionals to meet psychological needs alongside biological needs during nutritional interventions.

  18. Cl- channels of the gastric parietal cell that are active at low pH.

    Science.gov (United States)

    Cuppoletti, J; Baker, A M; Malinowska, D H

    1993-06-01

    HCl secretion across mammalian gastric parietal cell apical membrane may involve Cl- channels. H(+)-K(+)-ATPase-containing membranes isolated from gastric mucosa of histamine-stimulated rabbits were fused to planar lipid bilayers. Channels were recorded with symmetric 800 mM CsCl solutions, pH 7.4. A linear current-voltage (I-V) relationship was obtained, and conductance was 28 +/- 1 pS at 800 mM CsCl. Conductance was 6.9 +/- 2 pS at 150 mM CsCl. Reversal potential was +22 mV with a fivefold cis-trans CsCl concentration gradient, indicating that the channel was anion selective with a discrimination ratio of 6:1 for Cl- over Cs+. Anion selectivity of the channel was I- > Cl- > or = Br- > NO3-, and gluconate was impermeant. Channels obtained at pH 7.4 persisted when pH of medium bathing the trans side of the bilayer (pHtrans) was reduced to pH 3, without a change in conductance, linearity of I-V relationship, or ion selectivity. In contrast, asymmetric reduction of pH of medium bathing the cis side of the bilayer from 7.4 to 3 always resulted in loss of channel activity. At pH 7.4, open probability (Po) of the channel was voltage dependent, i.e., predominantly open at +80 mV but mainly closed at -80 mV. In contrast, with low pHtrans, channel Po at -80 mV was increased 3.5-fold. The Cl- channel was Ca2+ indifferent. In absence of ionophores, ion selectivity for support of H(+)-K(+)-ATPase activity and H+ transport was consistent with that exhibited by the channel and could be limited by substitution with NO3-, whereas maximal H(+)-K(+)-ATPase activity was indifferent to anion present, demonstrating that anion transport can be rate limiting. Cl- channels with similar characteristics (conductance, linear I-V relationship, and ion selectivity) were also present in H(+)-K(+)-ATPase-containing vesicles isolated from resting (cimetidine-treated) gastric mucosa, exhibiting at -80 mV a pH-independent approximately 3.5-fold lower Po than stimulated vesicle channels. At -80 m

  19. The relationship between physical activity, volitional skills and weight loss maintenance in Danish obese individuals

    DEFF Research Database (Denmark)

    Elsborg, Peter; Pfister, Gertrud Ursula; Helge, Jørn Wulff

    . However, the relationship between weight loss maintenance and volitional skills has not been investigated yet. This study investigates if volitional skills are related to the amount of physical activity in obese individuals, and their relationship with actual successful weight loss. Methods: Participants...... to exercise. Results: Success was determined by the difference between an individual’s highest ever and current BMI. Analyses indicated that the successful weight maintainers’ possessed significantly higher values in the volitional skills initiative, general self-confidence, volitional self......-confidence and coping with failure and lower values in lack of energy, postponing training, and avoiding effort than unsuccessful weight maintainers. Furthermore, the successful were significantly more physically active. Higher physical activity was related to higher positive lower negative values in a total of 9...

  20. Active Urbanization and Channel Adjustment in Apple Creek, Appleton, WI

    Science.gov (United States)

    Clark, J. J.

    2002-12-01

    Headwaters of the Apple Creek watershed have been and continue to be rapidly developed as part of the City of Appleton's long-term growth plan. Concurrent with early development, and prior to development over the past 4 years, two regional stormwater management facilities were constructed. Cross-sectional surveys and core transects were used to determine channel response to urbanization mitigated by stormwater management. The reach immediately downstream of the first pond complex has a narrow, but well established, wooded riparian zone and has not changed in size or shape over the past two years. An engineered reach approximately one mile downstream, however has exhibited widespread bed aggradation. Cross-sectional area decreased an average of 51% over the past four years. Despite the use of sediment and erosion control BMPs, sediment concentrations exceeding 1000 mg/L during base flow are not uncommon downstream of construction sites adjacent to the stream. The artificially widened channel, a reduction in stream gradient, and the backwater effect from downstream ponds caused much of this sediment to remain within the engineered reach. It is estimated that approximately 21,000 Mg of sediment is stored in this mile-long reach. As this sediment migrates downstream, the forebay of the second set of stormwater ponds will begin to fill, reducing storage capacity and thereby limiting its effectiveness in mitigating peak discharges and sequestering nutrients.

  1. Zinc-dependent multi-conductance channel activity in mitochondria isolated from ischemic brain.

    Science.gov (United States)

    Bonanni, Laura; Chachar, Mushtaque; Jover-Mengual, Teresa; Li, Hongmei; Jones, Adrienne; Yokota, Hidenori; Ofengeim, Dimitry; Flannery, Richard J; Miyawaki, Takahiro; Cho, Chang-Hoon; Polster, Brian M; Pypaert, Marc; Hardwick, J Marie; Sensi, Stefano L; Zukin, R Suzanne; Jonas, Elizabeth A

    2006-06-21

    Transient global ischemia is a neuronal insult that induces delayed cell death. A hallmark event in the early post-ischemic period is enhanced permeability of mitochondrial membranes. The precise mechanisms by which mitochondrial function is disrupted are, as yet, unclear. Here we show that global ischemia promotes alterations in mitochondrial membrane contact points, a rise in intramitochondrial Zn2+, and activation of large, multi-conductance channels in mitochondrial outer membranes by 1 h after insult. Mitochondrial channel activity was associated with enhanced protease activity and proteolytic cleavage of BCL-xL to generate its pro-death counterpart, deltaN-BCL-xL. The findings implicate deltaN-BCL-xL in large, multi-conductance channel activity. Consistent with this, large channel activity was mimicked by introduction of recombinant deltaN-BCL-xL to control mitochondria and blocked by introduction of a functional BCL-xL antibody to post-ischemic mitochondria via the patch pipette. Channel activity was also inhibited by nicotinamide adenine dinucleotide, indicative of a role for the voltage-dependent anion channel (VDAC) of the outer mitochondrial membrane. In vivo administration of the membrane-impermeant Zn2+ chelator CaEDTA before ischemia or in vitro application of the membrane-permeant Zn2+ chelator tetrakis-(2-pyridylmethyl) ethylenediamine attenuated channel activity, suggesting a requirement for Zn2+. These findings reveal a novel mechanism by which ischemic insults disrupt the functional integrity of the outer mitochondrial membrane and implicate deltaN-BCL-xL and VDAC in the large, Zn2+-dependent mitochondrial channels observed in post-ischemic hippocampal mitochondria.

  2. Mechanisms of Rose Bengal inhibition on SecA ATPase and ion channel activities.

    Science.gov (United States)

    Hsieh, Ying-Hsin; Huang, Ying-Ju; Jin, Jin-Shan; Yu, Liyan; Yang, Hsiuchin; Jiang, Chun; Wang, Binghe; Tai, Phang C

    2014-11-14

    SecA is an essential protein possessing ATPase activity in bacterial protein translocation for which Rose Bengal (RB) is the first reported sub-micromolar inhibitor in ATPase activity and protein translocation. Here, we examined the mechanisms of inhibition on various forms of SecA ATPase by conventional enzymatic assays, and by monitoring the SecA-dependent channel activity in the semi-physiological system in cells. We build on the previous observation that SecA with liposomes form active protein-conducting channels in the oocytes. Such ion channel activity is enhanced by purified Escherichia coli SecYEG-SecDF·YajC liposome complexes. Inhibition by RB could be monitored, providing correlation of in vitro activity and intact cell functionality. In this work, we found the intrinsic SecA ATPase is inhibited by RB competitively at low ATP concentration, and non-competitively at high ATP concentrations while the translocation ATPase with precursors and SecYEG is inhibited non-competitively by RB. The Inhibition by RB on SecA channel activity in the oocytes with exogenous ATP-Mg(2+), mimicking translocation ATPase activity, is also non-competitive. The non-competitive inhibition on channel activity has also been observed with SecA from other bacteria which otherwise would be difficult to examine without the cognate precursors and membranes. Copyright © 2014 Elsevier Inc. All rights reserved.

  3. Dysfunctional Hyperpolarization-Activated Cyclic Nucleotide-gated Ion Channels in Cardiac Diseases

    Directory of Open Access Journals (Sweden)

    Xiaoqi Zhao

    Full Text Available Abstract Hyperpolarization-activated cyclic nucleotide-gated (HCN channels are reverse voltage-dependent, and their activation depends on the hyperpolarization of the membrane and may be directly or indirectly regulated by the cyclic adenosine monophosphate (cAMP or other signal-transduction cascades. The distribution, quantity and activation states of HCN channels differ in tissues throughout the body. Evidence exhibits that HCN channels play critical roles in the generation and conduction of the electrical impulse and the physiopathological process of some cardiac diseases. They may constitute promising drug targets in the treatment of these cardiac diseases. Pharmacological treatment targeting HCN channels is of benefit to these cardiac conditions.

  4. ACTIVITY THEORY APPLIED AT CHANNEL EXPANSIONS IN SMALL AND MEDIUM ENTERPRISES

    Directory of Open Access Journals (Sweden)

    Siw Lundqvist

    2017-06-01

    Full Text Available Today’s commonly carried out channel expansions of commerce could be both costly and problematic to manage. Especially for small and medium-sized enterprises (SMEs that often suffer from a lack of digital competence, time and monetary resources in generally. Still, these transitions would be necessary to carry out because of customer demands and expectations concerning 24/7 availability, and access to digital commerce alternatives. Scarce resources are important reasons to search for how to carry out channel expansions with minimized problems. Activity theory (AT focuses on the whole in order to detect problems that hinder successful outcomes. Hence, this theory was applied to prior findings, from a project about SME’s channel expansions, highlighting several problems that could appear during these activities. Implications for research foremost involve issues connected to the use of AT; implications for practice particularly concern if and how AT could be used to support channel broadening activities.

  5. A new approach to physical activity maintenance: Rationale, design, and baseline data from the Keep Active Minnesota trial

    Directory of Open Access Journals (Sweden)

    Crain A Lauren

    2008-07-01

    Full Text Available Abstract Background Since many individuals who initiate physical activity programs are highly likely to return to a sedentary lifestyle, innovative strategies to efforts to increase the number of physically active older adults who successfully maintain beneficial levels of PA for a substantial length of time are needed. Methods/Design The Keep Active Minnesota Trial is a randomized controlled trial of an interactive phone- and mail-based intervention to help 50–70 year old adults who have recently increased their physical activity level, maintain that activity level over a 24-month period in comparison to usual care. Baseline, 6, 12, and 24 month measurement occurred via phone surveys with kilocalories expended per week in total and moderate-to-vigorous physical activity (CHAMPS Questionnaire as the primary outcome measures. Secondary outcomes include hypothesized mediators of physical activity change (e.g., physical activity enjoyment, self-efficacy, physical activity self-concept, body mass index, and depression. Seven day accelerometry data were collected on a sub-sample of participants at baseline and 24-month follow-up. Discussion The Keep Active Minnesota study offers an innovative approach to the perennial problem of physical activity relapse; by focusing explicitly on physical activity maintenance, the intervention holds considerable promise for modifying the typical relapse curve. Moreover, if shown to be efficacious, the use of phone- and mail-based intervention delivery offers potential for widespread dissemination. Trial registration ClinicalTrials.gov Identifier: NCT00283452.

  6. Identification and characterization of Ca2+-activated K+ channels in granulosa cells of the human ovary

    Directory of Open Access Journals (Sweden)

    Berg Ulrike

    2009-04-01

    Full Text Available Abstract Background Granulosa cells (GCs represent a major endocrine compartment of the ovary producing sex steroid hormones. Recently, we identified in human GCs a Ca2+-activated K+ channel (KCa of big conductance (BKCa, which is involved in steroidogenesis. This channel is activated by intraovarian signalling molecules (e.g. acetylcholine via raised intracellular Ca2+ levels. In this study, we aimed at characterizing 1. expression and functions of KCa channels (including BKCa beta-subunits, and 2. biophysical properties of BKCa channels. Methods GCs were obtained from in vitro-fertilization patients and cultured. Expression of mRNA was determined by standard RT-PCR and protein expression in human ovarian slices was detected by immunohistochemistry. Progesterone production was measured in cell culture supernatants using ELISAs. Single channels were recorded in the inside-out configuration of the patch-clamp technique. Results We identified two KCa types in human GCs, the intermediate- (IK and the small-conductance KCa (SK. Their functionality was concluded from attenuation of human chorionic gonadotropin-stimulated progesterone production by KCa blockers (TRAM-34, apamin. Functional IK channels were also demonstrated by electrophysiological recording of single KCa channels with distinctive features. Both, IK and BKCa channels were found to be simultaneously active in individual GCs. In agreement with functional data, we identified mRNAs encoding IK, SK1, SK2 and SK3 in human GCs and proteins of IK and SK2 in corresponding human ovarian cells. Molecular characterization of the BKCa channel revealed the presence of mRNAs encoding several BKCa beta-subunits (beta2, beta3, beta4 in human GCs. The multitude of beta-subunits detected might contribute to variations in Ca2+ dependence of individual BKCa channels which we observed in electrophysiological recordings. Conclusion Functional and molecular studies indicate the presence of active IK and SK

  7. Activation gating kinetics of GIRK channels are mediated by cytoplasmic residues adjacent to transmembrane domains.

    Science.gov (United States)

    Sadja, Rona; Reuveny, Eitan

    2009-01-01

    G-protein-coupled inwardly rectifying potassium channels (GIRK/Kir3.x) are involved in neurotransmission-mediated reduction of excitability. The gating mechanism following G protein activation of these channels likely proceeds from movement of inner transmembrane helices to allow K(+) ions movement through the pore of the channel. There is limited understanding of how the binding of G-protein betagamma subunits to cytoplasmic regions of the channel transduces the signal to the transmembrane regions. In this study, we examined the molecular basis that governs the activation kinetics of these channels, using a chimeric approach. We identified two regions as being important in determining the kinetics of activation. One region is the bottom of the outer transmembrane helix (TM1) and the cytoplasmic domain immediately adjacent (the slide helix); and the second region is the bottom of the inner transmembrane helix (TM2) and the cytoplasmic domain immediately adjacent. Interestingly, both of these regions are sufficient in mediating the kinetics of fast activation gating. This result suggests that there is a cooperative movement of either one of these domains to allow fast and efficient activation gating of GIRK channels.

  8. PWSCC Preventive Maintenance Activities for Alloy 600 in Japanese PWR Plants

    International Nuclear Information System (INIS)

    Yamamoto, K.; Sugimoto, N.; Onishi, K.; Okimura, K.

    2012-01-01

    Because many nuclear plants have been in operation for ages, the importance of preventive maintenance technologies is getting higher. One conspicuous problem found in pressurized water reactor (PWR) plants is the primary water stress corrosion cracking (PWSCC) observed in Alloy 600 (a kind of high nickel based alloy) parts. Alloy 600 was used for butt welds between low alloy steel and stainless steel of nozzles of Reactor Vessel (RV), Steam Generator (SG), and Pressurizer (Pz). As PWSCC occurred at these parts may cause Loss of Coolant Accident (LOCA), preventive maintenance is necessary. PWSCC is considered to be caused by a mixture of three elements: high residual tensile stress on surface, material (Alloy 600) and environment. PWSCC can be prevented by improving one of the elements. MHI has been developing stress improvement methods, for example, Water Jet Peening (WJP), Shot Peening by Ultrasonic vibration (USP), and Laser Stress Improvement Process (L-SIP). According to the situation, appropriate method is applied for each part. WJP has been applied for RV nozzles of a lot of plants in Japan. However PWSCC was observed in RV nozzles during the inspection before WJP in recent years, MHI developed the Advanced INLAY system to improve the material from Alloy 600 to Alloy 690. Alloy 600 on the inner surface of the nozzles is removed and welding with Alloy 690 is performed. In addition, heat treatments for the nozzles are difficult for its structural situation, so ambient temperature temper bead welding technique for RV nozzles was developed to make the heat treatments unnecessary. This paper describes countermeasures against PWSCC and introduces the maintenance activities performed in Japan. (author)

  9. Up-Regulatory Effects of Curcumin on Large Conductance Ca2+-Activated K+ Channels

    Science.gov (United States)

    Hei, Hongya; Li, Fangping; Wang, Yunman; Peng, Wen; Zhang, Xuemei

    2015-01-01

    Large conductance Ca2+-activated potassium channels (BK) are targets for research that explores therapeutic means to various diseases, owing to the roles of the channels in mediating multiple physiological processes in various cells and tissues. We investigated the pharmacological effects of curcumin, a compound isolated from the herb Curcuma longa, on BK channels. As recorded by whole-cell patch-clamp, curcumin increased BK (α) and BK (α+β1) currents in transfected HEK293 cells as well as the current density of BK in A7r5 smooth muscle cells in a dose-dependent manner. By incubating with curcumin for 24 hours, the current density of exogenous BK (α) in HEK293 cells and the endogenous BK in A7r5 cells were both enhanced notably, though the steady-state activation of the channels did not shift significantly, except for BK (α+β1). Curcumin up-regulated the BK protein expression without changing its mRNA level in A7r5 cells. The surface expression and the half-life of BK channels were also increased by curcumin in HEK293 cells. These effects of curcumin were abolished by MG-132, a proteasome inhibitor. Curcumin also increased ERK 1/2 phosphorylation, while inhibiting ERK by U0126 attenuated the curcumin-induced up-regulation of BK protein expression. We also observed that the curcumin-induced relaxation in the isolated rat aortic rings was significantly attenuated by paxilline, a BK channel specific blocker. These results show that curcumin enhances the activity of the BK channels by interacting with BK directly as well as enhancing BK protein expression through inhibiting proteasomal degradation and activating ERK signaling pathway. The findings suggest that curcumin is a potential BK channel activator and provide novel insight into its complicated pharmacological effects and the underlying mechanisms. PMID:26672753

  10. Up-Regulatory Effects of Curcumin on Large Conductance Ca2+-Activated K+ Channels.

    Directory of Open Access Journals (Sweden)

    Qijing Chen

    Full Text Available Large conductance Ca2+-activated potassium channels (BK are targets for research that explores therapeutic means to various diseases, owing to the roles of the channels in mediating multiple physiological processes in various cells and tissues. We investigated the pharmacological effects of curcumin, a compound isolated from the herb Curcuma longa, on BK channels. As recorded by whole-cell patch-clamp, curcumin increased BK (α and BK (α+β1 currents in transfected HEK293 cells as well as the current density of BK in A7r5 smooth muscle cells in a dose-dependent manner. By incubating with curcumin for 24 hours, the current density of exogenous BK (α in HEK293 cells and the endogenous BK in A7r5 cells were both enhanced notably, though the steady-state activation of the channels did not shift significantly, except for BK (α+β1. Curcumin up-regulated the BK protein expression without changing its mRNA level in A7r5 cells. The surface expression and the half-life of BK channels were also increased by curcumin in HEK293 cells. These effects of curcumin were abolished by MG-132, a proteasome inhibitor. Curcumin also increased ERK 1/2 phosphorylation, while inhibiting ERK by U0126 attenuated the curcumin-induced up-regulation of BK protein expression. We also observed that the curcumin-induced relaxation in the isolated rat aortic rings was significantly attenuated by paxilline, a BK channel specific blocker. These results show that curcumin enhances the activity of the BK channels by interacting with BK directly as well as enhancing BK protein expression through inhibiting proteasomal degradation and activating ERK signaling pathway. The findings suggest that curcumin is a potential BK channel activator and provide novel insight into its complicated pharmacological effects and the underlying mechanisms.

  11. Piezo proteins are pore-forming subunits of mechanically activated channels.

    Science.gov (United States)

    Coste, Bertrand; Xiao, Bailong; Santos, Jose S; Syeda, Ruhma; Grandl, Jörg; Spencer, Kathryn S; Kim, Sung Eun; Schmidt, Manuela; Mathur, Jayanti; Dubin, Adrienne E; Montal, Mauricio; Patapoutian, Ardem

    2012-02-19

    Mechanotransduction has an important role in physiology. Biological processes including sensing touch and sound waves require as-yet-unidentified cation channels that detect pressure. Mouse Piezo1 (MmPiezo1) and MmPiezo2 (also called Fam38a and Fam38b, respectively) induce mechanically activated cationic currents in cells; however, it is unknown whether Piezo proteins are pore-forming ion channels or modulate ion channels. Here we show that Drosophila melanogaster Piezo (DmPiezo, also called CG8486) also induces mechanically activated currents in cells, but through channels with remarkably distinct pore properties including sensitivity to the pore blocker ruthenium red and single channel conductances. MmPiezo1 assembles as a ∼1.2-million-dalton homo-oligomer, with no evidence of other proteins in this complex. Purified MmPiezo1 reconstituted into asymmetric lipid bilayers and liposomes forms ruthenium-red-sensitive ion channels. These data demonstrate that Piezo proteins are an evolutionarily conserved ion channel family involved in mechanotransduction.

  12. Purification of charybdotoxine, a specific inhibitor of the high-conductance Ca2+-activated K+ channel

    International Nuclear Information System (INIS)

    Smith, C.; Phillips, M.; Miller, C.

    1986-01-01

    Charybdotoxim is a high-affinity specific inhibitor of the high-conductance Ca 2+ -activated K + channel found in the plasma membranes of many vertebrate cell types. Using Ca 2+ -activated K + channels reconstituted into planar lipid bilayer membranes as an assay, the authors have purified the toxin from the venom of the scorpion Leiurus quinquestriatus by a two-step procedure involving chromatofocusing on SP-Sephadex, followed by reversed-phase high-performance liquid chromatography. Charybdotoxin is shown to be a highly basic protein with a mass of 10 kDa. Under the standard assay conditions, the purified toxin inhibits the Ca 2+ -activated K + channel with an apparent dissociation constant of 3.5 nM. The protein is unusually stable, with inhibitory potency being insensitive to boiling or exposure to organic solvents. The toxin's activity is sensitive to chymotrypsin treatment and to acylation of lysine groups. The protein may be radioiodinated without loss of activity

  13. Impact on a fish assemblage of the maintenance dredging of a navigation channel in a tropical coastal ecosystem

    Directory of Open Access Journals (Sweden)

    Demarques Ribeiro da Silva Junior

    2012-03-01

    Full Text Available Dredging and dredge-spoil disposal are among the major problems in coastal management. Many of the scientific contributions concerning the impacts of this practice are based on the study of sessile organisms and subtropical environments. We evaluated changes in the composition and abundance of a fish assemblage resulting from dredging and sediment disposal at the mouth and in the adjacent waters of the Caravelas River on the north-eastern coast of Brazil. Samples were collected in two directly impacted and three adjacent areas. Differences among stations were not significant, but the dredged site had the least diverse station, as expected. The stations farthest from the directly impacted areas apparently were not influenced by the coastal work, thus suggesting localised effects. The contribution of the present study is particularly important because of the study area's proximity to others that have high conservation value such as mangrove forests and coral reefs, and the relevance of the subject given the continuing dredging activity.A dragagem e descarte de sedimento se destacam como atividades que geram grandes distúrbios aos ecossistemas marinhos e, consequentemente tornam-se um desafio ao manejo e ordenamento costeiro. Grande parte dos estudos que abordam seus impactos é baseada em pesquisas com organismos sésseis e em ambientes temperados, criando uma lacuna no entendimento de seus efeitos sobre a ictiofauna, principalmente nas regiões tropicais. No presente estudo foram avaliadas as alterações na composição e abundância da comunidade de peixes sob influência da dragagem e descarte de sedimento na foz do Rio Caravelas, costa da região Nordeste do Brasil. As amostras foram obtidas em duas estações diretamente afetadas e em três outras áreas adjacentes. Não houve diferença significativa na diversidade média obtida para cada estação, porém a estação correspondente à área dragada apresentou o menor valor dentre as

  14. Multiple-channel detection of cellular activities by ion-sensitive transistors

    Science.gov (United States)

    Machida, Satoru; Shimada, Hideto; Motoyama, Yumi

    2018-04-01

    An ion-sensitive field-effect transistor to record cellular activities was demonstrated. This field-effect transistor (bio transistor) includes cultured cells on the gate insulator instead of gate electrode. The bio transistor converts a change in potential underneath the cells into variation of the drain current when ion channels open. The bio transistor has high detection sensitivity to even minute variations in potential utilizing a subthreshold swing region. To open ion channels, a reagent solution (acetylcholine) was added to a human-originating cell cultured on the bio transistor. The drain current was successfully decreased with the addition of acetylcholine. Moreover, we attempted to detect the opening of ion channels using a multiple-channel measurement circuit containing several bio transistors. As a consequence, the drain current distinctly decreased only after the addition of acetylcholine. We confirmed that this measurement system including bio transistors enables to observation of cellular activities sensitively and simultaneously.

  15. Activation of human IK and SK Ca2+ -activated K+ channels by NS309 (6,7-dichloro-1H-indole-2,3-dione 3-oxime)

    DEFF Research Database (Denmark)

    Strøbaek, Dorte; Teuber, Lene; Jørgensen, Tino D

    2004-01-01

    We have identified and characterized the compound NS309 (6,7-dichloro-1H-indole-2,3-dione 3-oxime) as a potent activator of human Ca2+ -activated K+ channels of SK and IK types, whereas it is devoid of effect on BK type channels. IK- and SK-channels have previously been reported to be activated...

  16. Hydralazine-induced vasodilation involves opening of high conductance Ca2+-activated K+ channels

    DEFF Research Database (Denmark)

    Bang, Lone; Nielsen-Kudsk, J E; Gruhn, N

    1998-01-01

    The purpose of this study was to investigate whether high conductance Ca2+-activated K+ channels (BK(Ca)) are mediating the vasodilator action of hydralazine. In isolated porcine coronary arteries, hydralazine (1-300 microM), like the K+ channel opener levcromakalim, preferentially relaxed......M) suppressed this response by 82% (P opening of BK(Ca) takes part in the mechanism whereby...

  17. Cholesterol regulates HERG K+ channel activation by increasing phospholipase C β1 expression.

    Science.gov (United States)

    Chun, Yoon Sun; Oh, Hyun Geun; Park, Myoung Kyu; Cho, Hana; Chung, Sungkwon

    2013-01-01

    Human ether-a-go-go-related gene (HERG) K(+) channel underlies the rapidly activating delayed rectifier K(+) conductance (IKr) during normal cardiac repolarization. Also, it may regulate excitability in many neuronal cells. Recently, we showed that enrichment of cell membrane with cholesterol inhibits HERG channels by reducing the levels of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] due to the activation of phospholipase C (PLC). In this study, we further explored the effect of cholesterol enrichment on HERG channel kinetics. When membrane cholesterol level was mildly increased in human embryonic kidney (HEK) 293 cells expressing HERG channel, the inactivation and deactivation kinetics of HERG current were not affected, but the activation rate was significantly decelerated at all voltages tested. The application of PtdIns(4,5)P2 or inhibitor for PLC prevented the effect of cholesterol enrichment, while the presence of antibody against PtdIns(4,5)P2 in pipette solution mimicked the effect of cholesterol enrichment. These results indicate that the effect of cholesterol enrichment on HERG channel is due to the depletion of PtdIns(4,5)P2. We also found that cholesterol enrichment significantly increases the expression of β1 and β3 isoforms of PLC (PLCβ1, PLCβ3) in the membrane. Since the effects of cholesterol enrichment on HERG channel were prevented by inhibiting transcription or by inhibiting PLCβ1 expression, we conclude that increased PLCβ1 expression leads to the deceleration of HERG channel activation rate via downregulation of PtdIns(4,5)P2. These results confirm a crosstalk between two plasma membrane-enriched lipids, cholesterol and PtdIns(4,5)P2, in the regulation of HERG channels.

  18. Fatty acid oxidation is required for active and quiescent brown adipose tissue maintenance and thermogenic programing.

    Science.gov (United States)

    Gonzalez-Hurtado, Elsie; Lee, Jieun; Choi, Joseph; Wolfgang, Michael J

    2018-01-01

    To determine the role of fatty acid oxidation on the cellular, molecular, and physiologic response of brown adipose tissue to disparate paradigms of chronic thermogenic stimulation. Mice with an adipose-specific loss of Carnitine Palmitoyltransferase 2 (Cpt2 A-/- ), that lack mitochondrial long chain fatty acid β-oxidation, were subjected to environmental and pharmacologic interventions known to promote thermogenic programming in adipose tissue. Chronic administration of β3-adrenergic (CL-316243) or thyroid hormone (GC-1) agonists induced a loss of BAT morphology and UCP1 expression in Cpt2 A-/- mice. Fatty acid oxidation was also required for the browning of white adipose tissue (WAT) and the induction of UCP1 in WAT. In contrast, chronic cold (15 °C) stimulation induced UCP1 and thermogenic programming in both control and Cpt2 A-/- adipose tissue albeit to a lesser extent in Cpt2 A-/- mice. However, thermoneutral housing also induced the loss of UCP1 and BAT morphology in Cpt2 A-/- mice. Therefore, adipose fatty acid oxidation is required for both the acute agonist-induced activation of BAT and the maintenance of quiescent BAT. Consistent with this data, Cpt2 A-/- BAT exhibited increased macrophage infiltration, inflammation and fibrosis irrespective of BAT activation. Finally, obese Cpt2 A-/- mice housed at thermoneutrality exhibited a loss of interscapular BAT and were refractory to β3-adrenergic-induced energy expenditure and weight loss. Mitochondrial long chain fatty acid β-oxidation is critical for the maintenance of the brown adipocyte phenotype both during times of activation and quiescence. Copyright © 2017 The Authors. Published by Elsevier GmbH.. All rights reserved.

  19. Fatty acid oxidation is required for active and quiescent brown adipose tissue maintenance and thermogenic programing

    Directory of Open Access Journals (Sweden)

    Elsie Gonzalez-Hurtado

    2018-01-01

    Full Text Available Objective: To determine the role of fatty acid oxidation on the cellular, molecular, and physiologic response of brown adipose tissue to disparate paradigms of chronic thermogenic stimulation. Methods: Mice with an adipose-specific loss of Carnitine Palmitoyltransferase 2 (Cpt2A−/−, that lack mitochondrial long chain fatty acid β-oxidation, were subjected to environmental and pharmacologic interventions known to promote thermogenic programming in adipose tissue. Results: Chronic administration of β3-adrenergic (CL-316243 or thyroid hormone (GC-1 agonists induced a loss of BAT morphology and UCP1 expression in Cpt2A−/− mice. Fatty acid oxidation was also required for the browning of white adipose tissue (WAT and the induction of UCP1 in WAT. In contrast, chronic cold (15 °C stimulation induced UCP1 and thermogenic programming in both control and Cpt2A−/− adipose tissue albeit to a lesser extent in Cpt2A−/− mice. However, thermoneutral housing also induced the loss of UCP1 and BAT morphology in Cpt2A−/− mice. Therefore, adipose fatty acid oxidation is required for both the acute agonist-induced activation of BAT and the maintenance of quiescent BAT. Consistent with this data, Cpt2A−/− BAT exhibited increased macrophage infiltration, inflammation and fibrosis irrespective of BAT activation. Finally, obese Cpt2A−/− mice housed at thermoneutrality exhibited a loss of interscapular BAT and were refractory to β3-adrenergic-induced energy expenditure and weight loss. Conclusion: Mitochondrial long chain fatty acid β-oxidation is critical for the maintenance of the brown adipocyte phenotype both during times of activation and quiescence. Keywords: Fatty acid oxidation, Brown adipose tissue, Cold induced thermogenesis, Adrenergic signaling, Adipose macrophage

  20. Deuteron and neutron induced activation in the Eveda accelerator materials: implications for the accelerator maintenance

    Energy Technology Data Exchange (ETDEWEB)

    Garcia, M.; Sanz, J.; Garcia, N.; Cabellos, O. [Madrid Univ. Politecnica, C/ Jose Gutierrez Abascal, lnstituto de Fusion Nuclear (Spain); Sauvan, R. [Universidad Nacional de Educacion a Distancia (UNED), Madrid (Spain); Moreno, C.; Sedano, L.A. [CIEMAT-Centro de Investigaciones Energeticas Medioambientales y Tecnologicas, Association Euratom-CIEMAT, Madrid (Spain)

    2007-07-01

    Full text of publication follows: The IFMIF (International Fusion Materials Irradiation Facility) is an accelerator-based DLi neutron source designed to test fusion reactor candidate materials for high fluence neutrons. Before deciding IFMIF construction, an engineering design and associated experimental data acquisition, defined as EVEDA, has been proposed. Along the EVEDA accelerator, deuteron beam losses collide with the accelerator materials, producing activation and consequent radiations responsible of dose. Calculation of the dose rates in the EVEDA accelerator room is necessary in order to analyze the feasibility for manual maintenance. Dose rates due to the activation produced by the deuteron beam losses interaction with the accelerator materials, will be calculated with the ACAB activation code, using EAF2007 library for deuteron activation cross-sections. Also, dose rates from the activation induced by the neutron source produced by the interaction of deuteron beam losses with the accelerator materials and the deuterium implanted in the structural lattice, will be calculated with the SRIM2006, TMAP7, DROSG2000/NEUYIE, MCNPX and ACAB codes. All calculations will be done for the EVEDA accelerator with the room temperature DTL structure, which is based on copper cavities for the DTL. Some calculations will be done for the superconducting DTL structure, based on niobium cavities for the DTL working at cryogenic temperature. Final analysis will show the dominant mechanisms and major radionuclides contributing to the surface dose rates. (authors)

  1. Instrumentation maintenance

    International Nuclear Information System (INIS)

    Mack, D.A.

    1976-09-01

    It is essential to any research activity that accurate and efficient measurements be made for the experimental parameters under consideration for each individual experiment or test. Satisfactory measurements in turn depend upon having the necessary instruments and the capability of ensuring that they are performing within their intended specifications. This latter requirement can only be achieved by providing an adequate maintenance facility, staffed with personnel competent to understand the problems associated with instrument adjustment and repair. The Instrument Repair Shop at the Lawrence Berkeley Laboratory is designed to achieve this end. The organization, staffing and operation of this system is discussed. Maintenance policy should be based on studies of (1) preventive vs. catastrophic maintenance, (2) records indicating when equipment should be replaced rather than repaired and (3) priorities established to indicate the order in which equipment should be repaired. Upon establishing a workable maintenance policy, the staff should be instructed so that they may provide appropriate scheduled preventive maintenance, calibration and corrective procedures, and emergency repairs. The education, training and experience of the maintenance staff is discussed along with the organization for an efficient operation. The layout of the various repair shops is described in the light of laboratory space and financial constraints

  2. ALTERNATIVE EQUATIONS FOR DYNAMIC BEHAVIOR OF IONIC CHANNEL ACTIVATION AND INACTIVATION GATES

    Directory of Open Access Journals (Sweden)

    Mahmut ÖZER

    2003-03-01

    Full Text Available In this paper, alternative equations for dynamics of ionic channel activation and inactivation gates are proposed based on the path probability method. Dynamic behavior of a voltage-gated ionic channel is modeled by the conventional Hodgkin-Huxley (H-H mathematical formalism. In that model, conductance of the channel is defined in terms of activation and inactivation gates. Dynamics of the activation and inactivation gates is modeled by first-order differential equations dependent on the gate variable and the membrane potential. In the new approach proposed in this study, dynamic behavior of activation and inactivation gates is modeled by a firstorder differential equation dependent on internal energy and membrane potential by using the path probability method which is widely used in statistical physics. The new model doesn't require the time constant and steadystate values which are used explicitly in the H-H model. The numerical results show validity of the proposed method.

  3. The impact of self-efficacy on physical activity maintenance in patients with hip osteoarthritis - a mixed methods study.

    Science.gov (United States)

    Hammer, Nanna Maria; Bieler, Theresa; Beyer, Nina; Midtgaard, Julie

    2016-08-01

    Understanding motivational factors related to physical activity (PA) maintenance is essential in promoting long-term exercise benefits. This study explored the impact of self-efficacy (SE) on post-intervention PA maintenance in patients with hip osteoarthritis. An SE-theory based mixed-methods sub-study of a trial investigating the effects of 4 months supervised exercise in patients with hip osteoarthritis. Questionnaire data (n = 52; baseline and 12 months) on PA and SE (Arthritis Self-Efficacy Scale, ASES, score-range 10-100) were analysed (Mann-Whitney test) for differences in characteristics of maintainers and non-maintainers. Semi-structured individual interviews (n = 15; at 12-months follow-up) were analysed using directed content analysis. Compared to non-maintainers (n = 9; 17%) maintainers (n = 31; 60%) had improved (p exercise outcome expectations and obligation towards the study influenced maintenance. SE contributes to understanding of post-intervention PA maintenance in patients with hip osteoarthritis. However, disease-related factors and clinical trial participation appears significant too. Implications for Rehabilitation Patients' perceived self-efficacy for physical activity contributes to the understanding of post-intervention physical activity maintenance in patients with hip osteoarthritis. Practitioners may benefit from incorporating the self-efficacy theory in the planning and execution of exercise interventions to promote post-intervention physical activity maintenance and long term health benefits. Post-intervention physical activity maintenance may be increased by focussing on the patients' exercise self-efficacy through verbal persuasion and support, disease-specific information and information on normal physiological responses to exercise combined with an individualised training progression to support experiences of success and achievement of desired outcomes.

  4. Dietary and physical activity behaviors among adults successful at weight loss maintenance

    Directory of Open Access Journals (Sweden)

    Gillespie Cathleen

    2006-07-01

    Full Text Available Abstract Background There is limited population-based data on behavioral factors found to be important for successful weight loss maintenance among adults. Methods Data from the 2004 Styles surveys, mailed to U.S. adults aged ≥18 years were used to examine the difference in selected weight loss strategies and attitudes among persons who reported successful weight loss attempts (lost weight and able to keep it off and persons who were not successful (previous attempts to lose weight were unsuccessful or they could not keep the lost weight off. Behaviors examined included modification of diet, leisure-time and sports activities, and self-monitoring, and barriers to weight management. Results Among adults who reported losing weight or trying to lose weight, 31.0% had been successful at both losing weight and maintenance after weight loss. Successful weight loss status differed by sex, age, and current weight status. Assessment of reported weight loss strategies, found that exercising ≥30 minutes/day and adding physical activity to daily life were significantly higher among successful versus unsuccessful weight losers. Individuals who were successful at weight loss and maintenance were less likely to use over-the-counter diet products than those who were unsuccessful at weight loss. Significantly more successful versus unsuccessful weight losers reported that on most days of the week they planned meals (35.9% vs. 24.9%, tracked calories (17.7% vs. 8.8%, tracked fat (16.4% vs. 6.6%, and measured food on plate (15.9% vs. 6.7%. Successful losers were also more likely to weigh themselves daily (20.3% vs. 11.0%. There were a significantly higher proportion of successful losers who reported lifting weights (19.0% versus unsuccessful (10.9%. The odds of being a successful weight loser were 48%–76% lower for those reporting exercise weight control barriers were influencing factors (e.g., no time, too tired to exercise, no one to exercise with, too

  5. Formation of activation mechanism for investment maintenance management of gas industry enterprises

    Directory of Open Access Journals (Sweden)

    Вікторія Валеріївна Чорній

    2016-11-01

    Full Text Available The problematic aspects of the existing management mechanism of investment management of gas industry enterprises are analyzed in the article. The ways of their elimination are suggested in accordance with the Energy Strategy of Ukraine – 2035 at different levels of its operation. The ways of improving the investment management mechanism of the enterprises in this industry on three levels are proved. Activation mechanism for investment maintenance management of gas industry enterprises is developed as result of research. Its implementation will allow enterprises of the industry to increase its investment attractiveness and effectively carry out the procedure for enterprises’ entry to the market of initial public offering to raise the required amount of investment resources for the modernization of investigated industry.

  6. Hybrid Particle Swarm Optimization for Hybrid Flowshop Scheduling Problem with Maintenance Activities

    Science.gov (United States)

    Li, Jun-qing; Pan, Quan-ke; Mao, Kun

    2014-01-01

    A hybrid algorithm which combines particle swarm optimization (PSO) and iterated local search (ILS) is proposed for solving the hybrid flowshop scheduling (HFS) problem with preventive maintenance (PM) activities. In the proposed algorithm, different crossover operators and mutation operators are investigated. In addition, an efficient multiple insert mutation operator is developed for enhancing the searching ability of the algorithm. Furthermore, an ILS-based local search procedure is embedded in the algorithm to improve the exploitation ability of the proposed algorithm. The detailed experimental parameter for the canonical PSO is tuning. The proposed algorithm is tested on the variation of 77 Carlier and Néron's benchmark problems. Detailed comparisons with the present efficient algorithms, including hGA, ILS, PSO, and IG, verify the efficiency and effectiveness of the proposed algorithm. PMID:24883414

  7. Hybrid Particle Swarm Optimization for Hybrid Flowshop Scheduling Problem with Maintenance Activities

    Directory of Open Access Journals (Sweden)

    Jun-qing Li

    2014-01-01

    Full Text Available A hybrid algorithm which combines particle swarm optimization (PSO and iterated local search (ILS is proposed for solving the hybrid flowshop scheduling (HFS problem with preventive maintenance (PM activities. In the proposed algorithm, different crossover operators and mutation operators are investigated. In addition, an efficient multiple insert mutation operator is developed for enhancing the searching ability of the algorithm. Furthermore, an ILS-based local search procedure is embedded in the algorithm to improve the exploitation ability of the proposed algorithm. The detailed experimental parameter for the canonical PSO is tuning. The proposed algorithm is tested on the variation of 77 Carlier and Néron’s benchmark problems. Detailed comparisons with the present efficient algorithms, including hGA, ILS, PSO, and IG, verify the efficiency and effectiveness of the proposed algorithm.

  8. Hybrid particle swarm optimization for hybrid flowshop scheduling problem with maintenance activities.

    Science.gov (United States)

    Li, Jun-qing; Pan, Quan-ke; Mao, Kun

    2014-01-01

    A hybrid algorithm which combines particle swarm optimization (PSO) and iterated local search (ILS) is proposed for solving the hybrid flowshop scheduling (HFS) problem with preventive maintenance (PM) activities. In the proposed algorithm, different crossover operators and mutation operators are investigated. In addition, an efficient multiple insert mutation operator is developed for enhancing the searching ability of the algorithm. Furthermore, an ILS-based local search procedure is embedded in the algorithm to improve the exploitation ability of the proposed algorithm. The detailed experimental parameter for the canonical PSO is tuning. The proposed algorithm is tested on the variation of 77 Carlier and Néron's benchmark problems. Detailed comparisons with the present efficient algorithms, including hGA, ILS, PSO, and IG, verify the efficiency and effectiveness of the proposed algorithm.

  9. Slick (Kcnt2 Sodium-Activated Potassium Channels Limit Peptidergic Nociceptor Excitability and Hyperalgesia

    Directory of Open Access Journals (Sweden)

    Danielle L Tomasello

    2017-09-01

    Full Text Available The Slick (Kcnt2 sodium-activated potassium (K Na channel is a rapidly gating and weakly voltage-dependent and sodium-dependent potassium channel with no clearly defined physiological function. Within the dorsal root ganglia (DRGs, we show Slick channels are exclusively expressed in small-sized and medium-sized calcitonin gene–related peptide (CGRP-containing DRG neurons, and a pool of channels are localized to large dense-core vesicles (LDCV-containing CGRP. We stimulated DRG neurons for CGRP release and found Slick channels contained within CGRP-positive LDCV translocated to the neuronal membrane. Behavioral studies in Slick knockout (KO mice indicated increased basal heat detection and exacerbated thermal hyperalgesia compared with wild-type littermate controls during neuropathic and chronic inflammatory pain. Electrophysiologic recordings of DRG neurons from Slick KO mice revealed that Slick channels contribute to outward current, propensity to fire action potentials (APs, and to AP properties. Our data suggest that Slick channels restrain the excitability of CGRP-containing neurons, diminishing pain behavior after inflammation and injury.

  10. Ca2+ Channel Re-localization to Plasma-Membrane Microdomains Strengthens Activation of Ca2+-Dependent Nuclear Gene Expression

    Directory of Open Access Journals (Sweden)

    Krishna Samanta

    2015-07-01

    Full Text Available In polarized cells or cells with complex geometry, clustering of plasma-membrane (PM ion channels is an effective mechanism for eliciting spatially restricted signals. However, channel clustering is also seen in cells with relatively simple topology, suggesting it fulfills a more fundamental role in cell biology than simply orchestrating compartmentalized responses. Here, we have compared the ability of store-operated Ca2+ release-activated Ca2+ (CRAC channels confined to PM microdomains with a similar number of dispersed CRAC channels to activate transcription factors, which subsequently increase nuclear gene expression. For similar levels of channel activity, we find that channel confinement is considerably more effective in stimulating gene expression. Our results identify a long-range signaling advantage to the tight evolutionary conservation of channel clustering and reveal that CRAC channel aggregation increases the strength, fidelity, and reliability of the general process of excitation-transcription coupling.

  11. Inhibition of T cell proliferation by selective block of Ca(2+)-activated K(+) channels

    DEFF Research Database (Denmark)

    Jensen, B S; Odum, Niels; Jorgensen, N K

    1999-01-01

    cell activation and proliferation has been investigated by using various blockers of IK channels. The Ca(2+)-activated K(+) current in human T cells is shown by the whole-cell voltage-clamp technique to be highly sensitive to clotrimazole, charybdotoxin, and nitrendipine, but not to ketoconazole...

  12. 76 FR 12967 - Agency Information Collection Activities; Proposed Collection; Comment Request; Channels of Trade...

    Science.gov (United States)

    2011-03-09

    ...] Agency Information Collection Activities; Proposed Collection; Comment Request; Channels of Trade Policy...)--Extension The Food Quality Protection Act of 1996 (FQPA), which amended the Federal Insecticide, Fungicide... Burden \\1\\ Annual Activity No. of frequency per Total annual Hours per Total hours respondents response...

  13. Cloning, functional expression, and characterization of a PKA-activated gastric Cl- channel.

    Science.gov (United States)

    Malinowska, D H; Kupert, E Y; Bahinski, A; Sherry, A M; Cuppoletti, J

    1995-01-01

    cDNA encoding a Cl- channel was isolated from a rabbit gastric library, sequenced, and expressed in Xenopus oocytes. The predicted protein (898 amino acids, relative molecular mass 98,433 Da) was overall 93% similar to the rat brain ClC-2 Cl- channel. However, a 151-amino acid stretch toward the COOH-terminus was 74% similar to ClC-2 with six amino acids deleted. Two new potential protein kinase A (PKA) phosphorylation sites (also protein kinase C phosphorylation sites) were introduced. cRNA-injected Xenopus oocytes expressed a Cl- channel that was active at pHtrans 3 and had a linear current-voltage (I-V) curve and a slope conductance of 29 +/- 1 pS at 800 mM CsCl. A fivefold Cl- gradient caused a rightward shift in the I-V curve with a reversal potential of +30 +/- 3 mV, indicating anion selectivity. The selectivity was I- > Cl- > NO3-. The native and recombinant Cl- channel were both activated in vitro by PKA catalytic subunit and ATP. The electrophysiological and regulatory properties of the cloned and the native channel were similar. The cloned protein may be the Cl- channel involved in gastric HCl secretion.

  14. The Activation Effect of Hainantoxin-I, a Peptide Toxin from the Chinese Spider, Ornithoctonus hainana, on Intermediate-Conductance Ca2+-Activated K+ Channels

    Directory of Open Access Journals (Sweden)

    Pengfei Huang

    2014-08-01

    Full Text Available Intermediate-conductance Ca2+-activated K+ (IK channels are calcium/calmodulin-regulated voltage-independent K+ channels. Activation of IK currents is important in vessel and respiratory tissues, rendering the channels potential drug targets. A variety of small organic molecules have been synthesized and found to be potent activators of IK channels. However, the poor selectivity of these molecules limits their therapeutic value. Venom-derived peptides usually block their targets with high specificity. Therefore, we searched for novel peptide activators of IK channels by testing a series of toxins from spiders. Using electrophysiological experiments, we identified hainantoxin-I (HNTX-I as an IK-channel activator. HNTX-I has little effect on voltage-gated Na+ and Ca2+ channels from rat dorsal root ganglion neurons and on the heterologous expression of voltage-gated rapidly activating delayed rectifier K+ channels (human ether-à-go-go-related gene; human ERG in HEK293T cells. Only 35.2% ± 0.4% of the currents were activated in SK channels, and there was no effect on BK channels. We demonstrated that HNTX-I was not a phrenic nerve conduction blocker or acutely toxic. This is believed to be the first report of a peptide activator effect on IK channels. Our study suggests that the activity and selectivity of HNTX-I on IK channels make HNTX-I a promising template for designing new drugs for cardiovascular diseases.

  15. Impedance spectroscopy of micro-Droplets reveals activation of Bacterial Mechanosensitive Channels in Hypotonic Solutions

    Science.gov (United States)

    Ebrahimi, Aida; Alam, Muhammad A.

    Rapid detection of bacterial pathogens is of great importance in healthcare, food safety, environmental monitoring, and homeland security. Most bacterial detection platforms rely on binary fission (i.e. cell growth) to reach a threshold cell population that can be resolved by the sensing method. Since cell division depends on the bacteria type, the detection time of such methods can vary from hours to days. In contrast, in this work, we show that bacteria cells can be detected within minutes by relying on activation of specific protein channels, i.e. mechanosensitive channels (MS channels). When cells are exposed to hypotonic solutions, MS channels allow efflux of solutes to the external solution which leads to release the excessive membrane tension. Release of the cytoplasmic solutes, in turn, results in increase of the electrical conductance measured by droplet-based impedance sensing. The approach can be an effective technique for fast, pre-screening of bacterial contamination at ultra-low concentration.

  16. The maintenance of elevated active chlorine levels in the Antarctic lower stratosphere through HCl null cycles

    Science.gov (United States)

    Müller, Rolf; Grooß, Jens-Uwe; Mannan Zafar, Abdul; Robrecht, Sabine; Lehmann, Ralph

    2018-03-01

    The Antarctic ozone hole arises from ozone destruction driven by elevated levels of ozone destroying (active) chlorine in Antarctic spring. These elevated levels of active chlorine have to be formed first and then maintained throughout the period of ozone destruction. It is a matter of debate how this maintenance of active chlorine is brought about in Antarctic spring, when the rate of formation of HCl (considered to be the main chlorine deactivation mechanism in Antarctica) is extremely high. Here we show that in the heart of the ozone hole (16-18 km or 85-55 hPa, in the core of the vortex), high levels of active chlorine are maintained by effective chemical cycles (referred to as HCl null cycles hereafter). In these cycles, the formation of HCl is balanced by immediate reactivation, i.e. by immediate reformation of active chlorine. Under these conditions, polar stratospheric clouds sequester HNO3 and thereby cause NO2 concentrations to be low. These HCl null cycles allow active chlorine levels to be maintained in the Antarctic lower stratosphere and thus rapid ozone destruction to occur. For the observed almost complete activation of stratospheric chlorine in the lower stratosphere, the heterogeneous reaction HCl + HOCl is essential; the production of HOCl occurs via HO2 + ClO, with the HO2 resulting from CH2O photolysis. These results are important for assessing the impact of changes of the future stratospheric composition on the recovery of the ozone hole. Our simulations indicate that, in the lower stratosphere, future increased methane concentrations will not lead to enhanced chlorine deactivation (through the reaction CH4 + Cl → HCl + CH3) and that extreme ozone destruction to levels below ≈ 0.1 ppm will occur until mid-century.

  17. Differential distribution of the sodium-activated potassium channels slick and slack in mouse brain.

    Science.gov (United States)

    Rizzi, Sandra; Knaus, Hans-Günther; Schwarzer, Christoph

    2016-07-01

    The sodium-activated potassium channels Slick (Slo2.1, KCNT2) and Slack (Slo2.2, KCNT1) are high-conductance potassium channels of the Slo family. In neurons, Slick and Slack channels are involved in the generation of slow afterhyperpolarization, in the regulation of firing patterns, and in setting and stabilizing the resting membrane potential. The distribution and subcellular localization of Slick and Slack channels in the mouse brain have not yet been established in detail. The present study addresses this issue through in situ hybridization and immunohistochemistry. Both channels were widely distributed and exhibited distinct distribution patterns. However, in some brain regions, their expression overlapped. Intense Slick channel immunoreactivity was observed in processes, varicosities, and neuronal cell bodies of the olfactory bulb, granular zones of cortical regions, hippocampus, amygdala, lateral septal nuclei, certain hypothalamic and midbrain nuclei, and several regions of the brainstem. The Slack channel showed primarily a diffuse immunostaining pattern, and labeling of cell somata and processes was observed only occasionally. The highest Slack channel expression was detected in the olfactory bulb, lateral septal nuclei, basal ganglia, and distinct areas of the midbrain, brainstem, and cerebellar cortex. In addition, comparing our data obtained from mouse brain with a previously published study on rat brain revealed some differences in the expression and distribution of Slick and Slack channels in these species. J. Comp. Neurol. 524:2093-2116, 2016. © 2015 The Authors The Journal of Comparative Neurology Published by Wiley Periodicals, Inc. © 2015 The Authors The Journal of Comparative Neurology Published by Wiley Periodicals, Inc.

