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Sample records for 20-110 kv network

  1. Choice of the Optimum Cross-Sections for 20-110-330 kV Overhead Lines Under Market Conditions / 20-110-330 Kv Gaisvadu Līniju Optimālo Šķērsgriezumu Izvēle Tirgus Apstākļos

    NASA Astrophysics Data System (ADS)

    Guseva, S.; Petrichenko, L.

    2012-12-01

    Approach is proposed to the choice of overhead line cross-sections that would be optimal under market conditions. For this purpose, in the design practice and at network reconstruction two methods exist: of the economic current density, and of the economic intervals. As of now, none of them works properly, since the cost of electrical materials, of building and construction as well as the total investments in a line are determined by the market prices. Therefore, a modified economic intervals’ method has been developed that can work under market conditions. The method allows also considering the discrete character of standard cross-sections for the lines and concrete technical and technically-economic parameters. For practical application of the proposed method, universal economic nomograms for 20-110-330 kV overhead lines have been calculated and constructed. In the paper, a user interface is proposed for realization of the modified method in Matlab environment. Viens no pilsētas elektriskā tīkla ekonomiskuma paaugstināšanas būtiskiem veidiem ir tīkla racionālā projektēšana. Elektriskā tīkla projekta ekonomiskuma kritērijs ir minimālās summārās ikgadējās izmaksas pie vienādām peļņas vērtībām projekta variantos. Elektropārvādes līniju optimālo šķērsgriezumu izvēle un summāro ikgadējo izmaksu minimums to izbūvei tiek nodrošināts ar ekonomisko intervālu metodes pielietojumu. Sakarā ar ekonomiskās politikas izmaiņām valstī ir izstrādāta un tiek piedāvāta ekonomisko intervālu metodes adaptācija tirgus ekonomikas apstākļiem. Tiek piedāvāts algoritms elektropārvades gaisvadu līniju optimālo šķērsgriezumu izvēlei. Metodes praktiskai realizācijai izstrādātas ekonomisko intervālu nomogrammas. Tiek izveidots lietotāja grafiskais interfeiss Matlab programmēšanas vidē. Ar šīs programmas palīdzību ir konstruētas universālās strāvu ekonomisko intervālu nomogrammas 20-110-330 kV gaisvadu l

  2. 21 CFR 20.110 - Data and information about Food and Drug Administration employees.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 1 2010-04-01 2010-04-01 false Data and information about Food and Drug Administration employees. 20.110 Section 20.110 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH... Data and information about Food and Drug Administration employees. (a) The name, title, grade,...

  3. 21 CFR 20.110 - Data and information about Food and Drug Administration employees.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 1 2011-04-01 2011-04-01 false Data and information about Food and Drug Administration employees. 20.110 Section 20.110 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH... Data and information about Food and Drug Administration employees. (a) The name, title, grade,...

  4. 31 CFR 20.110 - Are any of my Federal assistance awards exempt from this part?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 31 Money and Finance: Treasury 1 2014-07-01 2014-07-01 false Are any of my Federal assistance awards exempt from this part? 20.110 Section 20.110 Money and Finance: Treasury Office of the Secretary of the Treasury GOVERNMENTWIDE REQUIREMENTS FOR DRUG-FREE WORKPLACE (FINANCIAL ASSISTANCE)...

  5. 31 CFR 20.110 - Are any of my Federal assistance awards exempt from this part?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 31 Money and Finance: Treasury 1 2013-07-01 2013-07-01 false Are any of my Federal assistance awards exempt from this part? 20.110 Section 20.110 Money and Finance: Treasury Office of the Secretary of the Treasury GOVERNMENTWIDE REQUIREMENTS FOR DRUG-FREE WORKPLACE (FINANCIAL ASSISTANCE)...

  6. 31 CFR 20.110 - Are any of my Federal assistance awards exempt from this part?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 31 Money and Finance: Treasury 1 2011-07-01 2011-07-01 false Are any of my Federal assistance awards exempt from this part? 20.110 Section 20.110 Money and Finance: Treasury Office of the Secretary of the Treasury GOVERNMENTWIDE REQUIREMENTS FOR DRUG-FREE WORKPLACE (FINANCIAL ASSISTANCE)...

  7. 31 CFR 20.110 - Are any of my Federal assistance awards exempt from this part?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 31 Money and Finance: Treasury 1 2010-07-01 2010-07-01 false Are any of my Federal assistance awards exempt from this part? 20.110 Section 20.110 Money and Finance: Treasury Office of the Secretary of the Treasury GOVERNMENTWIDE REQUIREMENTS FOR DRUG-FREE WORKPLACE (FINANCIAL ASSISTANCE)...

  8. 31 CFR 20.110 - Are any of my Federal assistance awards exempt from this part?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 31 Money and Finance: Treasury 1 2012-07-01 2012-07-01 false Are any of my Federal assistance awards exempt from this part? 20.110 Section 20.110 Money and Finance: Treasury Office of the Secretary of the Treasury GOVERNMENTWIDE REQUIREMENTS FOR DRUG-FREE WORKPLACE (FINANCIAL ASSISTANCE)...