  18. Plasmin in Nephrotic Urine Activates the Epithelial Sodium Channel

    DEFF Research Database (Denmark)

    Svenningsen, Per; Bistrup, Claus; Friis, Ulla G

    2009-01-01

    stimulated amiloride-sensitive transepithelial sodium transport in M-1 cells and increased amiloride-sensitive whole-cell currents in Xenopus laevis oocytes heterologously expressing ENaC. Activation of ENaC by plasmin involved cleavage and release of an inhibitory peptide from the ENaC gamma subunit...

  19. Sediment transport in an active erodible channel bend

    Indian Academy of Sciences (India)

    Local variation of sediment transport is primarily controlled by active bank erosion, land spur and sand bar formation. Vertical distribution of suspended sediment concentration follows a power function with normalized depth. Average bed-material concentration at the reach level is computed from observed sediment profiles, ...

  20. Ion channel activity of membrane vesicles released from sea urchin sperm during the acrosome reaction

    International Nuclear Information System (INIS)

    Schulz, Joseph R.; Vega-Beltran, Jose L. de la; Beltran, Carmen; Vacquier, Victor D.; Darszon, Alberto

    2004-01-01

    The sperm acrosome reaction (AR) involves ion channel activation. In sea urchin sperm, the AR requires Ca 2+ and Na + influx and K + and H + efflux. During the AR, the plasma membrane fuses with the acrosomal vesicle membrane forming hybrid membrane vesicles that are released from sperm into the medium. This paper reports the isolation and preliminary characterization of these acrosome reaction vesicles (ARVs), using synaptosome-associated protein of 25 kDa (SNAP-25) as a marker. Isolated ARVs have a unique protein composition. The exocytosis regulatory proteins vesicle-associated membrane protein and SNAP-25 are inside ARVs, as judged by protease protection experiments, and membrane associated based on Triton X-114 partitioning. ARVs fused with planar bilayers display three main types of single channel activity. The most frequently recorded channel is cationic, weakly voltage dependent and has a low open probability that increases with negative potentials. This channel is activated by cAMP, blocked by Ba 2+ , and has a PK + /PNa + selectivity of 4.5. ARVs represent a novel membrane preparation suitable to deepen our understanding of ion channel activity in the AR and during fertilization

  1. Dendritic calcium channels and their activation by synaptic signals in auditory coincidence detector neurons.

    Science.gov (United States)

    Blackmer, Trillium; Kuo, Sidney P; Bender, Kevin J; Apostolides, Pierre F; Trussell, Laurence O

    2009-08-01

    The avian nucleus laminaris (NL) encodes the azimuthal location of low-frequency sound sources by detecting the coincidence of binaural signals. Accurate coincidence detection requires precise developmental regulation of the lengths of the fine, bitufted dendrites that characterize neurons in NL. Such regulation has been suggested to be driven by local, synaptically mediated, dendritic signals such as Ca(2+). We examined Ca(2+) signaling through patch clamp and ion imaging experiments in slices containing nucleus laminaris from embryonic chicks. Voltage-clamp recordings of neurons located in the NL showed the presence of large Ca(2+) currents of two types, a low voltage-activated, fast inactivating Ni(2+) sensitive channel resembling mammalian T-type channels, and a high voltage-activated, slowly inactivating Cd(2+) sensitive channel. Two-photon Ca(2+) imaging showed that both channel types were concentrated on dendrites, even at their distal tips. Single action potentials triggered synaptically or by somatic current injection immediately elevated Ca(2+) throughout the entire cell. Ca(2+) signals triggered by subthreshold synaptic activity were highly localized. Thus when electrical activity is suprathreshold, Ca(2+) channels ensure that Ca(2+) rises in all dendrites, even those that are synaptically inactive.

  2. Intermolecular Interactions in the TMEM16A Dimer Controlling Channel Activity.

    Science.gov (United States)

    Scudieri, Paolo; Musante, Ilaria; Gianotti, Ambra; Moran, Oscar; Galietta, Luis J V

    2016-12-08

    TMEM16A and TMEM16B are plasma membrane proteins with Ca 2+ -dependent Cl - channel function. By replacing the carboxy-terminus of TMEM16A with the equivalent region of TMEM16B, we obtained channels with potentiation of channel activity. Progressive shortening of the chimeric region restricted the "activating domain" to a short sequence close to the last transmembrane domain and led to TMEM16A channels with high activity at very low intracellular Ca 2+ concentrations. To elucidate the molecular mechanism underlying this effect, we carried out experiments based on double chimeras, Forster resonance energy transfer, and intermolecular cross-linking. We also modeled TMEM16A structure using the Nectria haematococca TMEM16 protein as template. Our results indicate that the enhanced activity in chimeric channels is due to altered interaction between the carboxy-terminus and the first intracellular loop in the TMEM16A homo-dimer. Mimicking this perturbation with a small molecule could be the basis for a pharmacological stimulation of TMEM16A-dependent Cl - transport.

  3. Osteopontin activates the diabetes-associated potassium channel TALK-1 in pancreatic β-cells.

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    Matthew T Dickerson

    Full Text Available Glucose-stimulated insulin secretion (GSIS relies on β-cell Ca2+ influx, which is modulated by the two-pore-domain K+ (K2P channel, TALK-1. A gain-of-function polymorphism in KCNK16, the gene encoding TALK-1, increases risk for developing type-2 diabetes. While TALK-1 serves an important role in modulating GSIS, the regulatory mechanism(s that control β-cell TALK-1 channels are unknown. Therefore, we employed a membrane-specific yeast two-hybrid (MYTH assay to identify TALK-1-interacting proteins in human islets, which will assist in determining signaling modalities that modulate TALK-1 function. Twenty-one proteins from a human islet cDNA library interacted with TALK-1. Some of these interactions increased TALK-1 activity, including intracellular osteopontin (iOPN. Intracellular OPN is highly expressed in β-cells and is upregulated under pre-diabetic conditions to help maintain normal β-cell function; however, the functional role of iOPN in β-cells is poorly understood. We found that iOPN colocalized with TALK-1 in pancreatic sections and coimmunoprecipitated with human islet TALK-1 channels. As human β-cells express two K+ channel-forming variants of TALK-1, regulation of these TALK-1 variants by iOPN was assessed. At physiological voltages iOPN activated TALK-1 transcript variant 3 channels but not TALK-1 transcript variant 2 channels. Activation of TALK-1 channels by iOPN also hyperpolarized resting membrane potential (Vm in HEK293 cells and in primary mouse β-cells. Intracellular OPN was also knocked down in β-cells to test its effect on β-cell TALK-1 channel activity. Reducing β-cell iOPN significantly decreased TALK-1 K+ currents and increased glucose-stimulated Ca2+ influx. Importantly, iOPN did not affect the function of other K2P channels or alter Ca2+ influx into TALK-1 deficient β-cells. These results reveal the first protein interactions with the TALK-1 channel and found that an interaction with iOPN increased

  4. Visual short-term memory: activity supporting encoding and maintenance in retinotopic visual cortex.

    Science.gov (United States)

    Sneve, Markus H; Alnæs, Dag; Endestad, Tor; Greenlee, Mark W; Magnussen, Svein

    2012-10-15

    Recent studies have demonstrated that retinotopic cortex maintains information about visual stimuli during retention intervals. However, the process by which transient stimulus-evoked sensory responses are transformed into enduring memory representations is unknown. Here, using fMRI and short-term visual memory tasks optimized for univariate and multivariate analysis approaches, we report differential involvement of human retinotopic areas during memory encoding of the low-level visual feature orientation. All visual areas show weaker responses when memory encoding processes are interrupted, possibly due to effects in orientation-sensitive primary visual cortex (V1) propagating across extrastriate areas. Furthermore, intermediate areas in both dorsal (V3a/b) and ventral (LO1/2) streams are significantly more active during memory encoding compared with non-memory (active and passive) processing of the same stimulus material. These effects in intermediate visual cortex are also observed during memory encoding of a different stimulus feature (spatial frequency), suggesting that these areas are involved in encoding processes on a higher level of representation. Using pattern-classification techniques to probe the representational content in visual cortex during delay periods, we further demonstrate that simply initiating memory encoding is not sufficient to produce long-lasting memory traces. Rather, active maintenance appears to underlie the observed memory-specific patterns of information in retinotopic cortex. Copyright © 2012 Elsevier Inc. All rights reserved.

  5. CAMKII activation is not required for maintenance of learning-induced enhancement of neuronal excitability.

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    Ori Liraz

    Full Text Available Pyramidal neurons in the piriform cortex from olfactory-discrimination trained rats show enhanced intrinsic neuronal excitability that lasts for several days after learning. Such enhanced intrinsic excitability is mediated by long-term reduction in the post-burst after-hyperpolarization (AHP which is generated by repetitive spike firing. AHP reduction is due to decreased conductance of a calcium-dependent potassium current, the sI(AHP. We have previously shown that learning-induced AHP reduction is maintained by persistent protein kinase C (PKC and extracellular regulated kinase (ERK activation. However, the molecular machinery underlying this long-lasting modulation of intrinsic excitability is yet to be fully described. Here we examine whether the CaMKII, which is known to be crucial in learning, memory and synaptic plasticity processes, is instrumental for the maintenance of learning-induced AHP reduction. KN93, that selectively blocks CaMKII autophosphorylation at Thr286, reduced the AHP in neurons from trained and control rat to the same extent. Consequently, the differences in AHP amplitude and neuronal adaptation between neurons from trained rats and controls remained. Accordingly, the level of activated CaMKII was similar in pirifrom cortex samples taken form trained and control rats. Our data show that although CaMKII modulates the amplitude of AHP of pyramidal neurons in the piriform cortex, its activation is not required for maintaining learning-induced enhancement of neuronal excitability.

  6. Effects of functional physical activity on the maintenance of motor function in Alzheimer's disease

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    Laís Fajersztajn

    Full Text Available Abstract It is widely known that older adults, even frail individuals, can improve their physical function using appropriately targeted exercise. Nevertheless, older adults with Alzheimer's disease (AD have been excluded from the majority of studies on exercise. The functional-task physical activity program is based on activities of daily living, and may be suited for elderly people with AD because it focuses on the maintenance and stimulation of preserved abilities. In addition, session costs are substantially reduced by adopting a group approach. Furthermore, the group approach may improve the social interaction of the demented patient. Objectives: To determine whether a functional-task physical activity program in groups can maintain motor function in elderly with AD. Methods: 10 elderly diagnosed with mild or moderate AD were assigned into one of two groups: subjects with and without intervention. The intervention consisted of a 12-week function-task physical activity program in groups. Measurements: activities of daily living (Katz and Lawton & Brody questionnaires, mobility (Timed Up and Go Test, Timed Up and Go manual Test and Timed Up and Go Cognitive Test, cognition (Mini-Mental State Examination, behavioral disturbances (Neuropsychiatric Inventory I-brief and functional balance (Berg Balance Scale. Results: A statistically significant difference between the two groups was found regarding the functional balance mean change measured by Berg scale score (p=0.046. A significant improvement of 1.60 points (95%CI[0.22;2.98] was observed in the intervention group on this scale, while the non-intervention group showed -0.40 points (95%CI[-1.78;0.98], no change. Conclusions: It is possible to treat mild and moderate Alzheimer's patients using a group approach. The functional task physical activity program was efficient in functional balance improvement and also appeared to prevent mobility decline.

  7. Channel-forming activities in the glycosomal fraction from the bloodstream form of Trypanosoma brucei.

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    Melisa Gualdron-López

    Full Text Available BACKGROUND: Glycosomes are a specialized form of peroxisomes (microbodies present in unicellular eukaryotes that belong to the Kinetoplastea order, such as Trypanosoma and Leishmania species, parasitic protists causing severe diseases of livestock and humans in subtropical and tropical countries. The organelles harbour most enzymes of the glycolytic pathway that is responsible for substrate-level ATP production in the cell. Glycolysis is essential for bloodstream-form Trypanosoma brucei and enzymes comprising this pathway have been validated as drug targets. Glycosomes are surrounded by a single membrane. How glycolytic metabolites are transported across the glycosomal membrane is unclear. METHODS/PRINCIPAL FINDINGS: We hypothesized that glycosomal membrane, similarly to membranes of yeast and mammalian peroxisomes, contains channel-forming proteins involved in the selective transfer of metabolites. To verify this prediction, we isolated a glycosomal fraction from bloodstream-form T. brucei and reconstituted solubilized membrane proteins into planar lipid bilayers. The electrophysiological characteristics of the channels were studied using multiple channel recording and single channel analysis. Three main channel-forming activities were detected with current amplitudes 70-80 pA, 20-25 pA, and 8-11 pA, respectively (holding potential +10 mV and 3.0 M KCl as an electrolyte. All channels were in fully open state in a range of voltages ±150 mV and showed no sub-conductance transitions. The channel with current amplitude 20-25 pA is anion-selective (P(K+/P(Cl-∼0.31, while the other two types of channels are slightly selective for cations (P(K+/P(Cl- ratios ∼1.15 and ∼1.27 for the high- and low-conductance channels, respectively. The anion-selective channel showed an intrinsic current rectification that may suggest a functional asymmetry of the channel's pore. CONCLUSIONS/SIGNIFICANCE: These results indicate that the membrane of glycosomes

  8. Effects of the small molecule HERG activator NS1643 on Kv11.3 channels.

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    Arne Bilet

    Full Text Available NS1643 is one of the small molecule HERG (Kv11.1 channel activators and has also been found to increase erg2 (Kv11.2 currents. We now investigated whether NS1643 is also able to act as an activator of Kv11.3 (erg3 channels expressed in CHO cells. Activation of rat Kv11.3 current occurred in a dose-dependent manner and maximal current increasing effects were obtained with 10 µM NS1643. At this concentration, steady-state outward current increased by about 80% and the current increase was associated with a significant shift in the voltage dependence of activation to more negative potentials by about 15 mV. In addition, activation kinetics were accelerated, whereas deactivation was slowed. There was no significant effect on the kinetics of inactivation and recovery from inactivation. The strong current-activating agonistic effect of NS1643 did not result from a shift in the voltage dependence of Kv11.3 channel inactivation and was independent from external Na(+ or Ca(2+. At the higher concentration of 20 µM, NS1643 induced clearly less current increase. The left shift in the voltage dependence of activation reversed and the voltage sensitivity of activation dramatically decreased along with a slowing of Kv11.3 channel activation. These data show that, in comparison to other Kv11 family members, NS1643 exerts distinct effects on Kv11.3 channels with especially pronounced partial antagonistic effects at higher concentration.

  9. SENSITIVE EFFECTS OF POTASSIUM AND CALCIUM CHANNEL BLOCKING AND ATP-SENSITIVE POTASSIUM CHANNEL ACTIVATORS ON SEMINAL VESICLE SMOOTH MUSCLE CONTRACTIONS

    Directory of Open Access Journals (Sweden)

    H SADRAEI

    2000-12-01

    Full Text Available Background. Seminal vesicle smooth muscle contraction is mediated through sympathetic and parasympathetic neurons activity. Although seminal vesicle plays an important role in male fertility, but little attention is given to mechanism involved in contraction of this organ.
    Methods. In this study effects of drugs which activate ATP - sensitive K channels and blockers of K and Ca channels were examined on contraction of guinea - pig isolated seminal vesicle due to electrical filled stimulation (EFS, noradrenaline, carbachol and KCI.
    Results. The K channel blocker tetraethyl ammonium potentate the EFS responses at all frequencies, while, the ATP - sensitive K channel inhibitor glibenclamide and the K channel opener levcromakalim, diazoxide, minoxidil and Ca channel blocker nifedipine all had relaxant effect on guinea - pig seminal vesicle.
    Discussion. This study indicate that activities of K and Ca channels is important in regulation of seminal vesicle contraction due to nerve stimulation, noradrenaline or carbachol.

  10. Intracellular long-chain acyl CoAs activate TRPV1 channels.

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    Yi Yu

    Full Text Available TRPV1 channels are an important class of membrane proteins that play an integral role in the regulation of intracellular cations such as calcium in many different tissue types. The anionic phospholipid phosphatidylinositol 4,5-bisphosphate (PIP2 is a known positive modulator of TRPV1 channels and the negatively charged phosphate groups interact with several basic amino acid residues in the proximal C-terminal TRP domain of the TRPV1 channel. We and other groups have shown that physiological sub-micromolar levels of long-chain acyl CoAs (LC-CoAs, another ubiquitous anionic lipid, can also act as positive modulators of ion channels and exchangers. Therefore, we investigated whether TRPV1 channel activity is similarly regulated by LC-CoAs. Our results show that LC-CoAs are potent activators of the TRPV1 channel and interact with the same PIP2-binding residues in TRPV1. In contrast to PIP2, LC-CoA modulation of TRPV1 is independent of Ca2+i, acting in an acyl side-chain saturation and chain-length dependent manner. Elevation of LC-CoAs in intact Jurkat T-cells leads to significant increases in agonist-induced Ca2+i levels. Our novel findings indicate that LC-CoAs represent a new fundamental mechanism for regulation of TRPV1 channel activity that may play a role in diverse cell types under physiological and pathophysiological conditions that alter fatty acid transport and metabolism such as obesity and diabetes.

  11. A chimeric prokaryotic pentameric ligand–gated channel reveals distinct pathways of activation

    Science.gov (United States)

    Schmandt, Nicolaus; Velisetty, Phanindra; Chalamalasetti, Sreevatsa V.; Stein, Richard A.; Bonner, Ross; Talley, Lauren; Parker, Mark D.; Mchaourab, Hassane S.; Yee, Vivien C.; Lodowski, David T.

    2015-01-01

    Recent high resolution structures of several pentameric ligand–gated ion channels have provided unprecedented details of their molecular architecture. However, the conformational dynamics and structural rearrangements that underlie gating and allosteric modulation remain poorly understood. We used a combination of electrophysiology, double electron–electron resonance (DEER) spectroscopy, and x-ray crystallography to investigate activation mechanisms in a novel functional chimera with the extracellular domain (ECD) of amine-gated Erwinia chrysanthemi ligand–gated ion channel, which is activated by primary amines, and the transmembrane domain of Gloeobacter violaceus ligand–gated ion channel, which is activated by protons. We found that the chimera was independently gated by primary amines and by protons. The crystal structure of the chimera in its resting state, at pH 7.0 and in the absence of primary amines, revealed a closed-pore conformation and an ECD that is twisted with respect to the transmembrane region. Amine- and pH-induced conformational changes measured by DEER spectroscopy showed that the chimera exhibits a dual mode of gating that preserves the distinct conformational changes of the parent channels. Collectively, our findings shed light on both conserved and divergent features of gating mechanisms in this class of channels, and will facilitate the design of better allosteric modulators. PMID:26415570

  12. Leisure-time activity is an important determinant of long-term weight maintenance after weight loss in the Sibutramine Trial on Obesity Reduction and Maintenance (STORM trial)

    DEFF Research Database (Denmark)

    van Baak, M. A.; van Mil, E.; Astrup, A.

    2003-01-01

    The success rate of long-term maintenance of weight loss in obese patients is usually low. To improve the success rate, determinants of long-term weight maintenance must be identified.......The success rate of long-term maintenance of weight loss in obese patients is usually low. To improve the success rate, determinants of long-term weight maintenance must be identified....

  13. Spontaneous motor activity during the development and maintenance of diet-induced obesity in the rat.

    Science.gov (United States)

    Levin, B E

    1991-09-01

    More than 80% of most daily spontaneous activities (assessed in an Omnitech activity monitor) occurred during the last hour of light and 12 h of the dark phase in 8 chow-fed male Sprague-Dawley rats. Thirty additional rats were, therefore, monitored over this 13-h period to assess the relationship of activity to the development and maintenance of diet-induced obesity (DIO) on a diet high in energy, fat and sucrose (CM diet). Nine of 20 rats became obese after 3 months on the CM diet, with 71% greater weight gain than 10 chow-fed controls. Eleven of 20 rats were diet resistant (DR), gaining the same amount of weight as chow-fed rats. Neither initial activity levels nor initial body weights on chow (Period I) differed significantly across retrospectively identified groups. After 3 months on CM diet or chow (Period II), as well as after an additional 3 months after CM diet-fed rats returned to chow (Period III), there were significant inverse correlations (r = -.606 to -.370) between body weight at the time of testing and various measures of movement in the horizontal plane. There was no relationship to dietary content nor consistent correlations of body weight or diet group to vertical movements, an indirect measure of ingestive behavior. Patterns of time spent in the vertical position were significantly different for DIO vs. DR rats in Period III, however. Thus, differences in food intake and metabolic efficiency, rather than differences in nocturnal activity, are probably responsible for the greater weight gain in DIO-prone rats placed on CM diet.(ABSTRACT TRUNCATED AT 250 WORDS)

  14. Inhibition of parathyroid hormone release by maitotoxin, a calcium channel activator

    International Nuclear Information System (INIS)

    Fitzpatrick, L.A.; Yasumoto, T.; Aurbach, G.D.

    1989-01-01

    Maitotoxin, a toxin derived from a marine dinoflagellate, is a potent activator of voltage-sensitive calcium channels. To further test the hypothesis that inhibition of PTH secretion by calcium is mediated via a calcium channel we studied the effect of maitotoxin on dispersed bovine parathyroid cells. Maitotoxin inhibited PTH release in a dose-dependent fashion, and inhibition was maximal at 1 ng/ml. Chelation of extracellular calcium by EGTA blocked the inhibition of PTH by maitotoxin. Maitotoxin enhanced the effects of the dihydropyridine calcium channel agonist (+)202-791 and increased the rate of radiocalcium uptake in parathyroid cells. Pertussis toxin, which ADP-ribosylates and inactivates a guanine nucleotide regulatory protein that interacts with calcium channels in the parathyroid cell, did not affect the inhibition of PTH secretion by maitotoxin. Maitotoxin, by its action on calcium channels allows entry of extracellular calcium and inhibits PTH release. Our results suggest that calcium channels are involved in the release of PTH. Inhibition of PTH release by maitotoxin is not sensitive to pertussis toxin, suggesting that maitotoxin may act distal to the site interacting with a guanine nucleotide regulatory protein, or maitotoxin could interact with other ions or second messengers to inhibit PTH release

  15. Anoctamin Calcium-Activated Chloride Channels May Modulate Inhibitory Transmission in the Cerebellar Cortex.

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    Weiping Zhang

    Full Text Available Calcium-activated chloride channels of the anoctamin (alias TMEM16 protein family fulfill critical functions in epithelial fluid transport, smooth muscle contraction and sensory signal processing. Little is known, however, about their contribution to information processing in the central nervous system. Here we examined the recent finding that a calcium-dependent chloride conductance impacts on GABAergic synaptic inhibition in Purkinje cells of the cerebellum. We asked whether anoctamin channels may underlie this chloride conductance. We identified two anoctamin channel proteins, ANO1 and ANO2, in the cerebellar cortex. ANO1 was expressed in inhibitory interneurons of the molecular layer and the granule cell layer. Both channels were expressed in Purkinje cells but, while ANO1 appeared to be retained in the cell body, ANO2 was targeted to the dendritic tree. Functional studies confirmed that ANO2 was involved in a calcium-dependent mode of ionic plasticity that reduces the efficacy of GABAergic synapses. ANO2 channels attenuated GABAergic transmission by increasing the postsynaptic chloride concentration, hence reducing the driving force for chloride influx. Our data suggest that ANO2 channels are involved in a Ca2+-dependent regulation of synaptic weight in GABAergic inhibition. Thus, in balance with the chloride extrusion mechanism via the co-transporter KCC2, ANO2 appears to regulate ionic plasticity in the cerebellum.

  16. Deep brain stimulation of the subthalamic nucleus alters frontal activity during spatial working memory maintenance of patients with Parkinson's disease.

    Science.gov (United States)

    Mayer, Jutta S; Neimat, Joseph; Folley, Bradley S; Bourne, Sarah K; Konrad, Peter E; Charles, David; Park, Sohee

    2016-08-01

    Deep brain stimulation (DBS) of the subthalamic nucleus (STN) improves the motor symptoms of Parkinson's disease (PD). The STN may represent an important relay station not only in the motor but also the associative cortico-striato-thalamocortical pathway. Therefore, STN stimulation may alter cognitive functions, such as working memory (WM). We examined cortical effects of STN-DBS on WM in early PD patients using functional near-infrared spectroscopy. The effects of dopaminergic medication on WM were also examined. Lateral frontal activity during WM maintenance was greater when patients were taking dopaminergic medication. STN-DBS led to a trend-level worsening of WM performance, accompanied by increased lateral frontal activity during WM maintenance. These findings suggest that STN-DBS in PD might lead to functional modifications of the basal ganglia-thalamocortical pathway during WM maintenance.

  17. Large-conductance Ca2+-activated K+ channel β1-subunit knockout mice are not hypertensive

    Science.gov (United States)

    Garver, Hannah; Galligan, James J.; Fink, Gregory D.

    2011-01-01

    Large-conductance Ca2+-activated K+ (BK) channels are composed of pore-forming α-subunits and accessory β1-subunits that modulate Ca2+ sensitivity. BK channels regulate arterial myogenic tone and renal Na+ clearance/K+ reabsorption. Previous studies using indirect or short-term blood pressure measurements found that BK channel β1-subunit knockout (BK β1-KO) mice were hypertensive. We evaluated 24-h mean arterial pressure (MAP) and heart rate in BK β1-KO mice using radiotelemetry. BK β1-KO mice did not have a higher 24-h average MAP when compared with wild-type (WT) mice, although MAP was ∼10 mmHg higher at night. The dose-dependent peak declines in MAP by nifedipine were only slightly larger in BK β1-KO mice. In BK β1-KO mice, giving 1% NaCl to mice to drink for 7 days caused a transient (5 days) elevation of MAP (∼5 mmHg); MAP returned to pre-saline levels by day 6. BK β1-KO mesenteric arteries in vitro demonstrated diminished contractile responses to paxilline, increased reactivity to Bay K 8644 and norepinephrine (NE), and maintained relaxation to isoproterenol. Paxilline and Bay K 8644 did not constrict WT or BK β1-KO mesenteric veins (MV). BK β1-subunits are not expressed in MV. The results indicate that BK β1-KO mice are not hypertensive on normal or high-salt intake. BK channel deficiency increases arterial reactivity to NE and L-type Ca2+ channel function in vitro, but the L-type Ca2+ channel modulation of MAP is not altered in BK β1-KO mice. BK and L-type Ca2+ channels do not modulate murine venous tone. It appears that selective loss of BK channel function in arteries only is not sufficient to cause sustained hypertension. PMID:21131476

  18. Ion channel signaling influences cellular proliferation and phagocyte activity during axolotl tail regeneration.

    Science.gov (United States)

    Franklin, Brandon M; Voss, S Randal; Osborn, Jeffrey L

    2017-08-01

    Little is known about the potential for ion channels to regulate cellular behaviors during tissue regeneration. Here, we utilized an amphibian tail regeneration assay coupled with a chemical genetic screen to identify ion channel antagonists that altered critical cellular processes during regeneration. Inhibition of multiple ion channels either partially (anoctamin1/Tmem16a, anoctamin2/Tmem16b, K V 2.1, K V 2.2, L-type Ca V channels and H/K ATPases) or completely (GlyR, GABA A R, K V 1.5 and SERCA pumps) inhibited tail regeneration. Partial inhibition of tail regeneration by blocking the calcium activated chloride channels, anoctamin1&2, was associated with a reduction of cellular proliferation in tail muscle and mesenchymal regions. Inhibition of anoctamin 1/2 also altered the post-amputation transcriptional response of p44/42 MAPK signaling pathway genes, including decreased expression of erk1/erk2. We also found that complete inhibition via voltage gated K + channel blockade was associated with diminished phagocyte recruitment to the amputation site. The identification of H + pumps as required for axolotl tail regeneration supports findings in Xenopus and Planaria models, and more generally, the conservation of ion channels as regulators of tissue regeneration. This study provides a preliminary framework for an in-depth investigation of the mechanistic role of ion channels and their potential involvement in regulating cellular proliferation and other processes essential to wound healing, appendage regeneration, and tissue repair. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Apparent intermediate K conductance channel hyposmotic activation in human lens epithelial cells.

    Science.gov (United States)

    Lauf, Peter K; Misri, Sandeep; Chimote, Ameet A; Adragna, Norma C

    2008-03-01

    This study explores the nature of K fluxes in human lens epithelial cells (LECs) in hyposmotic solutions. Total ion fluxes, Na-K pump, Cl-dependent Na-K-2Cl (NKCC), K-Cl (KCC) cotransport, and K channels were determined by 85Rb uptake and cell K (Kc) by atomic absorption spectrophotometry, and cell water gravimetrically after exposure to ouabain +/- bumetanide (Na-K pump and NKCC inhibitors), and ion channel inhibitors in varying osmolalities with Na, K, or methyl-d-glucamine and Cl, sulfamate, or nitrate. Reverse transcriptase polymerase chain reaction (RT-PCR), Western blot analyses, and immunochemistry were also performed. In isosmotic (300 mosM) media approximately 90% of the total Rb influx occurred through the Na-K pump and NKCC and approximately 10% through KCC and a residual leak. Hyposmotic media (150 mosM) decreased K(c) by a 16-fold higher K permeability and cell water, but failed to inactivate NKCC and activate KCC. Sucrose replacement or extracellular K to >57 mM, but not Rb or Cs, in hyposmotic media prevented Kc and water loss. Rb influx equaled Kc loss, both blocked by clotrimazole (IC50 approximately 25 microM) and partially by 1-[(2-chlorophenyl) diphenylmethyl]-1H-pyrazole (TRAM-34) inhibitors of the IK channel KCa3.1 but not by other K channel or connexin hemichannel blockers. Of several anion channel blockers (dihydro-indenyl)oxy]alkanoic acid (DIOA), 4-2(butyl-6,7-dichloro-2-cyclopentylindan-1-on-5-yl)oxybutyric acid (DCPIB), and phloretin totally or partially inhibited Kc loss and Rb influx, respectively. RT-PCR and immunochemistry confirmed the presence of KCa3.1 channels, aside of the KCC1, KCC2, KCC3 and KCC4 isoforms. Apparently, IK channels, possibly in parallel with volume-sensitive outwardly rectifying Cl channels, effect regulatory volume decrease in LECs.

  20. Ethanol affects network activity in cultured rat hippocampus: mediation by potassium channels.

    Directory of Open Access Journals (Sweden)

    Eduard Korkotian

    Full Text Available The effects of ethanol on neuronal network activity were studied in dissociated cultures of rat hippocampus. Exposure to low (0.25-0.5% ethanol concentrations caused an increase in synchronized network spikes, and a decrease in the duration of individual spikes. Ethanol also caused an increase in rate of miniature spontaneous excitatory postsynaptic currents. Higher concentrations of ethanol eliminated network spikes. These effects were reversible upon wash. The effects of the high, but not the low ethanol were blocked by the GABA antagonist bicuculline. The enhancing action of low ethanol was blocked by apamin, an SK potassium channel antagonist, and mimicked by 1-EBIO, an SK channel opener. It is proposed that in cultured hippocampal networks low concentration of ethanol is associated with SK channel activity, rather than the GABAergic receptor.

  1. Evolutionarily conserved mechanisms for the selection and maintenance of behavioural activity.

    Science.gov (United States)

    Fiore, Vincenzo G; Dolan, Raymond J; Strausfeld, Nicholas J; Hirth, Frank

    2015-12-19

    Survival and reproduction entail the selection of adaptive behavioural repertoires. This selection manifests as phylogenetically acquired activities that depend on evolved nervous system circuitries. Lorenz and Tinbergen already postulated that heritable behaviours and their reliable performance are specified by genetically determined programs. Here we compare the functional anatomy of the insect central complex and vertebrate basal ganglia to illustrate their role in mediating selection and maintenance of adaptive behaviours. Comparative analyses reveal that central complex and basal ganglia circuitries share comparable lineage relationships within clusters of functionally integrated neurons. These clusters are specified by genetic mechanisms that link birth time and order to their neuronal identities and functions. Their subsequent connections and associated functions are characterized by similar mechanisms that implement dimensionality reduction and transition through attractor states, whereby spatially organized parallel-projecting loops integrate and convey sensorimotor representations that select and maintain behavioural activity. In both taxa, these neural systems are modulated by dopamine signalling that also mediates memory-like processes. The multiplicity of similarities between central complex and basal ganglia suggests evolutionarily conserved computational mechanisms for action selection. We speculate that these may have originated from ancestral ground pattern circuitries present in the brain of the last common ancestor of insects and vertebrates. © 2015 The Authors.

  2. Mechanosensitive channels are activated by stress in the actin stress fibres, and could be involved in gravity sensing in plants.

    Science.gov (United States)

    Tatsumi, H; Furuichi, T; Nakano, M; Toyota, M; Hayakawa, K; Sokabe, M; Iida, H

    2014-01-01

    Mechanosensitive (MS) channels are expressed in a variety of cells. The molecular and biophysical mechanism involved in the regulation of MS channel activities is a central interest in basic biology. MS channels are thought to play crucial roles in gravity sensing in plant cells. To date, two mechanisms have been proposed for MS channel activation. One is that tension development in the lipid bilayer directly activates MS channels. The second mechanism proposes that the cytoskeleton is involved in the channel activation, because MS channel activities are modulated by pharmacological treatments that affect the cytoskeleton. We tested whether tension in the cytoskeleton activates MS channels. Mammalian endothelial cells were microinjected with phalloidin-conjugated beads, which bound to stress fibres, and a traction force to the actin cytoskeleton was applied by dragging the beads with optical tweezers. MS channels were activated when the force was applied, demonstrating that a sub-pN force to the actin filaments activates a single MS channel. Plants may use a similar molecular mechanism in gravity sensing, since the cytoplasmic Ca(2+) concentration increase induced by changes in the gravity vector was attenuated by potential MS channel inhibitors, and by actin-disrupting drugs. These results support the idea that the tension increase in actin filaments by gravity-dependent sedimentation of amyloplasts activates MS Ca(2+) -permeable channels, which can be the molecular mechanism of a Ca(2+) concentration increase through gravistimulation. We review recent progress in the study of tension sensing by actin filaments and MS channels using advanced biophysical methods, and discuss their possible roles in gravisensing. © 2013 German Botanical Society and The Royal Botanical Society of the Netherlands.

  3. Activation of the Ca2+-sensing receptors increases currents through inward rectifier K+ channels via activation of phosphatidylinositol 4-kinase

    OpenAIRE

    Liu, Chung-Hung; Chang, Hsueh-Kai; Lee, Sue-Ping; Shieh, Ru-Chi

    2016-01-01

    Inward rectifier K+ channels are important for maintaining normal electrical function in many cell types. The proper function of these channels requires the presence of membrane phosphoinositide 4,5-bisphosphate (PIP2). Stimulation of the Ca2+-sensing receptor CaR, a pleiotropic G protein-coupled receptor, activates both Gq/11, which decreases PIP2, and phosphatidylinositol 4-kinase (PI-4-K), which, conversely, increases PIP2. How membrane PIP2 levels are regulated by CaR activation and wheth...

  4. Melatonin mediates vasodilation through both direct and indirect activation of BKCa channels.

    Science.gov (United States)

    Zhao, T; Zhang, H; Jin, C; Qiu, F; Wu, Y; Shi, L

    2017-10-01

    Melatonin, synthesized primarily by the pineal gland, is a neuroendocrine hormone with high membrane permeability. The vascular effects of melatonin, including vasoconstriction and vasodilation, have been demonstrated in numerous studies. However, the mechanisms underlying these effects are not fully understood. Large-conductance Ca 2+ -activated K + (BK Ca ) channels are expressed broadly on smooth muscle cells and play an important role in vascular tone regulation. This study explored the mechanisms of myocyte BK Ca channels and endothelial factors underlying the action of melatonin on the mesenteric arteries (MAs). Vascular contractility and patch-clamp studies were performed on myocytes of MAs from Wistar rats. Melatonin induced significant vasodilation on MAs. In the presence of N ω -nitro-l-arginine methyl ester (l-NAME), a potent endothelial oxide synthase (eNOS) inhibitor, melatonin elicited concentration-dependent relaxation, with lowered pIC 50 The effect of melatonin was significantly attenuated in the presence of BK Ca channel blocker iberiotoxin or MT1/MT2 receptor antagonist luzindole in both (+) l-NAME and (-) l-NAME groups. In the (+) l-NAME group, iberiotoxin caused a parallel rightward shift of the melatonin concentration-relaxation curve, with pIC 50 lower than that of luzindole. Both inside-out and cell-attached patch-clamp recordings showed that melatonin significantly increased the open probability, mean open time and voltage sensitivity of BK Ca channels. In a cell-attached patch-clamp configuration, the melatonin-induced enhancement of BK Ca channel activity was significantly suppressed by luzindole. These findings indicate that in addition to the activation of eNOS, melatonin-induced vasorelaxation of MAs is partially attributable to its direct (passing through the cell membrane) and indirect (via MT1/MT2 receptors) activation of the BK Ca channels on mesenteric arterial myocytes. © 2017 Society for Endocrinology.

  5. Active Sites of Spinoxin, a Potassium Channel Scorpion Toxin, Elucidated by Systematic Alanine Scanning.

    Science.gov (United States)

    Peigneur, Steve; Yamaguchi, Yoko; Kawano, Chihiro; Nose, Takeru; Nirthanan, Selvanayagam; Gopalakrishnakone, Ponnampalam; Tytgat, Jan; Sato, Kazuki

    2016-05-31

    Peptide toxins from scorpion venoms constitute the largest group of toxins that target the voltage-gated potassium channel (Kv). Spinoxin (SPX) isolated from the venom of scorpion Heterometrus spinifer is a 34-residue peptide neurotoxin cross-linked by four disulfide bridges. SPX is a potent inhibitor of Kv1.3 potassium channels (IC50 = 63 nM), which are considered to be valid molecular targets in the diagnostics and therapy of various autoimmune disorders and cancers. Here we synthesized 25 analogues of SPX and analyzed the role of each amino acid in SPX using alanine scanning to study its structure-function relationships. All synthetic analogues showed similar disulfide bond pairings and secondary structures as native SPX. Alanine replacements at Lys(23), Asn(26), and Lys(30) resulted in loss of activity against Kv1.3 potassium channels, whereas replacements at Arg(7), Met(14), Lys(27), and Tyr(32) also largely reduced inhibitory activity. These results suggest that the side chains of these amino acids in SPX play an important role in its interaction with Kv1.3 channels. In particular, Lys(23) appears to be a key residue that underpins Kv1.3 channel inhibition. Of these seven amino acid residues, four are basic amino acids, suggesting that the positive electrostatic potential on the surface of SPX is likely required for high affinity interaction with Kv1.3 channels. This study provides insight into the structure-function relationships of SPX with implications for the rational design of new lead compounds targeting potassium channels with high potency.

  6. Low voltage-activated calcium channels gate transmitter release at the dorsal root ganglion sandwich synapse.

    Science.gov (United States)

    Rozanski, Gabriela M; Nath, Arup R; Adams, Michael E; Stanley, Elise F

    2013-11-15

    A subpopulation of dorsal root ganglion (DRG) neurons are intimately attached in pairs and separated solely by thin satellite glial cell membrane septa. Stimulation of one neuron leads to transglial activation of its pair by a bi-, purinergic/glutamatergic synaptic pathway, a transmission mechanism that we term sandwich synapse (SS) transmission. Release of ATP from the stimulated neuron can be attributed to a classical mechanism involving Ca(2+) entry via voltage-gated calcium channels (CaV) but via an unknown channel type. Specific blockers and toxins ruled out CaV1, 2.1 and 2.2. Transmission was, however, blocked by a moderate depolarization (-50 mV) or low-concentration Ni(2+) (0.1 mM). Transmission persisted using a voltage pulse to -40 mV from a holding potential of -80 mV, confirming the involvement of a low voltage-activated channel type and limiting the candidate channel type to either CaV3.2 or a subpopulation of inactivation- and Ni(2+)-sensitive CaV2.3 channels. Resistance of the neuron calcium current and SS transmission to SNX482 argue against the latter. Hence, we conclude that inter-somatic transmission at the DRG SS is gated by CaV3.2 type calcium channels. The use of CaV3 family channels to gate transmission has important implications for the biological function of the DRG SS as information transfer would be predicted to occur not only in response to action potentials but also to sub-threshold membrane voltage oscillations. Thus, the SS synapse may serve as a homeostatic signalling mechanism between select neurons in the DRG and could play a role in abnormal sensation such as neuropathic pain.

  7. A comprehensive search for calcium binding sites critical for TMEM16A calcium-activated chloride channel activity

    Science.gov (United States)

    Tien, Jason; Peters, Christian J; Wong, Xiu Ming; Cheng, Tong; Jan, Yuh Nung; Jan, Lily Yeh; Yang, Huanghe

    2014-01-01

    TMEM16A forms calcium-activated chloride channels (CaCCs) that regulate physiological processes such as the secretions of airway epithelia and exocrine glands, the contraction of smooth muscles, and the excitability of neurons. Notwithstanding intense interest in the mechanism behind TMEM16A-CaCC calcium-dependent gating, comprehensive surveys to identify and characterize potential calcium sensors of this channel are still lacking. By aligning distantly related calcium-activated ion channels in the TMEM16 family and conducting systematic mutagenesis of all conserved acidic residues thought to be exposed to the cytoplasm, we identify four acidic amino acids as putative calcium-binding residues. Alterations of the charge, polarity, and size of amino acid side chains at these sites alter the ability of different divalent cations to activate the channel. Furthermore, TMEM16A mutant channels containing double cysteine substitutions at these residues are sensitive to the redox potential of the internal solution, providing evidence for their physical proximity and solvent accessibility. DOI: http://dx.doi.org/10.7554/eLife.02772.001 PMID:24980701

  8. Status of reliability in determining SDDR for manual maintenance activities in ITER: Quality assessment of relevant activation cross sections involved

    International Nuclear Information System (INIS)

    Garcia, R.; Garcia, M.; Pampin, R.; Sanz, J.

    2016-01-01

    Highlights: • Feasibility of manual maintenance activities in ITER port cell and port interspace. • Activation of relevant materials and components placed in the current ITER model. • Dominant radionuclides and pathways for shutdown dose rate in ITER. • Quality analysis of typically used EAF and TENDL activation libraries is performed. • EAF performance found as trustworthy with slight recommended improvements. - Abstract: This paper assesses the quality of the EAF-2007 and 2010 activation cross sections for relevant reactions in the determination of the Shutdown Dose Rate (SDDR) in the Port Cell (PC) and Port Interspace (PI) areas of ITER. For each of relevant ITER materials, dominant radionuclides responsible of SDDR and their production pathways are listed. This information comes from a review of the recent reports/papers about SDDR in ITER and own calculations. A total of 26 relevant pathways are found. The quality of these cross sections pathways is assessed following EAF validation procedure, and for those found as not validated last TENDL library versions have been investigated in order to check possible improvements when compared to EAF. The use of EAF libraries is found as trustworthy and it is recommended for the prediction of SDDR in the ITER PC and PI. However, 3 cross section reactions are considered for further improvement: Co59(n,2n)Co58, Cu63(n,g)Cu64 and Cr50(n,g)Cr51.

  9. Status of reliability in determining SDDR for manual maintenance activities in ITER: Quality assessment of relevant activation cross sections involved

    Energy Technology Data Exchange (ETDEWEB)

    Garcia, R., E-mail: rgarciam@ind.uned.es [UNED, Power Engineering Department, C/Juan del Rosal 12, 28040 Madrid (Spain); Garcia, M. [UNED, Power Engineering Department, C/Juan del Rosal 12, 28040 Madrid (Spain); Pampin, R. [F4E, Torres Diagonal Litoral B3, Barcelona (Spain); Sanz, J. [UNED, Power Engineering Department, C/Juan del Rosal 12, 28040 Madrid (Spain)

    2016-11-15

    Highlights: • Feasibility of manual maintenance activities in ITER port cell and port interspace. • Activation of relevant materials and components placed in the current ITER model. • Dominant radionuclides and pathways for shutdown dose rate in ITER. • Quality analysis of typically used EAF and TENDL activation libraries is performed. • EAF performance found as trustworthy with slight recommended improvements. - Abstract: This paper assesses the quality of the EAF-2007 and 2010 activation cross sections for relevant reactions in the determination of the Shutdown Dose Rate (SDDR) in the Port Cell (PC) and Port Interspace (PI) areas of ITER. For each of relevant ITER materials, dominant radionuclides responsible of SDDR and their production pathways are listed. This information comes from a review of the recent reports/papers about SDDR in ITER and own calculations. A total of 26 relevant pathways are found. The quality of these cross sections pathways is assessed following EAF validation procedure, and for those found as not validated last TENDL library versions have been investigated in order to check possible improvements when compared to EAF. The use of EAF libraries is found as trustworthy and it is recommended for the prediction of SDDR in the ITER PC and PI. However, 3 cross section reactions are considered for further improvement: Co59(n,2n)Co58, Cu63(n,g)Cu64 and Cr50(n,g)Cr51.

  10. Active Dendrites and Differential Distribution of Calcium Channels Enable Functional Compartmentalization of Golgi Cells.

    Science.gov (United States)

    Rudolph, Stephanie; Hull, Court; Regehr, Wade G

    2015-11-25

    Interneurons are essential to controlling excitability, timing, and synaptic integration in neuronal networks. Golgi cells (GoCs) serve these roles at the input layer of the cerebellar cortex by releasing GABA to inhibit granule cells (grcs). GoCs are excited by mossy fibers (MFs) and grcs and provide feedforward and feedback inhibition to grcs. Here we investigate two important aspects of GoC physiology: the properties of GoC dendrites and the role of calcium signaling in regulating GoC spontaneous activity. Although GoC dendrites are extensive, previous studies concluded they are devoid of voltage-gated ion channels. Hence, the current view holds that somatic voltage signals decay passively within GoC dendrites, and grc synapses onto distal dendrites are not amplified and are therefore ineffective at firing GoCs because of strong passive attenuation. Using whole-cell recording and calcium imaging in rat slices, we find that dendritic voltage-gated sodium channels allow somatic action potentials to activate voltage-gated calcium channels (VGCCs) along the entire dendritic length, with R-type and T-type VGCCs preferentially located distally. We show that R- and T-type VGCCs located in the dendrites can boost distal synaptic inputs and promote burst firing. Active dendrites are thus critical to the regulation of GoC activity, and consequently, to the processing of input to the cerebellar cortex. In contrast, we find that N-type channels are preferentially located near the soma, and control the frequency and pattern of spontaneous firing through their close association with calcium-activated potassium (KCa) channels. Thus, VGCC types are differentially distributed and serve specialized functions within GoCs. Interneurons are essential to neural processing because they modulate excitability, timing, and synaptic integration within circuits. At the input layer of the cerebellar cortex, a single type of interneuron, the Golgi cell (GoC), carries these functions. The

  11. Voltage-gated potassium channels regulate calcium-dependent pathways involved in human T lymphocyte activation.

    Science.gov (United States)

    Lin, C S; Boltz, R C; Blake, J T; Nguyen, M; Talento, A; Fischer, P A; Springer, M S; Sigal, N H; Slaughter, R S; Garcia, M L

    1993-03-01

    The role that potassium channels play in human T lymphocyte activation has been investigated by using specific potassium channel probes. Charybdotoxin (ChTX), a blocker of small conductance Ca(2+)-activated potassium channels (PK,Ca) and voltage-gated potassium channels (PK,V) that are present in human T cells, inhibits the activation of these cells. ChTX blocks T cell activation induced by signals (e.g., anti-CD2, anti-CD3, ionomycin) that elicit a rise in intracellular calcium ([Ca2+]i) by preventing the elevation of [Ca2+]i in a dose-dependent manner. However, ChTX has no effect on the activation pathways (e.g., anti-CD28, interleukin 2 [IL-2]) that are independent of a rise in [Ca2+]i. In the former case, both proliferative response and lymphokine production (IL-2 and interferon gamma) are inhibited by ChTX. The inhibitory effect of ChTX can be demonstrated when added simultaneously, or up to 4 h after the addition of the stimulants. Since ChTX inhibits both PK,Ca and PK,V, we investigated which channel is responsible for these immunosuppressive effects with the use of two other peptides, noxiustoxin (NxTX) and margatoxin (MgTX), which are specific for PK,V. These studies demonstrate that, similar to ChTX, both NxTX and MgTX inhibit lymphokine production and the rise in [Ca2+]i. Taken together, these data provide evidence that blockade of PK,V affects the Ca(2+)-dependent pathways involved in T lymphocyte proliferation and lymphokine production by diminishing the rise in [Ca2+]i that occurs upon T cell activation.

  12. Effects on atrial fibrillation in aged hypertensive rats by Ca(2+)-activated K(+) channel inhibition

    DEFF Research Database (Denmark)

    Diness, Jonas Goldin; Skibsbye, Lasse; Jespersen, Thomas

    2011-01-01

    We have shown previously that inhibition of small conductance Ca(2+)-activated K(+) (SK) channels is antiarrhythmic in models of acutely induced atrial fibrillation (AF). These models, however, do not take into account that AF derives from a wide range of predisposing factors, the most prevalent ...

  13. Increased anion channel activity is an unavoidable event in ozone-induced programmed cell death.

    Directory of Open Access Journals (Sweden)

    Takashi Kadono

    Full Text Available BACKGROUND: Ozone is a major secondary air pollutant often reaching high concentrations in urban areas under strong daylight, high temperature and stagnant high-pressure systems. Ozone in the troposphere is a pollutant that is harmful to the plant. PRINCIPAL FINDINGS: By exposing cells to a strong pulse of ozonized air, an acute cell death was observed in suspension cells of Arabidopsis thaliana used as a model. We demonstrated that O(3 treatment induced the activation of a plasma membrane anion channel that is an early prerequisite of O(3-induced cell death in A. thaliana. Our data further suggest interplay of anion channel activation with well known plant responses to O(3, Ca(2+ influx and NADPH-oxidase generated reactive oxygen species (ROS in mediating the oxidative cell death. This interplay might be fuelled by several mechanisms in addition to the direct ROS generation by O(3; namely, H(2O(2 generation by salicylic and abscisic acids. Anion channel activation was also shown to promote the accumulation of transcripts encoding vacuolar processing enzymes, a family of proteases previously reported to contribute to the disruption of vacuole integrity observed during programmed cell death. SIGNIFICANCE: Collectively, our data indicate that anion efflux is an early key component of morphological and biochemical events leading to O(3-induced programmed cell death. Because ion channels and more specifically anion channels assume a crucial position in cells, an understanding about the underlying role(s for ion channels in the signalling pathway leading to programmed cell death is a subject that warrants future investigation.

  14. Ginseng gintonin activates the human cardiac delayed rectifier K+ channel: involvement of Ca2+/calmodulin binding sites.

    Science.gov (United States)

    Choi, Sun-Hye; Lee, Byung-Hwan; Kim, Hyeon-Joong; Jung, Seok-Won; Kim, Hyun-Sook; Shin, Ho-Chul; Lee, Jun-Hee; Kim, Hyoung-Chun; Rhim, Hyewhon; Hwang, Sung-Hee; Ha, Tal Soo; Kim, Hyun-Ji; Cho, Hana; Nah, Seung-Yeol

    2014-09-01

    Gintonin, a novel, ginseng-derived G protein-coupled lysophosphatidic acid (LPA) receptor ligand, elicits [Ca(2+)]i transients in neuronal and non-neuronal cells via pertussis toxin-sensitive and pertussis toxin-insensitive G proteins. The slowly activating delayed rectifier K(+) (I(Ks)) channel is a cardiac K(+) channel composed of KCNQ1 and KCNE1 subunits. The C terminus of the KCNQ1 channel protein has two calmodulin-binding sites that are involved in regulating I(Ks) channels. In this study, we investigated the molecular mechanisms of gintonin-mediated activation of human I(Ks) channel activity by expressing human I(Ks) channels in Xenopus oocytes. We found that gintonin enhances IKs channel currents in concentration- and voltage-dependent manners. The EC50 for the I(Ks) channel was 0.05 ± 0.01 μg/ml. Gintonin-mediated activation of the I(Ks) channels was blocked by an LPA1/3 receptor antagonist, an active phospholipase C inhibitor, an IP3 receptor antagonist, and the calcium chelator BAPTA. Gintonin-mediated activation of both the I(Ks) channel was also blocked by the calmodulin (CaM) blocker calmidazolium. Mutations in the KCNQ1 [Ca(2+)]i/CaM-binding IQ motif sites (S373P, W392R, or R539W)blocked the action of gintonin on I(Ks) channel. However, gintonin had no effect on hERG K(+) channel activity. These results show that gintonin-mediated enhancement of I(Ks) channel currents is achieved through binding of the [Ca(2+)]i/CaM complex to the C terminus of KCNQ1 subunit.

  15. Requirement for PLK1 kinase activity in the maintenance of a robust spindle assembly checkpoint

    Directory of Open Access Journals (Sweden)

    Aisling O'Connor

    2016-01-01

    Full Text Available During mitotic arrest induced by microtubule targeting drugs, the weakening of the spindle assembly checkpoint (SAC allows cells to progress through the cell cycle without chromosome segregation occurring. PLK1 kinase plays a major role in mitosis and emerging evidence indicates that PLK1 is also involved in establishing the checkpoint and maintaining SAC signalling. However, mechanistically, the role of PLK1 in the SAC is not fully understood, with several recent reports indicating that it can cooperate with either one of the major checkpoint kinases, Aurora B or MPS1. In this study, we assess the role of PLK1 in SAC maintenance. We find that in nocodazole-arrested U2OS cells, PLK1 activity is continuously required for maintaining Aurora B protein localisation and activity at kinetochores. Consistent with published data we find that upon PLK1 inhibition, phosphoThr3-H3, a marker of Haspin activity, is reduced. Intriguingly, Aurora B inhibition causes PLK1 to relocalise from kinetochores into fewer and much larger foci, possibly due to incomplete recruitment of outer kinetochore proteins. Importantly, PLK1 inhibition, together with partial inhibition of Aurora B, allows efficient SAC override to occur. This phenotype is more pronounced than the phenotype observed by combining the same PLK1 inhibitors with partial MPS1 inhibition. We also find that PLK1 inhibition does not obviously cooperate with Haspin inhibition to promote SAC override. These results indicate that PLK1 is directly involved in maintaining efficient SAC signalling, possibly by cooperating in a positive feedback loop with Aurora B, and that partially redundant mechanisms exist which reinforce the SAC.

  16. Supporting activity engagement by family carers at home: maintenance of agency and personhood in dementia.

    Science.gov (United States)

    Chung, Pat Yin Fan; Ellis-Hill, Caroline; Coleman, Peter

    2017-12-01

    An explorative paper to describe how family carers, through the caregiving journey, reaffirm and promote the agency of people with dementia. Agency is an important concept in dementia care and is crucial to the promotion of wellbeing and the delivery of person-centred care. This article is based on one of the key findings of a study that explored family carers' experiences of engaging their relatives in daily activities in domestic settings. Following research governance and ethical approval, 30 in-depth interviews (initial and follow-up) were carried out with 15 resident-carers of people with dementia who were recruited via local community mental health teams. Then five focus groups were conducted with 21 participants accessed through carers support groups. Interviews and focus groups were transcribed, coded and analysed using a grounded theory method. Findings showed the process in which family carers encouraged and sustained a sense of autonomy and control (agency) in their relative's daily activities. Key strategies used by carers included: being non-judgemental; facilitating a sense of worth; taking calculated risks; maintaining the continuity of their relative's identity; enhancing a sense of connection with their relative's role and identity using enjoyable activities; preventing inactivity and attending to the bodily source of the agency. Lack of support for carers could ultimately pose a risk to the maintenance of the agency of people with dementia. This study provides a deeper insight into the process used by home carers to support the agency of people with dementia. This is essential if practitioners are to identify and develop more realistic intervention strategies and to work in effective partnership with family carers. The implications for the creation of dementia-friendly communities are discussed.

  17. Biochemical and structural analysis of the hyperpolarization-activated K(+) channel MVP.

    Science.gov (United States)

    Randich, Amelia M; Cuello, Luis G; Wanderling, Sherry S; Perozo, Eduardo

    2014-03-18

    In contrast to the majority of voltage-gated ion channels, hyperpolarization-activated channels remain closed at depolarizing potentials and are activated at hyperpolarizing potentials. The basis for this reverse polarity is thought to be a result of differences in the way the voltage-sensing domain (VSD) couples to the pore domain. In the absence of structural data, the molecular mechanism of this reverse polarity coupling remains poorly characterized. Here we report the characterization of the structure and local dynamics of the closed activation gate (lower S6 region) of MVP, a hyperpolarization-activated potassium channel from Methanococcus jannaschii, by electron paramagnetic resonance (EPR) spectroscopy. We show that a codon-optimized version of MVP has high expression levels in Escherichia coli, is purified as a stable tetramer, and exhibits expected voltage-dependent activity when reconstituted in liposomes. EPR analysis of the mid to lower S6 region revealed positions exhibiting strong spin-spin coupling, indicating that the activation gate of MVP is closed at 0 mV. A comparison of local environmental parameters along the activation gate for MVP and KcsA indicates that MVP adopts a different closed conformation. These structural details set the stage for future evaluations of reverse electromechanical coupling in MVP.

  18. Biochemical and Structural Analysis of the Hyperpolarization-Activated K+ Channel MVP

    Science.gov (United States)

    2015-01-01

    In contrast to the majority of voltage-gated ion channels, hyperpolarization-activated channels remain closed at depolarizing potentials and are activated at hyperpolarizing potentials. The basis for this reverse polarity is thought to be a result of differences in the way the voltage-sensing domain (VSD) couples to the pore domain. In the absence of structural data, the molecular mechanism of this reverse polarity coupling remains poorly characterized. Here we report the characterization of the structure and local dynamics of the closed activation gate (lower S6 region) of MVP, a hyperpolarization-activated potassium channel from Methanococcus jannaschii, by electron paramagnetic resonance (EPR) spectroscopy. We show that a codon-optimized version of MVP has high expression levels in Escherichia coli, is purified as a stable tetramer, and exhibits expected voltage-dependent activity when reconstituted in liposomes. EPR analysis of the mid to lower S6 region revealed positions exhibiting strong spin–spin coupling, indicating that the activation gate of MVP is closed at 0 mV. A comparison of local environmental parameters along the activation gate for MVP and KcsA indicates that MVP adopts a different closed conformation. These structural details set the stage for future evaluations of reverse electromechanical coupling in MVP. PMID:24490868

  19. Blockade of TRPM7 channel activity and cell death by inhibitors of 5-lipoxygenase.

    Directory of Open Access Journals (Sweden)

    Hsiang-Chin Chen

    2010-06-01

    Full Text Available TRPM7 is a ubiquitous divalent-selective ion channel with its own kinase domain. Recent studies have shown that suppression of TRPM7 protein expression by RNA interference increases resistance to ischemia-induced neuronal cell death in vivo and in vitro, making the channel a potentially attractive pharmacological target for molecular intervention. Here, we report the identification of the 5-lipoxygenase inhibitors, NDGA, AA861, and MK886, as potent blockers of the TRPM7 channel. Using a cell-based assay, application of these compounds prevented cell rounding caused by overexpression of TRPM7 in HEK-293 cells, whereas inhibitors of 12-lipoxygenase and 15-lipoxygenase did not prevent the change in cell morphology. Application of the 5-lipoxygenase inhibitors blocked heterologously expressed TRPM7 whole-cell currents without affecting the protein's expression level or its cell surface concentration. All three inhibitors were also effective in blocking the native TRPM7 current in HEK-293 cells. However, two other 5-lipoxygenase specific inhibitors, 5,6-dehydro-arachidonic acid and zileuton, were ineffective in suppressing TRPM7 channel activity. Targeted knockdown of 5-lipoxygenase did not reduce TRPM7 whole-cell currents. In addition, application of 5-hydroperoxyeicosatetraenoic acid (5-HPETE, the product of 5-lipoxygenase, or 5-HPETE's downstream metabolites, leukotriene B4 and leukotriene D4, did not stimulate TRPM7 channel activity. These data suggested that NDGA, AA861, and MK886 reduced the TRPM7 channel activity independent of their effect on 5-lipoxygenase activity. Application of AA861 and NDGA reduced cell death for cells overexpressing TRPM7 cultured in low extracellular divalent cations. Moreover, treatment of HEK-293 cells with AA861 increased cell resistance to apoptotic stimuli to a level similar to that obtained for cells in which TRPM7 was knocked down by RNA interference. In conclusion, NDGA, AA861, and MK886 are potent blockers of

  20. Proteolytic fragmentation of inositol 1,4,5-trisphosphate receptors: a novel mechanism regulating channel activity?

    Science.gov (United States)

    Wang, Liwei; Alzayady, Kamil J; Yule, David I

    2016-06-01

    Inositol 1,4,5-trisphosphate receptors (IP3 Rs) are a family of ubiquitously expressed intracellular Ca(2+) release channels. Regulation of channel activity by Ca(2+) , nucleotides, phosphorylation, protein binding partners and other cellular factors is thought to play a major role in defining the specific spatiotemporal characteristics of intracellular Ca(2+) signals. These properties are, in turn, believed pivotal for the selective and specific physiological activation of Ca(2+) -dependent effectors. IP3 Rs are also substrates for the intracellular cysteine proteases, calpain and caspase. Cleavage of the IP3 R has been proposed to play a role in apoptotic cell death by uncoupling regions important for IP3 binding from the channel domain, leaving an unregulated leaky Ca(2+) pore. Contrary to this hypothesis, we demonstrate following proteolysis that N- and C-termini of IP3 R1 remain associated, presumably through non-covalent interactions. Further, we show that complementary fragments of IP3 R1 assemble into tetrameric structures and retain their ability to be regulated robustly by IP3 . While peptide continuity is clearly not necessary for IP3 -gating of the channel, we propose that cleavage of the IP3 R peptide chain may alter other important regulatory events to modulate channel activity. In this scenario, stimulation of the cleaved IP3 R may support distinct spatiotemporal Ca(2+) signals and activation of specific effectors. Notably, in many adaptive physiological events, the non-apoptotic activities of caspase and calpain are demonstrated to be important, but the substrates of the proteases are poorly defined. We speculate that proteolytic fragmentation may represent a novel form of IP3 R regulation, which plays a role in varied adaptive physiological processes. © 2015 The Authors. The Journal of Physiology © 2015 The Physiological Society.

  1. cAMP-dependent kinase does not modulate the Slack sodium-activated potassium channel.

    Science.gov (United States)

    Nuwer, Megan O; Picchione, Kelly E; Bhattacharjee, Arin

    2009-09-01

    The Slack gene encodes a Na(+)-activated K(+) channel and is expressed in many different types of neurons. Like the prokaryotic Ca(2+)-gated K(+) channel MthK, Slack contains two 'regulator of K(+) conductance' (RCK) domains within its carboxy terminal, domains likely involved in Na(+) binding and channel gating. It also contains multiple consensus protein kinase C (PKC) and protein kinase A (PKA) phosphorylation sites and although regulated by protein kinase C (PKC) phosphorylation, modulation by PKA has not been determined. To test if PKA directly regulates Slack, nystatin-perforated patch whole-cell currents were recorded from a human embryonic kidney (HEK-293) cell line stably expressing Slack. Bath application of forskolin, an adenylate cyclase activator, caused a rapid and complete inhibition of Slack currents however, the inactive homolog of forskolin, 1,9-dideoxyforskolin caused a similar effect. In contrast, bath application of 8-bromo-cAMP did not affect the amplitude nor the activation kinetics of Slack currents. In excised inside-out patch recordings, direct application of the PKA catalytic subunit to patches did not affect the open probability of Slack channels nor was open probability affected by direct application of protein phosphatase 2B. Preincubation of cells with the protein kinase A inhibitor KT5720 also did not change current density. Finally, mutating the consensus phosphorylation site located between RCK domain 1 and domain 2 from serine to glutamate did not affect current activation kinetics. We conclude that unlike PKC, phosphorylation by PKA does not acutely modulate the function and gating activation kinetics of Slack channels.

  2. MODELING A COMPUTER APPLICATION FOR MANAGEMENT OF MAINTENANCE ACTIVITIES OF UNPAVED ROADS

    Directory of Open Access Journals (Sweden)

    Ludmília de Souza Dias

    2015-08-01

    Full Text Available ABSTRACTThis study presents a contribution to the modeling of a computer application employing a method of serviceability performance for unpaved roads, aiming the management of maintenance/restoration activities of the primary surface layer. The proposed methodology consisted of field inspections during dry (April to September and rainy (October to March periods, during which objective evaluations were performed to survey of defects and their densities and degrees of severity. To aid the functional classification of analyzed road sections and the determination of the defect with major influence on the serviceability of these roads, the method of serviceability performance proposed by Silva (2009was implemented in the Visual Basic for Applications (VBA language in Microsoft Excel software. With the use of the computer application proposed it was possible to identify among the defects analyzed in field, through the index of serviceability of the sampling unit per defect type (ISUdef, which one had the greatest influence on determining the relative serviceability index per road section (IST. The results allow us to conclude that the computer application Road achieved satisfactory results, since the objective evaluation criteria applied to road sections denotes consistency regarding their serviceability.

  3. Recent progress in the field of automated welding applied to maintenance activities

    International Nuclear Information System (INIS)

    Cullafroz, M.

    2004-01-01

    Automated and robot welding has 5 advantages compared to manual welding: -) under some conditions the automated circular welding does not require requalification testing as manual welding does, -) welding heads in robots have a reduced size compared to manual gears so they can enter and treat complex piping, -) by using an adequate viewing system the operator can be more than 10 meters away from the welding site which means that the radiation doses he receives is cut by a factor 1.5 to 2, -) whatever the configuration is, the deposition rate in automated welding stays high, the quality standard is steady and the risk of repairing is low, -) a gain in productivity if adequate equipment is used. In general, automated welding requires a TIG welding process and is applied in maintenance activities to: -) the main primary system and other circuits in stainless austenitic steels, -) the main secondary system and other circuits in low-percentage carbon steels, and -) the closure of spent fuel canisters. An application to the repairing of BWR's pipes is shown. (A.C.)

  4. Control of helium activity in the fuel reactor channels; Kontrola aktivnosti heliuma u tehnoloskim kanalima

    Energy Technology Data Exchange (ETDEWEB)

    Vidmar, M; Milosevic, M; Hadzic, S [Institute of Nuclear Sciences Boris Kidric, Reaktor RA, Vinca, Beograd (Yugoslavia)

    1961-02-15

    The objective of this task was to study the possibility of detecting a damaged fuel channel, and to introduce automated procedure for continuous control of reactor channels during operation. The existing control systems at the RA reactor (permanent control of heavy water and helium activity, radiation monitoring of heavy water and helium system, measurements of fire damp gas percent) are not sufficient for fast detection of fuel element failures. Since a 'hot' fuel channel cannot be removed from the core because it should be cooled in the core by heavy water circulation, it is not possible to prevent contamination of heavy water by fission products. It is concluded that it is not indispensable to detect the failed fuel element promptly, i.e. that tome is not a critical issue.

  5. Calcium-Activated Cl- Channel: Insights on the Molecular Identity in Epithelial Tissues.

    Science.gov (United States)

    Rottgen, Trey S; Nickerson, Andrew J; Rajendran, Vazhaikkurichi M

    2018-05-10

    Calcium-activated chloride secretion in epithelial tissues has been described for many years. However, the molecular identity of the channel responsible for the Ca 2+ -activated Cl − secretion in epithelial tissues has remained a mystery. More recently, TMEM16A has been identified as a new putative Ca 2+ -activated Cl − channel (CaCC). The primary goal of this article will be to review the characterization of TMEM16A, as it relates to the physical structure of the channel, as well as important residues that confer voltage and Ca 2+ -sensitivity of the channel. This review will also discuss the role of TMEM16A in epithelial physiology and potential associated-pathophysiology. This will include discussion of developed knockout models that have provided much needed insight on the functional localization of TMEM16A in several epithelial tissues. Finally, this review will examine the implications of the identification of TMEM16A as it pertains to potential novel therapies in several pathologies.

  6. Taurine activates delayed rectifier KV channels via a metabotropic pathway in retinal neurons

    Science.gov (United States)

    Bulley, Simon; Liu, Yufei; Ripps, Harris; Shen, Wen

    2013-01-01

    Taurine is one of the most abundant amino acids in the retina, throughout the CNS, and in heart and muscle cells. In keeping with its broad tissue distribution, taurine serves as a modulator of numerous basic processes, such as enzyme activity, cell development, myocardial function and cytoprotection. Despite this multitude of functional roles, the precise mechanism underlying taurine's actions has not yet been identified. In this study we report findings that indicate a novel role for taurine in the regulation of voltage-gated delayed rectifier potassium (KV) channels in retinal neurons by means of a metabotropic receptor pathway. The metabotropic taurine response was insensitive to the Cl− channel blockers, picrotoxin and strychnine, but it was inhibited by a specific serotonin 5-HT2A receptor antagonist, MDL11939. Moreover, we found that taurine enhanced KV channels via intracellular protein kinase C-mediated pathways. When 5-HT2A receptors were expressed in human embryonic kidney cells, taurine and AL34662, a non-specific 5-HT2 receptor activator, produced a similar regulation of KIR channels. In sum, this study provides new evidence that taurine activates a serotonin system, apparently via 5-HT2A receptors and related intracellular pathways. PMID:23045337

  7. Manpower development for safe operation of nuclear power plant. China. Advice on maintenance planning. Activity: 3.1.2-Task-06. Technical report

    International Nuclear Information System (INIS)

    Chihara, Y.; Imai, K.

    1996-01-01

    Regarding the matter of quality assurance related to maintenance activities, we provided appropriate advice on the details of maintenance technologies and on quality control for the nuclear power plant. The advice was provided based on our experience in Japan, with explanations made from specialists' points of view in the form of a lecture. This also included an exchange of questions and answers regarding our experiences with quality control and maintenance activities in Japan

  8. Prediction of attendance at fitness center: a comparison between the theory of planned behavior, the social cognitive theory, and the physical activity maintenance theory

    OpenAIRE

    Jekauc, Darko; Völkle, Manuel; Wagner, Matthias O.; Mess, Filip; Reiner, Miriam; Renner, Britta

    2015-01-01

    In the processes of physical activity (PA) maintenance specific predictors are effective, which differ from other stages of PA development. Recently, Physical Activity Maintenance Theory (PAMT) was specifically developed for prediction of PA maintenance. The aim of the present study was to evaluate the predictability of the future behavior by the PAMT and compare it with the Theory of Planned Behavior (TPB) and Social Cognitive Theory (SCT). Participation rate in a fitness center was observed...

  9. Antischistosomal activity of a calcium channel antagonist on schistosomula and adult Schistosoma mansoni worms

    Directory of Open Access Journals (Sweden)

    Vanessa Silva-Moraes

    2013-08-01

    Full Text Available Current schistosomiasis control strategies are largely based on chemotherapeutic agents and a limited number of drugs are available today. Praziquantel (PZQ is the only drug currently used in schistosomiasis control programs. Unfortunately, this drug shows poor efficacy in patients during the earliest infection phases. The effects of PZQ appear to operate on the voltage-operated Ca2+channels, which are located on the external Schistosoma mansoni membrane. Because some Ca2+channels have dihydropyridine drug class (a class that includes nifedipine sensitivity, an in vitro analysis using a calcium channel antagonist (clinically used for cardiovascular hypertension was performed to determine the antischistosomal effects of nifedipine on schistosomula and adult worm cultures. Nifedipine demonstrated antischistosomal activity against schistosomula and significantly reduced viability at all of the concentrations used alone or in combination with PZQ. In contrast, PZQ did not show significant efficacy when used alone. Adult worms were also affected by nifedipine after a 24 h incubation and exhibited impaired motility, several lesions on the tegument and intense contractility. These data support the idea of Ca2+channels subunits as drug targets and favour alternative therapeutic schemes when drug resistance has been reported. In this paper, strong arguments encouraging drug research are presented, with a focus on exploring schistosomal Ca2+channels.

  10. Single Channel Analysis of Isoflurane and Ethanol Enhancement of Taurine-Activated Glycine Receptors.

    Science.gov (United States)

    Kirson, Dean; Todorovic, Jelena; Mihic, S John

    2018-01-01

    The amino acid taurine is an endogenous ligand acting on glycine receptors (GlyRs), which is released by astrocytes in many brain regions, such as the nucleus accumbens and prefrontal cortex. Taurine is a partial agonist with an efficacy significantly lower than that of glycine. Allosteric modulators such as ethanol and isoflurane produce leftward shifts of glycine concentration-response curves but have no effects at saturating glycine concentrations. In contrast, in whole-cell electrophysiology studies these modulators increase the effects of saturating taurine concentrations. A number of possible mechanisms may explain these enhancing effects, including modulator effects on conductance, channel open times, or channel closed times. We used outside-out patch-clamp single channel electrophysiology to investigate the mechanism of action of 200 mM ethanol and 0.55 mM isoflurane in enhancing the effects of a saturating concentration of taurine. Neither modulator enhanced taurine-mediated conductance. Isoflurane increased the probability of channel opening. Isoflurane also increased the lifetimes of the two shortest open dwell times while both agents decreased the likelihood of occurrence of the longest-lived intracluster channel-closing events. The mechanism of enhancement of GlyR functioning by these modulators is dependent on the efficacy of the agonist activating the receptor and the concentration of agonist tested. Copyright © 2017 by The American Society for Pharmacology and Experimental Therapeutics.

  11. The role of NH2-terminal positive charges in the activity of inward rectifier KATP channels.

    Science.gov (United States)

    Cukras, C A; Jeliazkova, I; Nichols, C G

    2002-09-01

    Approximately half of the NH(2) terminus of inward rectifier (Kir) channels can be deleted without significant change in channel function, but activity is lost when more than approximately 30 conserved residues before the first membrane spanning domain (M1) are removed. Systematic replacement of the positive charges in the NH(2) terminus of Kir6.2 with alanine reveals several residues that affect channel function when neutralized. Certain mutations (R4A, R5A, R16A, R27A, R39A, K47A, R50A, R54A, K67A) change open probability, whereas an overlapping set of mutants (R16A, R27A, K39A, K47A, R50A, R54A, K67A) change ATP sensitivity. Further analysis of the latter set differentiates mutations that alter ATP sensitivity as a consequence of altered open state stability (R16A, K39A, K67A) from those that may affect ATP binding directly (K47A, R50A, R54A). The data help to define the structural determinants of Kir channel function, and suggest possible structural motifs within the NH(2) terminus, as well as the relationship of the NH(2) terminus with the extended cytoplasmic COOH terminus of the channel.

  12. Scheduling a maintenance activity under skills constraints to minimize total weighted tardiness and late tasks

    Directory of Open Access Journals (Sweden)

    Djalal Hedjazi

    2015-04-01

    Full Text Available Skill management is a key factor in improving effectiveness of industrial companies, notably their maintenance services. The problem considered in this paper concerns scheduling of maintenance tasks under resource (maintenance teams constraints. This problem is generally known as unrelated parallel machine scheduling. We consider the problem with a both objectives of minimizing total weighted tardiness (TWT and number of tardiness tasks. Our interest is focused particularly on solving this problem under skill constraints, which each resource has a skill level. So, we propose a new efficient heuristic to obtain an approximate solution for this NP-hard problem and demonstrate his effectiveness through computational experiments. This heuristic is designed for implementation in a static maintenance scheduling problem (with unequal release dates, processing times and resource skills, while minimizing objective functions aforementioned.

  13. Development of maintenance equipment for nuclear material fabrication equipment in a highly active hot cell

    Energy Technology Data Exchange (ETDEWEB)

    Park, J. J.; Yang, M. S.; Kim, K. H. and others

    2000-09-01

    This report presents the development of a maintenance system for a highly contaminated nuclear material handling equipment at a hot-cell. This maintenance system has mainly three subsystems - a gamma-radiation measurement module for detecting a gamma-radiation level and identifying its distribution in-situ, a dry-type decontamination device for cleaning up contaminated particles, and a maintenance chamber for isolating contaminated equipment. The mechanical design considerations, controller, capabilities and remote operation and manipulation of the maintenance system are described. Such subsystems developed were installed and tested in the IMEF (Irradiated Material Examination Facility) M6 hot-cell after mock-up tests and performed their specific tasks successfully.

  14. Development of maintenance equipment for nuclear material fabrication equipment in a highly active hot cell

    International Nuclear Information System (INIS)

    Park, J. J.; Yang, M. S.; Kim, K. H. and others

    2000-09-01

    This report presents the development of a maintenance system for a highly contaminated nuclear material handling equipment at a hot-cell. This maintenance system has mainly three subsystems - a gamma-radiation measurement module for detecting a gamma-radiation level and identifying its distribution in-situ, a dry-type decontamination device for cleaning up contaminated particles, and a maintenance chamber for isolating contaminated equipment. The mechanical design considerations, controller, capabilities and remote operation and manipulation of the maintenance system are described. Such subsystems developed were installed and tested in the IMEF (Irradiated Material Examination Facility) M6 hot-cell after mock-up tests and performed their specific tasks successfully

  15. Activation of the ALT pathway for telomere maintenance can affect other sequences in the human genome.

    Science.gov (United States)

    Jeyapalan, Jennie N; Varley, Helen; Foxon, Jenny L; Pollock, Raphael E; Jeffreys, Alec J; Henson, Jeremy D; Reddel, Roger R; Royle, Nicola J

    2005-07-01

    Immortal human cells maintain telomere length by the expression of telomerase or through the alternative lengthening of telomeres (ALT). The ALT mechanism involves a recombination-like process that allows the rapid elongation of shortened telomeres. However, it is not known whether activation of the ALT pathway affects other sequences in the genome. To address this we have investigated, in ALT-expressing cell lines and tumours, the stability of tandem repeat sequences known to mutate via homologous recombination in the human germline. We have shown extraordinary somatic instability in the human minisatellite MS32 (D1S8) in ALT-expressing (ALT+) but not in normal or telomerase-expressing cell lines. The MS32 mutation frequency varied across 15 ALT+ cell lines and was on average 55-fold greater than in ALT- cell lines. The MS32 minisatellite was also highly unstable in three of eight ALT+ soft tissue sarcomas, indicating that somatic destabilization occurs in vivo. The MS32 mutation rates estimated for two ALT+ cell lines were similar to that seen in the germline. However, the internal structures of ALT and germline mutant alleles are very different, indicating differences in the underlying mutation mechanisms. Five other hypervariable minisatellites did not show elevated instability in ALT-expressing cell lines, indicating that minisatellite destabilization is not universal. The elevation of MS32 instability upon activation of the ALT pathway and telomere length maintenance suggests there is overlap between the underlying processes that may be tractable through analysis of the D1S8 locus.

  16. Skin Redox Balance Maintenance: The Need for an Nrf2-Activator Delivery System

    Directory of Open Access Journals (Sweden)

    Maya Ben-Yehuda Greenwald

    2016-01-01

    Full Text Available The skin, being the largest organ of the body, functions as a barrier between our body and the environment. It is consistently exposed to various exogenous and endogenous stressors (e.g., air pollutants, ionizing and non-ionizing irradiation, toxins, mitochondrial metabolism, enzyme activity, inflammatory process, etc. producing reactive oxygen species (ROS and physical damage (e.g., wounds, sunburns also resulting in reactive oxygen species production. Although skin is equipped with an array of defense mechanisms to counteract reactive oxygen species, augmented exposure and continued reactive oxygen species might result in excessive oxidative stress leading to many skin disorders including inflammatory diseases, pigmenting disorders and some types of cutaneous malignancy. The nuclear factor erythroid 2-related factor 2 (Nrf2 is an emerging regulator of cellular resistance and of defensive enzymes such as the phase II enzymes. Induction of the Keap1–Nrf2 pathway may have a beneficial effect in the treatment of a large number of skin disorders by stimulating an endogenous defense mechanism. However, prolonged and enhanced activation of this pathway is detrimental and, thus, limits the therapeutic potential of Keap1–Nrf2 modulators. Here, we review the consequences of oxidative stress to the skin, and the defense mechanisms that skin is equipped with. We describe the challenges of maintaining skin redox balance and its impact on skin status and function. Finally, we suggest a novel strategy for maintenance of skin redox homeostasis by modulating the Keap1–Nrf2 pathway using nanotechnology-based delivery systems.

  17. Advanced maintenance research programs

    International Nuclear Information System (INIS)

    Marston, T.U.; Gelhaus, F.; Burke, R.

    1985-01-01

    The purpose of this paper is to provide the reader with an idea of the advanced maintenance research program at the Electric Power Research Institute (EPRI). A brief description of the maintenance-related activities is provided as a foundation for the advanced maintenance research projects. The projects can be divided into maintenance planning, preventive maintenance program development and implementation, predictive (or conditional) maintenance, and innovative maintenance techniques. The projects include hardware and software development, human factors considerations, and technology promotion and implementation. The advanced concepts include: the incorporation of artificial intelligence into outage planning; turbine and pump maintenance; rotating equipment monitoring and diagnostics with the aid of expert systems; and the development of mobile robots for nuclear power plant maintenance

  18. [G-protein potentiates the activation of TNF-alpha on calcium-activated potassium channel in ECV304].

    Science.gov (United States)

    Lin, L; Zheng, Y; Qu, J; Bao, G

    2000-06-01

    Observe the effect of tumor necrosis factor-alpha (TNF-alpha) on calcium-activated potassium channel in ECV304 and the possible involvement of G-protein mediation in the action of TNF-alpha. Using the cell-attached configuration of patch clamp technique. (1) the activity of high-conductance calcium-activated potassium channel (BKca) was recorded. Its conductance is (202.54 +/- 16.62) pS; (2) the activity of BKca was potentiated by 200 U/ml TNF-alpha; (3) G-protein would intensify this TNF-alpha activation. TNF-alpha acted on vascular endothelial cell ECV304 could rapidly activate the activity of BKca. Opening of BKca resulted in membrane hyper-polarization which could increase electro-chemical gradient for the resting Ca2+ influx and open leakage calcium channel, thus resting cytoplasmic free Ca2+ concentration could be elevated. G-protein may exert an important regulation in this process.

  19. Activation of KCNQ Channels Suppresses Spontaneous Activity in Dorsal Root Ganglion Neurons and Reduces Chronic Pain after Spinal Cord Injury.

    Science.gov (United States)

    Wu, Zizhen; Li, Lin; Xie, Fuhua; Du, Junhui; Zuo, Yan; Frost, Jeffrey A; Carlton, Susan M; Walters, Edgar T; Yang, Qing

    2017-03-15

    A majority of people who have sustained spinal cord injury (SCI) experience chronic pain after injury, and this pain is highly resistant to available treatments. Contusive SCI in rats at T10 results in hyperexcitability of primary sensory neurons, which contributes to chronic pain. KCNQ channels are widely expressed in nociceptive dorsal root ganglion (DRG) neurons, are important for controlling their excitability, and their activation has proven effective in reducing pain in peripheral nerve injury and inflammation models. The possibility that activators of KCNQ channels could be useful for treating SCI-induced chronic pain is strongly supported by the following findings. First, SCI, unlike peripheral nerve injury, failed to decrease the functional or biochemical expression of KCNQ channels in DRG as revealed by electrophysiology, real-time quantitative polymerase chain reaction, and Western blot; therefore, these channels remain available for pharmacological targeting of SCI pain. Second, treatment with retigabine, a specific KCNQ channel opener, profoundly decreased spontaneous activity in primary sensory neurons of SCI animals both in vitro and in vivo without changing the peripheral mechanical threshold. Third, retigabine reversed SCI-induced reflex hypersensitivity, adding to our previous demonstration that retigabine supports the conditioning of place preference after SCI (an operant measure of spontaneous pain). In contrast to SCI animals, naïve animals showed no effects of retigabine on reflex sensitivity or conditioned place preference by pairing with retigabine, indicating that a dose that blocks chronic pain-related behavior has no effect on normal pain sensitivity or motivational state. These results encourage the further exploration of U.S. Food and Drug Administration-approved KCNQ activators for treating SCI pain, as well as efforts to develop a new generation of KCNQ activators that lack central side effects.

  20. International Union of Basic and Clinical Pharmacology. C. Nomenclature and Properties of Calcium-Activated and Sodium-Activated Potassium Channels.

    Science.gov (United States)

    Kaczmarek, Leonard K; Aldrich, Richard W; Chandy, K George; Grissmer, Stephan; Wei, Aguan D; Wulff, Heike

    2017-01-01

    A subset of potassium channels is regulated primarily by changes in the cytoplasmic concentration of ions, including calcium, sodium, chloride, and protons. The eight members of this subfamily were originally all designated as calcium-activated channels. More recent studies have clarified the gating mechanisms for these channels and have documented that not all members are sensitive to calcium. This article describes the molecular relationships between these channels and provides an introduction to their functional properties. It also introduces a new nomenclature that differentiates between calcium- and sodium-activated potassium channels. Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics.

  1. Detection of TRPV4 channel current-like activity in Fawn Hooded hypertensive (FHH rat cerebral arterial muscle cells.

    Directory of Open Access Journals (Sweden)

    Debebe Gebremedhin

    Full Text Available The transient receptor potential vallinoid type 4 (TRPV4 is a calcium entry channel known to modulate vascular function by mediating endothelium-dependent vasodilation. The present study investigated if isolated cerebral arterial myocytes of the Fawn Hooded hypertensive (FHH rat, known to display exaggerated KCa channel current activity and impaired myogenic tone, express TRPV4 channels at the transcript and protein level and exhibit TRPV4-like single-channel cationic current activity. Reverse transcription polymerase chain reaction (RT-PCR, Western blot, and immunostaining analysis detected the expression of mRNA transcript and translated protein of TRPV4 channel in FHH rat cerebral arterial myocytes. Patch clamp recording of single-channel current activity identified the presence of a single-channel cationic current with unitary conductance of ~85 pS and ~96 pS at hyperpolarizing and depolarizing potentials, respectively, that was inhibited by the TRPV4 channel antagonist RN 1734 or HC 067074 and activated by the potent TRPV4 channel agonist GSK1016790A. Application of negative pressure via the interior of the patch pipette increased the NPo of the TRPV4-like single-channel cationic current recorded in cell-attached patches at a patch potential of 60 mV that was inhibited by prior application of the TRPV4 channel antagonist RN 1734 or HC 067047. Treatment with the TRPV4 channel agonist GSK1016790A caused concentration-dependent increase in the NPo of KCa single-channel current recorded in cell-attached patches of cerebral arterial myocytes at a patch potential of 40 mV, which was not influenced by pretreatment with the voltage-gated L-type Ca2+ channel blocker nifedipine or the T-type Ca2+ channel blocker Ni2+. These findings demonstrate that FHH rat cerebral arterial myocytes express mRNA transcript and translated protein for TRPV4 channel and display TRPV4-like single-channel cationic current activity that was stretch-sensitive and

  2. Functional characterization of neurotransmitter activation and modulation in a nematode model ligand-gated ion channel.

    Science.gov (United States)

    Heusser, Stephanie A; Yoluk, Özge; Klement, Göran; Riederer, Erika A; Lindahl, Erik; Howard, Rebecca J

    2016-07-01

    The superfamily of pentameric ligand-gated ion channels includes neurotransmitter receptors that mediate fast synaptic transmission in vertebrates, and are targets for drugs including alcohols, anesthetics, benzodiazepines, and anticonvulsants. However, the mechanisms of ion channel opening, gating, and modulation in these receptors leave many open questions, despite their pharmacological importance. Subtle conformational changes in both the extracellular and transmembrane domains are likely to influence channel opening, but have been difficult to characterize given the limited structural data available for human membrane proteins. Recent crystal structures of a modified Caenorhabditis elegans glutamate-gated chloride channel (GluCl) in multiple states offer an appealing model system for structure-function studies. However, the pharmacology of the crystallographic GluCl construct is not well established. To establish the functional relevance of this system, we used two-electrode voltage-clamp electrophysiology in Xenopus oocytes to characterize activation of crystallographic and native-like GluCl constructs by L-glutamate and ivermectin. We also tested modulation by ethanol and other anesthetic agents, and used site-directed mutagenesis to explore the role of a region of Loop F which was implicated in ligand gating by molecular dynamics simulations. Our findings indicate that the crystallographic construct functionally models concentration-dependent agonism and allosteric modulation of pharmacologically relevant receptors. Specific substitutions at residue Leu174 in loop F altered direct L-glutamate activation, consistent with computational evidence for this region's role in ligand binding. These insights demonstrate conservation of activation and modulation properties in this receptor family, and establish a framework for GluCl as a model system, including new possibilities for drug discovery. In this study, we elucidate the validity of a modified glutamate

  3. Anoctamin 9/TMEM16J is a cation channel activated by cAMP/PKA signal.

    Science.gov (United States)

    Kim, Hyungsup; Kim, Hyesu; Lee, Jesun; Lee, Byeongjun; Kim, Hee-Ryang; Jung, Jooyoung; Lee, Mi-Ock; Oh, Uhtaek

    2018-05-01

    Anoctamins (ANOs) are multifunctional membrane proteins that consist of 10 homologs. ANO1 (TMEM16A) and ANO2 (TMEM16B) are anion channels activated by intracellular calcium that meditate numerous physiological functions. ANO6 is a scramblase that redistributes phospholipids across the cell membrane. The other homologs are not well characterized. We found ANO9/TMEM16J is a cation channel activated by a cAMP-dependent protein kinase A (PKA). Intracellular cAMP-activated robust currents in whole cells expressing ANO9, which were inhibited by a PKA blocker. A cholera toxin that persistently stimulated adenylate cyclase activated ANO9 as did the application of PKA. The cAMP-induced ANO9 currents were permeable to cations. The cAMP-dependent ANO9 currents were augmented by intracellular Ca 2+ . Ano9 transcripts were predominant in the intestines. Human intestinal SW480 cells expressed high levels of Ano9 transcripts and showed PKA inhibitor-reversible cAMP-dependent currents. We conclude that ANO9 is a cation channel activated by a cAMP/PKA pathway and could play a role in intestine function. Copyright © 2017. Published by Elsevier Ltd.

  4. Antisense oligonucleotides suppress cell-volume-induced activation of chloride channels.

    Science.gov (United States)

    Gschwentner, M; Nagl, U O; Wöll, E; Schmarda, A; Ritter, M; Paulmichl, M

    1995-08-01

    Cell volume regulation is an essential feature of most cells. After swelling in hypotonic media, the simultaneous activation of potassium and chloride channels is believed to be the initial, time-determining step in cell volume regulation. The activation of both pathways is functionally linked and enables the cells to lose ions and water, subsequently leading to cell shrinkage and readjustment of the initial volume. NIH 3T3 fibroblasts efficiently regulate their volume after swelling and bear chloride channels that are activated by decreasing extracellular osmolarity. The chloride current elicited in these cells after swelling is reminiscent of the current found in oocytes expressing an outwardly rectifying chloride current termed ICln. Introduction of antisense oligodeoxynucleotides complementary to the first 30 nucleotides of the coding region of the ICln channel into NIH 3T3 fibroblasts suppresses the activation of the swelling-induced chloride current. The experiments directly demonstrate an unambiguous link between a volume-activated chloride current and a cloned protein involved in chloride transport.

  5. Inward-rectifying potassium (Kir) channels regulate pacemaker activity in spinal nociceptive circuits during early life

    Science.gov (United States)

    Li, Jie; Blankenship, Meredith L.; Baccei, Mark L.

    2013-01-01

    Pacemaker neurons in neonatal spinal nociceptive circuits generate intrinsic burst-firing and are distinguished by a lower “leak” membrane conductance compared to adjacent, non-bursting neurons. However, little is known about which subtypes of leak channels regulate the level of pacemaker activity within the developing rat superficial dorsal horn (SDH). Here we demonstrate that a hallmark feature of lamina I pacemaker neurons is a reduced conductance through inward-rectifying potassium (Kir) channels at physiological membrane potentials. Differences in the strength of inward rectification between pacemakers and non-pacemakers indicate the presence of functionally distinct Kir currents in these two populations at room temperature. However, Kir currents in both groups showed high sensitivity to block by extracellular Ba2+ (IC50 ~ 10 µM), which suggests the presence of ‘classical’ Kir (Kir2.x) channels in the neonatal SDH. The reduced Kir conductance within pacemakers is unlikely to be explained by an absence of particular Kir2.x isoforms, as immunohistochemical analysis revealed the expression of Kir2.1, Kir2.2 and Kir2.3 within spontaneously bursting neurons. Importantly, Ba2+ application unmasked rhythmic burst-firing in ~42% of non-bursting lamina I neurons, suggesting that pacemaker activity is a latent property of a sizeable population of SDH cells during early life. In addition, the prevalence of spontaneous burst-firing within lamina I was enhanced in the presence of high internal concentrations of free Mg2+, consistent with its documented ability to block Kir channels from the intracellular side. Collectively, the results indicate that Kir channels are key modulators of pacemaker activity in newborn central pain networks. PMID:23426663

  6. Molecular and functional expression of high conductance Ca 2+ activated K+ channels in the eel intestinal epithelium

    DEFF Research Database (Denmark)

    Lionetto, Maria G; Rizzello, Antonia; Giordano, Maria E

    2008-01-01

    Several types of K(+) channels have been identified in epithelial cells. Among them high conductance Ca(2+)-activated K(+) channels (BK channels) are of relevant importance for their involvement in regulatory volume decrease (RVD) response following hypotonic stress. The aim of the present work...... was to investigate the functional and molecular expression of BK in the eel intestine, which is a useful experimental model for cell volume regulation research. In the present paper using rat BK channel-specific primer, a RT-PCR signal of 696 pb cDNA was detected in eel intestine, whole nucleotide sequence showed...... high similarity (83%) to the alpha subunit of BK channel family. BK channel protein expression was verified by immunoblotting and confocal microscopy, while the functional role of BK channels in epithelial ion transport mechanisms and cell volume regulation was examined by electrophysiological...

  7. Activation of the Ca2+-sensing receptors increases currents through inward rectifier K+ channels via activation of phosphatidylinositol 4-kinase.

    Science.gov (United States)

    Liu, Chung-Hung; Chang, Hsueh-Kai; Lee, Sue-Ping; Shieh, Ru-Chi

    2016-11-01

    Inward rectifier K + channels are important for maintaining normal electrical function in many cell types. The proper function of these channels requires the presence of membrane phosphoinositide 4,5-bisphosphate (PIP 2 ). Stimulation of the Ca 2+ -sensing receptor CaR, a pleiotropic G protein-coupled receptor, activates both G q/11 , which decreases PIP 2 , and phosphatidylinositol 4-kinase (PI-4-K), which, conversely, increases PIP 2 . How membrane PIP 2 levels are regulated by CaR activation and whether these changes modulate inward rectifier K + are unknown. In this study, we found that activation of CaR by the allosteric agonist, NPSR568, increased inward rectifier K + current (I K1 ) in guinea pig ventricular myocytes and currents mediated by Kir2.1 channels exogenously expressed in HEK293T cells with a similar sensitivity. Moreover, using the fluorescent PIP 2 reporter tubby-R332H-cYFP to monitor PIP 2 levels, we found that CaR activation in HEK293T cells increased membrane PIP 2 concentrations. Pharmacological studies showed that both phospholipase C (PLC) and PI-4-K are activated by CaR stimulation with the latter played a dominant role in regulating membrane PIP 2 and, thus, Kir currents. These results provide the first direct evidence that CaR activation upregulates currents through inward rectifier K + channels by accelerating PIP 2 synthesis. The regulation of I K1 plays a critical role in the stability of the electrical properties of many excitable cells, including cardiac myocytes and neurons. Further, synthetic allosteric modulators that increase CaR activity have been used to treat hyperparathyroidism, and negative CaR modulators are of potential importance in the treatment of osteoporosis. Thus, our results provide further insight into the roles played by CaR in the cardiovascular system and are potentially valuable for heart disease treatment and drug safety.

  8. Inhibition of G protein-activated inwardly rectifying K+ channels by different classes of antidepressants.

    Directory of Open Access Journals (Sweden)

    Toru Kobayashi

    Full Text Available Various antidepressants are commonly used for the treatment of depression and several other neuropsychiatric disorders. In addition to their primary effects on serotonergic or noradrenergic neurotransmitter systems, antidepressants have been shown to interact with several receptors and ion channels. However, the molecular mechanisms that underlie the effects of antidepressants have not yet been sufficiently clarified. G protein-activated inwardly rectifying K(+ (GIRK, Kir3 channels play an important role in regulating neuronal excitability and heart rate, and GIRK channel modulation has been suggested to have therapeutic potential for several neuropsychiatric disorders and cardiac arrhythmias. In the present study, we investigated the effects of various classes of antidepressants on GIRK channels using the Xenopus oocyte expression assay. In oocytes injected with mRNA for GIRK1/GIRK2 or GIRK1/GIRK4 subunits, extracellular application of sertraline, duloxetine, and amoxapine effectively reduced GIRK currents, whereas nefazodone, venlafaxine, mianserin, and mirtazapine weakly inhibited GIRK currents even at toxic levels. The inhibitory effects were concentration-dependent, with various degrees of potency and effectiveness. Furthermore, the effects of sertraline were voltage-independent and time-independent during each voltage pulse, whereas the effects of duloxetine were voltage-dependent with weaker inhibition with negative membrane potentials and time-dependent with a gradual decrease in each voltage pulse. However, Kir2.1 channels were insensitive to all of the drugs. Moreover, the GIRK currents induced by ethanol were inhibited by sertraline but not by intracellularly applied sertraline. The present results suggest that GIRK channel inhibition may reveal a novel characteristic of the commonly used antidepressants, particularly sertraline, and contributes to some of the therapeutic effects and adverse effects.

  9. CNS sites activated by renal pelvic epithelial sodium channels (ENaCs) in response to hypertonic saline in awake rats.

    Science.gov (United States)

    Goodwill, Vanessa S; Terrill, Christopher; Hopewood, Ian; Loewy, Arthur D; Knuepfer, Mark M

    2017-05-01

    In some patients, renal nerve denervation has been reported to be an effective treatment for essential hypertension. Considerable evidence suggests that afferent renal nerves (ARN) and sodium balance play important roles in the development and maintenance of high blood pressure. ARN are sensitive to sodium concentrations in the renal pelvis. To better understand the role of ARN, we infused isotonic or hypertonic NaCl (308 or 500mOsm) into the left renal pelvis of conscious rats for two 2hours while recording arterial pressure and heart rate. Subsequently, brain tissue was analyzed for immunohistochemical detection of the protein Fos, a marker for neuronal activation. Fos-immunoreactive neurons were identified in numerous sites in the forebrain and brainstem. These areas included the nucleus tractus solitarius (NTS), the lateral parabrachial nucleus, the paraventricular nucleus of the hypothalamus (PVH) and the supraoptic nucleus (SON). The most effective stimulus was 500mOsm NaCl. Activation of these sites was attenuated or prevented by administration of benzamil (1μM) or amiloride (10μM) into the renal pelvis concomitantly with hypertonic saline. In anesthetized rats, infusion of hypertonic saline but not isotonic saline into the renal pelvis elevated ARN activity and this increase was attenuated by simultaneous infusion of benzamil or amiloride. We propose that renal pelvic epithelial sodium channels (ENaCs) play a role in activation of ARN and, via central visceral afferent circuits, this system modulates fluid volume and peripheral blood pressure. These pathways may contribute to the development of hypertension. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Regulatory oversight of maintenance activities at nuclear power plants in France

    International Nuclear Information System (INIS)

    Bouton, J.P.; Lallement, J.

    1997-01-01

    In France the nuclear safety authority sets out the main safety objectives but the operator remains the first responsible for the NPP safety. During the operation, operators have to demonstrate that the safety level remains the same as defined in the design studies. Maintenance contributes to meet this objective. The French regulation insists on the quality of the safety related components maintenance, especially in the 1984 order. All gap between results and requirements have to be analyzed by operator who provide feedback measures to avoid similar failure to occur. This gap have to be mentioned to regulators. The use of probabilistic safety analysis (PSA) and reliability centered maintenance (RCM) methods is not well developed in France to optimize maintenance. For the French regulatory, the major difficulties in the use of PSA are: The impossibility to detect unanticipated failure mode; the validity of the input data; the validity of model use, based on ''engineering judgement''. For the specific case of passive components inspection, such as the vessel of the primary circuit, the French regulator has already indicated to the operator that the optimization of maintenance by use of PSA cannot be used, for the following reasons: The safety analysis does not take into account the failure of the vessel; unanticipated failure modes have already damaged part of the primary circuit; the use of defense in depth concept requires a systematic detection of any defect on the vessel. (author)

  11. Distribution of rSlo Ca2+-activated K+ channels in rat astrocyte perivascular endfeet.

    Science.gov (United States)

    Price, Diana L; Ludwig, Jeffrey W; Mi, Huaiyu; Schwarz, Thomas L; Ellisman, Mark H

    2002-11-29

    Evidence that Ca(2+)-activated K(+) (K(Ca)) channels play a role in cell volume changes and K(+) homeostasis led to a prediction that astrocytes would have K(Ca) channels near blood vessels in order to maintain K(+) homeostasis. Consistent with this thinking the present study demonstrates that rSlo K(Ca) channels are in glial cells of the adult rat central nervous system (CNS) and highly localized to specializations of astrocytes associated with the brain vasculature. Using confocal and thin-section electron microscopic immunolabeling methods the distribution of rSlo was examined in adult rat brain. Strong rSlo immunolabeling was present around the vasculature of most brain regions. Examination of dye-filled hippocampal astrocytes revealed rSlo immunolabeling polarized in astrocytic endfeet. Ultrastructural analysis confirmed that the rSlo staining was concentrated in astrocytic endfeet ensheathing capillaries as well as abutting the pia mater. Immunostaining within the endfeet was predominantly distributed at the plasma membrane directly adjacent to either the vascular basal lamina or the pial surface. The distribution of the aquaporin-4 (AQP-4) water channel was also examined using dye-filled hippocampal astrocytes. In confirmation of earlier reports, intense AQP-4 immunolabeling was generally observed at the perimeter of blood vessels, and coincided with perivascular endfeet and rSlo labeling. We propose that rSlo K(Ca) channels, with their sensitivity to membrane depolarization and intracellular calcium, play a role in the K(+) modulation of cerebral blood flow. Additional knowledge of the molecular and cellular machinery present at perivascular endfeet may provide insight into the structural and functional molecular elements responsible for the neuronal activity-dependent regulation of cerebral blood flow. Copyright 2002 Elsevier Science B.V.

  12. Structure-function relation of phospholamban: modulation of channel activity as a potential regulator of SERCA activity.

    Directory of Open Access Journals (Sweden)

    Serena Smeazzetto

    Full Text Available Phospholamban (PLN is a small integral membrane protein, which binds and inhibits in a yet unknown fashion the Ca(2+-ATPase (SERCA in the sarcoplasmic reticulum. When reconstituted in planar lipid bilayers PLN exhibits ion channel activity with a low unitary conductance. From the effect of non-electrolyte polymers on this unitary conductance we estimate a narrow pore with a diameter of ca. 2.2 Å for this channel. This value is similar to that reported for the central pore in the structure of the PLN pentamer. Hence the PLN pentamer, which is in equilibrium with the monomer, is the most likely channel forming structure. Reconstituted PLN mutants, which either stabilize (K27A and R9C or destabilize (I47A the PLN pentamer and also phosphorylated PLN still generate the same unitary conductance of the wt/non-phosphorylated PLN. However the open probability of the phosphorylated PLN and of the R9C mutant is significantly lower than that of the respective wt/non-phosphorylated control. In the context of data on PLN/SERCA interaction and on Ca(2+ accumulation in the sarcoplasmic reticulum the present results are consistent with the view that PLN channel activity could participate in the balancing of charge during Ca(2+ uptake. A reduced total conductance of the K(+ transporting PLN by phosphorylation or by the R9C mutation may stimulate Ca(2+ uptake in the same way as an inhibition of K(+ channels in the SR membrane. The R9C-PLN mutation, a putative cause of dilated cardiomyopathy, might hence affect SERCA activity also via its inherent low open probability.