  9. Kv3.3 Channels Bind Hax-1 and Arp2/3 to Assemble a Stable Local Actin Network that Regulates Channel Gating.

    PubMed

    Zhang, Yalan; Zhang, Xiao-Feng; Fleming, Matthew R; Amiri, Anahita; El-Hassar, Lynda; Surguchev, Alexei A; Hyland, Callen; Jenkins, David P; Desai, Rooma; Brown, Maile R; Gazula, Valeswara-Rao; Waters, Michael F; Large, Charles H; Horvath, Tamas L; Navaratnam, Dhasakumar; Vaccarino, Flora M; Forscher, Paul; Kaczmarek, Leonard K

    2016-04-01

    Mutations in the Kv3.3 potassium channel (KCNC3) cause cerebellar neurodegeneration and impair auditory processing. The cytoplasmic C terminus of Kv3.3 contains a proline-rich domain conserved in proteins that activate actin nucleation through Arp2/3. We found that Kv3.3 recruits Arp2/3 to the plasma membrane, resulting in formation of a relatively stable cortical actin filament network resistant to cytochalasin D that inhibits fast barbed end actin assembly. These Kv3.3-associated actin structures are required to prevent very rapid N-type channel inactivation during short depolarizations of the plasma membrane. The effects of Kv3.3 on the actin cytoskeleton are mediated by the binding of the cytoplasmic C terminus of Kv3.3 to Hax-1, an anti-apoptotic protein that regulates actin nucleation through Arp2/3. A human Kv3.3 mutation within a conserved proline-rich domain produces channels that bind Hax-1 but are impaired in recruiting Arp2/3 to the plasma membrane, resulting in growth cones with deficient actin veils in stem cell-derived neurons. PMID:26997484

  10. Experimental investigations of overvoltages in 6kV station service cable networks of thermal power plants

    SciTech Connect

    Vukelja, P.I.; Naumov, R.M.; Drobnjak, G.V.; Mrvic, J.D.

    1996-12-31

    The paper presents the results of experimental investigations of overvoltages on 6kV isolated neutral station service cable networks of thermal power plants. The overvoltages were recorded with capacitive voltage measurement systems made at the Nikola Tesla Institute. Wideband capacitive voltage measurement systems recorded a flat response from below power frequencies to 10MHz. Investigations of overvoltages were performed for appearance and interruption of metal earth faults, intermittent earth faults, switching operation of HV motors switchgear, switching operation of transformers switchgear, and transfer of the network supply from one transformer to another. On the basis of these investigations, certain measures are proposed for limiting overvoltages and for the reliability of station service of thermal power plants.

  11. 50 CFR 20.110 - Seasons, limits, and other regulations for certain Federal Indian reservations, Indian Territory...

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... annual regulatory schedules for this section, see the List of CFR Sections Affected, which appears in the... BIRD HUNTING Annual Seasons, Limits, and Shooting Hours Schedules § 20.110 Seasons, limits, and...

  12. 50 CFR 20.110 - Seasons, limits, and other regulations for certain Federal Indian reservations, Indian Territory...

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... annual regulatory schedules for this section, see the List of CFR Sections Affected, which appears in the... BIRD HUNTING Annual Seasons, Limits, and Shooting Hours Schedules § 20.110 Seasons, limits, and...

  13. 50 CFR 20.110 - Seasons, limits, and other regulations for certain Federal Indian reservations, Indian Territory...

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... annual regulatory schedules for this section, see the List of CFR Sections Affected, which appears in the... BIRD HUNTING Annual Seasons, Limits, and Shooting Hours Schedules § 20.110 Seasons, limits, and...

  14. 50 CFR 20.110 - Seasons, limits, and other regulations for certain Federal Indian reservations, Indian Territory...

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... annual regulatory schedules for this section, see the List of CFR Sections Affected, which appears in the... BIRD HUNTING Annual Seasons, Limits, and Shooting Hours Schedules § 20.110 Seasons, limits, and...

  15. 50 CFR 20.110 - Seasons, limits, and other regulations for certain Federal Indian reservations, Indian Territory...

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... annual regulatory schedules for this section, see the List of CFR Sections Affected, which appears in the... BIRD HUNTING Annual Seasons, Limits, and Shooting Hours Schedules § 20.110 Seasons, limits, and...

  16. Kv5, Kv6, Kv8, and Kv9 subunits: No simple silent bystanders

    PubMed Central

    2016-01-01

    Members of the electrically silent voltage-gated K+ (Kv) subfamilies (Kv5, Kv6, Kv8, and Kv9, collectively identified as electrically silent voltage-gated K+ channel [KvS] subunits) do not form functional homotetrameric channels but assemble with Kv2 subunits into heterotetrameric Kv2/KvS channels with unique biophysical properties. Unlike the ubiquitously expressed Kv2 subunits, KvS subunits show a more restricted expression. This raises the possibility that Kv2/KvS heterotetramers have tissue-specific functions, making them potential targets for the development of novel therapeutic strategies. Here, I provide an overview of the expression of KvS subunits in different tissues and discuss their proposed role in various physiological and pathophysiological processes. This overview demonstrates the importance of KvS subunits and Kv2/KvS heterotetramers in vivo and the importance of considering KvS subunits and Kv2/KvS heterotetramers in the development of novel treatments. PMID:26755771