  13. Activity of Palythoa caribaeorum Venom on Voltage-Gated Ion Channels in Mammalian Superior Cervical Ganglion Neurons

    Directory of Open Access Journals (Sweden)

    Fernando Lazcano-Pérez

    2016-05-01

    Full Text Available The Zoanthids are an order of cnidarians whose venoms and toxins have been poorly studied. Palythoa caribaeorum is a zoanthid commonly found around the Mexican coastline. In this study, we tested the activity of P. caribaeorum venom on voltage-gated sodium channel (NaV1.7, voltage-gated calcium channel (CaV2.2, the A-type transient outward (IA and delayed rectifier (IDR currents of KV channels of the superior cervical ganglion (SCG neurons of the rat. These results showed that the venom reversibly delays the inactivation process of voltage-gated sodium channels and inhibits voltage-gated calcium and potassium channels in this mammalian model. The compounds responsible for these effects seem to be low molecular weight peptides. Together, these results provide evidence for the potential use of zoanthids as a novel source of cnidarian toxins active on voltage-gated ion channels.

  14. Activity of Palythoa caribaeorum Venom on Voltage-Gated Ion Channels in Mammalian Superior Cervical Ganglion Neurons.

    Science.gov (United States)

    Lazcano-Pérez, Fernando; Castro, Héctor; Arenas, Isabel; García, David E; González-Muñoz, Ricardo; Arreguín-Espinosa, Roberto

    2016-05-05

    The Zoanthids are an order of cnidarians whose venoms and toxins have been poorly studied. Palythoa caribaeorum is a zoanthid commonly found around the Mexican coastline. In this study, we tested the activity of P. caribaeorum venom on voltage-gated sodium channel (NaV1.7), voltage-gated calcium channel (CaV2.2), the A-type transient outward (IA) and delayed rectifier (IDR) currents of KV channels of the superior cervical ganglion (SCG) neurons of the rat. These results showed that the venom reversibly delays the inactivation process of voltage-gated sodium channels and inhibits voltage-gated calcium and potassium channels in this mammalian model. The compounds responsible for these effects seem to be low molecular weight peptides. Together, these results provide evidence for the potential use of zoanthids as a novel source of cnidarian toxins active on voltage-gated ion channels.

  15. Plant Maintenance. The Licensee's Viewpoint

    Energy Technology Data Exchange (ETDEWEB)

    Ungi, T. [Heysham 2 Power Station, Nuclear Electric LTD (United Kingdom)

    1997-07-01

    Plant maintenance is a very complex process which requires considerable effort from both within the maintenance process and also many support activities. It is important that the plant maintenance policy is translated into a maintenance programme which will define all relevant aspects of the maintenance. An aspect of this maintenance programme will be a maintenance catalogue which will define the maintenance activities to be carried out and at what frequencies. This paper is aimed at discussing the maintenance philosophy and resulting maintenance catalogues currently adopted in Nuclear Electric Ltd, and in particular at Heysham 2 Power Station. It goes on to consider whether these maintenance catalogues contain the optimum maintenance and if not should they be changed. If change is required, the process by which this change will be brought about is also discussed. (author)

  16. Strong activation of bile acid-sensitive ion channel (BASIC) by ursodeoxycholic acid

    Science.gov (United States)

    Wiemuth, Dominik; Sahin, Hacer; Lefèvre, Cathérine M.T.; Wasmuth, Hermann E.; Gründer, Stefan

    2013-01-01

    Bile acid-sensitive ion channel (BASIC) is a member of the DEG/ENaC gene family of unknown function. Rat BASIC (rBASIC) is inactive at rest. We have recently shown that cholangiocytes, the epithelial cells lining the bile ducts, are the main site of BASIC expression in the liver and identified bile acids, in particular hyo- and chenodeoxycholic acid, as agonists of rBASIC. Moreover, it seems that extracellular divalent cations stabilize the resting state of rBASIC, because removal of extracellular divalent cations opens the channel. In this addendum, we demonstrate that removal of extracellular divalent cations potentiates the activation of rBASIC by bile acids, suggesting an allosteric mechanism. Furthermore, we show that rBASIC is strongly activated by the anticholestatic bile acid ursodeoxycholic acid (UDCA), suggesting that BASIC might mediate part of the therapeutic effects of UDCA. PMID:23064163

  17. Development of EPICS channel access embedded ActiveX components for GUI development

    International Nuclear Information System (INIS)

    Roy, A.; Bhole, R.B.; Pal, S.

    2012-01-01

    The paper describes the integration of Experimental Physics and Industrial Control System (EPICS) Channel Access (CA) protocol and Microsoft ActiveX technology towards developing a generalize operator interface (OPI) building facility for Windows platform. EPICS is used as the development architecture of the control system in Superconducting Cyclotron (SCC). Considering the operators' familiarity and compatibility with third party software, it was decided to use MS-Windows platform at operator interface level in SCC during commission. Microsoft Visual Basic (VB) is used on trial basis as OPI building platform to incorporate user specific features e.g. file system access for data storage and analysis, user authentication at OPI level etc. A set of EPICS Channel Access embedded ActiveX components is developed to ease the programming complexity and reduce developmental time of the OPI for Windows platform. OPIs, developed using these components and containing hundreds of process parameters, are being used reliably over a considerable period of time. (author)

  18. Neutron field for activation experiments in horizontal channel of training reactor VR-1

    Czech Academy of Sciences Publication Activity Database

    Štefánik, Milan; Katovsky, K.; Vinš, M.; Šoltéš, J.; Závorka, L.

    2014-01-01

    Roč. 104, NOV (2014), s. 302-305 ISSN 0969-806X. [1st International Conference on Dosimetry and its Applications (ICDA). Prague, 23.6.2013-28.6.2013] R&D Projects: GA MŠk LG14004 Institutional support: RVO:61389005 Keywords : spectral index * neutron spectrometry * dosimetry-foils activation technique * irradiation channel * reaction rate * Gamma -spectroscopy Subject RIV: BG - Nuclear, Atomic and Molecular Physics, Colliders Impact factor: 1.380, year: 2014

  19. Interaction of a dinoflagellate neurotoxin with voltage-activated ion channels in a marine diatom.

    Science.gov (United States)

    Kitchen, Sheila A; Bourdelais, Andrea J; Taylor, Alison R

    2018-01-01

    The potent neurotoxins produced by the harmful algal bloom species Karenia brevis are activators of sodium voltage-gated channels (VGC) in animals, resulting in altered channel kinetics and membrane hyperexcitability. Recent biophysical and genomic evidence supports widespread presence of homologous sodium (Na + ) and calcium (Ca 2+ ) permeable VGCs in unicellular algae, including marine phytoplankton. We therefore hypothesized that VGCs of these phytoplankton may be an allelopathic target for waterborne neurotoxins produced by K. brevis blooms that could lead to ion channel dysfunction and disruption of signaling in a similar manner to animal Na + VGCs. We examined the interaction of brevetoxin-3 (PbTx-3), a K. brevis neurotoxin, with the Na + /Ca 2+ VGC of the non-toxic diatom Odontella sinensi s using electrophysiology. Single electrode current- and voltage- clamp recordings from O. sinensis in the presence of PbTx-3 were used to examine the toxin's effect on voltage gated Na + /Ca 2+ currents. In silico analysis was used to identify the putative PbTx binding site in the diatoms. We identified Na + /Ca 2+ VCG homologs from the transcriptomes and genomes of 12 diatoms, including three transcripts from O. sinensis and aligned them with site-5 of Na + VGCs, previously identified as the PbTx binding site in animals. Up to 1 µM PbTx had no effect on diatom resting membrane potential or membrane excitability. The kinetics of fast inward Na + /Ca 2+ currents that underlie diatom action potentials were also unaffected. However, the peak inward current was inhibited by 33%, delayed outward current was inhibited by 25%, and reversal potential of the currents shifted positive, indicating a change in permeability of the underlying channels. Sequence analysis showed a lack of conservation of the PbTx binding site in diatom VGC homologs, many of which share molecular features more similar to single-domain bacterial Na + /Ca 2+ VGCs than the 4-domain eukaryote channels

  20. Expression and activity of acid-sensing ion channels in the mouse anterior pituitary.

    Directory of Open Access Journals (Sweden)

    Jianyang Du

    Full Text Available Acid sensing ion channels (ASICs are proton-gated cation channels that are expressed in the nervous system and play an important role in fear learning and memory. The function of ASICs in the pituitary, an endocrine gland that contributes to emotions, is unknown. We sought to investigate which ASIC subunits were present in the pituitary and found mRNA expression for all ASIC isoforms, including ASIC1a, ASIC1b, ASIC2a, ASIC2b, ASIC3 and ASIC4. We also observed acid-evoked ASIC-like currents in isolated anterior pituitary cells that were absent in mice lacking ASIC1a. The biophysical properties and the responses to PcTx1, amiloride, Ca2+ and Zn2+ suggested that ASIC currents were mediated predominantly by heteromultimeric channels that contained ASIC1a and ASIC2a or ASIC2b. ASIC currents were also sensitive to FMRFamide (Phe-Met-Arg-Phe amide, suggesting that FMRFamide-like compounds might endogenously regulate pituitary ASICs. To determine whether ASICs might regulate pituitary cell function, we applied low pH and found that it increased the intracellular Ca2+ concentration. These data suggest that ASIC channels are present and functionally active in anterior pituitary cells and may therefore influence their function.

  1. Membrane-tethered peptides patterned after the TRP domain (TRPducins) selectively inhibit TRPV1 channel activity.

    Science.gov (United States)

    Valente, Pierluigi; Fernández-Carvajal, Asia; Camprubí-Robles, María; Gomis, Ana; Quirce, Susana; Viana, Félix; Fernández-Ballester, Gregorio; González-Ros, José M; Belmonte, Carlos; Planells-Cases, Rosa; Ferrer-Montiel, Antonio

    2011-05-01

    The transient receptor potential vanilloid 1 (TRPV1) channel is a thermosensory receptor implicated in diverse physiological and pathological processes. The TRP domain, a highly conserved region in the C terminus adjacent to the internal channel gate, is critical for subunit tetramerization and channel gating. Here, we show that cell-penetrating, membrane-anchored peptides patterned after this protein domain are moderate and selective TRPV1 antagonists both in vitro and in vivo, blocking receptor activity in intact rat primary sensory neurons and their peripheral axons with mean decline time of 30 min. The most potent lipopeptide, TRP-p5, blocked all modes of TRPV1 gating with micromolar efficacy (IC(50)100 μM). TRP-p5 did not affect the capsaicin sensitivity of the vanilloid receptor. Our data suggest that TRP-p5 interferes with protein-protein interactions at the level of the TRP domain that are essential for the "conformational" change that leads to gate opening. Therefore, these palmitoylated peptides, which we termed TRPducins, are noncompetitive, voltage-independent, sequence-specific TRPV1 blockers. Our findings indicate that TRPducin-like peptides may embody a novel molecular strategy that can be exploited to generate a selective pharmacological arsenal for the TRP superfamily of ion channels.

  2. Unfolding of a Temperature-Sensitive Domain Controls Voltage-Gated Channel Activation.

    Science.gov (United States)

    Arrigoni, Cristina; Rohaim, Ahmed; Shaya, David; Findeisen, Felix; Stein, Richard A; Nurva, Shailika Reddy; Mishra, Smriti; Mchaourab, Hassane S; Minor, Daniel L

    2016-02-25

    Voltage-gated ion channels (VGICs) are outfitted with diverse cytoplasmic domains that impact function. To examine how such elements may affect VGIC behavior, we addressed how the bacterial voltage-gated sodium channel (BacNa(V)) C-terminal cytoplasmic domain (CTD) affects function. Our studies show that the BacNa(V) CTD exerts a profound influence on gating through a temperature-dependent unfolding transition in a discrete cytoplasmic domain, the neck domain, proximal to the pore. Structural and functional studies establish that the BacNa(V) CTD comprises a bi-partite four-helix bundle that bears an unusual hydrophilic core whose integrity is central to the unfolding mechanism and that couples directly to the channel activation gate. Together, our findings define a general principle for how the widespread four-helix bundle cytoplasmic domain architecture can control VGIC responses, uncover a mechanism underlying the diverse BacNa(V) voltage dependencies, and demonstrate that a discrete domain can encode the temperature-dependent response of a channel. Copyright © 2016 Elsevier Inc. All rights reserved.

  3. Environmental effectiveness of GAEC cross-compliance standard 1.1c ‘Maintenance of farm channel networks and field convexity’ and economic evaluation of the competitiveness gap for farmers

    Directory of Open Access Journals (Sweden)

    Paolo Bazzoffi

    2016-02-01

    Full Text Available This paper shows the results of the monitoring carried out in three farms of the project MO.NA.CO. in order to verify the effectiveness of the cross-compliance standard 1.1c which obliges the farmer to the ‘Maintenance of farm channel networks and field convexity’ in order to ensure its efficiency and functionality in draining water. It was also examined the competitiveness gap induced to the agricultural enterprise by the application of the standard, that is to say the additional costs borne by the beneficiary of the single payment determined by cleaning farm collector channels. Effectiveness was determined by evaluating the degradation of soil structure at the end of winter, on flat fields  sown in autumn with winter wheat, in the two cases: a Factual (channels along the field edges not clogged and no waterlogging present on the cultivated soil b Counterfactual (channels clogged and waterlogging present on the cultivated soil. The monitoring confirmed a positive effect of the adoption of this standard on predisposing soil to the ideal conditions for the maintenance of the structure. Despite the statistical evidence found, it must be said that the change in the surface roughness factor was so small as not to take any practical significance in order to affirm that the functional maintenance of collectors channels have been effective in reducing erosion. Overall, the soils were unstructured and crusted at the end of the observation period. Indexes Icli, NTU, and DS show a structural fragility from medium to high for soils of the three monitoring farms. This explains the lack of appreciable differences in the soil roughness parameter, especially in relation to   heavy rains and long waterlogging periods in the cropping years of monitoring. The competitiveness gap induced by the application of this standard, amounted to 19.89±€ 6.35 ha-1 year-1. Atmospheric emission of CO2, was equal to 14.53±6.62 kg ha-1 year-1. It is considered

  4. Photocontrol of Voltage-Gated Ion Channel Activity by Azobenzene Trimethylammonium Bromide in Neonatal Rat Cardiomyocytes.

    Directory of Open Access Journals (Sweden)

    Sheyda R Frolova

    Full Text Available The ability of azobenzene trimethylammonium bromide (azoTAB to sensitize cardiac tissue excitability to light was recently reported. The dark, thermally relaxed trans- isomer of azoTAB suppressed spontaneous activity and excitation propagation speed, whereas the cis- isomer had no detectable effect on the electrical properties of cardiomyocyte monolayers. As the membrane potential of cardiac cells is mainly controlled by activity of voltage-gated ion channels, this study examined whether the sensitization effect of azoTAB was exerted primarily via the modulation of voltage-gated ion channel activity. The effects of trans- and cis- isomers of azoTAB on voltage-dependent sodium (INav, calcium (ICav, and potassium (IKv currents in isolated neonatal rat cardiomyocytes were investigated using the whole-cell patch-clamp technique. The experiments showed that azoTAB modulated ion currents, causing suppression of sodium (Na+ and calcium (Ca2+ currents and potentiation of net potassium (K+ currents. This finding confirms that azoTAB-effect on cardiac tissue excitability do indeed result from modulation of voltage-gated ion channels responsible for action potential.

  5. A proton-activated, outwardly rectifying chloride channel in human umbilical vein endothelial cells

    International Nuclear Information System (INIS)

    Ma Zhiyong; Zhang Wei; Chen Liang; Wang Rong; Kan Xiaohong; Sun Guizhen; Liu Chunxi; Li Li; Zhang Yun

    2008-01-01

    Extracellular acidic pH-activated chloride channel I Cl,acid , has been characterized in HEK 293 cells and mammalian cardiac myocytes. This study was designed to characterize I Cl,acid in human umbilical vein endothelial cells(HUVECs). The activation and deactivation of the current rapidly and repeatedly follows the change of the extracellular solution at pH 4.3, with the threshold pH 5.3. In addition, at very positive potentials, the current displays a time-dependent facilitation. pH-response relationship for I Cl,acid revealed that EC 50 is pH 4.764 with a threshold pH value of pH 5.3 and nH of 14.545. The current can be blocked by the Cl - channel inhibitor DIDS (100 μM). In summary, for the first time we report the presence of proton-activated, outwardly rectifying chloride channel in HUVECs. Because an acidic environment can develop in local myocardium under pathological conditions such as myocardial ischemia, I Cl,acid would play a role in regulation of EC function under these pathological conditions

  6. Antibodies to the extracellular pore loop of TRPM8 act as antagonists of channel activation.

    Directory of Open Access Journals (Sweden)

    Silke Miller

    Full Text Available The mammalian transient receptor potential melastatin channel 8 (TRPM8 is highly expressed in trigeminal and dorsal root ganglia. TRPM8 is activated by cold temperature or compounds that cause a cooling sensation, such as menthol or icilin. TRPM8 may play a role in cold hypersensitivity and hyperalgesia in various pain syndromes. Therefore, TRPM8 antagonists are pursued as therapeutics. In this study we explored the feasibility of blocking TRPM8 activation with antibodies. We report the functional characterization of a rabbit polyclonal antibody, ACC-049, directed against the third extracellular loop near the pore region of the human TRPM8 channel. ACC-049 acted as a full antagonist at recombinantly expressed human and rodent TRPM8 channels in cell based agonist-induced 45Ca2+ uptake assays. Further, several poly-and monoclonal antibodies that recognize the same region also blocked icilin activation of not only recombinantly expressed TRPM8, but also endogenous TRPM8 expressed in rat dorsal root ganglion neurons revealing the feasibility of generating monoclonal antibody antagonists. We conclude that antagonist antibodies are valuable tools to investigate TRPM8 function and may ultimately pave the way for development of therapeutic antibodies.

  7. Channel Power in Multi-Channel Environments

    NARCIS (Netherlands)

    M.G. Dekimpe (Marnik); B. Skiera (Bernd)

    2004-01-01

    textabstractIn the literature, little attention has been paid to instances where companies add an Internet channel to their direct channel portfolio. However, actively managing multiple sales channels requires knowing the customers’ channel preferences and the resulting channel power. Two key

  8. A Human Error Analysis Procedure for Identifying Potential Error Modes and Influencing Factors for Test and Maintenance Activities

    International Nuclear Information System (INIS)

    Kim, Jae Whan; Park, Jin Kyun

    2010-01-01

    Periodic or non-periodic test and maintenance (T and M) activities in large, complex systems such as nuclear power plants (NPPs) are essential for sustaining stable and safe operation of the systems. On the other hand, it also has been raised that human erroneous actions that might occur during T and M activities has the possibility of incurring unplanned reactor trips (RTs) or power derate, making safety-related systems unavailable, or making the reliability of components degraded. Contribution of human errors during normal and abnormal activities of NPPs to the unplanned RTs is known to be about 20% of the total events. This paper introduces a procedure for predictively analyzing human error potentials when maintenance personnel perform T and M tasks based on a work procedure or their work plan. This procedure helps plant maintenance team prepare for plausible human errors. The procedure to be introduced is focusing on the recurrent error forms (or modes) in execution-based errors such as wrong object, omission, too little, and wrong action

  9. Cryo-EM structures of the TMEM16A calcium-activated chloride channel.

    Science.gov (United States)

    Dang, Shangyu; Feng, Shengjie; Tien, Jason; Peters, Christian J; Bulkley, David; Lolicato, Marco; Zhao, Jianhua; Zuberbühler, Kathrin; Ye, Wenlei; Qi, Lijun; Chen, Tingxu; Craik, Charles S; Jan, Yuh Nung; Minor, Daniel L; Cheng, Yifan; Jan, Lily Yeh

    2017-12-21

    Calcium-activated chloride channels (CaCCs) encoded by TMEM16A control neuronal signalling, smooth muscle contraction, airway and exocrine gland secretion, and rhythmic movements of the gastrointestinal system. To understand how CaCCs mediate and control anion permeation to fulfil these physiological functions, knowledge of the mammalian TMEM16A structure and identification of its pore-lining residues are essential. TMEM16A forms a dimer with two pores. Previous CaCC structural analyses have relied on homology modelling of a homologue (nhTMEM16) from the fungus Nectria haematococca that functions primarily as a lipid scramblase, as well as subnanometre-resolution electron cryo-microscopy. Here we present de novo atomic structures of the transmembrane domains of mouse TMEM16A in nanodiscs and in lauryl maltose neopentyl glycol as determined by single-particle electron cryo-microscopy. These structures reveal the ion permeation pore and represent different functional states. The structure in lauryl maltose neopentyl glycol has one Ca 2+ ion resolved within each monomer with a constricted pore; this is likely to correspond to a closed state, because a CaCC with a single Ca 2+ occupancy requires membrane depolarization in order to open (C.J.P. et al., manuscript submitted). The structure in nanodiscs has two Ca 2+ ions per monomer and its pore is in a closed conformation; this probably reflects channel rundown, which is the gradual loss of channel activity that follows prolonged CaCC activation in 1 mM Ca 2+ . Our mutagenesis and electrophysiological studies, prompted by analyses of the structures, identified ten residues distributed along the pore that interact with permeant anions and affect anion selectivity, as well as seven pore-lining residues that cluster near pore constrictions and regulate channel gating. Together, these results clarify the basis of CaCC anion conduction.

  10. Extracellular acidosis activates ASIC-like channels in freshly isolated cerebral artery smooth muscle cells.

    Science.gov (United States)

    Chung, Wen-Shuo; Farley, Jerry M; Swenson, Alyssa; Barnard, John M; Hamilton, Gina; Chiposi, Rumbidzayi; Drummond, Heather A

    2010-05-01

    Recent studies suggest that certain acid-sensing ion channels (ASIC) are expressed in vascular smooth muscle cells (VSMCs) and are required for VSMC functions. However, electrophysiological evidence of ASIC channels in VSMCs is lacking. The purpose of this study was to test the hypothesis that isolated cerebral artery VSMCs express ASIC-like channels. To address this hypothesis, we used RT-PCR, Western blotting, immunolabeling, and conventional whole cell patch-clamp technique. We found extracellular H(+)-induced inward currents in 46% of cells tested (n = 58 of 126 VSMCs, pH 6.5-5.0). The percentage of responsive cells and the current amplitude increased as the external H(+) concentration increased (pH(6.0), n = 28/65 VSMCs responsive, mean current density = 8.1 +/- 1.2 pA/pF). Extracellular acidosis (pH(6.0)) shifted the whole cell reversal potential toward the Nernst potential of Na(+) (n = 6) and substitution of extracellular Na(+) by N-methyl-d-glucamine abolished the inward current (n = 6), indicating that Na(+) is a major charge carrier. The broad-spectrum ASIC blocker amiloride (20 microM) inhibited proton-induced currents to 16.5 +/- 8.7% of control (n = 6, pH(6.0)). Psalmotoxin 1 (PcTx1), an ASIC1a inhibitor and ASIC1b activator, had mixed effects: PcTx1 either 1) abolished H(+)-induced currents (11% of VSMCs, 5/45), 2) enhanced or promoted activation of H(+)-induced currents (76%, 34/45), or 3) failed to promote H(+) activation in nonresponsive VSMCs (13%, 6/45). These findings suggest that freshly dissociated cerebral artery VSMCs express ASIC-like channels, which are predominantly formed by ASIC1b.

  11. Lateral transport of solutes in microfluidic channels using electrochemically generated gradients in redox-active surfactants.

    Science.gov (United States)

    Liu, Xiaoyang; Abbott, Nicholas L

    2011-04-15

    We report principles for a continuous flow process that can separate solutes based on a driving force for selective transport that is generated by a lateral concentration gradient of a redox-active surfactant across a microfluidic channel. Microfluidic channels fabricated with gold electrodes lining each vertical wall were used to electrochemically generate concentration gradients of the redox-active surfactant 11-ferrocenylundecyl-trimethylammonium bromide (FTMA) in a direction perpendicular to the flow. The interactions of three solutes (a hydrophobic dye, 1-phenylazo-2-naphthylamine (yellow AB), an amphiphilic molecule, 2-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-pentanoyl)-1-hexadecanoyl-sn-glycero-3-phosphocholine (BODIPY C(5)-HPC), and an organic salt, 1-methylpyridinium-3-sulfonate (MPS)) with the lateral gradients in surfactant/micelle concentration were shown to drive the formation of solute-specific concentration gradients. Two distinct physical mechanisms were identified to lead to the solute concentration gradients: solubilization of solutes by micelles and differential adsorption of the solutes onto the walls of the microchannels in the presence of the surfactant concentration gradient. These two mechanisms were used to demonstrate delipidation of a mixture of BODIPY C(5)-HPC (lipid) and MPS and purification of BODIPY C(5)-HPC from a mixture of BODIPY C(5)-HPC and yellow AB. Overall, the results of this study demonstrate that lateral concentration gradients of redox-active surfactants formed within microfluidic channels can be used to transport solutes across the microfluidic channels in a solute-dependent manner. The approach employs electrical potentials (solutions having high ionic strength (>0.1M), and offers the basis of continuous processes for the purification or separation of solutes in microscale systems. © 2011 American Chemical Society

  12. Physical activity maintenance among Spanish-speaking Latinas in a randomized controlled trial of an Internet-based intervention.

    Science.gov (United States)

    Hartman, Sheri J; Dunsiger, Shira I; Bock, Beth C; Larsen, Britta A; Linke, Sarah; Pekmezi, Dori; Marquez, Becky; Gans, Kim M; Mendoza-Vasconez, Andrea S; Marcus, Bess H

    2017-06-01

    Spanish-speaking Latinas have some of the lowest rates of meeting physical activity guidelines in the U.S. and are at high risk for many related chronic diseases. The purpose of the current study was to examine the maintenance of a culturally and individually-tailored Internet-based physical activity intervention for Spanish-speaking Latinas. Inactive Latinas (N  =  205) were randomly assigned to a 6-month Tailored Physical Activity Internet Intervention or a Wellness Contact Control Internet Group, with a 6-month follow-up. Maintenance was measured by assessing group differences in minutes per week of self-reported and accelerometer measured moderate to vigorous physical activity (MVPA) at 12 months after baseline and changes in MVPA between the end of the active intervention (month 6) and the end of the study (month 12). Potential moderators of the intervention were also examined. Data were collected between 2011 and 2014, and were analyzed in 2015 at the University of California, San Diego. The Intervention Group engaged in significantly more minutes of MVPA per week than the Control Group at the end of the maintenance period for both self-reported (mean diff. = 30.68, SE = 11.27, p = .007) and accelerometer measured (mean diff. = 11.47, SE = 3.19, p = .01) MVPA. There were no significant between- or within-group changes in MVPA from month 6 to 12. Greater intervention effects were seen for those with lower BMI (BMI × intervention = -6.67, SE = 2.88, p = .02) and lower perceived places to walk to in their neighborhood (access × intervention = -43.25, SE = 19.07, p = .02), with a trend for less family support (social support × intervention = -3.49, SE = 2.05, p = .08). Acculturation, health literacy, and physical activity related psychosocial variables were not significant moderators of the intervention effect during the maintenance period. Findings from the current study support the efficacy of an Internet

  13. Termination of Vernakalant-Resistant Atrial Fibrillation by Inhibition of Small-Conductance Ca2+-Activated K+ Channels in Pigs

    DEFF Research Database (Denmark)

    Diness, Jonas Goldin; Skibsbye, Lasse; Simó-Vicens, Rafel

    2017-01-01

    Background Evidence has emerged that small-conductance Ca2+-activated K+ (SK) channels constitute a new target for treatment of atrial fibrillation (AF). SK channels are predominantly expressed in the atria as compared with the ventricles. Various marketed antiarrhythmic drugs are limited by vent...

  14. Distribution, expression and functional effects of small conductance Ca-activated potassium (SK) channels in rat myometrium.

    Science.gov (United States)

    Noble, Karen; Floyd, Rachel; Shmygol, Andre; Shmygol, Anatoly; Mobasheri, A; Wray, Susan

    2010-01-01

    Calcium-activated potassium channels are important in a variety of smooth muscles, contributing to excitability and contractility. In the myometrium previous work has focussed on the large conductance channels (BK), and the role of small conductance channels (SK) has received scant attention, despite the finding that over-expression of an SK channel isoform (SK3) results in uterine dysfunction and delayed parturition. This study therefore characterises the expression of the three SK channel isoforms (SK1-3) in rat myometrium throughout pregnancy and investigates their effect on cytosolic [Ca] and force and compares this with that of BK channels. Consistent expression of all SK isoform transcripts and clear immunostaining of SK1-3 was found. Inhibition of SK1-3 channels (apamin, scyllatoxin) significantly inhibited outward current, caused membrane depolarisation and elicited action potentials in previously quiescent cells. Apamin or scyllatoxin increased the amplitude of [Ca] and force in spontaneously contracting myometrial strips throughout gestation. The functional effect of SK inhibition was larger than that of BK channel inhibition. Thus we show for the first time that SK1-3 channels are expressed and translated throughout pregnancy and contribute to outward current, regulate membrane potential and hence Ca signals in pregnant rat myometrium. They contribute more to quiescence that BK channels. 2009 Elsevier Ltd. All rights reserved.

  15. Identification of potential novel interaction partners of the sodium-activated potassium channels Slick and Slack in mouse brain.

    Science.gov (United States)

    Rizzi, Sandra; Schwarzer, Christoph; Kremser, Leopold; Lindner, Herbert H; Knaus, Hans-Günther

    2015-12-01

    The sodium-activated potassium channels Slick (Slo2.1, KCNT2) and Slack (Slo2.2, KCNT1) are paralogous channels of the Slo family of high-conductance potassium channels. Slick and Slack channels are widely distributed in the mammalian CNS and they play a role in slow afterhyperpolarization, generation of depolarizing afterpotentials and in setting and stabilizing the resting potential. In the present study we used a combined approach of (co)-immunoprecipitation studies, Western blot analysis, double immunofluorescence and mass spectrometric sequencing in order to investigate protein-protein interactions of the Slick and Slack channels. The data strongly suggest that Slick and Slack channels co-assemble into identical cellular complexes. Double immunofluorescence experiments revealed that Slick and Slack channels co-localize in distinct mouse brain regions. Moreover, we identified the small cytoplasmic protein beta-synuclein and the transmembrane protein 263 (TMEM 263) as novel interaction partners of both, native Slick and Slack channels. In addition, the inactive dipeptidyl-peptidase (DPP 10) and the synapse associated protein 102 (SAP 102) were identified as constituents of the native Slick and Slack channel complexes in the mouse brain. This study presents new insights into protein-protein interactions of native Slick and Slack channels in the mouse brain.

  16. The antipsychotic drug loxapine is an opener of the sodium-activated potassium channel slack (Slo2.2).

    Science.gov (United States)

    Biton, B; Sethuramanujam, S; Picchione, Kelly E; Bhattacharjee, A; Khessibi, N; Chesney, F; Lanneau, C; Curet, O; Avenet, P

    2012-03-01

    Sodium-activated potassium (K(Na)) channels have been suggested to set the resting potential, to modulate slow after-hyperpolarizations, and to control bursting behavior or spike frequency adaptation (Trends Neurosci 28:422-428, 2005). One of the genes that encodes K(Na) channels is called Slack (Kcnt1, Slo2.2). Studies found that Slack channels were highly expressed in nociceptive dorsal root ganglion neurons and modulated their firing frequency (J Neurosci 30:14165-14172, 2010). Therefore, Slack channel openers are of significant interest as putative analgesic drugs. We screened the library of pharmacologically active compounds with recombinant human Slack channels expressed in Chinese hamster ovary cells, by using rubidium efflux measurements with atomic absorption spectrometry. Riluzole at 500 μM was used as a reference agonist. The antipsychotic drug loxapine and the anthelmintic drug niclosamide were both found to activate Slack channels, which was confirmed by using manual patch-clamp analyses (EC(50) = 4.4 μM and EC(50) = 2.9 μM, respectively). Psychotropic drugs structurally related to loxapine were also evaluated in patch-clamp experiments, but none was found to be as active as loxapine. Loxapine properties were confirmed at the single-channel level with recombinant rat Slack channels. In dorsal root ganglion neurons, loxapine was found to behave as an opener of native K(Na) channels and to increase the rheobase of action potential. This study identifies new K(Na) channel pharmacological tools, which will be useful for further Slack channel investigations.

  17. Increased expression of the auxiliary beta(2-subunit of ventricular L-type Ca(2+ channels leads to single-channel activity characteristic of heart failure.

    Directory of Open Access Journals (Sweden)

    Roger Hullin

    2007-03-01

    Full Text Available Increased activity of single ventricular L-type Ca(2+-channels (L-VDCC is a hallmark in human heart failure. Recent findings suggest differential modulation by several auxiliary beta-subunits as a possible explanation.By molecular and functional analyses of human and murine ventricles, we find that enhanced L-VDCC activity is accompanied by altered expression pattern of auxiliary L-VDCC beta-subunit gene products. In HEK293-cells we show differential modulation of single L-VDCC activity by coexpression of several human cardiac beta-subunits: Unlike beta(1 or beta(3 isoforms, beta(2a and beta(2b induce a high-activity channel behavior typical of failing myocytes. In accordance, beta(2-subunit mRNA and protein are up-regulated in failing human myocardium. In a model of heart failure we find that mice overexpressing the human cardiac Ca(V1.2 also reveal increased single-channel activity and sarcolemmal beta(2 expression when entering into the maladaptive stage of heart failure. Interestingly, these animals, when still young and non-failing ("Adaptive Phase", reveal the opposite phenotype, viz: reduced single-channel activity accompanied by lowered beta(2 expression. Additional evidence for the cause-effect relationship between beta(2-subunit expression and single L-VDCC activity is provided by newly engineered, double-transgenic mice bearing both constitutive Ca(V1.2 and inducible beta(2 cardiac overexpression. Here in non-failing hearts induction of beta(2-subunit overexpression mimicked the increase of single L-VDCC activity observed in murine and human chronic heart failure.Our study presents evidence of the pathobiochemical relevance of beta(2-subunits for the electrophysiological phenotype of cardiac L-VDCC and thus provides an explanation for the single L-VDCC gating observed in human and murine heart failure.

  18. General anesthetic octanol and related compounds activate wild-type and delF508 cystic fibrosis chloride channels

    OpenAIRE

    Marcet, Brice; Becq, Frédéric; Norez, Caroline; Delmas, Patrick; Verrier, Bernard

    2004-01-01

    Cystic fibrosis transmembrane conductance regulator (CFTR) Cl− channel is defective during cystic fibrosis (CF). Activators of the CFTR Cl− channel may be useful for therapy of CF. Here, we demonstrate that a range of general anesthetics like normal-alkanols (n-alkanols) and related compounds can stimulate the Cl− channel activity of wild-type CFTR and delF508-CFTR mutant.The effects of n-alkanols like octanol on CFTR activity were measured by iodide (125I) efflux and patch-clamp techniques o...

  19. TNF-α promotes cell survival through stimulation of K+ channel and NFκB activity in corneal epithelial cells

    International Nuclear Information System (INIS)

    Wang Ling; Reinach, Peter; Lu, Luo

    2005-01-01

    Tumor necrosis factor (TNF-α) in various cell types induces either cell death or mitogenesis through different signaling pathways. In the present study, we determined in human corneal epithelial cells how TNF-α also promotes cell survival. Human corneal epithelial (HCE) cells were cultured in DMEM/F-12 medium containing 10% FBS. TNF-α stimulation induced activation of a voltage-gated K + channel detected by measuring single channel activity using patch clamp techniques. The effect of TNF-α on downstream events included NFκB nuclear translocation and increases in DNA binding activities, but did not elicit ERK, JNK, or p38 limb signaling activation. TNF-α induced increases in p21 expression resulting in partial cell cycle attenuation in the G 1 phase. Cell cycle progression was also mapped by flow cytometer analysis. Blockade of TNF-α-induced K + channel activity effectively prevented NFκB nuclear translocation and binding to DNA, diminishing the cell-survival protective effect of TNF-α. In conclusion, TNF-α promotes survival of HCE cells through sequential stimulation of K + channel and NFκB activities. This response to TNF-α is dependent on stimulating K + channel activity because following suppression of K + channel activity TNF-α failed to activate NFκB nuclear translocation and binding to nuclear DNA

  20. Active Galactic Videos: A YouTube Channel for Astronomy Education and Outreach

    Science.gov (United States)

    Calahan, Jenny; Gibbs, Aidan; Hardegree-Ullman, Melody; Hardegree-Ullman, Michael; Impey, Chris David; Kevis, Charlotte; Lewter, Austin; Mauldin, Emmalee; McKee, Carolyn; Olmedo, Alejandro; Pereira, Victoria; Thomas, Melissa; Wenger, Matthew

    2018-01-01

    Active Galactic Videos is an astronomy-focused YouTube channel run by a team at the University of Arizona. The channel both produces astronomy-focused educational content for public audiences and opens a window into the world of professional astronomy by showcasing the work done at Steward Observatory and in Southern Arizona. The channel is mainly run by undergraduate students from a variety of backgrounds including: astronomy, education, film, music, english, and writing. In addition to providing educational content for public audiences, this project provides opportunities for undergraduate students to learn about astronomy content, general astronomy pedagogy, as well as science communication. This is done through developing the practical skills needed to take on the challenge of creating effective and engaging videos. Students write, film, score, direct, and edit each video while conscious of how each piece can affect the teaching/storytelling of the concept at hand. The team has produced various styles of video: presentational, interviews, musical/poetic, tours, and documentaries. In addition to YouTube, the Active Galactic Videos team maintains a social media presence on Facebook, Twitter, and Instagram. These help to widely distribute the content as well as to publicize the main Youtube channel. In addition to providing an overview of our educational work, we present 51 videos, or two year's, worth of online analytics that we are using to better understand our audience, to examine what videos have been popular and successful, and how people are accessing our content. We will present our experience in order to help others learn about improving astronomy education online, as well as astronomy communication and outreach in general.We acknowledge the Howard Hughes Medical Institute for grant support of this and related education initiatives

  1. Small-conductance Ca2+-activated potassium type 2 channels regulate the formation of contextual fear memory.

    Directory of Open Access Journals (Sweden)

    Saravana R K Murthy

    Full Text Available Small-conductance, Ca2+ activated K+ channels (SK channels are expressed at high levels in brain regions responsible for learning and memory. In the current study we characterized the contribution of SK2 channels to synaptic plasticity and to different phases of hippocampal memory formation. Selective SK2 antisense-treatment facilitated basal synaptic transmission and theta-burst induced LTP in hippocampal brain slices. Using the selective SK2 antagonist Lei-Dab7 or SK2 antisense probes, we found that hippocampal SK2 channels are critical during two different time windows: 1 blockade of SK2 channels before the training impaired fear memory, whereas, 2 blockade of SK2 channels immediately after the training enhanced contextual fear memory. We provided the evidence that the post-training cleavage of the SK2 channels was responsible for the observed bidirectional effect of SK2 channel blockade on memory consolidation. Thus, Lei-Dab7-injection before training impaired the C-terminal cleavage of SK2 channels, while Lei-Dab7 given immediately after training facilitated the C-terminal cleavage. Application of the synthetic peptide comprising a leucine-zipper domain of the C-terminal fragment to Jurkat cells impaired SK2 channel-mediated currents, indicating that the endogenously cleaved fragment might exert its effects on memory formation by blocking SK2 channel-mediated currents. Our present findings suggest that SK2 channel proteins contribute to synaptic plasticity and memory not only as ion channels but also by additionally generating a SK2 C-terminal fragment, involved in both processes. The modulation of fear memory by down-regulating SK2 C-terminal cleavage might have applicability in the treatment of anxiety disorders in which fear conditioning is enhanced.

  2. Bupivacaine inhibits large conductance, voltage- and Ca2+- activated K+ channels in human umbilical artery smooth muscle cells

    Science.gov (United States)

    Martín, Pedro; Enrique, Nicolás; Palomo, Ana R. Roldán; Rebolledo, Alejandro; Milesi, Veronica

    2012-01-01

    Bupivacaine is a local anesthetic compound belonging to the amino amide group. Its anesthetic effect is commonly related to its inhibitory effect on voltage-gated sodium channels. However, several studies have shown that this drug can also inhibit voltage-operated K+ channels by a different blocking mechanism. This could explain the observed contractile effects of bupivacaine on blood vessels. Up to now, there were no previous reports in the literature about bupivacaine effects on large conductance voltage- and Ca2+-activated K+ channels (BKCa). Using the patch-clamp technique, it is shown that bupivacaine inhibits single-channel and whole-cell K+ currents carried by BKCa channels in smooth muscle cells isolated from human umbilical artery (HUA). At the single-channel level bupivacaine produced, in a concentration- and voltage-dependent manner (IC50 324 µM at +80 mV), a reduction of single-channel current amplitude and induced a flickery mode of the open channel state. Bupivacaine (300 µM) can also block whole-cell K+ currents (~45% blockage) in which, under our working conditions, BKCa is the main component. This study presents a new inhibitory effect of bupivacaine on an ion channel involved in different cell functions. Hence, the inhibitory effect of bupivacaine on BKCa channel activity could affect different physiological functions where these channels are involved. Since bupivacaine is commonly used during labor and delivery, its effects on umbilical arteries, where this channel is highly expressed, should be taken into account. PMID:22688134

  3. PREFRONTAL CORTEX ACTIVATION DURING STORY ENCODING/RETRIEVAL: A MULTI-CHANNEL FUNCTIONAL NEAR-INFRARED SPECTROSCOPY STUDY

    Directory of Open Access Journals (Sweden)

    Sara eBasso Moro

    2013-12-01

    Full Text Available Encoding, storage and retrieval constitute three fundamental stages in information processing and memory. They allow for the creation of new memory traces, the maintenance and the consolidation of these traces over time, and the access and recover of the stored information from short or long-term memory. Functional near-infrared spectroscopy (fNIRS is a non-invasive neuroimaging technique that measures concentration changes of oxygenated-hemoglobin (O2Hb and deoxygenated-hemoglobin (HHb in cortical microcirculation blood vessels by means of the characteristic absorption spectra of hemoglobin in the near-infrared range. In the present study, we monitored, using a sixteen-channel fNIRS system, the hemodynamic response during the encoding and retrieval processes (EP and RP, respectively over the prefrontal cortex (PFC of thirteen healthy subjects (27.2±2.6 y. while were performing the Logical Memory Test (LMT of the Wechsler Memory Scale. A LMT-related PFC activation was expected; specifically, it was hypothesized a neural dissociation between EP and RP. The results showed a heterogeneous O2Hb/HHb response over the mapped area during the EP and the RP, with a O2Hb progressive and prominent increment in ventrolateral PFC since the beginning of the EP. During the RP a broader activation, including the ventrolateral PFC, the dorsolateral PFC and the frontopolar cortex, was observed. This could be explained by the different contributions of the PFC regions in the EP and the RP. Considering the fNIRS applicability for the hemodynamic monitoring during the LMT performance, this study has demonstrated that fNIRS could be utilized as a valuable clinical diagnostic tool, and that it has the potential to be adopted in patients with cognitive disorders or slight working memory deficits.

  4. Annual report on surveillance and maintenance activities at Oak Ridge National Laboratory, Oak Ridge, Tennessee, fiscal year 1996

    International Nuclear Information System (INIS)

    1996-11-01

    In fiscal year (FY) 1995, the sites and facilities from both the Remedial Action (RA) and Decontamination and Decommissioning (D and D) programs were combined to form the Oak Ridge National Laboratory (ORNL) Environmental Restoration (ER) Surveillance and Maintenance (S and M) Program. Surveillance and Maintenance activities were conducted throughout FY 1996 at the RA facilities. Overall, the RA S and M Program consists of approximately 650 acres that include 14 waste area groupings with approximately 200 sites. These sites include 46 major facilities, several leak and contaminated soil sites, 38 inactive tanks, approximately 50 environmental study areas and approximately 2,973 wells and boreholes. Site inspections were conducted at established frequencies on appropriate sites in the RA S and M Program in accordance with the established S and M FY 1996 Incentive Task Order (ITO)

  5. Activities for turbine maintenance: planning, implementation and evaluation of inspection results

    International Nuclear Information System (INIS)

    Azcue, J.; Sanchez, M. A.; Alvaro, M.

    2014-01-01

    The article will cover the most significant aspects of how Tecnatom carry out the turbine maintenance work, both from the point of view of the inspection itself as the pre-and post tasks associated. Thus, the issues that will be part of the article are the Manual Inspection Service (MISI) and the inspection database, ISI WEB. These tools are essential for planning field work and serve as a guide to run later turbine inspections in different plants. Following this introduction the inspection teams and the techniques used by Tecnatom will be described. finally, the article will contain a small mention about management studies of life and integrity analysis of the turbine that can be performed, as well as involvement in its safe operation that is derived from them. Ultimately, the intention is to provide a comprehensive approach to the issue in question, the turbine maintenance work, so that the reader can get an ideal of the whole process. (Author)

  6. Codes and standards in repair and replacement process. Importance of corrective action in maintenance activities

    International Nuclear Information System (INIS)

    Aoki, Takayuki; Takagi, Toshiyuki

    2015-01-01

    For creating a corrective measure plan for industrial plant components, it is very important to have a well-grounded basis. Therefore, this paper proposes a method for determining the following three elements of such a plan: an equipment to be repaired, a corrective measure method to be adopted, and a timing of its implementation using a rational approach. And the importance of corrective measure in plant maintenance is also discussed. (author)

  7. Search after new agents for hyperpolarization-activated and cyclic nucleotide-gated ion channels; Suche nach neuen Wirkstoffen fuer Hyperpolarisationsaktivierte und zyklisch Nukleotid-gesteuerte Ionenkanaele

    Energy Technology Data Exchange (ETDEWEB)

    Struenker, T.

    2005-12-01

    Rhythmic activity of single cells or cellular networks is a common feature of most organisms. Cellular rhythms govern the beating of the heart, cycles of sleep and wakefulness, breathing, and the release of hormones. The endogenous rhythmic activity of many neurons and cardiac relies on a complex interplay between several distinct ion channels. In particular, one type of ion channel plays a prominent role in the control of rhythmic electrical activity because it determines the frequency of the oscillations. The activity of the channels is thus setting the ''pace'' of the activity; therefore, these channels are often referred to as ''pacemaker'' channels. Despite their obvious physiological importance it hasn't been until a few years ago that the genes encoding pacemaker channels have been identified. Because both hyperpolarization and cyclic nucleotides are key elements that control their activity, pacemaker channels have now been designated hyperpolarization-activated and cyclic nucleotide-gated (HCN) channels. From a scientific as well as medical point of view, HCN channels are interesting drug targets. Only a few substances are known that specifically affect HCN channels. In the present study, a microtiter plate-based high throughput screening assay for HCN1 and HCN4 channels was developed. With this assay, known drugs for HCN channels were characterized. Subsequently, venoms of snails, spiders, scorpions, and snakes were screened for toxins affecting HCN channel activity. A few venoms were identified that possibly contain drugs that act on HCN channels. (orig.)

  8. Relative transmembrane segment rearrangements during BK channel activation resolved by structurally assigned fluorophore–quencher pairing

    Science.gov (United States)

    Pantazis, Antonios

    2012-01-01

    Voltage-activated proteins can sense, and respond to, changes in the electric field pervading the cell membrane by virtue of a transmembrane helix bundle, the voltage-sensing domain (VSD). Canonical VSDs consist of four transmembrane helices (S1–S4) of which S4 is considered a principal component because it possesses charged residues immersed in the electric field. Membrane depolarization compels the charges, and by extension S4, to rearrange with respect to the field. The VSD of large-conductance voltage- and Ca-activated K+ (BK) channels exhibits two salient inconsistencies from the canonical VSD model: (1) the BK channel VSD possesses an additional nonconserved transmembrane helix (S0); and (2) it exhibits a “decentralized” distribution of voltage-sensing charges, in helices S2 and S3, in addition to S4. Considering these unique features, the voltage-dependent rearrangements of the BK VSD could differ significantly from the standard model of VSD operation. To understand the mode of operation of this unique VSD, we have optically tracked the relative motions of the BK VSD transmembrane helices during activation, by manipulating the quenching environment of site-directed fluorescent labels with native and introduced Trp residues. Having previously reported that S0 and S4 diverge during activation, in this work we demonstrate that S4 also diverges from S1 and S2, whereas S2, compelled by its voltage-sensing charged residues, moves closer to S1. This information contributes spatial constraints for understanding the BK channel voltage-sensing process, revealing the structural rearrangements in a non-canonical VSD. PMID:22802360

  9. PRRT2 controls neuronal excitability by negatively modulating Na+ channel 1.2/1.6 activity.

    Science.gov (United States)

    Fruscione, Floriana; Valente, Pierluigi; Sterlini, Bruno; Romei, Alessandra; Baldassari, Simona; Fadda, Manuela; Prestigio, Cosimo; Giansante, Giorgia; Sartorelli, Jacopo; Rossi, Pia; Rubio, Alicia; Gambardella, Antonio; Nieus, Thierry; Broccoli, Vania; Fassio, Anna; Baldelli, Pietro; Corradi, Anna; Zara, Federico; Benfenati, Fabio

    2018-04-01

    See Lerche (doi:10.1093/brain/awy073) for a scientific commentary on this article.Proline-rich transmembrane protein 2 (PRRT2) is the causative gene for a heterogeneous group of familial paroxysmal neurological disorders that include seizures with onset in the first year of life (benign familial infantile seizures), paroxysmal kinesigenic dyskinesia or a combination of both. Most of the PRRT2 mutations are loss-of-function leading to haploinsufficiency and 80% of the patients carry the same frameshift mutation (c.649dupC; p.Arg217Profs*8), which leads to a premature stop codon. To model the disease and dissect the physiological role of PRRT2, we studied the phenotype of neurons differentiated from induced pluripotent stem cells from previously described heterozygous and homozygous siblings carrying the c.649dupC mutation. Single-cell patch-clamp experiments on induced pluripotent stem cell-derived neurons from homozygous patients showed increased Na+ currents that were fully rescued by expression of wild-type PRRT2. Closely similar electrophysiological features were observed in primary neurons obtained from the recently characterized PRRT2 knockout mouse. This phenotype was associated with an increased length of the axon initial segment and with markedly augmented spontaneous and evoked firing and bursting activities evaluated, at the network level, by multi-electrode array electrophysiology. Using HEK-293 cells stably expressing Nav channel subtypes, we demonstrated that the expression of PRRT2 decreases the membrane exposure and Na+ current of Nav1.2/Nav1.6, but not Nav1.1, channels. Moreover, PRRT2 directly interacted with Nav1.2/Nav1.6 channels and induced a negative shift in the voltage-dependence of inactivation and a slow-down in the recovery from inactivation. In addition, by co-immunoprecipitation assays, we showed that the PRRT2-Nav interaction also occurs in brain tissue. The study demonstrates that the lack of PRRT2 leads to a hyperactivity of voltage

  10. PRRT2 controls neuronal excitability by negatively modulating Na+ channel 1.2/1.6 activity

    Science.gov (United States)

    Fruscione, Floriana; Valente, Pierluigi; Sterlini, Bruno; Romei, Alessandra; Baldassari, Simona; Fadda, Manuela; Prestigio, Cosimo; Giansante, Giorgia; Sartorelli, Jacopo; Rossi, Pia; Rubio, Alicia; Gambardella, Antonio; Nieus, Thierry; Broccoli, Vania; Fassio, Anna; Baldelli, Pietro; Corradi, Anna; Zara, Federico

    2018-01-01

    Abstract See Lerche (doi:10.1093/brain/awy073) for a scientific commentary on this article. Proline-rich transmembrane protein 2 (PRRT2) is the causative gene for a heterogeneous group of familial paroxysmal neurological disorders that include seizures with onset in the first year of life (benign familial infantile seizures), paroxysmal kinesigenic dyskinesia or a combination of both. Most of the PRRT2 mutations are loss-of-function leading to haploinsufficiency and 80% of the patients carry the same frameshift mutation (c.649dupC; p.Arg217Profs*8), which leads to a premature stop codon. To model the disease and dissect the physiological role of PRRT2, we studied the phenotype of neurons differentiated from induced pluripotent stem cells from previously described heterozygous and homozygous siblings carrying the c.649dupC mutation. Single-cell patch-clamp experiments on induced pluripotent stem cell-derived neurons from homozygous patients showed increased Na+ currents that were fully rescued by expression of wild-type PRRT2. Closely similar electrophysiological features were observed in primary neurons obtained from the recently characterized PRRT2 knockout mouse. This phenotype was associated with an increased length of the axon initial segment and with markedly augmented spontaneous and evoked firing and bursting activities evaluated, at the network level, by multi-electrode array electrophysiology. Using HEK-293 cells stably expressing Nav channel subtypes, we demonstrated that the expression of PRRT2 decreases the membrane exposure and Na+ current of Nav1.2/Nav1.6, but not Nav1.1, channels. Moreover, PRRT2 directly interacted with Nav1.2/Nav1.6 channels and induced a negative shift in the voltage-dependence of inactivation and a slow-down in the recovery from inactivation. In addition, by co-immunoprecipitation assays, we showed that the PRRT2-Nav interaction also occurs in brain tissue. The study demonstrates that the lack of PRRT2 leads to a hyperactivity of

  11. Channel Morphology and Bed Sediment Characteristics Before and After Habitat Enhancement Activities in the Uridil Property, Platte River, Nebraska, Water Years 2005-2008

    Science.gov (United States)

    Kinzel, Paul J.

    2009-01-01

    Fluvial geomorphic data were collected by the United States Geological Survey from July 2005 to June 2008 (a time period within water years 2005 to 2008) to monitor the effects of habitat enhancement activities conducted in the Platte River Whooping Crane Maintenance Trust's Uridil Property, located along the Platte River, Nebraska. The activities involved the removal of vegetation and sand from the tops of high permanent islands and the placement of the sand into the active river channel. This strategy was intended to enhance habitat for migratory water birds by lowering the elevations of the high islands, thereby eliminating a visual obstruction for roosting birds. It was also thought that the bare sand on the lowered island surfaces could serve as potential habitat for nesting water birds. Lastly, the project supplied a local source of sediment to the river to test the hypothesis that this material could contribute to the formation of lower sandbars and potential nesting sites downstream. Topographic surveys on the islands and along river transects were used to quantify the volume of removed sand and track the storage and movement of the introduced sand downstream. Sediment samples were also collected to map the spatial distribution of river bed sediment sizes before and after the management activities. While the project lowered the elevation of high islands, observations of the sand addition indicated the relatively fine-grained sand that was placed in the active river channel was rapidly transported by the flowing water. Topographic measurements made 3 months after the sand addition along transects in the area of sediment addition showed net aggradation over measurements made in 2005. In the year following the sand addition, 2007, elevated river flows from local rain events generally were accompanied by net degradation along transects within the area of sediment addition. In the spring of 2008, a large magnitude flow event of approximately 360 cubic meters per

  12. Light-activated control of protein channel assembly mediated by membrane mechanics

    Science.gov (United States)

    Miller, David M.; Findlay, Heather E.; Ces, Oscar; Templer, Richard H.; Booth, Paula J.

    2016-12-01

    Photochemical processes provide versatile triggers of chemical reactions. Here, we use a photoactivated lipid switch to modulate the folding and assembly of a protein channel within a model biological membrane. In contrast to the information rich field of water-soluble protein folding, there is only a limited understanding of the assembly of proteins that are integral to biological membranes. It is however possible to exploit the foreboding hydrophobic lipid environment and control membrane protein folding via lipid bilayer mechanics. Mechanical properties such as lipid chain lateral pressure influence the insertion and folding of proteins in membranes, with different stages of folding having contrasting sensitivities to the bilayer properties. Studies to date have relied on altering bilayer properties through lipid compositional changes made at equilibrium, and thus can only be made before or after folding. We show that light-activation of photoisomerisable di-(5-[[4-(4-butylphenyl)azo]phenoxy]pentyl)phosphate (4-Azo-5P) lipids influences the folding and assembly of the pentameric bacterial mechanosensitive channel MscL. The use of a photochemical reaction enables the bilayer properties to be altered during folding, which is unprecedented. This mechanical manipulation during folding, allows for optimisation of different stages of the component insertion, folding and assembly steps within the same lipid system. The photochemical approach offers the potential to control channel assembly when generating synthetic devices that exploit the mechanosensitive protein as a nanovalve.

  13. Structural mechanism underlying capsaicin binding and activation of TRPV1 ion channel

    Science.gov (United States)

    Cheng, Wei; Yang, Wei; Yu, Peilin; Song, Zhenzhen; Yarov-Yarovoy, Vladimir; Zheng, Jie

    2015-01-01

    Capsaicin bestows spiciness by activating TRPV1 channel with exquisite potency and selectivity. Capsaicin-bound channel structure was previously resolved by cryo-EM at 4.2-to-4.5 Å resolution, however important details required for mechanistic understandings are unavailable: capsaicin was registered as a small electron density, reflecting neither its chemical structure nor specific ligand-channel interactions. We obtained the missing atomic-level details by iterative computation, which were confirmed by systematic site-specific functional tests. We observed that the bound capsaicin takes “tail-up, head-down” configurations. The vanillyl and amide groups form specific interactions to anchor its bound position, while the aliphatic tail may sample a range of conformations, making it invisible in cryo-EM images. Capsaicin stabilizes the open state by “pull-and-contact” interactions between the vanillyl group and the S4-S5 linker. Our study provided a structural mechanism for the agonistic function of capsaicin and its analogs, and demonstrated an effective approach to obtain atomic level information from cryo-EM structures. PMID:26053297

  14. Molecular, biophysical, and pharmacological properties of calcium-activated chloride channels.

    Science.gov (United States)

    Kamaleddin, Mohammad Amin

    2018-02-01

    Calcium-activated chloride channels (CaCCs) are a family of anionic transmembrane ion channels. They are mainly responsible for the movement of Cl - and other anions across the biological membranes, and they are widely expressed in different tissues. Since the Cl - flow into or out of the cell plays a crucial role in hyperpolarizing or depolarizing the cells, respectively, the impact of intracellular Ca 2+ concentration on these channels is attracting a lot of attentions. After summarizing the molecular, biophysical, and pharmacological properties of CaCCs, the role of CaCCs in normal cellular functions will be discussed, and I will emphasize how dysregulation of CaCCs in pathological conditions can account for different diseases. A better understanding of CaCCs and a pivotal regulatory role of Ca 2+ can shed more light on the therapeutic strategies for different neurological disorders that arise from chloride dysregulation, such as asthma, cystic fibrosis, and neuropathic pain. © 2017 Wiley Periodicals, Inc.

  15. Twenty-four-hour exposure to altered blood flow modifies endothelial Ca2+-activated K+ channels in rat mesenteric arteries

    DEFF Research Database (Denmark)

    Hilgers, Rob H P; Janssen, Ger M J; Fazzi, Gregorio E

    2010-01-01

    We tested the hypothesis that changes in arterial blood flow modify the function of endothelial Ca2+-activated K+ channels [calcium-activated K+ channel (K(Ca)), small-conductance calcium-activated K+ channel (SK3), and intermediate calcium-activated K+ channel (IK1)] before arterial structural...... remodeling. In rats, mesenteric arteries were exposed to increased [+90%, high flow (HF)] or reduced blood flow [-90%, low flow (LF)] and analyzed 24 h later. There were no detectable changes in arterial structure or in expression level of endothelial nitric-oxide synthase, SK3, or IK1. Arterial relaxing...... arteries, the balance between the NO/prostanoid versus EDHF response was unaltered. However, the contribution of IK1 to the EDHF response was enhanced, as indicated by a larger effect of TRAM-34 and a larger residual NS309-induced relaxation in the presence of UCL 1684. Reduction of blood flow selectively...

  16. The role of entropic potential in voltage activation and K+ transport through Kv 1.2 channels

    Science.gov (United States)

    Wawrzkiewicz-Jałowiecka, Agata; Grzywna, Zbigniew J.

    2018-03-01

    We analyze the entropic effects of inner pore geometry changes of Kv 1.2 channel during membrane depolarization and their implications for the rate of transmembrane transport of potassium ions. We base this on the idea that spatial confinements within the channel pore give rise to entropic barriers which can both effectively affect the stability of open macroconformation and influence channel's ability to conduct the potassium ions through the membrane. First, we calculate the differences in entropy between voltage-activated and resting states of the channel. As a template, we take a set of structures of channel pore in an open state at different membrane potentials generated in our previous research. The obtained results indicate that tendency to occupy open states at membrane depolarization is entropy facilitated. Second, we describe the differences in rates of K+ transport through the channel pore at different voltages based on the results of appropriate random walk simulations in entropic and electric potentials. The simulated single channel currents (I) suggest that the geometry changes during membrane depolarization are an important factor contributing to the observed flow of potassium ions through the channel. Nevertheless, the charge distribution within the channel pore (especially at the extracellular entrance) seems most prominent for the observed I/Imax relation at a qualitative level at analyzed voltages.

  17. Maintenance management systems

    International Nuclear Information System (INIS)

    Rohan, M. de

    1989-01-01

    This paper is concerned principally with Maintenance Management systems and their effective introduction into organisations. Maintenance improvement is basically a problem of managing the maintenance department in the broadest sense. Improvement does not only lie in the area of special techniques, systems or procedures; although they are valuable tools, but rather in a balanced attack, carefully guided by management. Over recent years, maintenance systems have received the major emphasis and in many instances the selection of the system has become a pre-occupation, whereas the importance of each maintenance function must be recognised and good management practices applied to all maintenance activities. The ingredients for success in the implementation of maintenance management systems are summarised as: having a management committee, clear objectives, project approach using project management techniques and an enthusiastic leader, user managed and data processing supported project, realistic budget and an understanding of the financial audit requirements. (author)

  18. Activation of mutated TRPA1 ion channel by resveratrol in human prostate cancer associated fibroblasts (CAF).

    Science.gov (United States)

    Vancauwenberghe, Eric; Noyer, Lucile; Derouiche, Sandra; Lemonnier, Loïc; Gosset, Pierre; Sadofsky, Laura R; Mariot, Pascal; Warnier, Marine; Bokhobza, Alexandre; Slomianny, Christian; Mauroy, Brigitte; Bonnal, Jean-Louis; Dewailly, Etienne; Delcourt, Philippe; Allart, Laurent; Desruelles, Emilie; Prevarskaya, Natalia; Roudbaraki, Morad

    2017-08-01

    Previous studies showed the effects of resveratrol (RES) on several cancer cells, including prostate cancer (PCa) cell apoptosis without taking into consideration the impact of the tumor microenvironment (TME). The TME is composed of cancer cells, endothelial cells, blood cells, and cancer-associated fibroblasts (CAF), the main source of growth factors. The latter cells might modify in the TME the impact of RES on tumor cells via secreted factors. Recent data clearly show the impact of CAF on cancer cells apoptosis resistance via secreted factors. However, the effects of RES on PCa CAF have not been studied so far. We have investigated here for the first time the effects of RES on the physiology of PCa CAF in the context of TME. Using a prostate cancer CAF cell line and primary cultures of CAF from prostate cancers, we show that RES activates the N-terminal mutated Transient Receptor Potential Ankyrin 1 (TRPA1) channel leading to an increase in intracellular calcium concentration and the expression and secretion of growth factors (HGF and VEGF) without inducing apoptosis in these cells. Interestingly, in the present work, we also show that when the prostate cancer cells were co-cultured with CAF, the RES-induced cancer cell apoptosis was reduced by 40%, an apoptosis reduction canceled in the presence of the TRPA1 channel inhibitors. The present work highlights CAF TRPA1 ion channels as a target for RES and the importance of the channel in the epithelial-stromal crosstalk in the TME leading to resistance to the RES-induced apoptosis. © 2017 Wiley Periodicals, Inc.

  19. Single amino acids in the carboxyl terminal domain of aquaporin-1 contribute to cGMP-dependent ion channel activation

    Directory of Open Access Journals (Sweden)

    Yool Andrea J

    2003-10-01

    Full Text Available Abstract Background Aquaporin-1 (AQP1 functions as an osmotic water channel and a gated cation channel. Activation of the AQP1 ion conductance by intracellular cGMP was hypothesized to involve the carboxyl (C- terminus, based on amino acid sequence alignments with cyclic-nucleotide-gated channels and cGMP-selective phosphodiesterases. Results Voltage clamp analyses of human AQP1 channels expressed in Xenopus oocytes demonstrated that the nitric oxide donor, sodium nitroprusside (SNP; 3–14 mM activated the ionic conductance response in a dose-dependent manner. Block of soluble guanylate cyclase prevented the response. Enzyme immunoassays confirmed a linear dose-dependent relationship between SNP and the resulting intracellular cGMP levels (up to 1700 fmol cGMP /oocyte at 14 mM SNP. Results here are the first to show that the efficacy of ion channel activation is decreased by mutations of AQP1 at conserved residues in the C-terminal domain (aspartate D237 and lysine K243. Conclusions These data support the idea that the limited amino acid sequence similarities found between three diverse classes of cGMP-binding proteins are significant to the function of AQP1 as a cGMP-gated ion channel, and provide direct evidence for the involvement of the AQP1 C-terminal domain in cGMP-mediated ion channel activation.

  20. Maintenance of a physically active lifestyle after pulmonary rehabilitation in patients with COPD: a qualitative study toward motivational factors.

    Science.gov (United States)

    Stewart, Kelly F J; Meis, Jessie J M; van de Bool, Coby; Janssen, Daisy J A; Kremers, Stef P J; Schols, Annemie M W J

    2014-09-01

    To explore determinants of behavior change maintenance of a physically active lifestyle in patients with chronic obstructive pulmonary disease (COPD) 8-11 months after completion of a 4-month outpatient pulmonary rehabilitation program. A qualitative descriptive study of semistructured interviews. Pulmonary rehabilitation assessment center. Patients with COPD. Semistructured interviews until data saturation, coded by 2 independent researchers. Patients were classified as responder (maintenance or improvement) or nonresponder (relapse or decrease), based on 3 quantitative variables reflecting exercise capacity (Constant Work Rate Test), health-related quality of life (Short-Form health survey [SF-36]), and self-management abilities (Self-Management Ability Scale [SMAS-30/Version 2]). Mean (SD) forced expiratory volume in the first second (FEV1) among interviewees was 52.5% (14.4%) predicted and the mean age was 63.5 years (range: 45-78). The group consisted of 15 responders and 7 nonresponders. Physical limitations reduced competence to engage in an active lifestyle and responders appeared to experience higher levels of perceived competence. Social support was found important and the experienced understanding from fellow patients made exercising together enjoyable. Particularly, responders expressed autonomous motivation and said they exercised because of the benefits they gain from it. Unexpectedly, only responders also experienced controlled motivation. Perceived competence and autonomous motivation are important determinants for maintenance of an active lifestyle in patients with COPD. In contrast to common theoretical assumptions, a certain threshold level of controlled motivation may remain important in maintaining a physically active lifestyle after a pulmonary rehabilitation program. Copyright © 2014 AMDA – The Society for Post-Acute and Long-Term Care Medicine. Published by Elsevier Inc. All rights reserved.

  1. Activation of the chemosensing transient receptor potential channel A1 (TRPA1) by alkylating agents.

    Science.gov (United States)

    Stenger, Bernhard; Zehfuss, Franziska; Mückter, Harald; Schmidt, Annette; Balszuweit, Frank; Schäfer, Eva; Büch, Thomas; Gudermann, Thomas; Thiermann, Horst; Steinritz, Dirk

    2015-09-01

    The transient receptor potential ankyrin 1 (TRPA1) cation channel is expressed in different tissues including skin, lung and neuronal tissue. Recent reports identified TRPA1 as a sensor for noxious substances, implicating a functional role in the molecular toxicology. TRPA1 is activated by various potentially harmful electrophilic substances. The chemical warfare agent sulfur mustard (SM) is a highly reactive alkylating agent that binds to numerous biological targets. Although SM is known for almost 200 years, detailed knowledge about the pathophysiology resulting from exposure is lacking. A specific therapy is not available. In this study, we investigated whether the alkylating agent 2-chloroethyl-ethylsulfide (CEES, a model substance for SM-promoted effects) and SM are able to activate TRPA1 channels. CEES induced a marked increase in the intracellular calcium concentration ([Ca(2+)]i) in TRPA1-expressing but not in TRPA1-negative cells. The TRP-channel blocker AP18 diminished the CEES-induced calcium influx. HEK293 cells permanently expressing TRPA1 were more sensitive toward cytotoxic effects of CEES compared with wild-type cells. At low CEES concentrations, CEES-induced cytotoxicity was prevented by AP18. Proof-of-concept experiments using SM resulted in a pronounced increase in [Ca(2+)]i in HEK293-A1-E cells. Human A549 lung epithelial cells, which express TRPA1 endogenously, reacted with a transient calcium influx in response to CEES exposure. The CEES-dependent calcium response was diminished by AP18. In summary, our results demonstrate that alkylating agents are able to activate TRPA1. Inhibition of TRPA1 counteracted cellular toxicity and could thus represent a feasible approach to mitigate SM-induced cell damage.

  2. Initial integration of accident safety, waste management, recycling, effluent, and maintenance considerations for low-activation materials

    International Nuclear Information System (INIS)

    Piet, S.J.; Herring, J.S.; Cheng, E.T.; Fetter, S.

    1991-01-01

    A true low-activation material should ideally achieve all of the following objectives: 1. The possible prompt dose at the site boundary from 100% release of the inventory should be <2 Sv (200 rem); hence, the design would be inherently safe in that no possible accident could result in prompt radiation fatalities. 2. The possible cancers from realistic releases should be limited such that the accident risk is <0.1%/yr of the existing background cancer risk to local residents. This includes consideration of elemental volatility. 3. The decay heat should be limited so that active mitigative measures are not needed to protect the investment from cooling transients; hence, the design would be passively safe with respect to decay heat. 4. Used materials could be either recycled or disposed of as near- surface waste. 5. Hands-on maintenance should be possible around coolant system piping and components such as the heat exchanger. 6. Effluent of activation products should be minor compared to the major challenge of limiting tritium effluents. The most recent studies in these areas are used to determine which individual elements and engineering materials are low activation. Grades from A (best) to G (worst) are given to each element in the areas of accident safety, recycling, and waste management. Structure/fluid combinations are examined for low-activation effluents and out-of-blanket maintenance. The lowest activation structural materials are silicon carbide, vanadium alloys, and ferritic steels. Impurities and minor alloying constituents must be carefully considered. The lowest activation coolants are helium, water, FLiBe, and lithium. The lowest activation breeders are lithium, lithium oxide, lithium silicate, and FLiBe. Designs focusing on these truly low-activation materials will help achieve the excellent safety and environmental potential of fusion energy

  3. Neuronal fast activating and meningeal silent modulatory BK channel splice variants cloned from rat

    DEFF Research Database (Denmark)

    Poulsen, Asser Nyander; Jansen-Olesen, Inger; Olesen, Jes

    2011-01-01

    The big conductance calcium-activated K(+) channel (BK) is involved in regulating neuron and smooth muscle cell excitability. Functional diversity of BK is generated by alpha-subunit splice variation and co-expression with beta subunits. Here, we present six different splice combinations cloned...... and RCK2 (4 aa at SS1) and upstream of the calcium "bowl" (27 aa at SS4). Two other truncated variants, X2(92) and X2(188), lacking the intracellular C-terminal (stop downstream of S6), were cloned from cerebral vascular/meningeal tissue. They appear non-functional as no current expression was observed...

  4. Two Marine Cyanobacterial Aplysiatoxin Polyketides, Neo-debromoaplysiatoxin A and B, with K+ Channel Inhibition Activity.

    Science.gov (United States)

    Han, Bing-Nan; Liang, Ting-Ting; Keen, Lawrence Jordan; Fan, Ting-Ting; Zhang, Xiao-Dan; Xu, Lin; Zhao, Qi; Wang, Shu-Ping; Lin, Hou-Wen

    2018-02-02

    The isolation and structure elucidation of two cyanobacterial debromoaplysiatoxin (DAT) analogues, neo-debromoaplysiatoxin A (1) and neo-debromoaplysiatoxin B (2), were reported and found to possess 6/10/6 and 6/6/6 fused-ring systems, respectively, which are rarely seen among aplysiatoxins. Both compounds exhibited potent blocking activity against Kv1.5 with IC 50 values of 6.94 ± 0.26 and 0.30 ± 0.05 μM, respectively. These findings suggest the potential of aplysiatoxin analogues in modulating ionic channels and also provide links between the DAT target, protein kinase C, and cell regulation.

  5. A unifying mechanism for cancer cell death through ion channel activation by HAMLET.

    Science.gov (United States)

    Storm, Petter; Klausen, Thomas Kjaer; Trulsson, Maria; Ho C S, James; Dosnon, Marion; Westergren, Tomas; Chao, Yinxia; Rydström, Anna; Yang, Henry; Pedersen, Stine Falsig; Svanborg, Catharina

    2013-01-01

    Ion channels and ion fluxes control many aspects of tissue homeostasis. During oncogenic transformation, critical ion channel functions may be perturbed but conserved tumor specific ion fluxes remain to be defined. Here we used the tumoricidal protein-lipid complex HAMLET as a probe to identify ion fluxes involved in tumor cell death. We show that HAMLET activates a non-selective cation current, which reached a magnitude of 2.74±0.88 nA within 1.43±0.13 min from HAMLET application. Rapid ion fluxes were essential for HAMLET-induced carcinoma cell death as inhibitors (amiloride, BaCl2), preventing the changes in free cellular Na(+) and K(+) concentrations also prevented essential steps accompanying carcinoma cell death, including changes in morphology, uptake, global transcription, and MAP kinase activation. Through global transcriptional analysis and phosphorylation arrays, a strong ion flux dependent p38 MAPK response was detected and inhibition of p38 signaling delayed HAMLET-induced death. Healthy, differentiated cells were resistant to HAMLET challenge, which was accompanied by innate immunity rather than p38-activation. The results suggest, for the first time, a unifying mechanism for the initiation of HAMLET's broad and rapid lethal effect on tumor cells. These findings are particularly significant in view of HAMLET's documented therapeutic efficacy in human studies and animal models. The results also suggest that HAMLET offers a two-tiered therapeutic approach, killing cancer cells while stimulating an innate immune response in surrounding healthy tissues.

  6. X-ray irradiation activates K+ channels via H2O2 signaling.

    Science.gov (United States)

    Gibhardt, Christine S; Roth, Bastian; Schroeder, Indra; Fuck, Sebastian; Becker, Patrick; Jakob, Burkhard; Fournier, Claudia; Moroni, Anna; Thiel, Gerhard

    2015-09-09

    Ionizing radiation is a universal tool in tumor therapy but may also cause secondary cancers or cell invasiveness. These negative side effects could be causally related to the human-intermediate-conductance Ca2+-activated-K+-channel (hIK), which is activated by X-ray irradiation and affects cell proliferation and migration. To analyze the signaling cascade downstream of ionizing radiation we use genetically encoded reporters for H2O2 (HyPer) and for the dominant redox-buffer glutathione (Grx1-roGFP2) to monitor with high spatial and temporal resolution, radiation-triggered excursions of H2O2 in A549 and HEK293 cells. The data show that challenging cells with ≥1 Gy X-rays or with UV-A laser micro-irradiation causes a rapid rise of H2O2 in the nucleus and in the cytosol. This rise, which is determined by the rate of H2O2 production and glutathione-buffering, is sufficient for triggering a signaling cascade that involves an elevation of cytosolic Ca2+ and eventually an activation of hIK channels.

  7. Loss of Sodium-Activated Potassium Channel Slack and FMRP Differentially Affect Social Behavior in Mice.

    Science.gov (United States)

    Bausch, Anne E; Ehinger, Rebekka; Straubinger, Julia; Zerfass, Patrick; Nann, Yvette; Lukowski, Robert

    2018-05-31

    The sodium-activated potassium channel Slack (Slo2.2) is widely expressed in central and peripheral neurons where it is supposed to shape firing properties important for neuronal excitability. Slack activity is enhanced by interaction with the Fragile-X-Mental-Retardation-Protein (FMRP) and loss of FMRP leads to decreased sodium-activated potassium currents in medial nucleus of the trapezoid body neurons of the Fmr1-knockout (KO) mouse representing a mouse model of the human Fragile-X-Syndrome (FXS) and autism. Autism is a frequent comorbidity of FXS, but it is unclear whether Slack is involved in autistic or related conditions of FXS in vivo. By applying a wide range of behavioral tests, we compared social and autism-related behaviors in Slack- and FMRP-deficient mice. In our hands, as expected, FMRP-deficiency causes autism-related behavioral changes in nesting and in a marble-burying test. In contrast, Slack-deficient males exhibited specific abnormalities in sociability in direct and indirect social interaction tests. Hence, we show for the first time that a proper Slack channel function is mandatory for normal social behavior in mice. Nevertheless, as deficits in social behaviors seem to occur independently from each other in FMRP and Slack null mutants, we conclude that Slack is not involved in the autistic phenotype of FMRP KO mice. Copyright © 2018 IBRO. Published by Elsevier Ltd. All rights reserved.

  8. Phosphorylation of Minichromosome Maintenance 3 (MCM3) by Checkpoint Kinase 1 (Chk1) Negatively Regulates DNA Replication and Checkpoint Activation.

    Science.gov (United States)

    Han, Xiangzi; Mayca Pozo, Franklin; Wisotsky, Jacob N; Wang, Benlian; Jacobberger, James W; Zhang, Youwei

    2015-05-08

    Mechanisms controlling DNA replication and replication checkpoint are critical for the maintenance of genome stability and the prevention or treatment of human cancers. Checkpoint kinase 1 (Chk1) is a key effector protein kinase that regulates the DNA damage response and replication checkpoint. The heterohexameric minichromosome maintenance (MCM) complex is the core component of mammalian DNA helicase and has been implicated in replication checkpoint activation. Here we report that Chk1 phosphorylates the MCM3 subunit of the MCM complex at Ser-205 under normal growth conditions. Mutating the Ser-205 of MCM3 to Ala increased the length of DNA replication track and shortened the S phase duration, indicating that Ser-205 phosphorylation negatively controls normal DNA replication. Upon replicative stress treatment, the inhibitory phosphorylation of MCM3 at Ser-205 was reduced, and this reduction was accompanied with the generation of single strand DNA, the key platform for ataxia telangiectasia mutated and Rad3-related (ATR) activation. As a result, the replication checkpoint is activated. Together, these data provide significant insights into the regulation of both normal DNA replication and replication checkpoint activation through the novel phosphorylation of MCM3 by Chk1. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  9. The action of blocking agents applied to the inner face of Ca(2+)-activated K+ channels from human erythrocytes.

    Science.gov (United States)

    Dunn, P M

    1998-09-15

    The actions of clotrimazole and cetiedil, two drugs known to inhibit the Gardos channel, have been studied on single intermediate conductance calcium-activated potassium (IKCa) channels in inside out patches from human red blood cells, and compared with those of TEA and Ba2+ applied to the cytoplasmic face of the membrane. TEA produced a fast block which was observed as a reduction in the amplitude of the single channel current. This effect was weakly voltage dependent with the fraction of the membrane potential sensed by TEA at its binding site (delta) of 0.18 and a Kd at 0 mV of 20.5 mM. Ba2+ was a very potent blocker of the channel, breaking the single channel activity up into bursts, inter-spersed with silent periods lasting several seconds. The effect of Ba2+ was very voltage sensitive, delta = 0.44, and a Kd at 0 mV of 0.15 microM. Clotrimazole applied to the inner face of the membrane at a concentration block resulting in bursts of channel activity separated by quiescent periods lasting many seconds. The effect of clotrimazole was mimicked by a quaternary derivative UCL 1559, in keeping with an action at the cytoplasmic face of the channel. A high concentration of cetiedil (100 microM) produced only a weak block of the channel. The kinetics of this action were very slow, with burst and inter-burst intervals lasting several minutes. While inhibition of the Gardos channel by cetiedil is unlikely to involve an intracellular site of action, if clotrimazole is able to penetrate the membrane, part of its effect may result from binding to an intracellular site on the channel.

  10. Calcium channel blocker prevents stress-induced activation of renin and aldosterone in conscious pig

    International Nuclear Information System (INIS)

    Ceremuzynski, L.K.; Klos, J.; Barcikowski, B.; Herbaczynska-Cedro, K.

    1991-01-01

    A considerable amount of data suggest the involvement of calcium-mediated processes in the activation of the renin-angiotensin-aldosterone (RAA) cascade. To investigate the effect of calcium-channel inhibition on the RAA system, the authors studied 21 conscious pigs. Blood renin and aldosterone levels increased by subjecting animals to 24 hours of immobilization stress. Renin and aldosterone levels were repeatedly measured by radioimmunoassay in blood samples taken periodically over 24 hours from a chronically implanted arterial cannula. Pretreatment of the animals (N = 11) with nisoldipine, 2 x 20 mg p.o. daily for 2 days before and on the day of immobilization, transiently attenuated the stress-induced increase of plasma renin activity and completely prevented the rise of aldosterone, as compared to nontreated controls (N = 10). The finding that nisoldipine suppresses RAA activation induced by a nonpharmacologic stimulus in the conscious intact animal may have clinical implications

  11. Building relationships for better maintenance.

    Science.gov (United States)

    Lee, Hackman Hon Yin; Scott, David

    2009-02-01

    Effective management of building maintenance is a vital ingredient in ensuring a high quality built environment for any building's occupiers. However, maintenance is not high on the list of priorities for most organisations. Communication between top management personnel looking at maintenance issues from a strategic standpoint and maintenance staff considering them at an operational level is often not as good as it should be. When planning maintenance activities maintenance personnel often draw too heavily on their technical experience and expertise without taking sufficient account of wider organisational objectives or consulting effectively with top management. Senior managers also regularly complain about lack of managerial input from maintenance departments. Such barriers contribute to communication difficulties between top management at a strategic level and maintenance personnel at an operational level. Identifying where the key differences lie in senior managers' and maintenance personnel's viewpoints on maintenance strategy can prove invaluable in achieving some convergence of opinions and optimising the efficiency of the overall building maintenance process.

  12. Inhibition of Calcium-Activated Chloride Channel ANO1/TMEM16A Suppresses Tumor Growth and Invasion in Human Lung Cancer.

    Directory of Open Access Journals (Sweden)

    Linghan Jia

    Full Text Available Lung cancer or pulmonary carcinoma is primarily derived from epithelial cells that are thin and line on the alveolar surfaces of the lung for gas exchange. ANO1/TMEM16A, initially identified from airway epithelial cells, is a member of Ca2+-activated Cl- channels (CaCCs that function to regulate epithelial secretion and cell volume for maintenance of ion and tissue homeostasis. ANO1/TMEM16A has recently been shown to be highly expressed in several epithelium originated carcinomas. However, the role of ANO1 in lung cancer remains unknown. In this study, we show that inhibition of calcium-activated chloride channel ANO1/TMEM16A suppresses tumor growth and invasion in human lung cancer. ANO1 is upregulated in different human lung cancer cell lines. Knocking-down ANO1 by small hairpin RNAs inhibited proliferation, migration and invasion of GLC82 and NCI-H520 cancel cells evaluated by CCK-8, would-healing, transwell and 3D soft agar assays. ANO1 protein is overexpressed in 77.3% cases of human lung adenocarcinoma tissues detected by immunohistochemistry. Furthermore, the tumor growth in nude mice implanted with GLC82 cells was significantly suppressed by ANO1 silencing. Taken together, our findings provide evidence that ANO1 overexpression contributes to tumor growth and invasion of lung cancer; and suppressing ANO1 overexpression may have therapeutic potential in lung cancer therapy.

  13. Anti-Convulsant Activity of Boerhaavia diffusa: Plausible Role of Calcium Channel Antagonism

    Directory of Open Access Journals (Sweden)

    Mandeep Kaur

    2011-01-01

    Full Text Available “Ethnopharmacological” use of roots of Boerhaavia diffusa (B. diffusa in the treatment of epilepsy in Nigerian folk medicine and reports showing the presence of a calcium channel antagonistic compound “liriodendrin” in its roots, led us to undertake the present study. The study was designed to investigate the methanolic root extract of B. diffusa and its different fractions including liriodendrin-rich fraction for exploring the possible role of liriodendrin in its anti-convulsant activity. Air-dried roots of B. diffusa were extracted with methanol by cold maceration. The methanol soluble fraction of extract thus obtained was successively extracted to obtain liriodendrin-rich fraction and two side fractions, that is, chloroform fraction and phenolic compound fraction. Anti-convulsant activity of methanolic extract (1000, 1500 and 2000 mg kg-1, intraperitoneally (i.p. and its different fractions, that is, liriodendrin-rich fraction (10, 20 and 40 mg kg-1, i.p., chloroform fraction (20 mg kg-1, i.p. and phenolic compound fraction (1 mg kg-1, i.p. were studied in pentylenetetrazol (PTZ-induced seizures (75 mg kg-1, i.p.. The crude methanolic extract of B. diffusa and only its liriodendrin-rich fraction showed a dose-dependent protection against PTZ-induced convulsions. The liriodendrin-rich fraction also showed significant protection against seizures induced by BAY k-8644. These findings reiterated the anti-convulsant activity of methanolic extract of B. diffusa roots. Furthermore, it can be concluded that the observed anti-convulsant activity was due to its calcium channel antagonistic action as this activity was retained only in the liodendrin-rich fraction, which has additionally been confirmed by significant anti-convulsant activity of liriodendrin-rich fraction in BAY k-8644-induced seizures.

  14. Receptor channel TRPC6 orchestrate the activation of human hepatic stellate cell under hypoxia condition

    International Nuclear Information System (INIS)

    Iyer, Soumya C; Kannan, Anbarasu; Gopal, Ashidha; Devaraj, Niranjali; Halagowder, Devaraj

    2015-01-01

    Hepatic stellate cells (HSCs), a specialized stromal cytotype have a great impact on the biological behaviors of liver diseases. Despite this fact, the underlying mechanism that regulates HSC still remains poorly understood. The aim of the present study was to understand the role of TRPC6 signaling in regulating the molecular mechanism of HSCs in response to hypoxia. In the present study we showed that under hypoxia condition, the upregulated Hypoxia Inducible Factor 1α (HIF1α) increases NICD activation, which in turn induces the expression of transient receptor potential channel 6 (TRPC6) in HSC line lx-2. TRPC6 causes a sustained elevation of intracellular calcium which is coupled with the activation of the calcineurin-nuclear factor of activated T-cell (NFAT) pathway which activates the synthesis of extracellular matrix proteins. TRPC6 also activates SMAD2/3 dependent TGF-β signaling in facilitating upregulated expression of αSMA and collagen. As activated HSCs may be a suitable target for HCC therapy and targeting these cells rather than the HCC cells may result in a greater response. Collectively, our studies indicate for the first time the detailed mechanism of activation of HSC through TRPC6 signaling and thus being a promising therapeutic target. - Highlights: • HIF1α increases NICD, induces TRPC6 in lx2 cells. • TRPC6 a novel regulator in the activation of HSC. • HSCs as target for HCC therapy

  15. Receptor channel TRPC6 orchestrate the activation of human hepatic stellate cell under hypoxia condition

    Energy Technology Data Exchange (ETDEWEB)

    Iyer, Soumya C, E-mail: chidambaram.soumya@gmail.com [Unit of Biochemistry, Department of Zoology, School of Life Sciences, University of Madras, Guindy Campus, Chennai 600025, Tamilnadu (India); Kannan, Anbarasu [Department of Biochemistry, University of Madras, Guindy Campus, Chennai 600025, Tamilnadu (India); Gopal, Ashidha [Unit of Biochemistry, Department of Zoology, School of Life Sciences, University of Madras, Guindy Campus, Chennai 600025, Tamilnadu (India); Devaraj, Niranjali [Department of Biochemistry, University of Madras, Guindy Campus, Chennai 600025, Tamilnadu (India); Halagowder, Devaraj [Unit of Biochemistry, Department of Zoology, School of Life Sciences, University of Madras, Guindy Campus, Chennai 600025, Tamilnadu (India)

    2015-08-01

    Hepatic stellate cells (HSCs), a specialized stromal cytotype have a great impact on the biological behaviors of liver diseases. Despite this fact, the underlying mechanism that regulates HSC still remains poorly understood. The aim of the present study was to understand the role of TRPC6 signaling in regulating the molecular mechanism of HSCs in response to hypoxia. In the present study we showed that under hypoxia condition, the upregulated Hypoxia Inducible Factor 1α (HIF1α) increases NICD activation, which in turn induces the expression of transient receptor potential channel 6 (TRPC6) in HSC line lx-2. TRPC6 causes a sustained elevation of intracellular calcium which is coupled with the activation of the calcineurin-nuclear factor of activated T-cell (NFAT) pathway which activates the synthesis of extracellular matrix proteins. TRPC6 also activates SMAD2/3 dependent TGF-β signaling in facilitating upregulated expression of αSMA and collagen. As activated HSCs may be a suitable target for HCC therapy and targeting these cells rather than the HCC cells may result in a greater response. Collectively, our studies indicate for the first time the detailed mechanism of activation of HSC through TRPC6 signaling and thus being a promising therapeutic target. - Highlights: • HIF1α increases NICD, induces TRPC6 in lx2 cells. • TRPC6 a novel regulator in the activation of HSC. • HSCs as target for HCC therapy.

  16. Annual summary report on the surveillance and maintenance activities for the Oak Rige National Laboratory Environmental Restoration Program for fiscal year 1995

    International Nuclear Information System (INIS)

    1995-11-01

    This Annual Summary Report on the Surveillance and Maintenance Activities for the Oak Ridge National Laboratory Environmental Restoration Program for Fiscal Year 1995 was prepared to communicate the accomplishments of the Program during fiscal year 1995. This work was performed under work breakdown structure element 1.4.12.6.1.14.20 (activity data sheet 3314, ''Remedial Action Surveillance and Maintenance''). Publication of this document meets the Life Cycle Baseline milestone date of November 30, 1995. This document provides the accomplishments for both the Remedial Action and Decontamination and Decommissioning Surveillance and Maintenance programs

  17. A framework for the design of a voice-activated, intelligent, and hypermedia-based aircraft maintenance manual

    Science.gov (United States)

    Patankar, Manoj Shashikant

    Federal Aviation Regulations require Aviation Maintenance Technicians (AMTs) to refer to approved maintenance manuals when performing maintenance on airworthy aircraft. Because these manuals are paper-based, larger the size of the aircraft, more cumbersome are the manuals. Federal Aviation Administration (FAA) recognized the difficulties associated with the use of large manuals and conducted studies on the use of electronic media as an alternative to the traditional paper format. However, these techniques do not employ any artificial intelligence technologies and the user interface is limited to either a keyboard or a stylus pen. The primary emphasis of this research was to design a generic framework that would allow future development of voice-activated, intelligent, and hypermedia-based aircraft maintenance manuals. A prototype (VIHAMS-Voice-activated, Intelligent, and Hypermedia-based Aircraft Maintenance System) was developed, as a secondary emphasis, using the design and development techniques that evolved from this research. An evolutionary software design approach was used to design the proposed framework and the structured rapid prototyping technique was used to produce the VIHAMS prototype. VoiceAssist by Creative Labs was used to provide the voice interface so that the users (AMTs) could keep their hands free to work on the aircraft while maintaining complete control over the computer through discrete voice commands. KnowledgePro for Windows sp{TM}, an expert system shell, provided "intelligence" to the prototype. As a result of this intelligence, the system provided expert guidance to the user. The core information contained in conventional manuals was available in a hypermedia format. The prototype's operating hardware included a notebook computer with a fully functional audio system. An external microphone and the built-in speaker served as the input and output devices (along with the color monitor), respectively. Federal Aviation Administration

  18. Noradrenergic mechanisms and high blood pressure maintenance in genetic hypertension: The role of Gi proteins and voltage-dependent calcium channels

    Czech Academy of Sciences Publication Activity Database

    Zicha, Josef; Pintérová, Mária; Líšková, Silvia; Dobešová, Zdenka; Kuneš, Jaroslav

    2007-01-01

    Roč. 29, č. 4 (2007), s. 229-229 ISSN 1064-1963. [International symposium on SHR /12./. 20.10.2006-21.10.2006, Kyoto] R&D Projects: GA MZd(CZ) NR7786 Institutional research plan: CEZ:AV0Z50110509 Keywords : genetic hypertension * noradrenergic mechanisms * Gi proteins * voltage-dependent calcium channels Subject RIV: FA - Cardiovascular Diseases incl. Cardiotharic Surgery

  19. A company-wide activity to grow safety culture in a maintenance department of nuclear power plant

    International Nuclear Information System (INIS)

    Fukui, Hirokazu; Sugiman, Toshio

    2008-01-01

    This study aims to conduct a field survey to collect specific instances of 'learning activity' that would help develop a safety culture in a maintenance department of nuclear power plant, with 'activity theory' as a theoretical base, and to derive a conceptual model that portrays these instances as part of comprehensive organizational activities. First, the field survey found instances where an accident that occurred a few years ago provided momentum; a company-wide activity is being implemented in order to fulfill the requests of field workers so that their requests for improvements are positively honored and budgets are allocated to measures designed to accommodate these requests. This paper calls this company-wide activity 'an activity to materialize the requests for improvements in the workplace' and considers it to be a typical instance of learning activity. Second, the field survey found that, in the workplace under discussion, a daily meeting was held in a unique way and a study session was conducted frequently and regularly. Last, we came up with a conceptual model for the entire aspect of organizational activities in which we could position the above instances as well as the results of our past field surveys. This model conceptualizes organizational activities from the two aspects of 'learning activity (primary learning activity)' and 'management activity) and characterizes an activity that would transform the existing organizational activities into ones that could not be even predicted as 'secondary learning activity'. 'An activity to materialize the requests for improvement in the workplace' was taken as falling under the category of secondary learning activity. (author)

  20. NS309 decreases rat detrusor smooth muscle membrane potential and phasic contractions by activating SK3 channels

    Science.gov (United States)

    Parajuli, Shankar P; Hristov, Kiril L; Soder, Rupal P; Kellett, Whitney F; Petkov, Georgi V

    2013-01-01

    Background and Purpose Overactive bladder (OAB) is often associated with abnormally increased detrusor smooth muscle (DSM) contractions. We used NS309, a selective and potent opener of the small or intermediate conductance Ca2+-activated K+ (SK or IK, respectively) channels, to evaluate how SK/IK channel activation modulates DSM function. Experimental Approach We employed single-cell RT-PCR, immunocytochemistry, whole cell patch-clamp in freshly isolated rat DSM cells and isometric tension recordings of isolated DSM strips to explore how the pharmacological activation of SK/IK channels with NS309 modulates DSM function. Key Results We detected SK3 but not SK1, SK2 or IK channels expression at both mRNA and protein levels by RT-PCR and immunocytochemistry in DSM single cells. NS309 (10 μM) significantly increased the whole cell SK currents and hyperpolarized DSM cell resting membrane potential. The NS309 hyperpolarizing effect was blocked by apamin, a selective SK channel inhibitor. NS309 inhibited the spontaneous phasic contraction amplitude, force, frequency, duration and tone of isolated DSM strips in a concentration-dependent manner. The inhibitory effect of NS309 on spontaneous phasic contractions was blocked by apamin but not by TRAM-34, indicating no functional role of the IK channels in rat DSM. NS309 also significantly inhibited the pharmacologically and electrical field stimulation-induced DSM contractions. Conclusions and Implications Our data reveal that SK3 channel is the main SK/IK subtype in rat DSM. Pharmacological activation of SK3 channels with NS309 decreases rat DSM cell excitability and contractility, suggesting that SK3 channels might be potential therapeutic targets to control OAB associated with detrusor overactivity. PMID:23145946

  1. Evaluation channel performance in multichannel environments

    NARCIS (Netherlands)

    Gensler, S.; Dekimpe, M.; Skiera, B.

    2007-01-01

    Evaluating channel performance is crucial for actively managing multiple sales channels, and requires understanding the customers' channel preferences. Two key components of channel performance are (i) the existing customers' intrinsic loyalty to a particular channel and (ii) the channel's ability

  2. Nanomolar bifenthrin alters synchronous Ca2+ oscillations and cortical neuron development independent of sodium channel activity.

    Science.gov (United States)

    Cao, Zhengyu; Cui, Yanjun; Nguyen, Hai M; Jenkins, David Paul; Wulff, Heike; Pessah, Isaac N

    2014-04-01

    Bifenthrin, a relatively stable type I pyrethroid that causes tremors and impairs motor activity in rodents, is broadly used. We investigated whether nanomolar bifenthrin alters synchronous Ca(2+) oscillations (SCOs) necessary for activity-dependent dendritic development. Primary mouse cortical neurons were cultured 8 or 9 days in vitro (DIV), loaded with the Ca(2+) indicator Fluo-4, and imaged using a Fluorescence Imaging Plate Reader Tetra. Acute exposure to bifenthrin rapidly increased the frequency of SCOs by 2.7-fold (EC50 = 58 nM) and decreased SCO amplitude by 36%. Changes in SCO properties were independent of modifications in voltage-gated sodium channels since 100 nM bifenthrin had no effect on the whole-cell Na(+) current, nor did it influence neuronal resting membrane potential. The L-type Ca(2+) channel blocker nifedipine failed to ameliorate bifenthrin-triggered SCO activity. By contrast, the metabotropic glutamate receptor (mGluR)5 antagonist MPEP [2-methyl-6-(phenylethynyl)pyridine] normalized bifenthrin-triggered increase in SCO frequency without altering baseline SCO activity, indicating that bifenthrin amplifies mGluR5 signaling independent of Na(+) channel modification. Competitive [AP-5; (-)-2-amino-5-phosphonopentanoic acid] and noncompetitive (dizocilpine, or MK-801 [(5S,10R)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate]) N-methyl-d-aspartate antagonists partially decreased both basal and bifenthrin-triggered SCO frequency increase. Bifenthrin-modified SCO rapidly enhanced the phosphorylation of cAMP response element-binding protein (CREB). Subacute (48 hours) exposure to bifenthrin commencing 2 DIV-enhanced neurite outgrowth and persistently increased SCO frequency and reduced SCO amplitude. Bifenthrin-stimulated neurite outgrowth and CREB phosphorylation were dependent on mGluR5 activity since MPEP normalized both responses. Collectively these data identify a new mechanism by which bifenthrin potently alters Ca(2

  3. Levcromakalim- and isoprenaline-induced relaxation of human isolated airways--role of the epithelium and of K+ channel activation.

    Science.gov (United States)

    Black, J L; Johnson, P R; McKay, K O; Carey, D; Armour, C L

    1994-06-01

    In this study we have investigated the mechanism of action of levcromakalim and isoprenaline in human isolated airways with respect to the K+ channels they activate and the possibility that these smooth muscle relaxants activate K+ channels on the airway epithelium. Mechanical removal of the epithelial layer (mean percentage of epithelium present 20 +/- 3%, n = 20 tissues) did not affect the relaxation responses to levcromakalim or isoprenaline, either in terms of maximal relaxation or sensitivity. Whilst having no effect on isoprenaline-induced relaxation, studied from basal tone, the ATP-sensitive K+ channel blocker BRL 31660 (10, 30 and 50 microM) reduced relaxation responses induced (from basal tone) by levcromakalim from 74 +/- 6% (of the maximal response to isoprenaline) to 48 +/- 12% (n = 7), 9 +/- 9% (n = 4) and 0 (n = 4), respectively. Charybdotoxin, a blocker of high conductance Ca(2+)-activated K+ channels, at concentrations of 30 and 100 nM, had no effect on either levcromakalim- or or isoprenaline-induced relaxation responses and yet charybdotoxin was active at KCa channels in outside-out patches of hippocampal granule cells. Moreover, tetraethylammonium (10 mM) inhibited neither isoprenaline- nor levcromakalim-induced relaxation. This study has demonstrated that the relaxation responses elicited in human bronchus to isoprenaline and levcromakalim are likely to be the result of direct effects on the smooth muscle with no contribution from epithelial receptors or K+ channels. The actions of levcromakalim appear to be mediated only via activation of KATP channels. Further, we have made the important observation that, under the experimental conditions of our study, isoprenaline does not activate the KCa channel to produce relaxation in human bronchus.

  4. Mechanisms underlying the cardiac pacemaker: the role of SK4 calcium-activated potassium channels.

    Science.gov (United States)

    Weisbrod, David; Khun, Shiraz Haron; Bueno, Hanna; Peretz, Asher; Attali, Bernard

    2016-01-01

    The proper expression and function of the cardiac pacemaker is a critical feature of heart physiology. The sinoatrial node (SAN) in human right atrium generates an electrical stimulation approximately 70 times per minute, which propagates from a conductive network to the myocardium leading to chamber contractions during the systoles. Although the SAN and other nodal conductive structures were identified more than a century ago, the mechanisms involved in the generation of cardiac automaticity remain highly debated. In this short review, we survey the current data related to the development of the human cardiac conduction system and the various mechanisms that have been proposed to underlie the pacemaker activity. We also present the human embryonic stem cell-derived cardiomyocyte system, which is used as a model for studying the pacemaker. Finally, we describe our latest characterization of the previously unrecognized role of the SK4 Ca(2+)-activated K(+) channel conductance in pacemaker cells. By exquisitely balancing the inward currents during the diastolic depolarization, the SK4 channels appear to play a crucial role in human cardiac automaticity.

  5. Proceedings of the fourth international conference on CANDU maintenance

    International Nuclear Information System (INIS)

    1997-01-01

    These proceedings record the information presented at the 4th International Conference on CANDU Maintenance held November 16-18,1997 in Toronto, Canada. The papers for these proceedings were prepared on component maintenance, human performance, steam generator leak detection, fuel channel inspections, rotating equipment maintenance, surveillance programs, inspection techniques, valve maintenance, steam generator repairs and performance, reactor aging management and preventative maintenance

  6. Proceedings of the fourth international conference on CANDU maintenance

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1997-07-01

    These proceedings record the information presented at the 4th International Conference on CANDU Maintenance held November 16-18,1997 in Toronto, Canada. The papers for these proceedings were prepared on component maintenance, human performance, steam generator leak detection, fuel channel inspections, rotating equipment maintenance, surveillance programs, inspection techniques, valve maintenance, steam generator repairs and performance, reactor aging management and preventative maintenance.

  7. Orientation of the calcium channel beta relative to the alpha(12.2 subunit is critical for its regulation of channel activity.

    Directory of Open Access Journals (Sweden)

    Iuliia Vitko

    Full Text Available BACKGROUND: The Ca(vbeta subunits of high voltage-activated Ca(2+ channels control the trafficking and biophysical properties of the alpha(1 subunit. The Ca(vbeta-alpha(1 interaction site has been mapped by crystallographic studies. Nevertheless, how this interaction leads to channel regulation has not been determined. One hypothesis is that betas regulate channel gating by modulating movements of IS6. A key requirement for this direct-coupling model is that the linker connecting IS6 to the alpha-interaction domain (AID be a rigid structure. METHODOLOGY/PRINCIPAL FINDINGS: The present study tests this hypothesis by altering the flexibility and orientation of this region in alpha(12.2, then testing for Ca(vbeta regulation using whole cell patch clamp electrophysiology. Flexibility was induced by replacement of the middle six amino acids of the IS6-AID linker with glycine (PG6. This mutation abolished beta2a and beta3 subunits ability to shift the voltage dependence of activation and inactivation, and the ability of beta2a to produce non-inactivating currents. Orientation of Ca(vbeta with respect to alpha(12.2 was altered by deletion of 1, 2, or 3 amino acids from the IS6-AID linker (Bdel1, Bdel2, Bdel3, respectively. Again, the ability of Ca(vbeta subunits to regulate these biophysical properties were totally abolished in the Bdel1 and Bdel3 mutants. Functional regulation by Ca(vbeta subunits was rescued in the Bdel2 mutant, indicating that this part of the linker forms beta-sheet. The orientation of beta with respect to alpha was confirmed by the bimolecular fluorescence complementation assay. CONCLUSIONS/SIGNIFICANCE: These results show that the orientation of the Ca(vbeta subunit relative to the alpha(12.2 subunit is critical, and suggests additional points of contact between these subunits are required for Ca(vbeta to regulate channel activity.

  8. Regulation of Substantia Nigra Pars Reticulata GABAergic Neuron Activity by H2O2 via Flufenamic Acid-Sensitive Channels and KATP Channels

    Science.gov (United States)

    Lee, Christian R.; Witkovsky, Paul; Rice, Margaret E.