  17. Ablation of Kv3.1 and Kv3.3 potassium channels disrupts thalamocortical oscillations in vitro and in vivo.

    PubMed

    Espinosa, Felipe; Torres-Vega, Miguel A; Marks, Gerald A; Joho, Rolf H

    2008-05-21

    The genes Kcnc1 and Kcnc3 encode the subunits for the fast-activating/fast-deactivating, voltage-gated potassium channels Kv3.1 and Kv3.3, which are expressed in several brain regions known to be involved in the regulation of the sleep-wake cycle. When these genes are genetically eliminated, Kv3.1/Kv3.3-deficient mice display severe sleep loss as a result of unstable slow-wave sleep. Within the thalamocortical circuitry, Kv3.1 and Kv3.3 subunits are highly expressed in the thalamic reticular nucleus (TRN), which is thought to act as a pacemaker at sleep onset and to be involved in slow oscillatory activity (spindle waves) during slow-wave sleep. We showed that in cortical electroencephalographic recordings of freely moving Kv3.1/Kv3.3-deficient mice, spectral power is reduced up to 70% at frequencies <15 Hz. In addition, the number of sleep spindles in vivo as well as rhythmic rebound firing of TRN neurons in vitro is diminished in mutant mice. Kv3.1/Kv3.3-deficient TRN neurons studied in vitro show approximately 60% increase in action potential duration and a reduction in high-frequency firing after depolarizing current injections and during rebound burst firing. The results support the hypothesis that altered electrophysiological properties of TRN neurons contribute to the reduced EEG power at slow frequencies in the thalamocortical network of Kv3-deficient mice. PMID:18495891

  18. Mechanosensitive gating of Kv channels.

    PubMed

    Morris, Catherine E; Prikryl, Emil A; Joós, Béla

    2015-01-01

    K-selective voltage-gated channels (Kv) are multi-conformation bilayer-embedded proteins whose mechanosensitive (MS) Popen(V) implies that at least one conformational transition requires the restructuring of the channel-bilayer interface. Unlike Morris and colleagues, who attributed MS-Kv responses to a cooperative V-dependent closed-closed expansion↔compaction transition near the open state, Mackinnon and colleagues invoke expansion during a V-independent closed↔open transition. With increasing membrane tension, they suggest, the closed↔open equilibrium constant, L, can increase >100-fold, thereby taking steady-state Popen from 0→1; "exquisite sensitivity to small…mechanical perturbations", they state, makes a Kv "as much a mechanosensitive…as…a voltage-dependent channel". Devised to explain successive gK(V) curves in excised patches where tension spontaneously increased until lysis, their L-based model falters in part because of an overlooked IK feature; with recovery from slow inactivation factored in, their g(V) datasets are fully explained by the earlier model (a MS V-dependent closed-closed transition, invariant L≥4). An L-based MS-Kv predicts neither known Kv time courses nor the distinctive MS responses of Kv-ILT. It predicts Kv densities (hence gating charge per V-sensor) several-fold different from established values. If opening depended on elevated tension (L-based model), standard gK(V) operation would be compromised by animal cells' membrane flaccidity. A MS V-dependent transition is, by contrast, unproblematic on all counts. Since these issues bear directly on recent findings that mechanically-modulated Kv channels subtly tune pain-related excitability in peripheral mechanoreceptor neurons we undertook excitability modeling (evoked action potentials). Kvs with MS V-dependent closed-closed transitions produce nuanced mechanically-modulated excitability whereas an L-based MS-Kv yields extreme, possibly excessive (physiologically

  19. Kv7 and Kv11 channels in myometrial regulation

    PubMed Central

    Greenwood, Iain A; Tribe, Rachel M

    2014-01-01

    Ion channels play a key role in defining myometrial contractility. Modulation of ion channel populations is proposed to underpin gestational changes in uterine contractility associated with the transition from uterine quiescence to active labour. Of the myriad ion channels present in the uterus, this article will focus upon potassium channels encoded by the KCNQ genes and ether-à-go-go-related (ERG) genes. Voltage-gated potassium channels encoded by KCNQ and ERG (termed Kv7 and Kv11, respectively) are accepted as major determinants of neuronal excitability and the duration of the cardiac action potential. However, there is now growing appreciation that these ion channels have a major functional impact in vascular and non-vascular smooth muscle. Moreover, Kv7 channels may be potential therapeutic targets for the treatment of preterm labour. PMID:24121285

  20. Mechanosensitive Gating of Kv Channels

    PubMed Central

    Morris, Catherine E.; Prikryl, Emil A.; Joós, Béla

    2015-01-01

    K-selective voltage-gated channels (Kv) are multi-conformation bilayer-embedded proteins whose mechanosensitive (MS) Popen(V) implies that at least one conformational transition requires the restructuring of the channel-bilayer interface. Unlike Morris and colleagues, who attributed MS-Kv responses to a cooperative V-dependent closed-closed expansion↔compaction transition near the open state, Mackinnon and colleagues invoke expansion during a V-independent closed↔open transition. With increasing membrane tension, they suggest, the closed↔open equilibrium constant, L, can increase >100-fold, thereby taking steady-state Popen from 0→1; “exquisite sensitivity to small…mechanical perturbations”, they state, makes a Kv “as much a mechanosensitive…as…a voltage-dependent channel”. Devised to explain successive gK(V) curves in excised patches where tension spontaneously increased until lysis, their L-based model falters in part because of an overlooked IK feature; with recovery from slow inactivation factored in, their g(V) datasets are fully explained by the earlier model (a MS V-dependent closed-closed transition, invariant L≥4). An L-based MS-Kv predicts neither known Kv time courses nor the distinctive MS responses of Kv-ILT. It predicts Kv densities (hence gating charge per V-sensor) several-fold different from established values. If opening depended on elevated tension (L-based model), standard gK(V) operation would be compromised by animal cells’ membrane flaccidity. A MS V-dependent transition is, by contrast, unproblematic on all counts. Since these issues bear directly on recent findings that mechanically-modulated Kv channels subtly tune pain-related excitability in peripheral mechanoreceptor neurons we undertook excitability modeling (evoked action potentials). Kvs with MS V-dependent closed-closed transitions produce nuanced mechanically-modulated excitability whereas an L-based MS-Kv yields extreme, possibly excessive

  1. Comparative Analysis of Various Superconducting and Non-Superconducting Fault Current Limiting Devices Designed for Operation in a 110 kV/100 MW Power Network

    NASA Astrophysics Data System (ADS)

    Kopylov, S. I.; Altov, V. A.; Balashov, N. N.; Ivanov, S. S.; Zheltov, V. V.; Zemerikin, V. D.