    2011-01-01

    Substantia nigra pars reticulata (SNr) GABAergic neurons are key output neurons of the basal ganglia. Given the role of these neurons in motor control, it is important to understand factors that regulate their firing rate and pattern. One potential regulator is hydrogen peroxide (H2O2), a reactive oxygen species that is increasingly recognized as a neuromodulator. We used whole-cell current clamp recordings of SNr GABAergic neurons in guinea-pig midbrain slices to determine how H2O2 affects the activity of these neurons and to explore the classes of ion channels underlying those effects. Elevation of H2O2 levels caused an increase in the spontaneous firing rate of SNr GABAergic neurons, whether by application of exogenous H2O2 or amplification of endogenous H2O2 through inhibition of glutathione peroxidase with mercaptosuccinate. This effect was reversed by flufenamic acid (FFA), implicating transient receptor potential (TRP) channels. Conversely, depletion of endogenous H2O2 by catalase, a peroxidase enzyme, decreased spontaneous firing rate and firing precision of SNr neurons, demonstrating tonic control of firing rate by H2O2. Elevation of H2O2 in the presence of FFA revealed an inhibition of tonic firing that was prevented by blockade of ATP-sensitive K+ (KATP) channels with glibenclamide. In contrast to guinea-pig SNr neurons, the dominant effect of H2O2 elevation in mouse SNr GABAergic neurons was hyperpolarization, indicating a species difference in H2O2-dependent regulation. Thus, H2O2 is an endogenous modulator of SNr GABAergic neurons, acting primarily through presumed TRP channels in guinea-pig SNr, with additional modulation via KATP channels to regulate SNr output. PMID:21503158

  9. Proteolytic activation of the epithelial sodium channel ENaC in preeclampsia examined with urinary exosomes

    DEFF Research Database (Denmark)

    Nielsen, Maria Ravn; Rytz, Mie; Frederiksen-Møller, Britta

    2015-01-01

    OBJECTIVES: Increased activity of the epithelial sodium channel (ENaC) in the kidneys may explain the coupling between proteinuria, edema, suppressed aldosterone and hypertension in preeclampsia. Preeclamptic women excrete plasminogen-plasmin in urine. In vitro, plasmin increases the activity...... as a positive control for the presence of collecting duct membrane. RESULTS: Urine plasmin-plasminogen/creatinine ratio was increased in the preeclampsia group (p... pregnancy and preeclampsia CONCLUSIONS: It is possible to examine collecting duct transport proteins in urine exosome from pregnant women including γ-ENaC, 2) Urine exosome fraction displays a variable pattern of γ-ENaC signal with a predominance of cleaved forms in both normal and preeclamptic women...

  10. The Effect of Cognitive Activity on Sleep Maintenance in a Subsequent Daytime Nap.

    Science.gov (United States)

    Arzilli, Cinzia; Cerasuolo, Mariangela; Conte, Francesca; Bittoni, Valentina; Gatteschi, Claudia; Albinni, Benedetta; Giganti, Fiorenza; Ficca, Gianluca

    2018-01-25

    The aim of this study is to assess the effects of a learning task on the characteristics of a subsequent daytime nap. Thirty-eight subjects were administered a control nap (C) and one preceded by a cognitive training session (TR). Relative to C, TR naps showed significantly increased sleep duration with decreased sleep latency, as well as significantly increased sleep efficiency due to reduced awakening frequency. Meaningful trends were also found toward an increase of Stage 2 sleep proportion and a reduction of Stage 1 sleep, percentage of wake after sleep onset (WASO), and frequency of state transitions. Our results indicate that presleep learning favors sleep propensity and maintenance, offering the possibility to explore planned cognitive training as a low-cost treatment for sleep impairments.

  11. An intermediate-conductance Ca2+-activated K+ channel is important for secretion in pancreatic duct cells

    DEFF Research Database (Denmark)

    Hayashi, Mikio; Wang, Jing; Hede, Susanne Edeling

    2012-01-01

    2; Slack; Slick; and an intermediate-conductance Ca(2+)-activated K(+) (IK) channel (K(Ca)3.1). The following functional studies were focused on the IK channel. 5,6-Dichloro-1-ethyl-1,3-dihydro-2H-benzimidazole-2-one (DC-EBIO), an activator of IK channel, increased equivalent short-circuit current...

  12. Dopamine suppresses neuronal activity of Helisoma B5 neurons via a D2-like receptor, activating PLC and K channels.

    Science.gov (United States)

    Zhong, L R; Artinian, L; Rehder, V

    2013-01-03

    Dopamine (DA) plays fundamental roles as a neurotransmitter and neuromodulator in the central nervous system. How DA modulates the electrical excitability of individual neurons to elicit various behaviors is of great interest in many systems. The buccal ganglion of the freshwater pond snail Helisoma trivolvis contains the neuronal circuitry for feeding and DA is known to modulate the feeding motor program in Helisoma. The buccal neuron B5 participates in the control of gut contractile activity and is surrounded by dopaminergic processes, which are expected to release DA. In order to study whether DA modulates the electrical activity of individual B5 neurons, we performed experiments on physically isolated B5 neurons in culture and on B5 neurons within the buccal ganglion in situ. We report that DA application elicited a strong hyperpolarization in both conditions and turned the electrical activity from a spontaneously firing state to an electrically silent state. Using the cell culture system, we demonstrated that the strong hyperpolarization was inhibited by the D2 receptor antagonist sulpiride and the phospholipase C (PLC) inhibitor U73122, indicating that DA affected the membrane potential of B5 neurons through the activation of a D2-like receptor and PLC. Further studies revealed that the DA-induced hyperpolarization was inhibited by the K channel blockers 4-aminopyridine and tetraethylammonium, suggesting that K channels might serve as the ultimate target of DA signaling. Through its modulatory effect on the electrical activity of B5 neurons, the release of DA in vivo may contribute to a neuronal output that results in a variable feeding motor program. Copyright © 2012 IBRO. Published by Elsevier Ltd. All rights reserved.

  13. Maintenance methods

    International Nuclear Information System (INIS)

    Sanchis, H.; Aucher, P.

    1990-01-01

    The maintenance method applied at the Hague is summarized. The method was developed in order to solve problems relating to: the different specialist fields, the need for homogeneity in the maintenance work, the equipment diversity, the increase of the materials used at the Hague's new facilities. The aim of the method is to create a knowhow formalism, to facilitate maintenance, to ensure the running of the operations and to improve the estimation of the maintenance cost. One of the method's difficulties is the demonstration of the profitability of the maintenance operations [fr

  14. FY 1994 annual summary report of the surveillance and maintenance activities for the Oak Ridge National Laboratory Environmental Restoration Program

    International Nuclear Information System (INIS)

    1994-11-01

    The Oak Ridge National Laboratory (ORNL) Environmental Restoration (ER) Surveillance and Maintenance (S and M) Program was initiated to manage former waste management and environmental research sites contaminated with radioactive materials and/or hazardous chemicals. The S and M Program is responsible for managing designated sites/facilities from the end of their operating lives until final disposition or site stabilization. To effectively manage and perform the various S and M Program responsibilities, five summary-level work breakdown structure (WBS) elements have been established: S and M Preliminary Investigations, Special Projects, Routine S and M, Inactive Groundwater Wells, and Project Management. Routine S and M activities were conducted as scheduled throughout fiscal years (FY) 1994 at applicable inactive waste management (WM) and other contaminated areas. Overall, the ER S and M Program maintains 47 facilities, performs vegetation maintenance on approximately 230 acres, maintains 54 inactive tanks, and provides overall site management on over 700 acres. In addition to the routine S and M activities, detailed site inspections were conducted at established frequencies on appropriate sites in the ER S and M Program. This document provides a summary of the FY 1994 ORNL ER S and M Program accomplishments

  15. Modeling the Hybrid Flow Shop Scheduling Problem Followed by an Assembly Stage Considering Aging Effects and Preventive Maintenance Activities

    Directory of Open Access Journals (Sweden)

    Seyyed Mohammad Hassan Hosseini

    2016-05-01

    Full Text Available Scheduling problem for the hybrid flow shop scheduling problem (HFSP followed by an assembly stage considering aging effects additional preventive and maintenance activities is studied in this paper. In this production system, a number of products of different kinds are produced. Each product is assembled with a set of several parts. The first stage is a hybrid flow shop to produce parts. All machines can process all kinds of parts in this stage but each machine can process only one part at the same time. The second stage is a single assembly machine or a single assembly team of workers. The aim is to schedule the parts on the machines and assembly sequence and also determine when the preventive maintenance activities get done in order to minimize the completion time of all products (makespan. A mathematical modeling is presented and its validation is shown by solving an example in small scale. Since this problem has been proved strongly NP-hard, in order to solve the problem in medium and large scale, four heuristic algorithms is proposed based on the Johnson’s algorithm. The numerical experiments are used to run the mathematical model and evaluate the performance of the proposed algorithms.

  16. Analysis of maintenance strategies

    International Nuclear Information System (INIS)

    Laakso, K.; Simola, K.

    1998-01-01

    The main topics of the presentation include: (1) an analysis model and methods to evaluate maintenance action programs and the support decision to make changes in them and (2) to understand the maintenance strategies in a systems perspective as a basis for future developments. The subproject showed how systematic models for maintenance analysis and decision support, utilising computerised and statistical tool packages, can be taken into use for evaluation and optimisation of maintenance of active systems from the safety and economic point of view

  17. Reduced Tonoplast Fast-Activating and Slow-Activating Channel Activity Is Essential for Conferring Salinity Tolerance in a Facultative Halophyte, Quinoa1[C][W][OA

    Science.gov (United States)

    Bonales-Alatorre, Edgar; Shabala, Sergey; Chen, Zhong-Hua; Pottosin, Igor

    2013-01-01

    Halophyte species implement a “salt-including” strategy, sequestering significant amounts of Na+ to cell vacuoles. This requires a reduction of passive Na+ leak from the vacuole. In this work, we used quinoa (Chenopodium quinoa) to investigate the ability of halophytes to regulate Na+-permeable slow-activating (SV) and fast-activating (FV) tonoplast channels, linking it with Na+ accumulation in mesophyll cells and salt bladders as well as leaf photosynthetic efficiency under salt stress. Our data indicate that young leaves rely on Na+ exclusion to salt bladders, whereas old ones, possessing far fewer salt bladders, depend almost exclusively on Na+ sequestration to mesophyll vacuoles. Moreover, although old leaves accumulate more Na+, this does not compromise their leaf photochemistry. FV and SV channels are slightly more permeable for K+ than for Na+, and vacuoles in young leaves express less FV current and with a density unchanged in plants subjected to high (400 mm NaCl) salinity. In old leaves, with an intrinsically lower density of the FV current, FV channel density decreases about 2-fold in plants grown under high salinity. In contrast, intrinsic activity of SV channels in vacuoles from young leaves is unchanged under salt stress. In vacuoles of old leaves, however, it is 2- and 7-fold lower in older compared with young leaves in control- and salt-grown plants, respectively. We conclude that the negative control of SV and FV tonoplast channel activity in old leaves reduces Na+ leak, thus enabling efficient sequestration of Na+ to their vacuoles. This enables optimal photosynthetic performance, conferring salinity tolerance in quinoa species. PMID:23624857

  18. ‘Sleepy’ inward rectifier channels in guinea-pig cardiomyocytes are activated only during strong hyperpolarization

    Science.gov (United States)

    Liu, Gong Xin; Daut, Jürgen

    2002-01-01

    K+ channels of isolated guinea-pig cardiomyocytes were studied using the patch-clamp technique. At transmembrane potentials between −120 and −220 mV we observed inward currents through an apparently novel channel. The novel channel was strongly rectifying, no outward currents could be recorded. Between −200 and −160 mV it had a slope conductance of 42.8 ± 3.0 pS (s.d.; n = 96). The open probability (Po) showed a sigmoid voltage dependence and reached a maximum of 0.93 at −200 mV, half-maximal activation was approximately −150 mV. The voltage dependence of Po was not affected by application of 50 μm isoproterenol. The open-time distribution could be described by a single exponential function, the mean open time ranged between 73.5 ms at −220 mV and 1.4 ms at −160 mV. At least two exponential components were required to fit the closed time distribution. Experiments with different external Na+, K+ and Cl− concentrations suggested that the novel channel is K+ selective. Extracellular Ba2+ ions gave rise to a voltage-dependent reduction in Po by inducing long closed states; Cs+ markedly reduced mean open time at −200 mV. In cell-attached recordings the novel channel frequently converted to a classical inward rectifier channel, and vice versa. This conversion was not voltage dependent. After excision of the patch, the novel channel always converted to a classical inward rectifier channel within 0–3 min. This conversion was not affected by intracellular Mg2+, phosphatidylinositol (4,5)-bisphosphate or spermine. Taken together, our findings suggest that the novel K+ channel represents a different ‘mode’ of the classical inward rectifier channel in which opening occurs only at very negative potentials. PMID:11897847

  19. Electron transfer activation of a second water channel for proton transport in [FeFe]-hydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    Sode, Olaseni; Voth, Gregory A., E-mail: gavoth@uchicago.edu [Department of Chemistry, James Franck Institute, Institute for Biophysical Dynamics, Computation Institute, The University of Chicago, Chicago, Illinois 60637, USA and Computing, Environment and Life Sciences, Argonne National Laboratory, Argonne, Illinois 60439 (United States)

    2014-12-14

    Hydrogenase enzymes are important because they can reversibly catalyze the production of molecular hydrogen. Proton transport mechanisms have been previously studied in residue pathways that lead to the active site of the enzyme via residues Cys299 and Ser319. The importance of this pathway and these residues has been previously exhibited through site-specific mutations, which were shown to interrupt the enzyme activity. It has been shown recently that a separate water channel (WC2) is coupled with electron transport to the active site of the [FeFe]-hydrogenase. The water-mediated proton transport mechanisms of the enzyme in different electronic states have been studied using the multistate empirical valence bond reactive molecular dynamics method, in order to understand any role WC2 may have in facilitating the residue pathway in bringing an additional proton to the enzyme active site. In a single electronic state A{sup 2−}, a water wire was formed through which protons can be transported with a low free energy barrier. The remaining electronic states were shown, however, to be highly unfavorable to proton transport in WC2. A double amino acid substitution is predicted to obstruct proton transport in electronic state A{sup 2-} by closing a cavity that could otherwise fill with water near the proximal Fe of the active site.

  20. Voltage Gated Calcium Channel Activation by Backpropagating Action Potentials Downregulates NMDAR Function

    Directory of Open Access Journals (Sweden)

    Anne-Kathrin Theis

    2018-04-01

    Full Text Available The majority of excitatory synapses are located on dendritic spines of cortical glutamatergic neurons. In spines, compartmentalized Ca2+ signals transduce electrical activity into specific long-term biochemical and structural changes. Action potentials (APs propagate back into the dendritic tree and activate voltage gated Ca2+ channels (VGCCs. For spines, this global mode of spine Ca2+ signaling is a direct biochemical feedback of suprathreshold neuronal activity. We previously demonstrated that backpropagating action potentials (bAPs result in long-term enhancement of spine VGCCs. This activity-dependent VGCC plasticity results in a large interspine variability of VGCC Ca2+ influx. Here, we investigate how spine VGCCs affect glutamatergic synaptic transmission. We combined electrophysiology, two-photon Ca2+ imaging and two-photon glutamate uncaging in acute brain slices from rats. T- and R-type VGCCs were the dominant depolarization-associated Ca2+conductances in dendritic spines of excitatory layer 2 neurons and do not affect synaptic excitatory postsynaptic potentials (EPSPs measured at the soma. Using two-photon glutamate uncaging, we compared the properties of glutamatergic synapses of single spines that express different levels of VGCCs. While VGCCs contributed to EPSP mediated Ca2+ influx, the amount of EPSP mediated Ca2+ influx is not determined by spine VGCC expression. On a longer timescale, the activation of VGCCs by bAP bursts results in downregulation of spine NMDAR function.

  1. Electron transfer activation of a second water channel for proton transport in [FeFe]-hydrogenase

    International Nuclear Information System (INIS)

    Sode, Olaseni; Voth, Gregory A.

    2014-01-01

    Hydrogenase enzymes are important because they can reversibly catalyze the production of molecular hydrogen. Proton transport mechanisms have been previously studied in residue pathways that lead to the active site of the enzyme via residues Cys299 and Ser319. The importance of this pathway and these residues has been previously exhibited through site-specific mutations, which were shown to interrupt the enzyme activity. It has been shown recently that a separate water channel (WC2) is coupled with electron transport to the active site of the [FeFe]-hydrogenase. The water-mediated proton transport mechanisms of the enzyme in different electronic states have been studied using the multistate empirical valence bond reactive molecular dynamics method, in order to understand any role WC2 may have in facilitating the residue pathway in bringing an additional proton to the enzyme active site. In a single electronic state A 2− , a water wire was formed through which protons can be transported with a low free energy barrier. The remaining electronic states were shown, however, to be highly unfavorable to proton transport in WC2. A double amino acid substitution is predicted to obstruct proton transport in electronic state A 2- by closing a cavity that could otherwise fill with water near the proximal Fe of the active site

  2. Determining k channel activation curves from k channel currents often requires the goldman-hodgkin-katz equation.

    Science.gov (United States)

    Clay, John R

    2009-01-01

    Potassium ion current in nerve membrane, I(K), has traditionally been described by I(K) = g(K)(V - E(K)), where g(K) is the K ion conductance, V is membrane potential and E(K) is the K(+) Nernst potential. This description has been unchallenged by most investigators in neuroscience since its introduction almost 60 years ago. The problem with the I(K) approximately (V - E(K)) 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(+) concentration is significantly higher than the extracellular concentration. Consequently, the slope conductance at potentials positive to E(K) cannot be the same as that for potentials negative to E(K), as the linear proportionality between I(K) and (V - E(K)) requires. Instead I(K) has a non-linear dependence on (V - E(K)) 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.

  3. 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.

  4. Syntectonic Mississippi River Channel Response: Integrating River Morphology and Seismic Imaging to Detect Active Faults

    Science.gov (United States)

    Magnani, M. B.

    2017-12-01

    Alluvial rivers, even great rivers such as the Mississippi, respond to hydrologic and geologic controls. Temporal variations of valley gradient can significantly alter channel morphology, as the river responds syntectonically to attain equilibrium. The river will alter its sinuosity, in an attempt to maintain a constant gradient on a surface that changes slope through time. Therefore, changes of river pattern can be the first clue that active tectonics is affecting an area of pattern change. Here I present geomorphological and seismic imaging evidence of a previously unknown fault crossing the Mississippi river south of the New Madrid seismic zone, between Caruthersville, Missouri and Osceola, Arkansas, and show that both datasets support Holocene fault movement, with the latest slip occurring in the last 200 years. High resolution marine seismic reflection data acquired along the Mississippi river imaged a NW-SE striking north-dipping fault displacing the base of the Quaternary alluvium by 15 m with reverse sense of movement. The fault consistently deforms the Tertiary, Cretaceous and Paleozoic formations. Historical river channel planforms dating back to 1765 reveal that the section of the river channel across the fault has been characterized by high sinuosity and steep projected-channel slope compared to adjacent river reaches. In particular, the reach across the fault experienced a cutoff in 1821, resulting in a temporary lowering of sinuosity followed by an increase between the survey of 1880 and 1915. Under the assumption that the change in sinuosity reflects river response to a valley slope change to maintain constant gradient, I use sinuosity through time to calculate the change in valley slope since 1880 and therefore to estimate the vertical displacement of the imaged fault in the past 200 years. Based on calculations so performed, the vertical offset of the fault is estimated to be 0.4 m, accrued since at least 1880. If the base of the river alluvium

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

    DEFF Research Database (Denmark)

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

    2008-01-01

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

  6. Pentachlorophenol-Induced Cytotoxic, Mitogenic, and Endocrine-Disrupting Activities in Channel Catfish, Ictalurus punctatus

    Directory of Open Access Journals (Sweden)

    Paul B. Tchounwou

    2004-09-01

    Full Text Available Pentachlorophenol (PCP is an organochlorine compound that has been widely used as a biocide in several industrial, agricultural, and domestic applications. Although it has been shown to induce systemic toxicity and carcinogenesis in several experimental studies, the literature is scarce regarding its toxic mechanisms of action at the cellular and molecular levels. Recent investigations in our laboratory have shown that PCP induces cytotoxicity and transcriptionally activates stress genes in human liver carcinoma (HepG2 cells [1]. In this research, we hypothesize that environmental exposure to PCP may trigger cytotoxic, mitogenic, and endocrine-disrupting activities in aquatic organisms including fish. To test this hypothesis, we carried out in vitro cultures of male channel catfish hepatocytes, and performed the fluorescein diacetate assay (FDA to assess for cell viability, and the Western Blot analysis to assess for vitellogenin expression following exposure to PCP. Data obtained from FDA experiments indicated a strong dose-response relationship with respect to PCP cytotoxicity. Upon 48 hrs of exposure, the chemical dose required to cause 50% reduction in cell viability (LD50 was computed to be 1,987.0 + 9.6 μg PCP/mL. The NOAEL and LOAEL were 62.5 + 10.3 μg PCP/mL and 125.0+15.2 μg PCP/mL, respectively. At lower levels of exposure, PCP was found to be mitogenic, showing a strong dose- and time-dependent response with regard to cell proliferation. Western Blot analysis demonstrated the potential of PCP to cause endocrine-disrupting activity, as evidenced by the up regulation of the 125-kDa vitellogenin protein the hepatocytes of male channel catfish.

  7. Perencanaan Kegiatan Maintenance Dengan Metode Reability Centered Maintenance (Rcm) II

    OpenAIRE

    Rachmad Hidayat; Nachnul Ansori; Ali Imron

    2010-01-01

    Maintenance Activity Planning by Reability Centered Maintenance (RCM) II Method. This research discussmaintenance activity by using RCM II method to determine failure function risk at compresor screw. Calculation isgiven to magnitude optimum time maintenance interval by considering the cost maintenance and the cost reparation.From the research results with RPN points out that critical component that needs to get main priority in givemaintenance on compresor screw are bust logistic on timeworn...

  8. Maintenance Plan for the Hanford Immobilized Low-Activity Tank Waste Performance Assessment

    International Nuclear Information System (INIS)

    MANN, F.M.

    2000-01-01

    The plan for maintaining the Hanford Immobilized Low-Activity Tank Waste Performance Assessment (PA) is described. The plan includes expected work on PA reviews and revisions, waste reports, monitoring, other operational activities, etc

  9. Activation, Permeability, and Inhibition of Astrocytic and Neuronal Large Pore (Hemi)channels

    DEFF Research Database (Denmark)

    Hansen, Daniel Bloch; Ye, Zu-Cheng; Calloe, Kirstine

    2014-01-01

    overlapping sensitivity to the inhibitors Brilliant Blue, gadolinium, and carbenoxolone. These results demonstrated isoform-specific characteristics among the large pore membrane channels; an open (hemi)channel is not a nonselective channel. With these isoform-specific properties in mind, we characterized...

  10. An improved ivermectin-activated chloride channel receptor for inhibiting electrical activity in defined neuronal populations

    DEFF Research Database (Denmark)

    Lynagh, Timothy Peter; Lynch, Joseph W

    2010-01-01

    The ability to silence the electrical activity of defined neuronal populations in vivo is dramatically advancing our understanding of brain function. This technology may eventually be useful clinically for treating a variety of neuropathological disorders caused by excessive neuronal activity...... conductance, homomeric expression, and human origin may render the F207A/A288G alpha1 glycine receptor an improved silencing receptor for neuroscientific and clinical purposes. As all known highly ivermectin-sensitive GluClRs contain an endogenous glycine residue at the corresponding location, this residue...

  11. Energetic performance is improved by specific activation of K+ fluxes through K(Ca) channels in heart mitochondria

    DEFF Research Database (Denmark)

    Aon, Miguel A; Cortassa, Sonia; Wei, An-Chi

    2009-01-01

    Mitochondrial volume regulation depends on K+ movement across the inner membrane and a mitochondrial Ca2+-dependent K+ channel (mitoK(Ca)) reportedly contributes to mitochondrial K+ uniporter activity. Here we utilize a novel K(Ca) channel activator, NS11021, to examine the role of mito...... similar nonspecific (toxin-insensitive) effects at high concentrations. The results indicate that activating K+ flux through mitoK(Ca) mediates a beneficial effect on energetics that depends on mitochondrial swelling with maintained DeltaPsi(m)....

  12. Synergistic activation of G protein-gated inwardly rectifying potassium channels by cholesterol and PI(4,5)P2.

    Science.gov (United States)

    Bukiya, Anna N; Rosenhouse-Dantsker, Avia

    2017-07-01

    G-protein gated inwardly rectifying potassium (GIRK or Kir3) channels play a major role in the control of the heart rate, and require the membrane phospholipid phosphatidylinositol-bis-phosphate (PI(4,5)P 2 ) for activation. Recently, we have shown that the activity of the heterotetrameric Kir3.1/Kir3.4 channel that underlies atrial K ACh currents was enhanced by cholesterol. Similarly, the activities of both the Kir3.4 homomer and its active pore mutant Kir3.4* (Kir3.4_S143T) were also enhanced by cholesterol. Here we employ planar lipid bilayers to investigate the crosstalk between PI(4,5)P 2 and cholesterol, and demonstrate that these two lipids act synergistically to activate Kir3.4* currents. Further studies using the Xenopus oocytes heterologous expression system suggest that PI(4,5)P 2 and cholesterol act via distinct binding sites. Whereas PI(4,5)P 2 binds to the cytosolic domain of the channel, the putative binding region of cholesterol is located at the center of the transmembrane domain overlapping the central glycine hinge region of the channel. Together, our data suggest that changes in the levels of two key membrane lipids - cholesterol and PI(4,5)P 2 - could act in concert to provide fine-tuning of Kir3 channel function. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Different expressions of high voltage-activated Ca2+ channel types in the rostral reticular thalamic nucleus of the absence epileptic WAG/Rij rat.

    NARCIS (Netherlands)

    Bovenkamp-Janssen, M.C. van de; Scheenen, W.J.J.M.; Kuijpers-Kwant, F.J.; Kozicz, L.T.; Veening, J.G.; Luijtelaar, E.L.J.M. van; McEnery, M.W.; Roubos, E.W.

    2004-01-01

    In the WAG/Rij rat, a model for human absence epilepsy, spike-wave discharges (SWD) and absence epileptic behavior develop after the age of 3 months. The rostral part of the reticular thalamic nucleus (rRTN) is involved in SWD. Ca(2+) channels play a central role in the initiation and maintenance of

  14. Designing Interventions that Last: A Classification of Environmental Behaviors in Relation to the Activities, Costs, and Effort Involved for Adoption and Maintenance

    Science.gov (United States)

    Moore, Harriet E.; Boldero, Jennifer

    2017-01-01

    Policy makers draw on behavioral research to design interventions that promote the voluntary adoption of environmental behavior in societies. Many environmental behaviors will only be effective if they are maintained over the long-term. In the context of climate change and concerns about future water security, behaviors that involve reducing energy consumption and improving water quality must be continued indefinitely to mitigate global warming and preserve scarce resources. Previous reviews of environmental behavior have focused exclusively on factors related to adoption. This review investigates the factors that influence both adoption and maintenance, and presents a classification of environmental behaviors in terms of the activities, costs, and effort required for both adoption and maintenance. Three categories of behavior are suggested. One-off behaviors involve performing an activity once, such as purchasing an energy efficient washing machine, or signing a petition. Continuous behaviors involve the performance of the same set of behaviors for adoption and for maintenance, such as curbside recycling. Dynamic behaviors involve the performance of different behaviors for adoption and maintenance, such as revegetation. Behaviors can also be classified into four categories related to cost and effort: those that involve little cost and effort for adoption and maintenance, those that involve moderate cost and effort for adoption and maintenance, those that involve a high cost or effort for adoption and less for maintenance, and those that involve less cost or effort for adoption and a higher amount for maintenance. In order to design interventions that last, policy makers should consider the factors that influence the maintenance as well as the adoption of environmental behaviors. PMID:29163265

  15. Social influence and adolescent health-related physical activity in structured and unstructured settings: role of channel and type.

    Science.gov (United States)

    Spink, Kevin S; Wilson, Kathleen S; Ulvick, Jocelyn

    2012-08-01

    Social influence channels (e.g., parents) and types (e.g., compliance) have each been related to physical activity independently, but little is known about how these two categories of influence may operate in combination. This study examined the relationships between various combinations of social influence and physical activity among youth across structured and unstructured settings. Adolescents (N=304), classified as high or low active, reported the social influence combinations they received for being active. Participants identified three channels and three types of influence associated with being active. For structured activity, compliance with peers and significant others predicted membership in the high active group (values of psocial influence, when examining health-related physical activity.

  16. KCa2 and KCa3 channels in learning and memory processes, and neurodegeneration

    Directory of Open Access Journals (Sweden)

    Els F. E. Kuiper

    2012-06-01

    Full Text Available Calcium-activated potassium (KCa channels are present throughout the central nervous system as well as many peripheral tissues. Activation of KCa channels is essential for maintenance of the neuronal membrane potential and was shown to underlie the afterhyperpolarization (AHP that regulates action potential firing and limits the firing frequency of repetitive action potentials. Different subtypes of KCa channels were anticipated on the basis of their physiological and pharmacological profiles, and cloning revealed two well defined but phylogenetic distantly related groups of channels. The group subject of this review includes both the small-conductance KCa2 channels (KCa2.1, KCa2.2, and KCa2.3 and the intermediate-conductance (KCa3.1 channel. These channels are activated by submicromolar intracellular Ca2+ concentrations and are voltage independent. Of all KCa channels only the KCa2 channels can be potently but differentially blocked by the bee-venom apamin. In the past few years modulation of KCa channel activation revealed new roles for KCa2 channels in controlling dendritic excitability, synaptic functioning and synaptic plasticity. Furthermore, KCa2 channels appeared to be involved in neurodegeneration, and learning and memory processes. In this review, we focus on the role of KCa2 and KCa3 channels in these latter mechanisms with emphasis on learning and memory, Alzheimer’s disease and on the interplay between neuroinflammation and different neurotransmitters/neuromodulators, their signalling components and KCa channel activation.

  17. What are the required maintenance and consolidation activities to run at design performance levels (injectors and LHC) until 2035?

    CERN Document Server

    Baird, S

    2014-01-01

    Assuming that Linac4 is connected to the PSB in LS2, we will outline the maintenance and basic consolidation works that will be needed to maintain design performance of the LHC and its Injector chain until 2035, with an overall reliability as good as that achieved in the first LHC operation period 2009 to 2013. Using these data we will estimate the shutdown schedule needed throughout this period to complete these maintenance and consolidation works. These estimates will also include the required radiation cool-down periods, time for system re-commissioning and testing as well as the time needed to restart the accelerator chain for LHC colliding beam operation. As some of the consolidation activities needed for the PS and SPS machines are related to the radia tion dose taken by the machine equipment (e.g., irradiated cable replacement and magnet renovation) the variation of these time estimates as a function of beam losses in the Injector chain will also be covered.

  18. Filament Activation in Response to Magnetic Flux Emergence and Cancellation in Filament Channels

    Science.gov (United States)

    Li, Ting; Zhang, Jun; Ji, Haisheng

    2015-06-01

    We conducted a comparative analysis of two filaments that showed a quite different activation in response to the flux emergence within the filament channels. The observations from the Solar Dynamics Observatory (SDO) and Global Oscillation Network Group (GONG) were made to analyze the two filaments on 2013 August 17 - 20 (SOL2013-08-17) and September 29 (SOL2013-09-29). The first event showed that the main body of the filament was separated into two parts when an active region (AR) emerged with a maximum magnetic flux of about 6.4×1021 Mx underlying the filament. The close neighborhood and common direction of the bright threads in the filament and the open AR fan loops suggest a similar magnetic connectivity of these two flux systems. The equilibrium of the filament was not destroyed three days after the start of the emergence of the AR. To our knowledge, similar observations have never been reported before. In the second event, the emerging flux occurred nearby a barb of the filament with a maximum magnetic flux of 4.2×1020 Mx, about one order of magnitude lower than that of the first event. Two patches of parasitic polarity in the vicinity of the barb merged, then cancelled with nearby network fields. About 20 hours after the onset of the emergence, the filament erupted. Our findings imply that the location of emerging flux within the filament channel is probably crucial to filament evolution. If the flux emergence appears nearby the barbs, it is highly likely that the emerging flux and the filament magnetic fields will cancel, which may lead to the eruption of the filament. The comparison of the two events shows that the emergence of a small AR may still not be enough to disrupt the stability of a filament system, and the actual eruption only occurs after the flux cancellation sets in.

  19. Characterization of the human pH- and PKA-activated ClC-2G(2 alpha) Cl- channel.

    Science.gov (United States)

    Sherry, A M; Stroffekova, K; Knapp, L M; Kupert, E Y; Cuppoletti, J; Malinowska, D H

    1997-08-01

    A ClC-2G(2 alpha) Cl- channel was identified to be present in human lung and stomach, and a partial cDNA for this Cl- channel was cloned from a human fetal lung library. A full-length expressible human ClC-2G(2 alpha) cDNA was constructed by ligation of mutagenized expressible rabbit ClC-2G(2 alpha) cDNA with the human lung ClC-2G(2 alpha) cDNA, expressed in oocytes, and characterized at the single-channel level. Adenosine 3',5'-cyclic monophosphate-dependent protein kinase (PKA) treatment increased the probability of opening of the channel (Po). After PKA activation, the channel exhibited a linear (r = 0.99) current-voltage curve with a slope conductance of 22.1 +/- 0.8 pS in symmetric 800 mM tetraethylammonium chloride (TEACl; pH 7.4). Under fivefold gradient conditions of TEACl, a reversal potential of +21.5 +/- 2.8 mV was measured demonstrating anion-to-cation discrimination. As previously demonstrated for the rabbit ClC-2G(2 alpha) Cl- channel, the human analog, hClC-2G(2 alpha), was active at pH 7.4 as well as when the pH of the extracellular face of the channel (trans side of the bilayer; pHtrans) was asymmetrically reduced to pH 3.0. The extent of PKA activation was dependent on pHtrans. With PKA treatment, Po increased fourfold with a pHtrans of 7.4 and eightfold with a pHtrans of 3.0. Effects of sequential PKA addition followed by pHtrans reduction on the same channel suggested that the PKA- and pH-dependent increases in channel Po were separable and cumulative. Northern analysis showed ClC-2G(2 alpha) mRNA to be present in human adult and fetal lung and adult stomach, and quantitative reverse transcriptase-polymerase chain reaction showed this channel to be present in the adult human lung and stomach at about one-half the level found in fetal lung. The findings of the present study suggest that the ClC-2G(2 alpha) Cl- channel may play an important role in Cl- transport in the fetal and adult human lung.

  20. Ion channeling

    International Nuclear Information System (INIS)

    Erramli, H.; Blondiaux, G.

    1994-01-01

    Channeling phenomenon was predicted, many years ago, by stark. The first channeling experiments were performed in 1963 by Davies and his coworkers. Parallely Robinson and Oen have investigated this process by simulating trajectories of ions in monocrystals. This technique has been combined with many methods like Rutherford Backscattering Spectrometry (R.B.S.), Particles Induced X-rays Emission (P.I.X.E) and online Nuclear Reaction (N.R.A.) to localize trace elements in the crystal or to determine crystalline quality. To use channeling for material characterization we need data about the stopping power of the incident particle in the channeled direction. The ratios of channeled to random stopping powers of silicon for irradiation in the direction have been investigated and compared to the available theoretical results. We describe few applications of ion channeling in the field of materials characterization. Special attention is given to ion channeling combined with Charged Particle Activation Analysis (C.P.A.A.) for studying the behaviour of oxygen atoms in Czochralski silicon lattices under the influence of internal gettering and in different gaseous atmospheres. Association between ion channeling and C.P.A.A was also utilised for studying the influence of the growing conditions on concentration and position of carbon atoms at trace levels in the MOVPE Ga sub (1-x) Al sub x lattice. 6 figs., 1 tab., 32 refs. (author)

  1. Maitotoxin Is a Potential Selective Activator of the Endogenous Transient Receptor Potential Canonical Type 1 Channel in Xenopus laevis Oocytes

    Directory of Open Access Journals (Sweden)

    Pedro L. Flores

    2017-06-01

    Full Text Available Maitotoxin (MTX is the most potent marine toxin known to date. It is responsible for a particular human intoxication syndrome called ciguatera fish poisoning (CFP. Several reports indicate that MTX is an activator of non-selective cation channels (NSCC in different cell types. The molecular identity of these channels is still an unresolved topic, and it has been proposed that the transient receptor potential (TRP channels are involved in this effect. In Xenopus laevis oocytes, MTX at picomolar (pM concentrations induces the activation of NSCC with functional and pharmacological properties that resemble the activity of TRP channels. The purpose of this study was to characterize the molecular identity of the TRP channel involved in the MTX response, using the small interference RNA (siRNA approach and the two-electrode voltage-clamp technique (TEVC. The injection of a specifically designed siRNA to silence the transient receptor potential canonical type 1 (TRPC1 protein expression abolished the MTX response. MTX had no effect on oocytes, even at doses 20-fold higher compared to cells without injection. Total mRNA and protein levels of TRPC1 were notably diminished. The TRPC4 siRNA did not change the MTX effect, even though it was important to note that the protein level was reduced by the silencing of TRPC4. Our results suggest that MTX could be a selective activator of TRPC1 channels in X. laevis oocytes and a useful pharmacological tool for further studies on these TRP channels.

  2. Phytochemicals from Ruta graveolens Activate TAS2R Bitter Taste Receptors and TRP Channels Involved in Gustation and Nociception

    Directory of Open Access Journals (Sweden)

    Giuseppe Mancuso

    2015-10-01

    Full Text Available Ruta graveolens (rue is a spontaneous plant in the Mediterranean area with a strong aroma and a very intense bitter taste, used in gastronomy and in folk medicine. From the leaves, stems and fruits of rue, we isolated rutin, rutamarin, three furanocoumarins, two quinolinic alkaloids, a dicoumarin and two long chain ketones. Bitter taste and chemesthetic properties have been evaluated by in vitro assays with twenty receptors of the TAS2R family and four TRP ion channels involved in gustation and nociception. Among the alkaloids, skimmianine was active as a specific agonist of T2R14, whereas kokusaginin did not activate any of the tested receptors. The furanocoumarins activates TAS2R10, 14, and 49 with different degrees of selectivity, as well as the TRPA1 somatosensory ion channel. Rutamarin is an agonist of TRPM5 and TRPV1 and a strong antagonist of TRPM8 ion channels.

  3. Phytochemicals from Ruta graveolens Activate TAS2R Bitter Taste Receptors and TRP Channels Involved in Gustation and Nociception.

    Science.gov (United States)

    Mancuso, Giuseppe; Borgonovo, Gigliola; Scaglioni, Leonardo; Bassoli, Angela

    2015-10-16

    Ruta graveolens (rue) is a spontaneous plant in the Mediterranean area with a strong aroma and a very intense bitter taste, used in gastronomy and in folk medicine. From the leaves, stems and fruits of rue, we isolated rutin, rutamarin, three furanocoumarins, two quinolinic alkaloids, a dicoumarin and two long chain ketones. Bitter taste and chemesthetic properties have been evaluated by in vitro assays with twenty receptors of the TAS2R family and four TRP ion channels involved in gustation and nociception. Among the alkaloids, skimmianine was active as a specific agonist of T2R14, whereas kokusaginin did not activate any of the tested receptors. The furanocoumarins activates TAS2R10, 14, and 49 with different degrees of selectivity, as well as the TRPA1 somatosensory ion channel. Rutamarin is an agonist of TRPM5 and TRPV1 and a strong antagonist of TRPM8 ion channels.

  4. Terbinafine is a novel and selective activator of the two-pore domain potassium channel TASK3.

    Science.gov (United States)

    Wright, Paul D; Veale, Emma L; McCoull, David; Tickle, David C; Large, Jonathan M; Ococks, Emma; Gothard, Gemma; Kettleborough, Catherine; Mathie, Alistair; Jerman, Jeffrey

    2017-11-04

    Two-pore domain potassium channels (K2Ps) are characterized by their four transmembrane domain and two-pore topology. They carry background (or leak) potassium current in a variety of cell types. Despite a number of important roles there is currently a lack of pharmacological tools with which to further probe K2P function. We have developed a cell-based thallium flux assay, using baculovirus delivered TASK3 (TWIK-related acid-sensitive K + channel 3, KCNK9, K2P9.1) with the aim of identifying novel, selective TASK3 activators. After screening a library of 1000 compounds, including drug-like and FDA approved molecules, we identified Terbinafine as an activator of TASK3. In a thallium flux assay a pEC50 of 6.2 ( ±0.12) was observed. When Terbinafine was screened against TASK2, TREK2, THIK1, TWIK1 and TRESK no activation was observed in thallium flux assays. Several analogues of Terbinafine were also purchased and structure activity relationships examined. To confirm Terbinafine's activation of TASK3 whole cell patch clamp electrophysiology was carried out and clear potentiation observed in both the wild type channel and the pathophysiological, Birk-Barel syndrome associated, G236R TASK3 mutant. No activity at TASK1 was observed in electrophysiology studies. In conclusion, we have identified the first selective activator of the two-pore domain potassium channel TASK3. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Maintenance of neural activities in torpid Rhinolophus ferrumequinum bats revealed by 2D gel-based proteome analysis.

    Science.gov (United States)

    Yin, Qiuyuan; Zhang, Yijian; Dong, Dong; Lei, Ming; Zhang, Shuyi; Liao, Chen-Chung; Pan, Yi-Hsuan

    2017-08-01

    Bats are the only mammals capable of self-powered flying. Many bat species hibernate in winter. A reversible control of cerebral activities is critical for bats to accommodate a repeated torpor-arousal cycle during hibernation. Little is known about the molecular mechanisms that regulate neuronal activities in torpid bats. In this study, Rhinolophus ferrumequinum bat brain proteins were fractionated, and their abundance in active and torpid states was compared. Results of 2D gel-based proteomics showed that 38% of identified proteins with a significant change in abundance are involved in synaptic vesicle recycling and cytoskeletal integrity. Changes in the abundance of proteins related to RNA splicing, proteostasis, redox homeostasis, mitochondrial function, and energy metabolism were also detected. In addition, the levels of GNAO1 (guanine nucleotide-binding protein G αo subunit), an important modulator of neuronal transmembrane signaling, were significantly increased in the insoluble protein fraction of torpid bats; this may be due to GNAO1 palmitoylation making it insoluble. Our data provide molecular evidence for the maintenance of neuronal activities in torpid bats and suggest that a reversible palmitoylation of the G protein plays a role in the regulation of neuronal activities during bat hibernation. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Sustained activation of STAT5 is essential for chromatin remodeling and maintenance of mammary-specific function

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Ren; Nelson, Celeste M.; Muschler, John L.; Veiseh, Mandana; Vonderhaar, Barbara K.; Bissell, Mina J.

    2009-06-03

    Epithelial cells, once dissociated and placed in two-dimensional (2D) cultures, rapidly lose tissue-specific functions. We showed previously that in addition to prolactin, signaling by laminin-111 was necessary to restore functional differentiation of mammary epithelia. Here, we elucidate two additional aspects of laminin-111 action. We show that in 2D cultures, the prolactin receptor is basolaterally localized and physically segregated from its apically placed ligand. Detachment of the cells exposes the receptor to ligation by prolactin leading to signal transducers and activators of transcription protein 5 (STAT5) activation, but only transiently and not sufficiently for induction of milk protein expression. We show that laminin-111 reorganizes mammary cells into polarized acini, allowing both the exposure of the prolactin receptor and sustained activation of STAT5. The use of constitutively active STAT5 constructs showed that the latter is necessary and sufficient for chromatin reorganization and {beta}-casein transcription. These results underscore the crucial role of continuous laminin signaling and polarized tissue architecture in maintenance of transcription factor activation, chromatin organization, and tissue-specific gene expression.

  7. Mechanical regulation of stem-cell differentiation by the stretch-activated Piezo channel.

    Science.gov (United States)

    He, Li; Si, Guangwei; Huang, Jiuhong; Samuel, Aravinthan D T; Perrimon, Norbert

    2018-03-01

    Somatic stem cells constantly adjust their self-renewal and lineage commitment by integrating various environmental cues to maintain tissue homeostasis. Although numerous chemical and biological signals have been identified that regulate stem-cell behaviour, whether stem cells can directly sense mechanical signals in vivo remains unclear. Here we show that mechanical stress regulates stem-cell differentiation in the adult Drosophila midgut through the stretch-activated ion channel Piezo. We find that Piezo is specifically expressed in previously unidentified enteroendocrine precursor cells, which have reduced proliferation ability and are destined to become enteroendocrine cells. Loss of Piezo activity reduces the generation of enteroendocrine cells in the adult midgut. In addition, ectopic expression of Piezo in all stem cells triggers both cell proliferation and enteroendocrine cell differentiation. Both the Piezo mutant and overexpression phenotypes can be rescued by manipulation of cytosolic Ca 2+ levels, and increases in cytosolic Ca 2+ resemble the Piezo overexpression phenotype, suggesting that Piezo functions through Ca 2+ signalling. Further studies suggest that Ca 2+ signalling promotes stem-cell proliferation and differentiation through separate pathways. Finally, Piezo is required for both mechanical activation of stem cells in a gut expansion assay and the increase of cytosolic Ca 2+ in response to direct mechanical stimulus in a gut compression assay. Thus, our study demonstrates the existence of a specific group of stem cells in the fly midgut that can directly sense mechanical signals through Piezo.

  8. Transient receptor potential A1 channel contributes to activation of the muscle reflex.

    Science.gov (United States)

    Koba, Satoshi; Hayes, Shawn G; Sinoway, Lawrence I

    2011-01-01

    This study was undertaken to elucidate the role played by transient receptor potential A1 channels (TRPA1) in activating the muscle reflex, a sympathoexcitatory drive originating in contracting muscle. First, we tested the hypothesis that stimulation of the TRPA1 located on muscle afferents reflexly increases sympathetic nerve activity. In decerebrate rats, allyl isothiocyanate, a TRPA1 agonist, was injected intra-arterially into the hindlimb muscle circulation. This led to a 33% increase in renal sympathetic nerve activity (RSNA). The effect of allyl isothiocyanate was a reflex because the response was prevented by sectioning the sciatic nerve. Second, we tested the hypothesis that blockade of TRPA1 reduces RSNA response to contraction. Thirty-second continuous static contraction of the hindlimb muscles, induced by electrical stimulation of the peripheral cut ends of L(4) and L(5) ventral roots, increased RSNA and blood pressure. The integrated RSNA during contraction was reduced by HC-030031, a TRPA1 antagonist, injected intra-arterially (163 ± 24 vs. 95 ± 21 arbitrary units, before vs. after HC-030031, P reflex. Increases in RSNA in response to injection into the muscle circulation of arachidonic acid, bradykinin, and diprotonated phosphate, which are metabolic by-products of contraction and stimulants of muscle afferents during contraction, were reduced by HC-030031. These observations suggest that the TRPA1 located on muscle afferents is part of the muscle reflex and further support the notion that arachidonic acid metabolites, bradykinin, and diprotonated phosphate are candidates for endogenous agonists of TRPA1.

  9. The long-term effects of switching from active intravenous bisphosphonate treatment to low-dose maintenance therapy in children with osteogenesis imperfecta.

    Science.gov (United States)

    Biggin, Andrew; Zheng, Linda; Briody, Julie N; Coorey, Craig P; Munns, Craig F

    2015-01-01

    Intravenous bisphosphonate therapy is the first-line treatment in moderate-to-severe osteogenesis imperfecta (OI), but there are varied treatment protocols with little data on long-term efficacy. This study evaluates the clinical outcomes when transitioning from active bisphosphonate treatment to maintenance therapy. A retrospective review was conducted on 17 patients before treatment, following active treatment (zoledronate 0.05 mg/kg 6-monthly or pamidronate 6-9 mg/kg/year) and after establishment on maintenance treatment for more than 2 years (zoledronate 0.025 mg/kg 6-monthly or pamidronate lean tissue mass increased during active treatment. These improvements were maintained during the period of maintenance treatment. Vertebral height improved in fractured thoracic vertebrae from pre-treatment to active therapy and improved further during maintenance treatment. Metacarpal cortical thickness and relative cortical area also increased over the treatment periods. Maintenance intravenous bisphosphonate therapy preserved the beneficial effects of active treatment at the doses stated above. Further studies are required to determine the optimal bisphosphonate treatment regimen in the management of children with OI. © 2015 S. Karger AG, Basel.

  10. A trolley mounted magazine for reactor maintenance

    Energy Technology Data Exchange (ETDEWEB)

    Brennan, P.J.; Madani, M.; Ridgway, G.H. [GE Hitachi Nuclear Energy Canada, Peterborough, Ontario (Canada)], E-mail: patrick1.brennan@ge.com, mehdi.madani@ge.com, guy.ridgway@ge.com; Lundy, E.; Knight, D. [IM and CS (Inspection, Maintenance and Commerical Services), Ontario Power Generation, Ajax, Ontario (Canada)], E-mail: erroll.lundy@opg.com, david.knight@opg.com

    2009-03-15

    This paper describes the design of a mechanism incorporating a rotary magazine to be mounted on a fuelling machine transport trolley for use at a Darlington reactor during a feeder replacement or maintenance outage. The magazine stores reactor channel maintenance components, such as channel isolation plugs and vented closure plugs, in twelve available magazine channels. Use of the magazine rather than a fuelling machine reduces the time required to transfer such components between the Central Service Area and reactor channels. Component transfers are accomplished by locking the fuelling machine onto one of the magazine channels and using a local controller to execute commands received from the fuel handling control system. (author)

  11. A trolley mounted magazine for reactor maintenance

    Energy Technology Data Exchange (ETDEWEB)

    Brennan, P.J.; Madani, M.; Ridgway, G.H., E-mail: patrick1.brennan@ge.com, E-mail: mehdi.madani@ge.com, E-mail: guy.ridgway@ge.com [GE Hitachi Nuclear Energy Canada, Peterborough, Ontario (Canada); Lundy, E.; Knight, D., E-mail: erroll.lundy@opg.com, E-mail: david.knight@opg.com [Ontario Power Generation, Inspection, Maintenance and Commercial Services, Ajax, Ontario (Canada)

    2008-07-01

    This paper describes the design of a mechanism incorporating a rotary magazine to be mounted on a fuelling machine transport trolley for use at a Darlington reactor during a feeder replacement or maintenance outage. The magazine stores reactor channel maintenance components, such as channel isolation plugs and vented closure plugs, in twelve available magazine channels. Use of the magazine rather than a fuelling machine reduces the time required to transfer such components between the Central Service Area and reactor channels. Component transfers are accomplished by locking the fuelling machine onto one of the magazine channels and using a local controller to execute commands received from the fuel handling control system. (author)

  12. A trolley mounted magazine for reactor maintenance

    International Nuclear Information System (INIS)

    Brennan, P.J.; Madani, M.; Ridgway, G.H.; Lundy, E.; Knight, D.

    2009-01-01

    This paper describes the design of a mechanism incorporating a rotary magazine to be mounted on a fuelling machine transport trolley for use at a Darlington reactor during a feeder replacement or maintenance outage. The magazine stores reactor channel maintenance components, such as channel isolation plugs and vented closure plugs, in twelve available magazine channels. Use of the magazine rather than a fuelling machine reduces the time required to transfer such components between the Central Service Area and reactor channels. Component transfers are accomplished by locking the fuelling machine onto one of the magazine channels and using a local controller to execute commands received from the fuel handling control system. (author)

  13. A trolley mounted magazine for reactor maintenance

    International Nuclear Information System (INIS)

    Brennan, P.J.; Madani, M.; Ridgway, G.H.; Lundy, E.; Knight, D.

    2008-01-01

    This paper describes the design of a mechanism incorporating a rotary magazine to be mounted on a fuelling machine transport trolley for use at a Darlington reactor during a feeder replacement or maintenance outage. The magazine stores reactor channel maintenance components, such as channel isolation plugs and vented closure plugs, in twelve available magazine channels. Use of the magazine rather than a fuelling machine reduces the time required to transfer such components between the Central Service Area and reactor channels. Component transfers are accomplished by locking the fuelling machine onto one of the magazine channels and using a local controller to execute commands received from the fuel handling control system. (author)

  14. Wave-induced Maintenance of Suspended Sediment Concentration during Slack in a Tidal Channel on a Sheltered Macro-tidal Flat, Gangwha Island, Korea

    Science.gov (United States)

    Lee, Guan-hong; Kang, KiRyong

    2018-05-01

    A field campaign was conducted to better understand the influence of wave action, in terms of turbulence and bed shear stress, on sediment resuspension and transport processes on a protected tidal flat. An H-frame was deployed in a tidal channel south of Gangwha Island for 6 tidal cycles during November 2006 with instrumentation including an Acoustic Doppler Velocimeter, an Acoustic Backscatter System, and an Optical Backscatter Sensor. During calm conditions, the current-induced shear was dominant and responsible for suspending sediments during the accelerating phases of flood and ebb. During the high-tide slack, both bed shear stress and suspended sediment concentration were reduced. The sediment flux was directed landward due to the scour-lag effect over a tidal cycle. On the other hand, when waves were stronger, the wave-induced turbulence appeared to keep sediments in suspension even during the high-tide slack, while the current-induced shear remained dominant during the accelerating phases of flood and ebb. The sediment flux under strong waves was directed offshore due to the sustained high suspended sediment concentration during the high-tide slack. Although strong waves can induce offshore sediment flux, infrequent events with strong waves are unlikely to alter the long-term accretion of the protected southern Gangwha tidal flats.

  15. Automated Methodologies for the Design of Flow Diagrams for Development and Maintenance Activities

    Science.gov (United States)

    Shivanand M., Handigund; Shweta, Bhat

    The Software Requirements Specification (SRS) of the organization is a text document prepared by strategic management incorporating the requirements of the organization. These requirements of ongoing business/ project development process involve the software tools, the hardware devices, the manual procedures, the application programs and the communication commands. These components are appropriately ordered for achieving the mission of the concerned process both in the project development and the ongoing business processes, in different flow diagrams viz. activity chart, workflow diagram, activity diagram, component diagram and deployment diagram. This paper proposes two generic, automatic methodologies for the design of various flow diagrams of (i) project development activities, (ii) ongoing business process. The methodologies also resolve the ensuing deadlocks in the flow diagrams and determine the critical paths for the activity chart. Though both methodologies are independent, each complements other in authenticating its correctness and completeness.

  16. Beta1 integrins activate a MAPK signalling pathway in neural stem cells that contributes to their maintenance

    DEFF Research Database (Denmark)

    Campos, Lia S; Leone, Dino P; Relvas, Joao B

    2004-01-01

    , signalling is required for neural stem cell maintenance, as assessed by neurosphere formation, and inhibition or genetic ablation of beta1 integrin using cre/lox technology reduces the level of MAPK activity. We conclude that integrins are therefore an important part of the signalling mechanisms that control......The emerging evidence that stem cells develop in specialised niches highlights the potential role of environmental factors in their regulation. Here we examine the role of beta1 integrin/extracellular matrix interactions in neural stem cells. We find high levels of beta1 integrin expression...... in the stem-cell containing regions of the embryonic CNS, with associated expression of the laminin alpha2 chain. Expression levels of laminin alpha2 are reduced in the postnatal CNS, but a population of cells expressing high levels of beta1 remains. Using neurospheres - aggregate cultures, derived from...

  17. [2,3-diphosphoglycerate level during the active and maintenance treatment of iron-deficiency anemia patients].

    Science.gov (United States)

    Iordanova, E; Dosheva, I; Lulcheva, F; Tsvetkova, N; Dobrev, K

    1985-01-01

    The objective of the present study was to obtain information about the duration of tissue hypoxia in patients with iron deficiency anemia. That fact is of importance for the determination of the duration of maintenance iron therapy. The level of 2,3-diphosphoglycerate was studied during the treatment, after the correction of anemic syndrome and after 60-day out-patient department treatment. The data obtained revealed that the level of 2,3-diphosphoglycerate was considerably elevated, as compared with the norm, before the treatment. After the active treatment and correction of anemic syndrome it was decreased, but remaining above the norm. By the 60th day of the out-patient department treatment the decrease continued and the level of 2,3-diphosphoglycerate approached the norm.

  18. CLEAN-ROADS project: air quality considerations after the application of a novel MDSS on winter road maintenance activities

    Science.gov (United States)

    Pretto, Ilaria; Malloci, Elisa; Tonidandel, Gabriele; Benedetti, Guido; Di Napoli, Claudia; Piazza, Andrea; Apolloni, Roberto; Cavaliere, Roberto

    2016-04-01

    With this poster we present the environmental benefit on air quality derived by the application of the CLEAN-ROADS pilot project. The CLEAN-ROADS project addresses the problem of the environmental pollution caused by de-icing salts during winter road maintenance activities in the Province of Trento (Italy). A demonstrative Maintenance Decision Support System (MDSS) has been developed in order to improve the intervention procedures of the road management service. Specifically it aims to optimize the efficiency of how available resources (e.g., salt consumption) are currently used while guaranteeing the current level of road safety. The CLEAN-ROADS project has been tested and validated on a test area located in a valley bottom (Adige Valley), where the highest optimization margins are to be expected. The project supports current road maintenance practices, which has proved to be reliable and accurate, with a new scalable and energy-efficient road monitoring system. This system is based on a network of road weather stations (road weather information system, RWIS) installed on the test route. It is capable to collect real-time data about the road conditions and to perform short-term and now-cast road weather forecasts, which actively integrate weather data and bulletins covering the target area [1]. This poster presents the results obtained from a three-year monitoring activity with the aim to (1) determine the impact of de-icing salts on air quality and (2) quantify the improvements obtained by the application of the CLEAN-ROADS project on air quality. The Ambient Air Quality and Cleaner Air for Europe Directive (2008/50/EC) states that contributions to exceedances of particulate matter PM10 limit values that are attributable to road winter salting may be subtracted when assessing compliance with air quality limit values, once provided that reasonable measures have been taken to lower concentrations [2]. As the de-icing salts used in road maintenance are mainly based

  19. Biological activity of the functional epitope of ciguatoxin fragment AB on the neuroblastoma sodium channel in tissue culture.

    Science.gov (United States)

    Hokama, Y; Chun, K E; Campora, C E; Higa, N; Suma, C; Hamajima, A; Isobe, M

    2006-01-01

    It is well established that the targeted receptor for ciguatoxin (CTX) in mammalian tissues is the sodium channel, affecting the influx of sodium into cells and altering the action potential and function of the cell. Since the syntheses of fragments of CTX has become available, our focus has been on the receptor functions of the west sphere AB and east sphere JKLM fragments using the neuroblastoma cell assay, guinea pig atrium assay, and the membrane immunobead assay (MIA). The data presented here suggest that the west sphere AB of the ciguatoxin molecule is the active portion and is responsible for the activation of the sodium channels. (c) 2006 Wiley-Liss, Inc.

  20. Extracts and compounds active on TRP ion channels from Waldheimia glabra, a ritual medicinal plant from Himalaya.

    Science.gov (United States)

    Giorgi, Annamaria; Bassoli, Angela; Borgonovo, Gigliola; Panseri, Sara; Manzo, Alessandra; Pentimalli, Daniela; Schiano Moriello, Aniello; De Petrocellis, Luciano

    2017-08-15

    Waldheimia glabra (Decne.) Regel is a wild plant from the Himalayan Mountains, commonly known as Smooth Ground Daisy. This plant is traditionally used by local populations in religious rituals (incense) or in traditional herbal medicine to treat skin diseases, headache, joint pain and fever. In literature few data are available on the investigation of this aromatic plant. The present work aims at deepening knowledge about the chemical composition of W. glabra extracts and incense, as well as its activity on TRP ion channels. Extracts and incense of W. glabra were analyzed by using HS-SPME GC/MS, GC/MS and NMR analysis. Tests on the activity of W. glabra extracts and isolated compounds (+)-ludartin 1 and B-ring-homo-tonghaosu 2 on TRP channels were also performed. Some extracts and pure compounds from W. glabra showed an interesting activity in terms of efficacy and potency on rat TRPA1, an ion channel involved in several sensory mechanisms, including pungency, environmental irritation and pain perception. Activity is discussed and compared with that of other known TRPA1 natural agonists with different chemical structures. All compounds showed only a negligible inhibition activity on rat TRPM8 ion channel. Our findings demonstrate that W. glabra is involved in the receptor activation mechanism and therefore represents a new natural product potentially useful in pharmaceutical and agrifood research. Copyright © 2017 Elsevier GmbH. All rights reserved.

  1. Cellular hyper-excitability caused by mutations that alter the activation process of voltage-gated sodium channels

    Directory of Open Access Journals (Sweden)

    Mohamed-Yassine eAMAROUCH

    2015-02-01

    Full Text Available Voltage-gated sodium channels (Nav are widely expressed as macro-molecular complexes in both excitable and non-excitable tissues. In excitable tissues, the upstroke of the action potential is the result of the passage of a large and rapid influx of sodium ions through these channels. NaV dysfunction has been associated with an increasingly wide range of neurological, muscular and cardiac disorders. The purpose of this review is to summarize the recently identified sodium channel mutations that are linked to hyper-excitability phenotypes and associated with the alteration of the activation process of voltage gated sodium channels. Indeed, several clinical manifestations that demonstrate an alteration of tissue excitability were recently shown to be strongly associated with the presence of mutations that affect the activation process of the voltage-gated sodium channels. These emerging genotype-phenotype correlations have expanded the clinical spectrum of sodium channelopathies to include disorders which feature a hyper-excitability phenotype that may or may not be associated with a cardiomyopathy. The p.I141V mutation in SCN4A and SCN5A, as well as its homologous p.I136V mutation in SCN9A, are interesting examples of mutations that have been linked to inherited hyperexcitability myotonia, exercise-induced polymorphic ventricular arrhythmias and erythromelalgia, respectively. Regardless of which sodium channel isoform is investigated, the substitution of the isoleucine to valine in the locus 141 induces similar modifications in the biophysical properties of the voltage-gated sodium channels by shifting the voltage-dependence of steady state activation towards more negative potentials.

  2. Hydrogen Sulfide Prevents Advanced Glycation End-Products Induced Activation of the Epithelial Sodium Channel

    Directory of Open Access Journals (Sweden)

    Qiushi Wang

    2015-01-01

    Full Text Available Advanced glycation end-products (AGEs are complex and heterogeneous compounds implicated in diabetes. Sodium reabsorption through the epithelial sodium channel (ENaC at the distal nephron plays an important role in diabetic hypertension. Here, we report that H2S antagonizes AGEs-induced ENaC activation in A6 cells. ENaC open probability (PO in A6 cells was significantly increased by exogenous AGEs and that this AGEs-induced ENaC activity was abolished by NaHS (a donor of H2S and TEMPOL. Incubating A6 cells with the catalase inhibitor 3-aminotriazole (3-AT mimicked the effects of AGEs on ENaC activity, but did not induce any additive effect. We found that the expression levels of catalase were significantly reduced by AGEs and both AGEs and 3-AT facilitated ROS uptake in A6 cells, which were significantly inhibited by NaHS. The specific PTEN and PI3K inhibitors, BPV(pic  and LY294002, influence ENaC activity in AGEs-pretreated A6 cells. Moreover, after removal of AGEs from AGEs-pretreated A6 cells for 72 hours, ENaC PO remained at a high level, suggesting that an AGEs-related “metabolic memory” may be involved in sodium homeostasis. Our data, for the first time, show that H2S prevents AGEs-induced ENaC activation by targeting the ROS/PI3K/PTEN pathway.

  3. OH vibrational activation and decay dynamics of CH4-OH entrance channel complexes

    International Nuclear Information System (INIS)

    Wheeler, Martyn D.; Tsiouris, Maria; Lester, Marsha I.; Lendvay, Gyoergy

    2000-01-01

    Infrared spectroscopy has been utilized to examine the structure and vibrational decay dynamics of CH 4 -OH complexes that have been stabilized in the entrance channel to the CH 4 +OH hydrogen abstraction reaction. Rotationally resolved infrared spectra of the CH 4 -OH complexes have been obtained in the OH fundamental and overtone regions using an IR-UV (infrared-ultraviolet) double-resonance technique. Pure OH stretching bands have been identified at 3563.45(5) and 6961.98(4) cm-1 (origins), along with combination bands involving the simultaneous excitation of OH stretching and intermolecular bending motions. The infrared spectra exhibit extensive homogeneous broadening arising from the rapid decay of vibrationally activated CH 4 -OH complexes due to vibrational relaxation and/or reaction. Lifetimes of 38(5) and 25(3) ps for CH 4 -OH prepared with one and two quanta of OH excitation, respectively, have been extracted from the infrared spectra. The nascent distribution of the OH products from vibrational predissociation has been evaluated by ultraviolet probe laser-induced fluorescence measurements. The dominant inelastic decay channel involves the transfer of one quantum of OH stretch to the pentad of CH 4 vibrational states with energies near 3000 cm-1. The experimental findings are compared with full collision studies of vibrationally excited OH with CH 4 . In addition, ab initio electronic structure calculations have been carried out to elucidate the minimum energy configuration of the CH 4 -OH complex. The calculations predict a C 3v geometry with the hydrogen of OH pointing toward one of four equivalent faces of the CH 4 tetrahedron, consistent with the analysis of the experimental infrared spectra. (c) 2000 American Institute of Physics

  4. Drosophila SLC5A11 Mediates Hunger by Regulating K(+) Channel Activity.

    Science.gov (United States)

    Park, Jin-Yong; Dus, Monica; Kim, Seonil; Abu, Farhan; Kanai, Makoto I; Rudy, Bernardo; Suh, Greg S B

    2016-08-08

    Hunger is a powerful drive that stimulates food intake. Yet, the mechanism that determines how the energy deficits that result in hunger are represented in the brain and promote feeding is not well understood. We previously described SLC5A11-a sodium/solute co-transporter-like-(or cupcake) in Drosophila melanogaster, which is required for the fly to select a nutritive sugar over a sweeter nonnutritive sugar after periods of food deprivation. SLC5A11 acts on approximately 12 pairs of ellipsoid body (EB) R4 neurons to trigger the selection of nutritive sugars, but the underlying mechanism is not understood. Here, we report that the excitability of SLC5A11-expressing EB R4 neurons increases dramatically during starvation and that this increase is abolished in the SLC5A11 mutation. Artificial activation of SLC5A11-expresssing neurons is sufficient to promote feeding and hunger-driven behaviors; silencing these neurons has the opposite effect. Notably, SLC5A11 transcript levels in the brain increase significantly when flies are starved and decrease shortly after starved flies are refed. Furthermore, expression of SLC5A11 is sufficient for promoting hunger-driven behaviors and enhancing the excitability of SLC5A11-expressing neurons. SLC5A11 inhibits the function of the Drosophila KCNQ potassium channel in a heterologous expression system. Accordingly, a knockdown of dKCNQ expression in SLC5A11-expressing neurons produces hunger-driven behaviors even in fed flies, mimicking the overexpression of SLC5A11. We propose that starvation increases SLC5A11 expression, which enhances the excitability of SLC5A11-expressing neurons by suppressing dKCNQ channels, thereby conferring the hunger state. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Cytoskeleton, L-type Ca2+ and stretch activated channels in injured skeletal muscle

    Directory of Open Access Journals (Sweden)

    Fabio Francini

    2013-07-01

    Full Text Available The extra-sarcomeric cytoskeleton (actin microfilaments and anchoring proteins is involved in maintaining the sarco-membrane stiffness and integrity and in turn the mechanical stability and function of the intra- and sub-sarcoplasmic proteins. Accordingly, it regulates Ca2+ entry through the L-type Ca2+ channels and the mechano-sensitivity of the stretch activated channels (SACs. Moreover, being intra-sarcomeric cytoskeleton bound to costameric proteins and other proteins of the sarcoplasma by intermediate filaments, as desmin, it integrates the properties of the sarcolemma with the skeletal muscle fibres contraction. The aim of this research was to compare the cytoskeleton, SACs and the ECC alterations in two different types of injured skeletal muscle fibres: by muscle denervation and mechanical overload (eccentric contraction. Experiments on denervation were made in isolated Soleus muscle of male Wistar rats; forced eccentric-contraction (EC injury was achieved in Extensor Digitorum Longus muscles of Swiss mice. The method employed conventional intracellular recording with microelectrodes inserted in a single fibre of an isolated skeletal muscle bundle. The state of cytoskeleton was evaluated by recording SAC currents and by evaluating the resting membrane potential (RMP value determined in current-clamp mode. The results demonstrated that in both injured skeletal muscle conditions the functionality of L-type Ca2+ current, ICa, was affected. In parallel, muscle fibres showed an increase of the resting membrane permeability and of the SAC current. These issues, together with a more depolarized RMP are an index of altered cytoskeleton. In conclusion, we found a symilar alteration of ICa, SAC and cytoskeleton in both injured skeletal muscle conditions.

  6. Mutation of I696 and W697 in the TRP box of vanilloid receptor subtype I modulates allosteric channel activation.

    Science.gov (United States)

    Gregorio-Teruel, Lucia; Valente, Pierluigi; González-Ros, José Manuel; Fernández-Ballester, Gregorio; Ferrer-Montiel, Antonio

    2014-03-01

    The transient receptor potential vanilloid receptor subtype I (TRPV1) channel acts as a polymodal sensory receptor gated by chemical and physical stimuli. Like other TRP channels, TRPV1 contains in its C terminus a short, conserved domain called the TRP box, which is necessary for channel gating. Substitution of two TRP box residues-I696 and W697-with Ala markedly affects TRPV1's response to all activating stimuli, which indicates that these two residues play a crucial role in channel gating. We systematically replaced I696 and W697 with 18 native l-amino acids (excluding cysteine) and evaluated the effect on voltage- and capsaicin-dependent gating. Mutation of I696 decreased channel activation by either voltage or capsaicin; furthermore, gating was only observed with substitution of hydrophobic amino acids. Substitution of W697 with any of the 18 amino acids abolished gating in response to depolarization alone, shifting the threshold to unreachable voltages, but not capsaicin-mediated gating. Moreover, vanilloid-activated responses of W697X mutants showed voltage-dependent gating along with a strong voltage-independent component. Analysis of the data using an allosteric model of activation indicates that mutation of I696 and W697 primarily affects the allosteric coupling constants of the ligand and voltage sensors to the channel pore. Together, our findings substantiate the notion that inter- and/or intrasubunit interactions at the level of the TRP box are critical for efficient coupling of stimulus sensing and gate opening. Perturbation of these interactions markedly reduces the efficacy and potency of the activating stimuli. Furthermore, our results identify these interactions as potential sites for pharmacological intervention.

  7. Preventative Maintenance.

    Science.gov (United States)

    Migliorino, James

    Boards of education must be convinced that spending money up front for preventive maintenance will, in the long run, save districts' tax dollars. A good program of preventive maintenance can minimize disruption of service; reduce repair costs, energy consumption, and overtime; improve labor productivity and system equipment reliability; handle…

  8. Acute stress enhances learning and memory by activating acid-sensing ion channels in rats.

    Science.gov (United States)

    Ye, Shunjie; Yang, Rong; Xiong, Qiuju; Yang, Youhua; Zhou, Lianying; Gong, Yeli; Li, Changlei; Ding, Zhenhan; Ye, Guohai; Xiong, Zhe

    2018-04-15

    Acute stress has been shown to enhance learning and memory ability, predominantly through the action of corticosteroid stress hormones. However, the valuable targets for promoting learning and memory induced by acute stress and the underlying molecular mechanisms remain unclear. Acid-sensing ion channels (ASICs) play an important role in central neuronal systems and involves in depression, synaptic plasticity and learning and memory. In the current study, we used a combination of electrophysiological and behavioral approaches in an effort to explore the effects of acute stress on ASICs. We found that corticosterone (CORT) induced by acute stress caused a potentiation of ASICs current via glucocorticoid receptors (GRs) not mineralocorticoid receptors (MRs). Meanwhile, CORT did not produce an increase of ASICs current by pretreated with GF109203X, an antagonist of protein kinase C (PKC), whereas CORT did result in a markedly enhancement of ASICs current by bryostatin 1, an agonist of PKC, suggesting that potentiation of ASICs function may be depended on PKC activating. More importantly, an antagonist of ASICs, amiloride (10 μM) reduced the performance of learning and memory induced by acute stress, which is further suggesting that ASICs as the key components involves in cognitive processes induced by acute stress. These results indicate that acute stress causes the enhancement of ASICs function by activating PKC signaling pathway, which leads to potentiated learning and memory. Copyright © 2018 Elsevier Inc. All rights reserved.

  9. Behavioural decisions of travel-time ratios for work, maintenance and leisure activities in the Netherlands.

    NARCIS (Netherlands)

    Susilo, Y.S.; Dijst, M.J.

    2010-01-01

    Using the 2005 Dutch National Travel Survey data-set this paper investigates the influences of socio-demographics, journey patterns and built environment factors on the ratio of travel time and activity duration that an individual spends when engaging in work, daily shopping, non-daily shopping

  10. Activation of acid-sensing ion channels by localized proton transient reveals their role in proton signaling.

    Science.gov (United States)

    Zeng, Wei-Zheng; Liu, Di-Shi; Liu, Lu; She, Liang; Wu, Long-Jun; Xu, Tian-Le

    2015-09-15

    Extracellular transients of pH alterations likely mediate signal transduction in the nervous system. Neuronal acid-sensing ion channels (ASICs) act as sensors for extracellular protons, but the mechanism underlying ASIC activation remains largely unknown. Here, we show that, following activation of a light-activated proton pump, Archaerhodopsin-3 (Arch), proton transients induced ASIC currents in both neurons and HEK293T cells co-expressing ASIC1a channels. Using chimera proteins that bridge Arch and ASIC1a by a glycine/serine linker, we found that successful coupling occurred within 15 nm distance. Furthermore, two-cell sniffer patch recording revealed that regulated release of protons through either Arch or voltage-gated proton channel Hv1 activated neighbouring cells expressing ASIC1a channels. Finally, computational modelling predicted the peak proton concentration at the intercellular interface to be at pH 6.7, which is acidic enough to activate ASICs in vivo. Our results highlight the pathophysiological role of proton signalling in the nervous system.

  11. Phosphatidylinositol (4,5)bisphosphate inhibits K+-efflux channel activity in NT1 tobacco cultured cells.

    Science.gov (United States)

    Ma, Xiaohong; Shor, Oded; Diminshtein, Sofia; Yu, Ling; Im, Yang Ju; Perera, Imara; Lomax, Aaron; Boss, Wendy F; Moran, Nava

    2009-02-01

    In the animal world, the regulation of ion channels by phosphoinositides (PIs) has been investigated extensively, demonstrating a wide range of channels controlled by phosphatidylinositol (4,5)bisphosphate (PtdInsP2). To understand PI regulation of plant ion channels, we examined the in planta effect of PtdInsP2 on the K+-efflux channel of tobacco (Nicotiana tabacum), NtORK (outward-rectifying K channel). We applied a patch clamp in the whole-cell configuration (with fixed "cytosolic" Ca2+ concentration and pH) to protoplasts isolated from cultured tobacco cells with genetically manipulated plasma membrane levels of PtdInsP2 and cellular inositol (1,4,5)trisphosphate: "Low PIs" had depressed levels of these PIs, and "High PIs" had elevated levels relative to controls. In all of these cells, K channel activity, reflected in the net, steady-state outward K+ currents (IK), was inversely related to the plasma membrane PtdInsP2 level. Consistent with this, short-term manipulations decreasing PtdInsP2 levels in the High PIs, such as pretreatment with the phytohormone abscisic acid (25 microM) or neutralizing the bath solution from pH 5.6 to pH 7, increased IK (i.e. NtORK activity). Moreover, increasing PtdInsP2 levels in controls or in abscisic acid-treated high-PI cells, using the specific PI-phospholipase C inhibitor U73122 (2.5-4 microM), decreased NtORK activity. In all cases, IK decreases stemmed largely from decreased maximum attainable NtORK channel conductance and partly from shifted voltage dependence of channel gating to more positive potentials, making it more difficult to activate the channels. These results are consistent with NtORK inhibition by the negatively charged PtdInsP2 in the internal plasma membrane leaflet. Such effects are likely to underlie PI signaling in intact plant cells.

  12. Urinary Proteolytic Activation of Renal Epithelial Na+ Channels in Chronic Heart Failure.

    Science.gov (United States)

    Zheng, Hong; Liu, Xuefei; Sharma, Neeru M; Li, Yulong; Pliquett, Rainer U; Patel, Kaushik P

    2016-01-01

    One of the key mechanisms involved in renal Na(+) retention in chronic heart failure (CHF) is activation of epithelial Na(+) channels (ENaC) in collecting tubules. Proteolytic cleavage has an important role in activating ENaC. We hypothesized that enhanced levels of proteases in renal tubular fluid activate ENaC, resulting in renal Na(+) retention in rats with CHF. CHF was produced by left coronary artery ligation in rats. By immunoblotting, we found that several urinary serine proteases were significantly increased in CHF rats compared with sham rats (fold increases: furin 6.7, prostasin 23.6, plasminogen 2.06, and plasmin 3.57 versus sham). Similar increases were observed in urinary samples from patients with CHF. Whole-cell patch clamp was conducted in cultured renal collecting duct M-1 cells to record Na(+) currents. Protease-rich urine (from rats and patients with CHF) significantly increased the Na(+) inward current in M-1 cells. Two weeks of protease inhibitor treatment significantly abrogated the enhanced diuretic and natriuretic responses to ENaC inhibitor benzamil in rats with CHF. Increased podocyte lesions were observed in the kidneys of rats with CHF by transmission electron microscopy. Consistent with these results, podocyte damage markers desmin and podocin expressions were also increased in rats with CHF (increased ≈2-folds). These findings suggest that podocyte damage may lead to increased proteases in the tubular fluid, which in turn contributes to the enhanced renal ENaC activity, providing a novel mechanistic insight for Na(+) retention commonly observed in CHF. © 2015 American Heart Association, Inc.

  13. Urinary proteolytic activation of renal epithelial Na+ channels in chronic heart failure

    Science.gov (United States)

    Zheng, Hong; Liu, Xuefei; Sharma, Neeru M.; Li, Yulong; Pliquett, Rainer U; Patel, Kaushik P.

    2015-01-01

    One of the key mechanisms involved in renal Na+ retention in chronic heart failure (CHF) is activation of epithelial Na+ channels (ENaC) in collecting tubules. Proteolytic cleavage has an important role in activating ENaC. We hypothesized that enhanced levels of proteases in renal tubular fluid activate ENaC resulting in renal Na+ retention in rats with CHF. CHF was produced by left coronary artery ligation in rats. By immunoblotting, we found that several urinary serine proteases were significantly increased in CHF rats compared to sham rats (fold increases: furin 6.7, prostasin 23.6, plasminogen 2.06 and plasmin 3.57 vs. sham). Similar increases were observed in urinary samples from patients with CHF. Whole-cell patch-clamp was conducted in cultured renal collecting duct M-1 cells to record Na+ currents. Protease-rich urine (from rats and patients with CHF) significantly increased the Na+ inward current in M-1 cells. Two weeks of protease inhibitor treatment significantly abrogated the enhanced diuretic and natriuretic responses to ENaC inhibitor benzamil in rats with CHF. Increased podocyte lesions were observed in the kidneys of rats with CHF by transmission electron microscopy. Consistent with these results, podocyte damage markers desmin and podocin expressions were also increased in rats with CHF (increased ~2 folds). These findings suggest that podocyte damage may lead to increased proteases in the tubular fluid which in turn contributes to the enhanced renal ENaC activity, providing a novel mechanistic insight for Na+ retention commonly observed in CHF. PMID:26628676

  14. Cholesterol up-regulates neuronal G protein-gated inwardly rectifying potassium (GIRK) channel activity in the hippocampus.

    Science.gov (United States)

    Bukiya, Anna N; Durdagi, Serdar; Noskov, Sergei; Rosenhouse-Dantsker, Avia

    2017-04-14

    Hypercholesterolemia is a well known risk factor for the development of neurodegenerative disease. However, the underlying mechanisms are mostly unknown. In recent years, it has become increasingly evident that cholesterol-driven effects on physiology and pathophysiology derive from its ability to alter the function of a variety of membrane proteins including ion channels. Yet, the effect of cholesterol on G protein-gated inwardly rectifying potassium (GIRK) channels expressed in the brain is unknown. GIRK channels mediate the actions of inhibitory brain neurotransmitters. As a result, loss of GIRK function can enhance neuron excitability, whereas gain of GIRK function can reduce neuronal activity. Here we show that in rats on a high-cholesterol diet, cholesterol levels in hippocampal neurons are increased. We also demonstrate that cholesterol plays a critical role in modulating neuronal GIRK currents. Specifically, cholesterol enrichment of rat hippocampal neurons resulted in enhanced channel activity. In accordance, elevated currents upon cholesterol enrichment were also observed in Xenopus oocytes expressing GIRK2 channels, the primary GIRK subunit expressed in the brain. Furthermore, using planar lipid bilayers, we show that although cholesterol did not affect the unitary conductance of GIRK2, it significantly enhanced the frequency of channel openings. Last, combining computational and functional approaches, we identified two putative cholesterol-binding sites in the transmembrane domain of GIRK2. These findings establish that cholesterol plays a critical role in modulating GIRK activity in the brain. Because up-regulation of GIRK function can reduce neuronal activity, our findings may lead to novel approaches for prevention and therapy of cholesterol-driven neurodegenerative disease. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  15. Regulatory inspection activities related to inspection planning, plant maintenance and assessment of safety. Proceedings of an international workshop

    Energy Technology Data Exchange (ETDEWEB)

    Van Binnebeek, J. J. [AIB-Vincotte Nuclear - AVN, Avenue du Roi, 157, B-1060 Brussels (Belgium); Aubrey, Richard; Grandame, Melvyn [Atomic Energy Control Board - AECB, P.O. Box 1046, Station B, 280 Slater Street, Ottawa, Ontario K1P 5S9 (Canada); Aro, Ilari [Finnish Centre for Radiation and Nuclear Safety - STUK, P.O. Box 14, FIN-00881 Helsinki (Finland); Balloffet, Yves [DRIRE Rhone Alpes, 146, rue Pierre Corneille, 69426 Lyon CEDEX 03 (France); Klonk, Hartmut [Bundesamt fuer Strahlenschutz - BfS, Federal Office for Radiation Protection, Postbox 10 01 49, 38201 Salzgitter 1 (Germany); Manzella, Pietro [A.N.P.A., Via V. Brancati, 48, 1-00144 Roma EUR (Italy); Koizumi, Hiroyoshi [Tech. Stan. Dept. - JAPEIC, Shin-Toranomon Bldg., 1-5-11, Akasaka, Minato-ku, Tokyo 107 (Japan); Bouvrie, E.C. des [Ministry of Social Affairs and Employment, Nuclear Safety Dept. KFD, P.O. Box 90804, 2509 LV The Hague (Netherlands); Forsberg, Staffan [Swedish Nuclear Power Inspectorate - SKI, Klarabergsviadukten 90, S-10658 Stockholm (Sweden); Lang, Hans-Guenter [Section Plant Coordination and Inspection, Swiss Federal Nuclear Safety Inspectorate - HSK, CH-5232 Villigen-HSK (Switzerland); Mehew, Robert; Warren, Thomas; Woodhouse, Paul [Health and Safety Executive - NII, St. Peter' s House, Balliol Road, Bootle, Merseyside L20 3LZ (United Kingdom); Gallo, Robert M. [Special Inspection Branch, US Nuclear Regulatory Commission - US NRC, Mail Stop 0-9A1, Washington, DC 20555 (United States); Campbell, Rob [International Atomic Energy Agency - IAEA, P.O. Box 100, A-1400 Vienna (International Atomic Energy Agency (IAEA))

    1997-07-01

    The NEA Committee on Nuclear Regulatory Activities (CNRA) believes that an essential factor in ensuring the safety of nuclear installations is the continuing exchange and analysis of technical information and data. To facilitate this exchange the Committee has established Working Groups and Groups of Experts in specialised topics. CNRA believes that safety inspections are a major element in the regulatory authority's efforts to ensure the safe operation of nuclear facilities. Considering the importance of these issues, the Committee has established a special Working Group on Inspection Practices (WGIP). The purpose of WGIP, is to facilitate the exchange of information and experience related to regulatory safety inspections between CNRA Member countries. This was the 3. international workshop held by the WGIP on regulatory inspection activities. The focus of this workshop was on 3 main topics; Inspection Planning, Plant Maintenance and Assessment of Safety. This document presents the proceedings from the workshop, including: workshop programme, results and conclusions, papers and presentations and the list of participants. The main purpose of the Workshop is to provide a forum of exchange of information on the regulatory inspection activities.

  16. Regulatory inspection activities related to inspection planning, plant maintenance and assessment of safety. Proceedings of an international workshop

    International Nuclear Information System (INIS)

    Van Binnebeek, J.J.; Aubrey, Richard; Grandame, Melvyn; Aro, Ilari; Balloffet, Yves; Klonk, Hartmut; Manzella, Pietro; Koizumi, Hiroyoshi; Bouvrie, E.C. des; Forsberg, Staffan; Lang, Hans-Guenter; Mehew, Robert; Warren, Thomas; Woodhouse, Paul; Gallo, Robert M.; Campbell, Rob; )

    1997-01-01

    The NEA Committee on Nuclear Regulatory Activities (CNRA) believes that an essential factor in ensuring the safety of nuclear installations is the continuing exchange and analysis of technical information and data. To facilitate this exchange the Committee has established Working Groups and Groups of Experts in specialised topics. CNRA believes that safety inspections are a major element in the regulatory authority's efforts to ensure the safe operation of nuclear facilities. Considering the importance of these issues, the Committee has established a special Working Group on Inspection Practices (WGIP). The purpose of WGIP, is to facilitate the exchange of information and experience related to regulatory safety inspections between CNRA Member countries. This was the 3. international workshop held by the WGIP on regulatory inspection activities. The focus of this workshop was on 3 main topics; Inspection Planning, Plant Maintenance and Assessment of Safety. This document presents the proceedings from the workshop, including: workshop programme, results and conclusions, papers and presentations and the list of participants. The main purpose of the Workshop is to provide a forum of exchange of information on the regulatory inspection activities

  17. Research and development strategy and maintenance engineering to strengthen the basis of ageing management. International activities

    International Nuclear Information System (INIS)

    Sekimura, Naoto

    2009-01-01

    Systematic development of information basis for database and knowledge-base has been performed in addition to the development of codes and standards by academic societies, regulatory bodies and industries through the intensive domestic safety research collaborations and international collaboration, through the continuous revision of Strategy Maps for Ageing Management and Safe Long Term Operation. Important international activities in IAEA and OECD/NEA especially for knowledge-base and extraction of commendable practices in ageing management are discussed. (author)

  18. Lessons Learned from Outsourcing the Pearl Harbor MK-48 Intermediate Maintenance Activity

    Science.gov (United States)

    2008-03-01

    transfer of ownership of assets. However, outsourcing is not simply a procurement decision. All organizations procure elements of their operations...systems, commercialization, and franchising (Office of Management and Budget, 2004). Strengths of Outsourcing Outsourcing is more efficient than...operating the IMA with organic resources vs . the cost of a comparable service provided by a contractor. Activity-based costing would also help managers

  19. EFFECTIVENESS OF DIFFERENT TRAINING PROGRAMS FOR PHYSICAL PERFORMANCE MAINTENANCE IN THE CONDITION OF LOW MOTOR ACTIVITY.

    Science.gov (United States)

    Fomina, E V; Uskov, K V

    Investigations made by the World Health Organization showed that lack. of physical activity and sedentary lifestyle are among ten major causes for death and disability. Typially, studies of the negative effects of reduced physical activity and their prevention face difficulties of providing standard conditions. These issues were obviated successfully in experiment MARS-500 on simulation of a space exploration mission. Human subjects were 6 men from Russia, France, Italy and China who spent 520 days in isolation. To prevent the negative effects of low physical activity, the volunteers performed resistive and cyclic exercises in different periods of isolation. The study was designed with two pauses in the training program. Physical performance was evaluated with an incremental loading test, maximal voluntary effort test (Russian-Austrian MDS resistive exercise system), and PWC-170 (bicycle ergometer). Level of physical performance of the subjects in this experiment never degraded to below baseline values. The proposed training system comprising different kinds of exercise ensured stability or improvement of subjects' physical performance as compared with baseline level.

  20. Proceedings of the third international conference on CANDU maintenance

    International Nuclear Information System (INIS)

    1995-01-01

    The third international conference on Candu maintenance included sessions on the following topics: predictive maintenance, reliability improvements, steam generator monitoring, tools and instrumentation, valve performance, fuel channel inspection and maintenance, steam generator maintenance, environmental qualification, predictive maintenance, instrumentation and control, steam generator cleaning, decontamination and radiation protection, inspection techniques, maintenance program strategies and valve packing experience, remote tooling/ robotics and fuel handling. The individual papers have been abstracted separately

  1. Proceedings of the third international conference on CANDU maintenance

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1996-12-31

    The third international conference on Candu maintenance included sessions on the following topics: predictive maintenance, reliability improvements, steam generator monitoring, tools and instrumentation, valve performance, fuel channel inspection and maintenance, steam generator maintenance, environmental qualification, predictive maintenance, instrumentation and control, steam generator cleaning, decontamination and radiation protection, inspection techniques, maintenance program strategies and valve packing experience, remote tooling/ robotics and fuel handling. The individual papers have been abstracted separately.

  2. Identification of the functional binding pocket for compounds targeting small-conductance Ca²⁺-activated potassium channels.

    Science.gov (United States)

    Zhang, Miao; Pascal, John M; Schumann, Marcel; Armen, Roger S; Zhang, Ji-Fang

    2012-01-01

    Small- and intermediate-conductance Ca(2+)-activated potassium channels, activated by Ca(2+)-bound calmodulin, have an important role in regulating membrane excitability. These channels are also linked to clinical abnormalities. A tremendous amount of effort has been devoted to developing small molecule compounds targeting these channels. However, these compounds often suffer from low potency and lack of selectivity, hindering their potential for clinical use. A key contributing factor is the lack of knowledge of the binding site(s) for these compounds. Here we demonstrate by X-ray crystallography that the binding pocket for the compounds of the 1-ethyl-2-benzimidazolinone (1-EBIO) class is located at the calmodulin-channel interface. We show that, based on structure data and molecular docking, mutations of the channel can effectively change the potency of these compounds. Our results provide insight into the molecular nature of the binding pocket and its contribution to the potency and selectivity of the compounds of the 1-EBIO class.

  3. Identification of the functional binding pocket for compounds targeting small-conductance Ca2+-activated potassium channels

    Science.gov (United States)

    Zhang, Miao; Pascal, John M.; Schumann, Marcel; Armen, Roger S.; Zhang, Ji-fang

    2012-01-01

    Small- and intermediate-conductance Ca2+-activated potassium channels, activated by Ca2+-bound calmodulin, play an important role in regulating membrane excitability. These channels are also linked to clinical abnormalities. A tremendous amount of effort has been devoted to developing small molecule compounds targeting these channels. However, these compounds often suffer from low potency and lack of selectivity, hindering their potentials for clinical use. A key contributing factor is the lack of knowledge of the binding site(s) for these compounds. Here we demonstrate by X-ray crystallography that the binding pocket for the compounds of the 1-EBIO class is located at the calmodulin-channel interface. We show that, based on structure data and molecular docking, mutations of the channel can effectively change the potency of these compounds. Our results provide insight into the molecular nature of the binding pocket and its contribution to the potency and selectivity of the compounds of the 1-EBIO class. PMID:22929778

  4. [Amplitude Changes of Low Frequency Fluctuation in Brain Spontaneous Nervous Activities Induced by Needling at Hand Taiyin Lung Channel].

    Science.gov (United States)

    Zhou, You-long; Su, Cheng-guo; Liu, Shou-fang; Jin, Xiang-yu; Duan, Yan-li; Chen, Xiao-yan; Zhao, Shu-hua; Wang, Quan-liang; Dang, Chang-lin

    2016-05-01

    To observe amplitude changes of low frequency fluctuation in brain spontaneous nervous activities induced by needling at Hand Taiyin Lung Channel, and to preliminarily explore the possible brain function network of Hand Taiyin Lung Channel. By using functional magnetic resonance imaging (fMRI), 16 healthy volunteers underwent resting-state scanning (R1) and scanning with retained acupuncture at Hand Taiyin Lung Channel (acupuncture, AP). Data of fMRI collected were statistically calculated using amplitude of low frequency fluctuations (ALFF). Under R1 significantly enhanced ALFF occurred in right precuneus, left inferior parietal lobule, bilateral superior temporal gyrus, bilateral middle frontal gyrus, left superior frontal gyrus, left inferior frontal gyrus, left medial frontal gyrus. Under AP significantly enhanced ALFF occurred in right precuneus, bilateral superior frontal gyrus, cerebellum, bilateral middle frontal gyrus, right medial frontal gyrus, and so on. Compared with R1, needing at Hand Taiyin Lung Channel could significantly enhance ALFF in right gyrus subcallosum and right inferior frontal gyrus. Significant decreased ALFF appeared in right postcentral gyrus, left precuneus, left superior temporal gyrus, left middle temporal gyrus, and so on. Needing at Hand Taiyin Lung Channel could significantly change fixed activities of cerebral cortex, especially in right subcallosal gyrus, right inferior frontal gyrus, and so on.

  5. Homogenization of independent inspection criteria of Quality Assurance in the maintenance activities; Homogeneizacion de criterios en la inspeccion independiente de Garantia de Calidad a las actividades de mantenimiento

    Energy Technology Data Exchange (ETDEWEB)

    Gomez Cardinanos, R.

    2012-07-01

    Among the official documents governing the operation of Spanish nuclear power plants is the Quality Manual and Quality Assurance. This manual gives a formal answer to the requirements of the regulations and the Nuclear Safety Council. To monitor the compliance of its requirements, the plants are controlled by independent organizations. Since 2007, in nuclear Sta. Maria de Garona is developed a project to improve the performance of the QA inspectors during maintenance activities. Results of the project have been edited data sheets on specific activities of maintenance. The project aims to incorporate sheets that help QA inspectors in their daily inspection.

  6. Insulin activates single amiloride-blockable Na channels in a distal nephron cell line (A6).

    Science.gov (United States)

    Marunaka, Y; Hagiwara, N; Tohda, H

    1992-09-01

    Using the patch-clamp technique, we studied the effect of insulin on an amiloride-blockable Na channel in the apical membrane of a distal nephron cell line (A6) cultured on permeable collagen films for 10-14 days. NPo (N, number of channels per patch membrane; Po, average value of open probability of individual channels in the patch) under baseline conditions was 0.88 +/- 0.12 (SE)(n = 17). After making cell-attached patches on the apical membrane which contained Na channels, insulin (1 mU/ml) was applied to the serosal bath. While maintaining the cell-attached patch, NPo significantly increased to 1.48 +/- 0.19 (n = 17; P less than 0.001) after 5-10 min of insulin application. The open probability of Na channels was 0.39 +/- 0.01 (n = 38) under baseline condition, and increased to 0.66 +/- 0.03 (n = 38, P less than 0.001) after addition of insulin. The baseline single-channel conductance was 4pS, and neither the single-channel conductance nor the current-voltage relationship was significantly changed by insulin. These results indicate that insulin increases Na absorption in the distal nephron by increasing the open probability of the amiloride-blockable Na channel.

  7. Hyperthyroidism enhances 5-HT-induced contraction of the rat pulmonary artery: role of calcium-activated chloride channel activation.

    Science.gov (United States)

    Oriowo, Mabayoje A; Oommen, Elsie; Khan, Islam

    2011-11-01

    Experimentally-induced hyperthyroidism in rodents is associated with signs and symptoms of pulmonary hypertension. The main objective of the present study was to investigate the effect of thyroxine-induced pulmonary hypertension on the contractile response of the pulmonary artery to 5-HT and the possible underlying signaling pathway. 5-HT concentration-dependently contracted artery segments from control and thyroxine-treated rats with pD(2) values of 5.04 ± 0.19 and 5.34 ± 0.14, respectively. The maximum response was significantly greater in artery segments from thyroxine-treated rats. Neither BW 723C86 (5-HT(2B)-receptor agonist) nor CP 93129 (5-HT(1B)-receptor agonist) contracted ring segments of the pulmonary artery from control and thyroxine-treated rats at concentrations up to 10(-4)M. There was no significant difference in the level of expression of 5-HT(2A)-receptor protein between the two groups. Ketanserin (3 × 10(-8)M) produced a rightward shift of the concentration-response curve to 5-HT in both groups with equal potency (-logK(B) values were 8.1 ± 0.2 and 7.9 ± 0.1 in control and thyroxine-treated rats, respectively). Nifedipine (10(-6)M) inhibited 5-HT-induced contractions in artery segments from control and thyroxine-treated rats and was more effective against 5-HT-induced contraction in artery segments for thyroxine-treated rats. The calcium-activated chloride channel blocker, niflumic acid (10(-4)M) also inhibited 5-HT-induced contractions in artery segments from control and thyroxine-treated rats and was more effective against 5-HT-induced contraction in artery segments for thyroxine-treated rats. It was concluded that hyperthyroidism enhanced 5-HT-induced contractions of the rat pulmonary artery by a mechanism involving increased activity of calcium-activated chloride channels. Copyright © 2011 Elsevier B.V. All rights reserved.

  8. Blockade of Ca2+-activated K+ channels in T cells: an option for the treatment of multiple sclerosis?

    DEFF Research Database (Denmark)

    Madsen, Lars Siim; Christophersen, Palle; Olesen, Søren-Peter

    2005-01-01

    Voltage- and Ca(2+)-dependent K(+) channels in the membrane of both T and B lymphocytes are important for the cellular immune response. In the current issue of the European Journal of Immunology, Reich et al. demonstrate that selective blockade of the intermediate-conductance Ca(2+)-activated K(+...

  9. Role of calcium-activated potassium channels with small conductance in bradykinin-induced vasodilation of porcine retinal arterioles

    DEFF Research Database (Denmark)

    Dalsgaard, Thomas; Kroigaard, Christel; Bek, Toke

    2009-01-01

    PURPOSE: Endothelial dysfunction and impaired vasodilation may be involved in the pathogenesis of retinal vascular diseases. In the present study, the mechanisms underlying bradykinin vasodilation were examined and whether calcium-activated potassium channels of small (SK(Ca)) and intermediate (IK...

  10. Alkali pH directly activates ATP-sensitive K+ channels and inhibits insulin secretion in beta-cells.

    Science.gov (United States)

    Manning Fox, Jocelyn E; Karaman, Gunce; Wheeler, Michael B

    2006-11-17

    Glucose stimulation of pancreatic beta-cells is reported to lead to sustained alkalization, while extracellular application of weak bases is reported to inhibit electrical activity and decrease insulin secretion. We hypothesize that beta-cell K(ATP) channel activity is modulated by alkaline pH. Using the excised patch-clamp technique, we demonstrate a direct stimulatory action of alkali pH on recombinant SUR1/Kir6.2 channels due to increased open probability. Bath application of alkali pH similarly activates native islet beta-cell K(ATP) channels, leading to an inhibition of action potentials, and hyperpolarization of membrane potential. In situ pancreatic perfusion confirms that these cellular effects of alkali pH are observable at a functional level, resulting in decreases in both phase 1 and phase 2 glucose-stimulated insulin secretion. Our data are the first to report a stimulatory effect of a range of alkali pH on K(ATP) channel activity and link this to downstream effects on islet beta-cell function.

  11. Improvements to executive function during exercise training predict maintenance of physical activity over the following year

    Directory of Open Access Journals (Sweden)

    John eBest

    2014-05-01

    Full Text Available Previous studies have shown that exercise training benefits cognitive, neural, and physical health markers in older adults. It is likely that these positive effects will diminish if participants return to sedentary lifestyles following training cessation. Theory posits that that the neurocognitive processes underlying self-regulation, namely executive function (EF, are important to maintaining positive health behaviors. Therefore, we examined whether better EF performance in older women would predict greater adherence to routine physical activity (PA over 1 year following a 12-month resistance exercise training randomized controlled trial. The study sample consisted of 125 community-dwelling women aged 65 to 75 years old. Our primary outcome measure was self-reported PA, as measured by the Physical Activity Scale for the Elderly (PASE, assessed on a monthly basis from month 13 to month 25. Executive function was assessed using the Stroop Test at baseline (month 0 and post-training (month 12. Latent growth curve analyses showed that, on average, PA decreased during the follow-up period but at a decelerating rate. Women who made greater improvements to EF during the training period showed better adherence to PA during the 1-year follow-up period (β = -.36, p .10. Overall, these findings suggest that improving EF plays an important role in whether older women maintain higher levels of PA following exercise training and that this association is only apparent after training when environmental support for PA is low.

  12. Neural activation patterns of successful episodic encoding: Reorganization during childhood, maintenance in old age

    Directory of Open Access Journals (Sweden)

    Yee Lee Shing

    2016-08-01

    Full Text Available The two-component framework of episodic memory (EM development posits that the contributions of medial temporal lobe (MTL and prefrontal cortex (PFC to successful encoding differ across the lifespan. To test the framework’s hypotheses, we compared subsequent memory effects (SME of 10–12 year-old children, younger adults, and older adults using functional magnetic resonance imaging (fMRI. Memory was probed by cued recall, and SME were defined as regional activation differences during encoding between subsequently correctly recalled versus omitted items. In MTL areas, children’s SME did not differ in magnitude from those of younger and older adults. In contrast, children’s SME in PFC were weaker than the corresponding SME in younger and older adults, in line with the hypothesis that PFC contributes less to successful encoding in childhood. Differences in SME between younger and older adults were negligible. The present results suggest that, among individuals with high memory functioning, the neural circuitry contributing to successful episodic encoding is reorganized from middle childhood to adulthood. Successful episodic encoding in later adulthood, however, is characterized by the ability to maintain the activation patterns that emerged in young adulthood.

  13. Maintenance of stability in γ spectrometric system of low active and environmental samples - a practical experience

    International Nuclear Information System (INIS)

    Ravishankar, R.; Bandyopadhyay, T.; Sarkar, P.K.

    2011-01-01

    Particle Accelerators are becoming part of the society with more and more medical and Industrial types are added every year in addition to research type of accelerators. The outflow of materials to the public domain from such accelerator facilities need to checked carefully and must be released after ensuring the activities of such materials should not exceed the regulatory limits. Health Physics Unit, VECC is involved in analyzing food product samples, seized samples which are suspected to contain Uranium etc and other environmental samples in addition to analyzing radioactive materials evolved from Operational Health Physics work. Most of these analyses involve γ Spectrometric Systems of high efficiency and high resolution types. The efficacy of the analysis and results depends on various parameters of the spectrometric system. The electrical noise from the power supply system and other noises picked up, even in the range of a few milli volts range, have been found to affect the stability of the system. These effects may not be present initially during installation but may creep in due course due to various reasons including weather conditions, wear and tear etc. Unless these problems are attended in regular intervals, the stability of the spectrometric systems and hence the results of analysis of the low active and environmental samples, will not be satisfactory. The work describes the practical problems faced by Health Physics Unit, the methods employed in identifying the problems, the necessary remedial measures taken, the final outcome in the stability and the procedures framed in order to avoid in future. (author)

  14. Breast cancer survivors involved in vigorous team physical activity: psychosocial correlates of maintenance participation.

    Science.gov (United States)

    Culos-Reed, S Nicole; Shields, Christopher; Brawley, Lawrence R

    2005-07-01

    Physical activity is increasingly being promoted as a means to achieve both physical and psychological benefits for cancer survivors. For women with breast cancer, one sport growing in popularity is dragon boating. The purpose of the present investigation was to examine the psychosocial correlates of dragon boat participation over the course of a season. Six crews completed the baseline (early-season) assessment (n = 109) and late-season assessments (n = 56). The self-report questionnaire completed at both time points included an assessment of the theory of planned behaviour variables, quality of life, cohesion, and physical activity levels. A prospective examination of the TPB variables revealed attitude at early season as the only significant predictor of behavioural intentions 12 weeks later at late season (R2 adjusted = 0.27, p cohesive at a level similar to that for female sport teams among the asymptomatic population. As well, participants' health-related quality of life was similar to normal, healthy women of similar age for both mental and physical health. Copyright 2004 John Wiley & Sons, Ltd.

  15. Compartmentalized beta subunit distribution determines characteristics and ethanol sensitivity of somatic, dendritic, and terminal large-conductance calcium-activated potassium channels in the rat central nervous system.