    As it is known one of the most promising fault current limiting (FCL) devices for high-power electric networks can be the so-called transformer type superconducting fault current limiter (SFCL) with the primary winding connected to the load in series and the secondary one shortened by a fast-acting circuit-breaker. These devices when made of conventional materials can be very large and expensive - e.g., for a 100 MW circuit under protection the total mass of copper winding conductors can exceed 15 tons and the heat losses in a normal operating mode can be more than 200 kW. Therefore, using of high-temperature superconductors (HTSC) can be a solution which can sufficiently improve the mass, geometrical and operational characteristics of an FCL. Unlike other superconducting AC devices, the magnetic field in SFCL does not exceed 0.1 - 0.2 T what allows using HTSC windings even at a comparatively high level of AC losses existing nowadays. In this paper is performed a comparative analysis of various designs of SCFL with the non-superconducting FCL. It has been shown that the former have a mass by an order of magnitude lower than the latter and the rate of lowering of heat losses in a normal operating mode is the same. The equalization of costs of both designs is expected to be reached within the nearest 3 - 5 five years.

  2. Kv1.3/Kv1.5 heteromeric channels compromise pharmacological responses in macrophages

    SciTech Connect

    Villalonga, Nuria; Escalada, Artur; Vicente, Ruben; Sanchez-Tillo, Ester; Celada, Antonio; Solsona, Carles; Felipe, Antonio . E-mail: afelipe@ub.edu

    2007-01-26

    Voltage-dependent K{sup +} (Kv) channels are involved in the immune response. Kv1.3 is highly expressed in activated macrophages and T-effector memory cells of autoimmune disease patients. Macrophages are actively involved in T-cell activation by cytokine production and antigen presentation. However, unlike T-cells, macrophages express Kv1.5, which is resistant to Kv1.3-drugs. We demonstrate that mononuclear phagocytes express different Kv1.3/Kv1.5 ratios, leading to biophysically and pharmacologically distinct channels. Therefore, Kv1.3-based treatments to alter physiological responses, such as proliferation and activation, are impaired by Kv1.5 expression. The presence of Kv1.5 in the macrophagic lineage should be taken into account when designing Kv1.3-based therapies.

  3. Auxiliary KCNE subunits modulate both homotetrameric Kv2.1 and heterotetrameric Kv2.1/Kv6.4 channels

    PubMed Central

    David, Jens-Peter; Stas, Jeroen I.; Schmitt, Nicole; Bocksteins, Elke

    2015-01-01

    The diversity of the voltage-gated K+ (Kv) channel subfamily Kv2 is increased by interactions with auxiliary β-subunits and by assembly with members of the modulatory so-called silent Kv subfamilies (Kv5-Kv6 and Kv8-Kv9). However, it has not yet been investigated whether these two types of modulating subunits can associate within and modify a single channel complex simultaneously. Here, we demonstrate that the transmembrane β-subunit KCNE5 modifies the Kv2.1/Kv6.4 current extensively, whereas KCNE2 and KCNE4 only exert minor effects. Co-expression of KCNE5 with Kv2.1 and Kv6.4 did not alter the Kv2.1/Kv6.4 current density but modulated the biophysical properties significantly; KCNE5 accelerated the activation, slowed the deactivation and steepened the slope of the voltage-dependence of the Kv2.1/Kv6.4 inactivation by accelerating recovery of the closed-state inactivation. In contrast, KCNE5 reduced the current density ~2-fold without affecting the biophysical properties of Kv2.1 homotetramers. Co-localization of Kv2.1, Kv6.4 and KCNE5 was demonstrated with immunocytochemistry and formation of Kv2.1/Kv6.4/KCNE5 and Kv2.1/KCNE5 complexes was confirmed by Fluorescence Resonance Energy Transfer experiments performed in HEK293 cells. These results suggest that a triple complex consisting of Kv2.1, Kv6.4 and KCNE5 subunits can be formed. In vivo, formation of such tripartite Kv2.1/Kv6.4/KCNE5 channel complexes might contribute to tissue-specific fine-tuning of excitability. PMID:26242757

  4. A compact 100 kV high voltage glycol capacitor

    NASA Astrophysics Data System (ADS)

    Wang, Langning; Liu, Jinliang; Feng, Jiahuai

    2015-01-01

    A high voltage capacitor is described in this paper. The capacitor uses glycerol as energy storage medium, has a large capacitance close to 1 nF, can hold off voltages of up to 100 kV for μs charging time. Allowing for low inductance, the capacitor electrode is designed as coaxial structure, which is different from the common structure of the ceramic capacitor. With a steady capacitance at different frequencies and a high hold-off voltage of up to 100 kV, the glycol capacitor design provides a potential substitute for the ceramic capacitors in pulse-forming network modulator to generate high voltage pulses with a width longer than 100 ns.

  5. Differential Protein Kinase C-dependent Modulation of Kv7.4 and Kv7.5 Subunits of Vascular Kv7 Channels*

    PubMed Central

    Brueggemann, Lioubov I.; Mackie, Alexander R.; Cribbs, Leanne L.; Freda, Jessica; Tripathi, Abhishek; Majetschak, Matthias; Byron, Kenneth L.

    2014-01-01

    The Kv7 family (Kv7.1–7.5) of voltage-activated potassium channels contributes to the maintenance of resting membrane potential in excitable cells. Previously, we provided pharmacological and electrophysiological evidence that Kv7.4 and Kv7.5 form predominantly heteromeric channels and that Kv7 activity is regulated by protein kinase C (PKC) in response to vasoconstrictors in vascular smooth muscle cells. Direct evidence for Kv7.4/7.5 heteromer formation, however, is lacking. Furthermore, it remains to be determined whether both subunits are regulated by PKC. Utilizing proximity ligation assays to visualize single molecule interactions, we now show that Kv7.4/Kv.7.5 heteromers are endogenously expressed in vascular smooth muscle cells. Introduction of dominant-negative Kv7.4 and Kv7.5 subunits in mesenteric artery myocytes reduced endogenous Kv7 currents by 84 and 76%, respectively. Expression of an inducible protein kinase Cα (PKCα) translocation system revealed that PKCα activation is sufficient to suppress endogenous Kv7 currents in A7r5 rat aortic and mesenteric artery smooth muscle cells. Arginine vasopressin (100 and 500 pm) and the PKC activator phorbol 12-myristate 13-acetate (1 nm) each inhibited human (h) Kv7.5 and hKv7.4/7.5, but not hKv7.4 channels expressed in A7r5 cells. A decrease in hKv7.5 and hKv7.4/7.5 current densities was associated with an increase in PKC-dependent phosphorylation of the channel proteins. These findings provide further evidence for a differential regulation of Kv7.4 and Kv7.5 channel subunits by PKC-dependent phosphorylation and new mechanistic insights into the role of heteromeric subunit assembly for regulation of vascular Kv7 channels. PMID:24297175

  6. The voltage-dependent K+ channels Kv1.3 and Kv1.5 in human cancer

    PubMed Central

    Comes, Núria; Bielanska, Joanna; Vallejo-Gracia, Albert; Serrano-Albarrás, Antonio; Marruecos, Laura; Gómez, Diana; Soler, Concepció; Condom, Enric; Ramón y Cajal, Santiago; Hernández-Losa, Javier; Ferreres, Joan C.; Felipe, Antonio

    2013-01-01

    Voltage-dependent K+ channels (Kv) are involved in a number of physiological processes, including immunomodulation, cell volume regulation, apoptosis as well as differentiation. Some Kv channels participate in the proliferation and migration of normal and tumor cells, contributing to metastasis. Altered expression of Kv1.3 and Kv1.5 channels has been found in several types of tumors and cancer cells. In general, while the expression of Kv1.3 apparently exhibits no clear pattern, Kv1.5 is induced in many of the analyzed metastatic tissues. Interestingly, evidence indicates that Kv1.5 channel shows inversed correlation with malignancy in some gliomas and non-Hodgkin's lymphomas. However, Kv1.3 and Kv1.5 are similarly remodeled in some cancers. For instance, expression of Kv1.3 and Kv1.5 correlates with a certain grade of tumorigenicity in muscle sarcomas. Differential remodeling of Kv1.3 and Kv1.5 expression in human cancers may indicate their role in tumor growth and their importance as potential tumor markers. However, despite of this increasing body of information, which considers Kv1.3 and Kv1.5 as emerging tumoral markers, further research must be performed to reach any conclusion. In this review, we summarize what it has been lately documented about Kv1.3 and Kv1.5 channels in human cancer. PMID:24133455

  7. Differential Activation of Vascular Smooth Muscle Kv7.4, Kv7.5, and Kv7.4/7.5 Channels by ML213 and ICA-069673

    PubMed Central

    Brueggemann, Lyubov I.; Haick, Jennifer M.; Cribbs, Leanne L.

    2014-01-01

    Recent research suggests that smooth muscle cells express Kv7.4 and Kv7.5 voltage-activated potassium channels, which contribute to maintenance of their resting membrane voltage. New pharmacologic activators of Kv7 channels, ML213 (N-mesitybicyclo[2.2.1]heptane-2-carboxamide) and ICA-069673 N-(6-chloropyridin-3-yl)-3,4-difluorobenzamide), have been reported to discriminate among channels formed from different Kv7 subtypes. We compared the effects of ML213 and ICA-069673 on homomeric human Kv7.4, Kv7.5, and heteromeric Kv7.4/7.5 channels exogenously expressed in A7r5 vascular smooth muscle cells. We found that, despite its previous description as a selective activator of Kv7.2 and Kv7.4, ML213 significantly increased the maximum conductance of homomeric Kv7.4 and Kv7.5, as well as heteromeric Kv7.4/7.5 channels, and induced a negative shift of their activation curves. Current deactivation rates decreased in the presence of the ML213 (10 μM) for all three channel combinations. Mutants of Kv7.4 (W242L) and Kv7.5 (W235L), previously found to be insensitive to another Kv7 channel activator, retigabine, were also insensitive to ML213 (10 μM). In contrast to ML213, ICA-069673 robustly activated Kv7.4 channels but was significantly less effective on homomeric Kv7.5 channels. Heteromeric Kv7.4/7.5 channels displayed intermediate responses to ICA-069673. In each case, ICA-069673 induced a negative shift of the activation curves without significantly increasing maximal conductance. Current deactivation rates decreased in the presence of ICA-069673 in a subunit-specific manner. Kv7.4 W242L responded to ICA-069673-like wild-type Kv7.4, but a Kv7.4 F143A mutant was much less sensitive to ICA-069673. Based on these results, ML213 and ICA-069673 likely bind to different sites and are differentially selective among Kv7.4, Kv7.5, and Kv7.4/7.5 channel subtypes. PMID:24944189