    Science.gov (United States)

    Wynne, P M; Puig, S I; Martin, G E; Treistman, S N

    2009-06-01

    Neurons are highly differentiated and polarized cells, whose various functions depend upon the compartmentalization of ion channels. The rat hypothalamic-neurohypophysial system (HNS), in which cell bodies and dendrites reside in the hypothalamus, physically separated from their nerve terminals in the neurohypophysis, provides a particularly powerful preparation in which to study the distribution and regional properties of ion channel proteins. Using electrophysiological and immunohistochemical techniques, we characterized the large-conductance calcium-activated potassium (BK) channel in each of the three primary compartments (soma, dendrite, and terminal) of HNS neurons. We found that dendritic BK channels, in common with somatic channels but in contrast to nerve terminal channels, are insensitive to iberiotoxin. Furthermore, analysis of dendritic BK channel gating kinetics indicates that they, like somatic channels, have fast activation kinetics, in contrast to the slow gating of terminal channels. Dendritic and somatic channels are also more sensitive to calcium and have a greater conductance than terminal channels. Finally, although terminal BK channels are highly potentiated by ethanol, somatic and dendritic channels are insensitive to the drug. The biophysical and pharmacological properties of somatic and dendritic versus nerve terminal channels are consistent with the characteristics of exogenously expressed alphabeta1 versus alphabeta4 channels, respectively. Therefore, one possible explanation for our findings is a selective distribution of auxiliary beta1 subunits to the somatic and dendritic compartments and beta4 to the terminal compartment. This hypothesis is supported immunohistochemically by the appearance of distinct punctate beta1 or beta4 channel clusters in the membrane of somatic and dendritic or nerve terminal compartments, respectively.

  16. Antispasmodic and Antidiarrheal Activities of Valeriana hardwickii Wall. Rhizome Are Putatively Mediated through Calcium Channel Blockade.

    Science.gov (United States)

    Bashir, Samra; Memon, Raafia; Gilani, Anwar H

    2011-01-01

    Valeriana hardwickii is indigenous to Pakistan, Burma and Ceylon, where it is traditionally being used as an antispasmodic and antidiarrheal, besides its culinary use as spice. The aim of this paper was to provide pharmacological validation to these medicinal uses. The crude aqueous-methanolic extract of Valeriana hardwickii rhizome (Vh.Cr) was studied on isolated rabbit jejunum and castor oil-induced diarrhea in mice for spasmolytic and antidiarrheal properties, respectively. Vh.Cr caused concentration-dependent (0.01-1 mg/mL) relaxation of spontaneous contractions in isolated rabbit jejunum and inhibited K(+)-induced contractions (0.01-0.3 mg/mL), similar to verapamil, suggestive of calcium channel blockade (CCB). The CCB effect was confirmed when pretreatment of the jejunum preparations with Vh.Cr produced a concentration-dependent (0.03-0.1 mg/mL) rightward shift in the Ca(++) concentration-response curves, as caused by verapamil. Vh.Cr exhibited dose-dependent (100-300 mg/kg) protection against castor oil-induced diarrhea in mice. Loperamide, a standard antidiarrheal drug, similarly prevented the diarrhea. These data indicate the presence of CCB effect in the extract of Valeriana hardwickii rhizome, possibly mediating its antispasmodic and antidiarrheal activities and provide a scientific base for its traditional use in hyperactive gut disorders.

  17. Antispasmodic and Antidiarrheal Activities of Valeriana hardwickii Wall. Rhizome Are Putatively Mediated through Calcium Channel Blockade

    Directory of Open Access Journals (Sweden)

    Samra Bashir

    2011-01-01

    Full Text Available Valeriana hardwickii is indigenous to Pakistan, Burma and Ceylon, where it is traditionally being used as an antispasmodic and antidiarrheal, besides its culinary use as spice. The aim of this paper was to provide pharmacological validation to these medicinal uses. The crude aqueous-methanolic extract of Valeriana hardwickii rhizome (Vh.Cr was studied on isolated rabbit jejunum and castor oil-induced diarrhea in mice for spasmolytic and antidiarrheal properties, respectively. Vh.Cr caused concentration-dependent (0.01–1 mg/mL relaxation of spontaneous contractions in isolated rabbit jejunum and inhibited K+-induced contractions (0.01–0.3 mg/mL, similar to verapamil, suggestive of calcium channel blockade (CCB. The CCB effect was confirmed when pretreatment of the jejunum preparations with Vh.Cr produced a concentration-dependent (0.03–0.1 mg/mL rightward shift in the Ca++ concentration-response curves, as caused by verapamil. Vh.Cr exhibited dose-dependent (100–300 mg/kg protection against castor oil-induced diarrhea in mice. Loperamide, a standard antidiarrheal drug, similarly prevented the diarrhea. These data indicate the presence of CCB effect in the extract of Valeriana hardwickii rhizome, possibly mediating its antispasmodic and antidiarrheal activities and provide a scientific base for its traditional use in hyperactive gut disorders.

  18. Antispasmodic activity of Symplocos paniculata is mediated through opening of ATP-dependent K+ channel

    Directory of Open Access Journals (Sweden)

    Khalid Hussain Janbaz

    2016-06-01

    Full Text Available Symplocos paniculata is a medicinal plant used by native healers to manage gastrointestinal ailments. The crude methanolic extract of S. paniculata was screened pharmacologically both in vitro and in vivo for the validation of its therapeutic potential. It suppressed the spontaneous activity of isolated rabbit jejunum preparations and also caused inhibition of the low K+ (20 mM- induced spastic contractions in isolated rabbit jejunum preparations in a manner comparable to cromakalim. The relaxant effect was found to be blocked following glibenclamide exposure of the isolated tissue preparations similar to cromakalim, suggesting that observed response was likely to be mediated through opening of ATP dependent K+ channels. Following oral administration to mice provided protection against castor oil-induced diarrhea in a manner similar to loperamide. The plant material was found safe in toxicity study up to oral dose of 8 g/kg in mice. Hence, present study provides a scientific basis for the vernacular use of S. paniculata in gastro-intestinal system.

  19. Activation of the TASK-2 channel after cell swelling is dependent on tyrosine phosphorylation

    DEFF Research Database (Denmark)

    Kirkegaard, Signe Skyum; Lambert, Ian Henry; Gammeltoft, Steen

    2010-01-01

    (K,vol) indicating that inhibition of RVD reflects inhibition of TASK-2. We find that in EATC the tyrosine kinase inhibitor genistein inhibits RVD by 90%, and that the tyrosine phosphatase inhibitor monoperoxo(picolinato)-oxo-vanadate(V) [mpV(pic)] shifted the volume set point for inactivation of the channel...... to a lower cell volume. Swelling-activated K(+) efflux was impaired by genistein and the Src kinase family inhibitor 4-amino-5-(4-chloro-phenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP2) and enhanced by the tyrosine phosphatase inhibitor mpV(pic). With the use of the TASK-2 inhibitor clofilium......, it is demonstrated that mpV(pic) increased the volume-sensitive part of the K(+) efflux 1.3 times. To exclude K(+) efflux via a KCl cotransporter, cellular Cl(-) was substituted with NO(3)(-). Also under these conditions K(+) efflux was completely blocked by genistein. Thus tyrosine kinases seem to be involved...

  20. Synthetic ciguatoxins selectively activate Nav1.8-derived chimeric sodium channels expressed in HEK293 cells.

    Science.gov (United States)

    Yamaoka, Kaoru; Inoue, Masayuki; Miyazaki, Keisuke; Hirama, Masahiro; Kondo, Chie; Kinoshita, Eiji; Miyoshi, Hiroshi; Seyama, Issei

    2009-03-20

    The synthetic ciguatoxin CTX3C has been shown to activate tetrodotoxin (TTX)-sensitive sodium channels (Na(v)1.2, Na(v)1.4, and Na(v)1.5) by accelerating activation kinetics and shifting the activation curve toward hyperpolarization (Yamaoka, K., Inoue, M., Miyahara, H., Miyazaki, K., and Hirama, M. (2004) Br. J. Pharmacol. 142, 879-889). In this study, we further explored the effects of CTX3C on the TTX-resistant sodium channel Na(v)1.8. TTX-resistant channels have been shown to be involved in transducing pain and related sensations (Akopian, A. N., Sivilotti, L., and Wood, J. N. (1996) Nature 379, 257-262). Thus, we hypothesized that ciguatoxin-induced activation of the Na(v)1.8 current would account for the neurological symptoms of ciguatera poisoning. We found that 0.1 mum CTX3C preferentially affected the activation process of the Na(v)1.8 channel compared with those of the Na(v)1.2 and Na(v)1.4 channels. Importantly, without stimulation, 0.1 mum CTX3C induced a large leakage current (I (L)). The conductance of the I (L) calculated relative to the maximum conductance (G (max)) was 10 times larger than that of Na(v)1.2 or Na(v)1.4. To determine the molecular domain of Na(v)1.8 responsible for conferring higher sensitivity to CTX3C, we made two chimeric constructs from Na(v)1.4 and Na(v)1.8. Chimeras containing the N-terminal half of Na(v)1.8 exhibited a large response similar to wild-type Na(v)1.8, indicating that the region conferring high sensitivity to ciguatoxin action is located in the D1 or D2 domains.

  1. Synthetic Ciguatoxins Selectively Activate Nav1.8-derived Chimeric Sodium Channels Expressed in HEK293 Cells*

    Science.gov (United States)

    Yamaoka, Kaoru; Inoue, Masayuki; Miyazaki, Keisuke; Hirama, Masahiro; Kondo, Chie; Kinoshita, Eiji; Miyoshi, Hiroshi; Seyama, Issei

    2009-01-01

    The synthetic ciguatoxin CTX3C has been shown to activate tetrodotoxin (TTX)-sensitive sodium channels (Nav1.2, Nav1.4, and Nav1.5) by accelerating activation kinetics and shifting the activation curve toward hyperpolarization (Yamaoka, K., Inoue, M., Miyahara, H., Miyazaki, K., and Hirama, M. (2004) Br. J. Pharmacol. 142, 879–889). In this study, we further explored the effects of CTX3C on the TTX-resistant sodium channel Nav1.8. TTX-resistant channels have been shown to be involved in transducing pain and related sensations (Akopian, A. N., Sivilotti, L., and Wood, J. N. (1996) Nature 379, 257–262). Thus, we hypothesized that ciguatoxin-induced activation of the Nav1.8 current would account for the neurological symptoms of ciguatera poisoning. We found that 0.1 μm CTX3C preferentially affected the activation process of the Nav1.8 channel compared with those of the Nav1.2 and Nav1.4 channels. Importantly, without stimulation, 0.1 μm CTX3C induced a large leakage current (IL). The conductance of the IL calculated relative to the maximum conductance (Gmax) was 10 times larger than that of Nav1.2 or Nav1.4. To determine the molecular domain of Nav1.8 responsible for conferring higher sensitivity to CTX3C, we made two chimeric constructs from Nav1.4 and Nav1.8. Chimeras containing the N-terminal half of Nav1.8 exhibited a large response similar to wild-type Nav1.8, indicating that the region conferring high sensitivity to ciguatoxin action is located in the D1 or D2 domains. PMID:19164297

  2. The N-terminal domain of Slack determines the formation and trafficking of Slick/Slack heteromeric sodium-activated potassium channels.

    Science.gov (United States)

    Chen, Haijun; Kronengold, Jack; Yan, Yangyang; Gazula, Valeswara-Rao; Brown, Maile R; Ma, Liqun; Ferreira, Gonzalo; Yang, Youshan; Bhattacharjee, Arin; Sigworth, Fred J; Salkoff, Larry; Kaczmarek, Leonard K

    2009-04-29

    Potassium channels activated by intracellular Na(+) ions (K(Na)) play several distinct roles in regulating the firing patterns of neurons, and, at the single channel level, their properties are quite diverse. Two known genes, Slick and Slack, encode K(Na) channels. We have now found that Slick and Slack subunits coassemble to form heteromeric channels that differ from the homomers in their unitary conductance, kinetic behavior, subcellular localization, and response to activation of protein kinase C. Heteromer formation requires the N-terminal domain of Slack-B, one of the alternative splice variants of the Slack channel. This cytoplasmic N-terminal domain of Slack-B also facilitates the localization of heteromeric K(Na) channels to the plasma membrane. Immunocytochemical studies indicate that Slick and Slack-B subunits are coexpressed in many central neurons. Our findings provide a molecular explanation for some of the diversity in reported properties of neuronal K(Na) channels.

  3. Maintenance optimization after RCM

    International Nuclear Information System (INIS)

    Doyle, E.K.; Lee, C.-G.; Cho, D.

    2005-01-01

    Variant forms of RCM (Reliability Centered Maintenance) have been the maintenance optimizing tools of choice in industry for the last 20 years. Several such optimization techniques have been implemented at the Bruce Nuclear Station. Further cost refinement of the Station preventive maintenance strategy whereby decisions are based on statistical analysis of historical failure data are now being evaluated. The evaluation includes a requirement to demonstrate that earlier optimization projects have long term positive impacts. This proved to be a significant challenge. Eventually a methodology was developed using Crowe/AMSAA (Army Materials Systems Analysis Activity) plots to justify expenditures on further optimization efforts. (authors)

  4. Nuclear power plants maintenance

    International Nuclear Information System (INIS)

    Anon.

    1988-01-01

    Nuclear power plants maintenance now appears as an important factor contributing to the competitivity of nuclea energy. The articles published in this issue describe the way maintenance has been organized in France and how it led to an actual industrial activity developing and providing products and services. An information note about Georges Besse uranium enrichment plant (Eurodif) recalls that maintenance has become a main data not only for power plants but for all nuclear industry installations. (The second part of this dossier will be published in the next issue: vol. 1 January-February 1989) [fr

  5. Initiating statistical maintenance optimization

    International Nuclear Information System (INIS)

    Doyle, E. Kevin; Tuomi, Vesa; Rowley, Ian

    2007-01-01

    Since the 1980 s maintenance optimization has been centered around various formulations of Reliability Centered Maintenance (RCM). Several such optimization techniques have been implemented at the Bruce Nuclear Station. Further cost refinement of the Station preventive maintenance strategy includes evaluation of statistical optimization techniques. A review of successful pilot efforts in this direction is provided as well as initial work with graphical analysis. The present situation reguarding data sourcing, the principle impediment to use of stochastic methods in previous years, is discussed. The use of Crowe/AMSAA (Army Materials Systems Analysis Activity) plots is demonstrated from the point of view of justifying expenditures in optimization efforts. (author)

  6. Na+K+-ATPase activity and K+ channels differently contribute to vascular relaxation in male and female rats.

    Directory of Open Access Journals (Sweden)

    Fernanda Moura Vargas Dias

    Full Text Available Gender associated differences in vascular reactivity regulation might contribute to the low incidence of cardiovascular disease in women. Cardiovascular protection is suggested to depend on female sex hormones' effects on endothelial function and vascular tone regulation. We tested the hypothesis that potassium (K+ channels and Na+K+-ATPase may be involved in the gender-based vascular reactivity differences. Aortic rings from female and male rats were used to examine the involvement of K+ channels and Na+K+-ATPase in vascular reactivity. Acetylcholine (ACh-induced relaxation was analyzed in the presence of L-NAME (100 µM and the following K+ channels blockers: tetraethylammonium (TEA, 2 mM, 4-aminopyridine (4-AP, 5 mM, iberiotoxin (IbTX, 30 nM, apamin (0.5 µM and charybdotoxin (ChTX, 0.1 µM. The ACh-induced relaxation sensitivity was greater in the female group. After incubation with 4-AP the ACh-dependent relaxation was reduced in both groups. However, the dAUC was greater in males, suggesting that the voltage-dependent K+ channel (Kv participates more in males. Inhibition of the three types of Ca2+-activated K+ channels induced a greater reduction in Rmax in females than in males. The functional activity of the Na+K+-ATPase was evaluated by KCl-induced relaxation after L-NAME and OUA incubation. OUA reduced K+-induced relaxation in female and male groups, however, it was greater in males, suggesting a greater Na+K+-ATPase functional activity. L-NAME reduced K+-induced relaxation only in the female group, suggesting that nitric oxide (NO participates more in their functional Na+K+-ATPase activity. These results suggest that the K+ channels involved in the gender-based vascular relaxation differences are the large conductance Ca2+-activated K+ channels (BKCa in females and Kv in males and in the K+-induced relaxation and the Na+K+-ATPase vascular functional activity is greater in males.

  7. Maintenance Mentor

    National Research Council Canada - National Science Library

    Jacobs, John

    2003-01-01

    Maintenance Mentor (MXM) is a research effort conducted by a joint AFRL/HESR and Northrop Grumman Information Technology team to identify the basic, high-level requirements necessary for improving flight line diagnostic capabilities...

  8. Acid sensing ion channel (ASIC) inhibitors exhibit anxiolytic-like activity in preclinical pharmacological models.

    Science.gov (United States)

    Dwyer, Jason M; Rizzo, Stacey J Sukoff; Neal, Sarah J; Lin, Qian; Jow, Flora; Arias, Robert L; Rosenzweig-Lipson, Sharon; Dunlop, John; Beyer, Chad E

    2009-03-01

    Acid sensing ion channels (ASICs) are proton-gated ion channels located in the central and peripheral nervous systems. Of particular interest is ASIC1a, which is located in areas associated with fear and anxiety behaviors. Recent reports suggest a role for ASIC1a in preclinical models of fear conditioning and anxiety. The present experiments evaluated various ASIC inhibitors in preclinical models of autonomic and behavioral parameters of anxiety. In addition, neurochemical studies evaluated the effects of an ASIC inhibitor (A-317567) on neurotransmitter levels in the amygdala. In electrophysiological studies using hippocampal primary neuronal cultures, three ASIC inhibitors (PcTX-1, A-317567, and amiloride) produced concentration-dependent inhibition of acid-evoked currents. In the stress-induced hyperthermia model, acute administration of psalmotoxin 1 (PcTX-1; 10-56 ng, i.c.v.), A-317567 (0.1-1.0 mg/kg, i.p.), and amiloride (10-100 mg/kg, i.p.) prevented stress-induced elevations in core body temperature. In the four-plate test, acute treatment with PcTX-1 (10-56 ng, i.c.v.) and A-317567 (0.01-0.1 mg/kg, i.p.), but not amiloride (3-100 mg/kg, i.p.), produced dose-dependent and significant increases in the number of punished crossings relative to vehicle-treated animals. Additionally, PcTX-1 (56-178 ng, i.c.v.), A-317567 (0.1-10 mg/kg, i.p.), and amiloride (10-100 mg/kg, i.p.) lacked significant anxiolytic-like activity in the elevated zero maze. In neurochemical studies, an infusion of A-317567 (100 microM) into the amygdala significantly elevated the extracellular levels of GABA, but not glutamate, in this brain region. These findings demonstrate that ASIC inhibition produces anxiolytic-like effects in some behavioral models and indicate a potential role for GABAergic mechanisms to underlie these anxiolytic-like effects.

  9. MARKETING CHANNELS

    Directory of Open Access Journals (Sweden)

    Ljiljana Stošić Mihajlović

    2014-07-01

    Full Text Available Marketing channel is a set of entities and institutions, completion of distribution and marketing activities, attend the efficient and effective networking of producers and consumers. Marketing channels include the total flows of goods, money and information taking place between the institutions in the system of marketing, establishing a connection between them. The functions of the exchange, the physical supply and service activities, inherent in the system of marketing and trade. They represent paths which products and services are moving after the production, which will ultimately end up buying and eating by the user.

  10. Proteoglycans, ion channels and cell-matrix adhesion

    DEFF Research Database (Denmark)

    Mitsou, Ioli; Multhaupt, Hinke A.B.; Couchman, John R.

    2017-01-01

    , maintenance, repair and disease.The cytoplasmic domains of syndecans, while having no intrinsic kinase activity, can nevertheless signal through binding proteins.All syndecans appear to be connected to the actin cytoskeleton and can therefore contribute to cell adhesion, notably to the ECM and migration.......Recent data now suggest that syndecans can regulate stretchactivated ion channels.The structure and function of the syndecans and the ion channels are reviewed here, along with an analysis of ion channel functions in cell-matrix adhesion.This area sheds new light on the syndecans, not least since evidence...

  11. Role of volume-regulated and calcium-activated anion channels in cell volume homeostasis, cancer and drug resistance

    DEFF Research Database (Denmark)

    Hoffmann, Else Kay; Sørensen, Belinda Halling; Sauter, Daniel Rafael Peter

    2015-01-01

    to be an essential component of both VRAC and VSOAC. Reduced VRAC and VSOAC activities are seen in drug resistant cancer cells. ANO1 is a calcium-activated chloride channel expressed on the plasma membrane of e.g. secretory epithelia. ANO1 is amplified and highly expressed in a large number of carcinomas. The gene...... functions as well as their role in cancer and drug resistance....

  12. Active Channel Reservation for Coexistence Mechanism (ACROS) for IEEE 802.15.4 and IEEE 802.11

    Science.gov (United States)

    Shin, Soo Young; Woo, Dong Hyuk; Lee, Jong Wook; Park, Hong Seong; Kwon, Wook Hyun

    In this paper, a coexistence mechanism between IEEE 802.15.4 and IEEE 802.11b, Active Channel Reservation for cOexiStence (ACROS), is proposed. The key idea underlining ACROS is to reserve the channel for IEEE 802.15.4 transmission, where IEEE 802.11 transmissions are forbidden. The request-to-send (RTS)/clear-to send (CTS) mechanism within IEEE 802.11 is used to reserve a channel. The proposed ACROS mechanism is implemented into a PC based prototype. The embedded version of ACROS is also developed to mitigate the timing drift problem in the PC-based ACROS. The efficiency of ACROS is shown using the throughput and packet error rate achieved in actual experiments.

  13. Solubilization, partial purification, and reconstitution of glutamate- and N-methyl-D-aspartate-activated cation channels from brain synaptic membranes

    International Nuclear Information System (INIS)

    Ly, A.M.; Michaelis, E.K.

    1991-01-01

    L-Glutamate-activated cation channel proteins from rat brain synaptic membranes were solubilized, partially purified, and reconstituted into liposomes. Optimal conditions for solubilization and reconstitution included treatment of the membranes with nonionic detergents in the presence of neutral phospholipids plus glycerol. Quench-flow procedures were developed to characterize the rapid kinetics of ion flux induced by receptor agonists. [ 14 C]Methylamine, a cation that permeates through the open channel of both vertebrate and invertebrate glutamate receptors, was used to measure the activity of glutamate receptor-ion channel complexes in reconstituted liposomes. L-Glutamate caused an increase in the rate of [ 14 C]methylamine influx into liposomes reconstituted with either solubilized membrane proteins or partially purified glutamate-binding proteins. Of the major glutamate receptor agonists, only N-methyl-D-aspartate activated cation fluxes in liposomes reconstituted with glutamate-binding proteins. In liposomes reconstituted with glutamate-binding proteins, N-methyl-D-aspartate- or glutamate-induced influx of NA + led to a transient increase in the influx of the lipid-permeable anion probe S 14 CN - . These results indicate the functional reconstitution of N-methyl-D-aspartate-sensitive glutamate receptors and the role of the ∼69-kDa protein in the function of these ion channels

  14. KCNN Genes that Encode Small-Conductance Ca2+-Activated K+ Channels Influence Alcohol and Drug Addiction.

    Science.gov (United States)

    Padula, Audrey E; Griffin, William C; Lopez, Marcelo F; Nimitvilai, Sudarat; Cannady, Reginald; McGuier, Natalie S; Chesler, Elissa J; Miles, Michael F; Williams, Robert W; Randall, Patrick K; Woodward, John J; Becker, Howard C; Mulholland, Patrick J

    2015-07-01

    Small-conductance Ca(2+)-activated K(+) (KCa2) channels control neuronal excitability and synaptic plasticity, and have been implicated in substance abuse. However, it is unknown if genes that encode KCa2 channels (KCNN1-3) influence alcohol and drug addiction. In the present study, an integrative functional genomics approach shows that genetic datasets for alcohol, nicotine, and illicit drugs contain the family of KCNN genes. Alcohol preference and dependence QTLs contain KCNN2 and KCNN3, and Kcnn3 transcript levels in the nucleus accumbens (NAc) of genetically diverse BXD strains of mice predicted voluntary alcohol consumption. Transcript levels of Kcnn3 in the NAc negatively correlated with alcohol intake levels in BXD strains, and alcohol dependence enhanced the strength of this association. Microinjections of the KCa2 channel inhibitor apamin into the NAc increased alcohol intake in control C57BL/6J mice, while spontaneous seizures developed in alcohol-dependent mice following apamin injection. Consistent with this finding, alcohol dependence enhanced the intrinsic excitability of medium spiny neurons in the NAc core and reduced the function and protein expression of KCa2 channels in the NAc. Altogether, these data implicate the family of KCNN genes in alcohol, nicotine, and drug addiction, and identify KCNN3 as a mediator of voluntary and excessive alcohol consumption. KCa2.3 channels represent a promising novel target in the pharmacogenetic treatment of alcohol and drug addiction.

  15. Inhibition of G-Protein-Activated Inwardly Rectifying K+ Channels by the Selective Norepinephrine Reuptake Inhibitors Atomoxetine and Reboxetine

    Science.gov (United States)

    Kobayashi, Toru; Washiyama, Kazuo; Ikeda, Kazutaka

    2010-01-01

    Atomoxetine and reboxetine are commonly used as selective norepinephrine reuptake inhibitors (NRIs) for the treatment of attention-deficit/hyperactivity disorder and depression, respectively. Furthermore, recent studies have suggested that NRIs may be useful for the treatment of several other psychiatric disorders. However, the molecular mechanisms underlying the various effects of NRIs have not yet been sufficiently clarified. G-protein-activated inwardly rectifying K+ (GIRK or Kir3) channels have an important function in regulating neuronal excitability and heart rate, and GIRK channel modulation has been suggested to be a potential treatment for several neuropsychiatric disorders and cardiac arrhythmias. In this study, we investigated the effects of atomoxetine and reboxetine on GIRK channels using the Xenopus oocyte expression assay. In oocytes injected with mRNA for GIRK1/GIRK2, GIRK2, or GIRK1/GIRK4 subunits, extracellular application of atomoxetine or reboxetine reversibly reduced GIRK currents. The inhibitory effects were concentration-dependent, but voltage-independent, and time-independent during each voltage pulse. However, Kir1.1 and Kir2.1 channels were insensitive to atomoxetine and reboxetine. Atomoxetine and reboxetine also inhibited GIRK currents induced by activation of cloned A1 adenosine receptors or by intracellularly applied GTPγS, a nonhydrolyzable GTP analogue. Furthermore, the GIRK currents induced by ethanol were concentration-dependently inhibited by extracellularly applied atomoxetine but not by intracellularly applied atomoxetine. The present results suggest that atomoxetine and reboxetine inhibit brain- and cardiac-type GIRK channels, revealing a novel characteristic of clinically used NRIs. GIRK channel inhibition may contribute to some of the therapeutic effects of NRIs and adverse side effects related to nervous system and heart function. PMID:20393461

  16. General anesthetic octanol and related compounds activate wild-type and delF508 cystic fibrosis chloride channels.

    Science.gov (United States)

    Marcet, Brice; Becq, Frédéric; Norez, Caroline; Delmas, Patrick; Verrier, Bernard

    2004-03-01

    1. Cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel is defective during cystic fibrosis (CF). Activators of the CFTR Cl(-) channel may be useful for therapy of CF. Here, we demonstrate that a range of general anesthetics like normal-alkanols (n-alkanols) and related compounds can stimulate the Cl(-) channel activity of wild-type CFTR and delF508-CFTR mutant. 2. The effects of n-alkanols like octanol on CFTR activity were measured by iodide ((125)I) efflux and patch-clamp techniques on three distinct cellular models: (1). CFTR-expressing Chinese hamster ovary cells, (2). human airway Calu-3 epithelial cells and (3). human airway JME/CF15 epithelial cells which express the delF508-CFTR mutant. 3. Our data show for the first time that n-alkanols activate both wild-type CFTR and delF508-CFTR mutant. Octanol stimulated (125)I efflux in a dose-dependent manner in CFTR-expressing cells (wild-type and delF508) but not in cell lines lacking CFTR. (125)I efflux and Cl(-) currents induced by octanol were blocked by glibenclamide but insensitive to 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid, as expected for a CFTR Cl(-) current. 4. CFTR activation by octanol was neither due to cell-to-cell uncoupling properties of octanol nor to an intracellular cAMP increase. CFTR activation by octanol requires phosphorylation by protein kinase-A (PKA) since it was prevented by H-89, a PKA inhibitor. 5. n-Alkanols chain length was an important determinant for channel activation, with rank order of potencies: 1-heptanoloctanoloctanol<1-decanol. Our findings may be of valuable interest for developing novel therapeutic strategies for CF.

  17. Darlington Station outage - a maintenance perspective

    International Nuclear Information System (INIS)

    Plourde, J.; Marczak, J.; Stone, M.; Myers, R.; Sutton, K.

    1997-01-01

    Ontario Hydro's Darlington Nuclear Generating Station (4x881MW(e)net) has carried out its first station outage since full commercial operation. The outage presented challenges to the organization in terms of outage planning, support, management, and safe execution within the constraints of schedule, budget and resources. This paper will focus on the success of the outage maintenance program, identifying the major work programs - a vacuum structure and containment outage, an emergency service water system outage, an emergency coolant injection system outage, intake channel inspections, low pressure service water inspections, and significant outage maintenance work on each of the four reactor units. Planning for the outage was initiated early in anticipation of this important milestone in the station's life. Detailed safety reviews - nuclear, radiation, and conventional - were conducted in support of the planned maintenance program. System lineup and work protection were provided by the Station Operator work group. Work protection permitry was initiated well in advance of the outage. Station maintenance staff resources were bolstered in support of the outage to ensure program execution could be maintained within the schedule. Training programs were in place to ensure that expectations were clear and that high standards would be maintained. Materials management issues in support of maintenance activities were given high priority to ensure no delays to the planned work. Station management review and monitoring in preparation for and during the outage ensured that staff priorities remained focused. Lessons learned from the outage execution are being formalized in maintenance procedures and outage management procedures, and shared with the nuclear community. (author)

  18. Burst activity and ultrafast activation kinetics of CaV1.3 Ca²⁺ channels support presynaptic activity in adult gerbil hair cell ribbon synapses.

    Science.gov (United States)

    Zampini, Valeria; Johnson, Stuart L; Franz, Christoph; Knipper, Marlies; Holley, Matthew C; Magistretti, Jacopo; Masetto, Sergio; Marcotti, Walter

    2013-08-15

    Auditory information transfer to afferent neurons relies on precise triggering of neurotransmitter release at the inner hair cell (IHC) ribbon synapses by Ca²⁺ entry through CaV1.3 Ca²⁺ channels. Despite the crucial role of CaV1.3 Ca²⁺ channels in governing synaptic vesicle fusion, their elementary properties in adult mammals remain unknown. Using near-physiological recording conditions we investigated Ca²⁺ channel activity in adult gerbil IHCs. We found that Ca²⁺ channels are partially active at the IHC resting membrane potential (-60 mV). At -20 mV, the large majority (>70%) of Ca²⁺ channel first openings occurred with an estimated delay of about 50 μs in physiological conditions, with a mean open time of 0.5 ms. Similar to other ribbon synapses, Ca²⁺ channels in IHCs showed a low mean open probability (0.21 at -20 mV), but this increased significantly (up to 0.91) when Ca²⁺ channel activity switched to a bursting modality. We propose that IHC Ca²⁺ channels are sufficiently rapid to transmit fast signals of sound onset and support phase-locking. Short-latency Ca²⁺ channel opening coupled to multivesicular release would ensure precise and reliable signal transmission at the IHC ribbon synapse.

  19. [3H]PN200-110 and [3H]ryanodine binding and reconstitution of ion channel activity with skeletal muscle membranes

    International Nuclear Information System (INIS)

    Hamilton, S.L.; Alvarez, R.M.; Fill, M.; Hawkes, M.J.; Brush, K.L.; Schilling, W.P.; Stefani, E.

    1989-01-01

    Skeletal muscle membranes derived either from the tubular (T) network or from the sarcoplasmic reticulum (SR) were characterized with respect to the binding of the dihydropyridine, [ 3 H]PN200-110, and the alkaloid, [ 3 H]ryanodine; polypeptide composition; and ion channel activity. Conditions for optimizing the binding of these radioligands are discussed. A bilayer pulsing technique is described and is used to examine the channels present in these membranes. Fusion of T-tubule membranes into bilayers revealed the presence of chloride channels and dihydropyridine-sensitive calcium channels with three distinct conductances. The dihydropyridine-sensitive channels were further characterized with respect to their voltage dependence. Pulsing experiments indicated that two different populations of dihydropyridine-sensitive channels existed. Fusion of heavy SR vesicles revealed three different ion channels; the putative calcium release channel, a potassium channel, and a chloride channel. Thus, this fractionation procedure provides T-tubules and SR membranes which, with radioligand binding and single channel recording techniques, provide a useful tool to study the characteristics of skeletal muscle ion channels and their possible role in excitation-contraction coupling

  20. Inhibition of TRPA1 channel activity in sensory neurons by the glial cell line-derived neurotrophic factor family member, artemin

    Directory of Open Access Journals (Sweden)

    Wang Shenglan

    2011-05-01

    Full Text Available Abstract Background The transient receptor potential (TRP channel subtype A1 (TRPA1 is known to be expressed on sensory neurons and respond to changes in temperature, pH and local application of certain noxious chemicals such as allyl isothiocyanate (AITC. Artemin is a neuronal survival and differentiation factor and belongs to the glial cell line-derived neurotrophic factor (GDNF family. Both TRPA1 and artemin have been reported to be involved in pathological pain initiation and maintenance. In the present study, using whole-cell patch clamp recording technique, in situ hybridization and behavioral analyses, we examined the functional interaction between TRPA1 and artemin. Results We found that 85.8 ± 1.9% of TRPA1-expressing neurons also expressed GDNF family receptor alpha 3 (GFR α3, and 87.5 ± 4.1% of GFRα3-expressing neurons were TRPA1-positive. In whole-cell patch clamp analysis, a short-term treatment of 100 ng/ml artemin significantly suppressed the AITC-induced TRPA1 currents. A concentration-response curve of AITC resulting from the effect of artemin showed that this inhibition did not change EC50 but did lower the AITC-induced maximum response. In addition, pre-treatment of artemin significantly suppressed the number of paw lifts induced by intraplantar injection of AITC, as well as the formalin-induced pain behaviors. Conclusions These findings that a short-term application of artemin inhibits the TRPA1 channel's activity and the sequential pain behaviors suggest a role of artemin in regulation of sensory neurons.

  1. Antiarrhythmic effect of either negative modulation or blockade of small conductance Ca2+ activated K+ channels on ventricular fibrillation in guinea pig Langendorff perfused heart

    DEFF Research Database (Denmark)

    Diness, Jonas Goldin; Kirchhoff, Jeppe Egedal; Sheykhzade, Majid

    2015-01-01

    During recent years small conductance Ca activated K (SK) channels have been reported to play a role in cardiac electrophysiology. SK channels seem to be expressed in atria and ventricles but from a functional perspective atrial activity is predominant. A general notion seems to be that cardiac S...

  2. Highly selective water channel activity measured by voltage clamp: analysis of planar lipid bilayers reconstituted with purified AqpZ.

    Science.gov (United States)

    Pohl, P; Saparov, S M; Borgnia, M J; Agre, P

    2001-08-14

    Aquaporins are membrane channels selectively permeated by water or water plus glycerol. Conflicting reports have described ion conductance associated with some water channels, raising the question of whether ion conductance is a general property of the aquaporin family. To clarify this question, a defined system was developed to simultaneously measure water permeability and ion conductance. The Escherichia coli water channel aquaporin-Z (AqpZ) was studied, because it is a highly stable tetramer. Planar lipid bilayers were formed from unilamellar vesicles containing purified AqpZ. The hydraulic conductivity of bilayers made from the total extract of E. coli lipids increased 3-fold if reconstituted with AqpZ, but electric conductance was unchanged. No channel activity was detected under voltage-clamp conditions, indicating that less than one in 10(9) transport events is electrogenic. Microelectrode measurements were simultaneously undertaken adjacent to the membrane. Changes in sodium concentration profiles accompanying transmembrane water flow permitted calculation of the activation energies: 14 kcal/mol for protein-free lipid bilayers and 4 kcal/mol for lipid bilayers containing AqpZ. Neither the water permeability nor the electric conductivity exhibited voltage dependence. This sensitive system demonstrated that AqpZ is permeated by water but not charged ions and should permit direct analyses of putative electrogenic properties of other aquaporins.

  3. Renovascular BK(Ca) channels are not activated in vivo under resting conditions and during agonist stimulation

    DEFF Research Database (Denmark)

    Magnusson, Linda; Sørensen, Charlotte Mehlin; Braunstein, Thomas Hartig

    2006-01-01

    We investigated the role of large-conductance Ca(2+)-activated K(+) (BK(Ca)) channels for the basal renal vascular tone in vivo. Furthermore, the possible buffering by BK(Ca) of the vasoconstriction elicited by angiotensin II (ANG II) or norepinephrine (NE) was investigated. The possible activation.......3 nmol/min) did not have any effect. Renal injection of ANG II (1-4 ng) or NE (10-40 ng) produced a transient decrease in RBF. These responses were not affected by preinfusion of TEA or IBT. Renal infusion of the BK(Ca) opener NS-1619 (90.0 nmol/min) did not affect basal RBF or the response to NE......, there is no indication for a major role for BK(Ca) channels in the control of basal renal tone in vivo. Furthermore, BK(Ca) channels do not have a buffering effect on the rat renal vascular responses to ANG II and NE. The fact that NS-1619 attenuates the ANG II response indicates that the renal vascular BK(Ca) channels...

  4. Synthesis of Rh/Macro-Porous Alumina Over Micro-Channel Plate and Its Catalytic Activity Tests for Diesel Reforming.

    Science.gov (United States)

    Seong, Yeon Baek; Kim, Yong Sul; Park, No-Kuk; Lee, Tae Jin

    2015-11-01

    Macro-porous Al2O3 as the catalytic support material was synthesized using colloidal polystyrene spheres over a micro-channel plate. The colloidal polystyrene spheres were used as a template for the production of an ordered macro porous material using an alumina nitrate solution as the precursor for Al2O3. The close-packed colloidal crystal array template method was applied to the formulation of ordered macro-porous Al2O3 used as a catalytic support material over a micro-channel plate. The solvent in the mixture solution, which also contained the colloidal polystyrene solution, aluminum nitrate solution and the precursor of the catalytic active materials (Rh), was evaporated in a vacuum oven at 50 degrees C. The ordered polystyrene spheres and aluminum salt of the solid state were deposited over a micro channel plate, and macro-porous Al2O3 was formed after calcination at 600 degrees C to remove the polystyrene spheres. The catalytic activity of the Rh/macro-porous alumina supported over the micro-channel plate was tested for diesel reforming.

  5. Managed maintenance, the next step in power plant maintenance

    International Nuclear Information System (INIS)

    Butterworth, G.; Anderson, T.M.

    1984-01-01

    The Westinghouse Nuclear Services Integration Division managed maintenance services are described. Essential to the management and control of a total plant maintenance programme is the development of a comprehensive maintenance specification. During recent years Westinghouse has jointly developed total plant engineering-based maintenance specifications with a number of utilities. The process employed and the experience to date are described. To efficiently implement the maintenance programme Westinghouse has developed a computer software program specifically designed for day to day use at the power plant by maintenance personnel. This program retains an equipment maintenance history, schedules maintenance activities, issues work orders and performs a number of sophisticated analyses of the maintenance backlog and forecast, equipment failure rates, etc. The functions of this software program are described and details of Westinghouse efforts to support the utilities in reducing outage times through development of predefined outage plans for critical report maintenance activities are given. Also described is the experience gained in the training of specialized maintenance personnel, employing competency-based training techniques and equipment mock-ups, and the benefits experienced, in terms of improved quality and productivity of maintenance performed. The success experienced with these methods has caused Westinghouse to expand the use of these training techniques to the more routine skill areas of power plant maintenance. A significant reduction in the operating costs of nuclear power plants will only be brought about by a significant improvement in the quality of maintenance. Westinghouse intends to effect this change by expanding its international service capabilities and to make major investments in order to promote technological developments in the area of power plant maintenance. (author)

  6. The Chromatin Remodeler BPTF Activates a Stemness Gene-Expression Program Essential for the Maintenance of Adult Hematopoietic Stem Cells.

    Science.gov (United States)

    Xu, Bowen; Cai, Ling; Butler, Jason M; Chen, Dongliang; Lu, Xiongdong; Allison, David F; Lu, Rui; Rafii, Shahin; Parker, Joel S; Zheng, Deyou; Wang, Gang Greg

    2018-03-13

    Self-renewal and differentiation of adult stem cells are tightly regulated partly through configuration of chromatin structure by chromatin remodelers. Using knockout mice, we here demonstrate that bromodomain PHD finger transcription factor (BPTF), a component of the nucleosome remodeling factor (NURF) chromatin-remodeling complex, is essential for maintaining the population size of hematopoietic stem/progenitor cells (HSPCs), including long-term hematopoietic stem cells (HSCs). Bptf-deficient HSCs are defective in reconstituted hematopoiesis, and hematopoietic-specific knockout of Bptf caused profound defects including bone marrow failure and anemia. Genome-wide transcriptome profiling revealed that BPTF loss caused downregulation of HSC-specific gene-expression programs, which contain several master transcription factors (Meis1, Pbx1, Mn1, and Lmo2) required for HSC maintenance and self-renewal. Furthermore, we show that BPTF potentiates the chromatin accessibility of key HSC "stemness" genes. These results demonstrate an essential requirement of the chromatin remodeler BPTF and NURF for activation of "stemness" gene-expression programs and proper function of adult HSCs. Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.

  7. Carboxyl-terminal Truncations of ClC-Kb Abolish Channel Activation by Barttin Via Modified Common Gating and Trafficking*

    Science.gov (United States)

    Stölting, Gabriel; Bungert-Plümke, Stefanie; Franzen, Arne; Fahlke, Christoph

    2015-01-01

    ClC-K chloride channels are crucial for auditory transduction and urine concentration. Mutations in CLCNKB, the gene encoding the renal chloride channel hClC-Kb, cause Bartter syndrome type III, a human genetic condition characterized by polyuria, hypokalemia, and alkalosis. In recent years, several Bartter syndrome-associated mutations have been described that result in truncations of the intracellular carboxyl terminus of hClC-Kb. We here used a combination of whole-cell patch clamp, confocal imaging, co-immunoprecipitation, and surface biotinylation to study the functional consequences of a frequent CLCNKB mutation that creates a premature stop codon at Trp-610. We found that W610X leaves the association of hClC-Kb and the accessory subunit barttin unaffected, but impairs its regulation by barttin. W610X attenuates hClC-Kb surface membrane insertion. Moreover, W610X results in hClC-Kb channel opening in the absence of barttin and prevents further barttin-mediated activation. To describe how the carboxyl terminus modifies the regulation by barttin we used V166E rClC-K1. V166E rClC-K1 is active without barttin and exhibits prominent, barttin-regulated voltage-dependent gating. Electrophysiological characterization of truncated V166E rClC-K1 demonstrated that the distal carboxyl terminus is necessary for slow cooperative gating. Since barttin modifies this particular gating process, channels lacking the distal carboxyl-terminal domain are no longer regulated by the accessory subunit. Our results demonstrate that the carboxyl terminus of hClC-Kb is not part of the binding site for barttin, but functionally modifies the interplay with barttin. The loss-of-activation of truncated hClC-Kb channels in heterologous expression systems fully explains the reduced basolateral chloride conductance in affected kidneys and the clinical symptoms of Bartter syndrome patients. PMID:26453302

  8. People's response to the promotion of the effective use of nuclear power plants. From the viewpoint of favoring providing information about maintenance activities

    International Nuclear Information System (INIS)

    Sakai, Yukimi; Ueda, Yoshitaka; Goto, Manabu

    2010-01-01

    In order to elucidate people's response to the promotion of the effective use of nuclear power plants by high-reliability maintenance, we conducted a questionnaire survey targeting 306 respondents living in the cities of Kyoto, Osaka and Kobe. The questionnaire comprises information about maintenance contents and regulatory system concerning extension of plant operation cycle, long life operation, and plant power-up, not as a measure for safety but a measure for promoting effective use of power plants. It was found that a high number of respondents held a positive attitude towards the promotion of the effective use of plants, even if they tend to be anxious about safety problems. Moreover, the study suggests that no enhance the feeling of safety, information about maintenance activities and regulatory system was more effective than information about actual technical possibility. (author)

  9. Preventive and Predictive Maintenance, Warehousing of Spares, Periodic Testing and In-Service Inspection Activities at the Nigerian Research Reactor-1 Facility

    Energy Technology Data Exchange (ETDEWEB)

    Yusuf, I.; Mati, A. A.; Dewu, B. B.M., [Centre for Energy Research and Training, Ahmadu Bello University, Zaria (Nigeria)

    2014-08-15

    The Nigerian Research Reactor–1, or NIRR-1, is sited at Centre for Energy Research and Training, Ahmadu Bello University, Zaria, Nigeria. Activities on preventive or routine maintenance have been institutionalized since the commissioning of the reactor in February 2004. This has grossly reduced the rates of corrective maintenance activities and helped the reactor management a great deal in predicting failure rates of reactor components and other auxiliary units. Routine maintenance of systems and components are being carried out on a weekly, quarterly and annual basis based on manufacturer’s recommendations, which have been reviewed and improved over the years. The paper presents the implementation of maintenance activities in NIRR-1 from its initial criticality in 2004 till today and the new scheme for periodic testing and in-service-inspection developed after an IAEA Integrated Safety Assessment of Research Reactors mission. The measures put in place are envisaged to reduce the negative impact of ageing on NIRR-1 and its auxiliary systems. (author)

  10. Which Socio-Ecological Factors Associate with a Switch to or Maintenance of Active and Passive Transport during the Transition from Primary to Secondary School?

    Science.gov (United States)

    Vanwolleghem, Griet; Van Dyck, Delfien; De Meester, Femke; De Bourdeaudhuij, Ilse; Cardon, Greet; Gheysen, Freja

    2016-01-01

    The aim was to investigate which individual, psychosocial and physical neighborhood environmental factors associate with children's switch to or maintenance of active/passive transport to school and to leisure time destinations during the transition from primary to secondary school. Children (n = 313) filled out a questionnaire in the last year of primary school and 2 years later to assess socio-demographic characteristics and self-reported transport. One of their parents completed a questionnaire to assess parental perceptions of psychosocial and physical neighborhood environmental factors. The increase of the home-school distance was significantly associated with children's switch to or maintenance of passive transport to school compared to a switch to (OR = 0.81; p = 0.03) and maintenance (OR = 0.87; p = 0.03) of active transport to school. Low SES was associated with children's switch to active transport to school compared to maintenance of active transport (OR = 3.67; p = 0.07). For transport to leisure time destinations, other factors such as parental perceived neighborhood safety from traffic and crime (OR = 2.78; p = 0.004), a positive social norm (OR = 1.49; p = 0.08), positive attitudes (OR = 1.39; p = 0.08) (i.e. more benefits, less barriers) towards their children's physical activity and poor walking/cycling facilities in the neighborhood (OR = 0.70; p = 0.06) were associated with children's maintenance of active transport to leisure time destinations compared to a switch to or maintenance of passive transport. This longitudinal study can give directions for interventions promoting children's active transport during the transition to secondary school. It is necessary to promote different possibilities at primary school for children to use active transport when going to secondary school. Walking/cycling a part of the home-school trip can be a possible solution for children who will be living at non-feasible distances from secondary school. Providing safe

  11. Activation of endothelial and epithelial K(Ca) 2.3 calcium-activated potassium channels by NS309 relaxes human small pulmonary arteries and bronchioles

    DEFF Research Database (Denmark)

    Kroigaard, Christel; Dalsgaard, Thomas; Nielsen, Gorm

    2012-01-01

    BACKGROUND AND PURPOSE: Small (K(Ca) 2) and intermediate (K(Ca) 3.1) conductance calcium-activated potassium channels (K(Ca) ) may contribute to both epithelium- and endothelium-dependent relaxations, but this has not been established in human pulmonary arteries and bronchioles. Therefore, we inv...... targets for treatment of pulmonary hypertension and chronic obstructive pulmonary disease....

  12. Dioxin-induced acute cardiac mitochondrial oxidative damage and increased activity of ATP-sensitive potassium channels in Wistar rats

    International Nuclear Information System (INIS)

    Pereira, Susana P.; Pereira, Gonçalo C.; Pereira, Cláudia V.; Carvalho, Filipa S.; Cordeiro, Marília H.; Mota, Paula C.; Ramalho-Santos, João; Moreno, António J.; Oliveira, Paulo J.

    2013-01-01

    The environmental dioxin 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is classified as a Group 1 human carcinogen and teratogenic agent. We hypothesize that TCDD-induced oxidative stress may also interfere with mitochondrial ATP-sensitive potassium channels (mitoKATP), which are known to regulate and to be regulated by mitochondrial redox state. We investigated the effects of an acute treatment of male Wistar rats with TCDD (50 μg/kg i.p.) and measured the regulation of cardiac mitoKATP. While the function of cardiac mitochondria was slightly depressed, mitoKATP activity was 52% higher in animals treated with TCDD. The same effects were not observed in liver mitochondria isolated from the same animals. Our data also shows that regulation of mitochondrial ROS production by mitoKATP activity is different in both groups. To our knowledge, this is the first report to show that TCDD increases mitoKATP activity in the heart, which may counteract the increased oxidative stress caused by the dioxin during acute exposure. -- Highlights: •Acute TCDD treatment of Wistar rats causes cardiac oxidative stress. •Acute TCDD treatment causes cardiac mitochondrial alterations. •Mitochondrial liver vs. heart alterations are distinct. •TCDD treatment resulted in altered activity of cardiac mitochondrial K-ATP channels. -- Dioxin alters the regulation of cardiac mitochondrial ATP-sensitive potassium channels and disturbs mitochondrial physiology

  13. Inhibitory actions by ibandronate sodium, a nitrogen-containing bisphosphonate, on calcium-activated potassium channels in Madin–Darby canine kidney cells

    Directory of Open Access Journals (Sweden)

    Sheng-Nan Wu

    2015-01-01

    Full Text Available The nitrogen-containing bisphosphonates used for management of the patients with osteoporosis were reported to influence the function of renal tubular cells. However, how nitrogen-containing bisphosphates exert any effects on ion currents remains controversial. The effects of ibandronate (Iban, a nitrogen-containing bisphosphonate, on ionic channels, including two types of Ca2+-activated K+ (KCa channels, namely, large-conductance KCa (BKCa and intermediate-conductance KCa (IKCa channels, were investigated in Madin–Darby canine kidney (MDCK cells. In whole-cell current recordings, Iban suppressed the amplitude of voltage-gated K+ current elicited by long ramp pulse. Addition of Iban caused a reduction of BKCa channels accompanied by a right shift in the activation curve of BKCa channels, despite no change in single-channel conductance. Ca2+ sensitivity of these channels was modified in the presence of this compound; however, the magnitude of Iban-mediated decrease in BKCa-channel activity under membrane stretch with different negative pressure remained unchanged. Iban suppressed the probability of BKCa-channel openings linked primarily to a shortening in the slow component of mean open time in these channels. The dissociation constant needed for Iban-mediated suppression of mean open time in MDCK cells was 12.2 μM. Additionally, cell exposure to Iban suppressed the activity of IKCa channels, and DC-EBIO or 9-phenanthrol effectively reversed its suppression. Under current-clamp configuration, Iban depolarized the cells and DC-EBIO or PF573228 reversed its depolarizing effect. Taken together, the inhibitory action of Iban on KCa-channel activity may contribute to the underlying mechanism of pharmacological or toxicological actions of Iban and its structurally similar bisphosphonates on renal tubular cells occurring in vivo.

  14. Activity of the Service for maintenance of the electronic equipment of the RA reactor in 1976, report - Annex IV

    International Nuclear Information System (INIS)

    Milosevic, M.

    1976-01-01

    Within the organizational structure of the RA reactor staff, the Service for instrumentation maintenance has the following tasks: maintenance of the existing electronic equipment; participating in experiments planning and preparation of electronic equipment; purchasing new equipment, spare parts and components; construction of new equipment for internal needs; implementation of new equipment. Basic instrumentation of the reactor facility includes: control and protection system, and dosimetry system [sr

  15. Fluorescence-tracking of activation gating in human ERG channels reveals rapid S4 movement and slow pore opening.

    Directory of Open Access Journals (Sweden)

    Zeineb Es-Salah-Lamoureux

    2010-05-01

    Full Text Available hERG channels are physiologically important ion channels which mediate cardiac repolarization as a result of their unusual gating properties. These are very slow activation compared with other mammalian voltage-gated potassium channels, and extremely rapid inactivation. The mechanism of slow activation is not well understood and is investigated here using fluorescence as a direct measure of S4 movement and pore opening.Tetramethylrhodamine-5-maleimide (TMRM fluorescence at