  8. Kv7 channels regulate pairwise spiking covariability in health and disease.

    PubMed

    Ocker, Gabriel Koch; Doiron, Brent

    2014-07-15

    Low-threshold M currents are mediated by the Kv7 family of potassium channels. Kv7 channels are important regulators of spiking activity, having a direct influence on the firing rate, spike time variability, and filter properties of neurons. How Kv7 channels affect the joint spiking activity of populations of neurons is an important and open area of study. Using a combination of computational simulations and analytic calculations, we show that the activation of Kv7 conductances reduces the covariability between spike trains of pairs of neurons driven by common inputs. This reduction is beyond that explained by the lowering of firing rates and involves an active cancellation of common fluctuations in the membrane potentials of the cell pair. Our theory shows that the excess covariance reduction is due to a Kv7-induced shift from low-pass to band-pass filtering of the single neuron spike train response. Dysfunction of Kv7 conductances is related to a number of neurological diseases characterized by both elevated firing rates and increased network-wide correlations. We show how changes in the activation or strength of Kv7 conductances give rise to excess correlations that cannot be compensated for by synaptic scaling or homeostatic modulation of passive membrane properties. In contrast, modulation of Kv7 activation parameters consistent with pharmacological treatments for certain hyperactivity disorders can restore normal firing rates and spiking correlations. Our results provide key insights into how regulation of a ubiquitous potassium channel class can control the coordination of population spiking activity. PMID:24790164

  9. Conserved Negative Charges in the N-terminal Tetramerization Domain Mediate Efficient Assembly of Kv2.1 and Kv2.1/Kv6.4 Channels*

    PubMed Central

    Bocksteins, Elke; Labro, Alain J.; Mayeur, Evy; Bruyns, Tine; Timmermans, Jean-Pierre; Adriaensen, Dirk; Snyders, Dirk J.

    2009-01-01

    Voltage-gated potassium (Kv) channels are transmembrane tetramers of individual α-subunits. Eight different Shaker-related Kv subfamilies have been identified in which the tetramerization domain T1, located on the intracellular N terminus, facilitates and controls the assembly of both homo- and heterotetrameric channels. Only the Kv2 α-subunits are able to form heterotetramers with members of the silent Kv subfamilies (Kv5, Kv6, Kv8, and Kv9). The T1 domain contains two subdomains, A and B box, which presumably determine subfamily specificity by preventing incompatible subunits to assemble. In contrast, little is known about the involvement of the A/B linker sequence. Both Kv2 and silent Kv subfamilies contain a fully conserved and negatively charged sequence (CDD) in this linker that is lacking in the other subfamilies. Neutralizing these aspartates in Kv2.1 by mutating them to alanines did not affect the gating properties, but reduced the current density moderately. However, charge reversal arginine substitutions strongly reduced the current density of these homotetrameric mutant Kv2.1 channels and immunocytochemistry confirmed the reduced expression at the plasma membrane. Förster resonance energy transfer measurements using confocal microscopy showed that the latter was not due to impaired trafficking, but to a failure to assemble the tetramer. This was further confirmed with co-immunoprecipitation experiments. The corresponding arginine substitution in Kv6.4 prevented its heterotetrameric interaction with Kv2.1. These results indicate that these aspartates (especially the first one) in the A/B box linker of the T1 domain are required for efficient assembly of both homotetrameric Kv2.1 and heterotetrameric Kv2.1/silent Kv6.4 channels. PMID:19717558

  10. Functional analysis of Kv1.2 and paddle chimera Kv channels in planar lipid bilayers

    PubMed Central

    Tao, Xiao; MacKinnon, Roderick

    2010-01-01

    Summary Voltage-dependent K+ channels play key roles in shaping electrical signaling in both excitable as well as non-excitable cells. These channels open and close in response to the voltage changes across the cell membrane. Many studies have been carried out in order to understand the voltage sensing mechanism. Our laboratory recently determined the atomic structures of a mammalian voltage-dependent K+ channel Kv1.2 and a mutant of Kv1.2 named the ‘paddle-chimera’ channel, in which the voltage sensor paddle was transferred from Kv2.1 to Kv1.2. These two structures provide atomic descriptions of voltage-dependent channels with unprecedented clarity. Until now the functional integrity of these two channels biosynthesized in yeast cells have not been assessed. Here we report the electrophysiological and pharmacological properties of Kv1.2 and the paddle chimera channels in planar lipid bilayers. We demonstrate that Pichia yeast produce ‘normally functioning’ mammalian voltage-dependent K+ channels with qualitatively similar features to the Shaker K+ channel in the absence of the N-terminal inactivation gate, and that the paddle chimera mutant channel functions as well as Kv1.2. We find, however, that in several respects the Kv1.2 channel exhibits functional properties that are distinct from Kv1.2 channels reported in the literature. PMID:18638484

  11. Problems of operating the 500-kV outdoor electrical equipment of the Sayano-Shushenskoe hydroelectric station

    SciTech Connect

    Mitrofanov, A.N.

    1994-10-01

    The Sayano-Shushenskoe hydroelectric station is part of the Siberian interconnected power system. Power is transmitted to the grid by two 500 kV transmission lines. The site characteristics made nearby placement of conventional 500 kV switchyard equipment impossible. The original plans were to place the switchyard 35 km away, but the 500 kV lines to the yard would have passed through mountain regions with intense thunderstorm activity. Because of the unique design of the generators, the number of lightning-induced disconnects was substantial. For greater reliability, 500 kV equipment suitable for placement at the sight was designed, and the overall electrical layout of the yard was revised by reconstructing the middle network of the 4/3 scheme to three networks with two switches per connection. Sectioning of the collecting bus systems was necessary for a further increase in the reliability of the yard.

  12. Redistribution of Kv1 and Kv7 enhances neuronal excitability during structural axon initial segment plasticity

    PubMed Central

    Kuba, Hiroshi; Yamada, Rei; Ishiguro, Go; Adachi, Ryota

    2015-01-01

    Structural plasticity of the axon initial segment (AIS), the trigger zone of neurons, is a powerful means for regulating neuronal activity. Here, we show that AIS plasticity is not limited to structural changes; it also occurs as changes in ion-channel expression, which substantially augments the efficacy of regulation. In the avian cochlear nucleus, depriving afferent inputs by removing cochlea elongated the AIS, and simultaneously switched the dominant Kv channels at the AIS from Kv1.1 to Kv7.2. Due to the slow activation kinetics of Kv7.2, the redistribution of the Kv channels reduced the shunting conductance at the elongated AIS during the initiation of action potentials and effectively enhanced the excitability of the deprived neurons. The results indicate that the functional plasticity of the AIS works cooperatively with the structural plasticity and compensates for the loss of afferent inputs to maintain the homeostasis of auditory circuits after hearing loss by cochlea removal. PMID:26581625

  13. Loss of 115 kV Power

    SciTech Connect

    Smith, J.A.

    2001-08-22

    This report discusses the postulated loss of 115 kV power. Continuous electrical power to Savannah River Plant reactors is necessary to maintain water flow for heat removal and essential monitoring and control. Should power supplied to the plant 115 kV system from offsite be lost, on-site generation is sufficient to maintain all reactors in a safe shutdown mode for an indefinite period. Should on-site generators for the 115 kV grid also be lost, diesel-electric generators within each reactor building are also sufficient to maintain safe shutdown for a finite period. In all cases DC power for necessary monitoring and control would be available from battery systems with AC converter backup.

  14. The 1200-kV system study

    NASA Astrophysics Data System (ADS)

    1982-02-01

    Performance requirements for a 1200 kV circuit breaker for use in a gas-insulated substation were defined and a performance analysis of a 1200 kV insulator/disconnect switch for gas-insulated substation performed. The performance requirements included: insulation requirements of 60 Hz, switching surge and lightning, both phase to ground and across the open breaker; prospective transient recovery voltage requirements related to fault current interrupting; an opening scheme capable of limiting phase to ground and across the breaker overvoltages resulting from line dropping and out of phase switching; a closing scheme capable of limiting line overvoltages during reclosing to 1.5 pu (2 percent level). The investigation of each of these items and of lightning characteristics and effects is discussed. The 1200 kV disconnect switch study analyzed bus energizing, bus de-energizing, transformer de-energizing, and the effect of hysteresis losses. Results from these analyses are presented.

  15. Polyunsaturated Fatty Acids Modify the Gating of Kv Channels

    PubMed Central

    Moreno, Cristina; Macias, Alvaro; Prieto, Angela; De La Cruz, Alicia; Valenzuela, Carmen

    2012-01-01

    Polyunsaturated fatty acids (PUFAs) have been reported to exhibit antiarrhythmic properties, which are attributed to their capability to modulate ion channels. This PUFAs ability has been reported to be due to their effects on the gating properties of ion channels. In the present review, we will focus on the role of PUFAs on the gating of two Kv channels, Kv1.5 and Kv11.1. Kv1.5 channels are blocked by n−3 PUFAs of marine [docosahexaenoic acid (DHA) and eicosapentaenoic acid] and plant origin (alpha-linolenic acid, ALA) at physiological concentrations. The blockade of Kv1.5 channels by PUFAs steeply increased in the range of membrane potentials coinciding with those of Kv1.5 channel activation, suggesting that PUFAs-channel binding may derive a significant fraction of its voltage sensitivity through the coupling to channel gating. A similar shift in the activation voltage was noted for the effects of n–6 arachidonic acid (AA) and DHA on Kv1.1, Kv1.2, and Kv11.1 channels. PUFAs-Kv1.5 channel interaction is time-dependent, producing a fast decay of the current upon depolarization. Thus, Kv1.5 channel opening is a prerequisite for the PUFA-channel interaction. Similar to the Kv1.5 channels, the blockade of Kv11.1 channels by AA and DHA steeply increased in the range of membrane potentials that coincided with the range of Kv11.1 channel activation, suggesting that the PUFAs-Kv channel interactions are also coupled to channel gating. Furthermore, AA regulates the inactivation process in other Kv channels, introducing a fast voltage-dependent inactivation in non-inactivating Kv channels. These results have been explained within the framework that AA closes voltage-dependent potassium channels by inducing conformational changes in the selectivity filter, suggesting that Kv channel gating is lipid dependent. PMID:22973228

  16. KV1 and KV3 Potassium Channels Identified at Presynaptic Terminals of the Corticostriatal Synapses in Rat

    PubMed Central

    Meneses, David; Vega, Ana V.; Torres-Cruz, Francisco Miguel; Barral, Jaime

    2016-01-01

    In the last years it has been increasingly clear that KV-channel activity modulates neurotransmitter release. The subcellular localization and composition of potassium channels are crucial to understanding its influence on neurotransmitter release. To investigate the role of KV in corticostriatal synapses modulation, we combined extracellular recording of population-spike and pharmacological blockage with specific and nonspecific blockers to identify several families of KV channels. We induced paired-pulse facilitation (PPF) and studied the changes in paired-pulse ratio (PPR) before and after the addition of specific KV blockers to determine whether particular KV subtypes were located pre- or postsynaptically. Initially, the presence of KV channels was tested by exposing brain slices to tetraethylammonium or 4-aminopyridine; in both cases we observed a decrease in PPR that was dose dependent. Further experiments with tityustoxin, margatoxin, hongotoxin, agitoxin, dendrotoxin, and BDS-I toxins all rendered a reduction in PPR. In contrast heteropodatoxin and phrixotoxin had no effect. Our results reveal that corticostriatal presynaptic KV channels have a complex stoichiometry, including heterologous combinations KV1.1, KV1.2, KV1.3, and KV1.6 isoforms, as well as KV3.4, but not KV4 channels. The variety of KV channels offers a wide spectrum of possibilities to regulate neurotransmitter release, providing fine-tuning mechanisms to modulate synaptic strength. PMID:27379187

  17. KV1 and KV3 Potassium Channels Identified at Presynaptic Terminals of the Corticostriatal Synapses in Rat.

    PubMed

    Meneses, David; Vega, Ana V; Torres-Cruz, Francisco Miguel; Barral, Jaime

    2016-01-01

    In the last years it has been increasingly clear that KV-channel activity modulates neurotransmitter release. The subcellular localization and composition of potassium channels are crucial to understanding its influence on neurotransmitter release. To investigate the role of KV in corticostriatal synapses modulation, we combined extracellular recording of population-spike and pharmacological blockage with specific and nonspecific blockers to identify several families of KV channels. We induced paired-pulse facilitation (PPF) and studied the changes in paired-pulse ratio (PPR) before and after the addition of specific KV blockers to determine whether particular KV subtypes were located pre- or postsynaptically. Initially, the presence of KV channels was tested by exposing brain slices to tetraethylammonium or 4-aminopyridine; in both cases we observed a decrease in PPR that was dose dependent. Further experiments with tityustoxin, margatoxin, hongotoxin, agitoxin, dendrotoxin, and BDS-I toxins all rendered a reduction in PPR. In contrast heteropodatoxin and phrixotoxin had no effect. Our results reveal that corticostriatal presynaptic KV channels have a complex stoichiometry, including heterologous combinations KV1.1, KV1.2, KV1.3, and KV1.6 isoforms, as well as KV3.4, but not KV4 channels. The variety of KV channels offers a wide spectrum of possibilities to regulate neurotransmitter release, providing fine-tuning mechanisms to modulate synaptic strength. PMID:27379187

  18. 600 kV modulator design for the SLAC Next Linear Collider Test Accelerator

    SciTech Connect

    Harris, K.; de Lamare, J.; Nesterov, V.; Cassel, R.

    1992-07-01

    Preliminary design for the SLAC Next Linear Collider Test Accelerator (NLCTA) requires a pulse power source to produce a 600 kV, 600 A, 1.4 {mu}s, 0.1% flat top pulse with rise and fall times of approximately 100 ns to power an X-Band klystron with a microperveance of 1.25 at {approx} 100 MW peak RF power. The design goals for the modulator, including those previously listed, are peak modulator pulse power of 340 MW operating at 120 Hz. A three-stage darlington pulse-forming network, which produces a >100 kV, 1.4 {mu}s pulse, is coupled to the klystron load through a 6:1 pulse transformer. Careful consideration of the transformer leakage inductance, klystron capacitance, system layout, and component choice is necessary to produce the very fast rise and fall times at 600 kV operating continuously at 120 Hz.

  19. 1200-kV system study

    SciTech Connect

    Not Available

    1982-02-01

    The 1200kV Systems Study Project consisted of two major tasks: definition of performance requirements for a 1200kV circuit breaker for use in a gas-insulated substation; and performance analysis of a 1200 kV insolator/disconnect switch for gas-insulated substation. For the first task, the performance requirements included: insulation requirements - 60 Hz, switching surge and lightning, both phase to ground and across the open breaker; prospective transient recovery voltage requirements related to fault current interrupting; an opening scheme capable of limiting phase to ground and across the breaker overvoltages resulting from line dropping and out of phase switching; a closing scheme capable of limiting line overvoltages during reclosing to 1.5 pu (2 percent level). The investigation of each of these items and of lightning characteristics and effects is discussed. The 1200 kV disconnect switch study analyzed bus energizing, bus de-energizing, transformer de-energizing, and the effect of hysteresis losses. Results from these analyses are presented. (LCL)

  20. Inactivation Gating of Kv4 Potassium Channels

    PubMed Central

    Jerng, Henry H.; Shahidullah, Mohammad; Covarrubias, Manuel

    1999-01-01

    Kv4 channels represent the main class of brain A-type K+ channels that operate in the subthreshold range of membrane potentials (Serodio, P., E. Vega-Saenz de Miera, and B. Rudy. 1996. J. Neurophysiol. 75:2174– 2179), and their function depends critically on inactivation gating. A previous study suggested that the cytoplasmic NH2- and COOH-terminal domains of Kv4.1 channels act in concert to determine the fast phase of the complex time course of macroscopic inactivation (Jerng, H.H., and M. Covarrubias. 1997. Biophys. J. 72:163–174). To investigate the structural basis of slow inactivation gating of these channels, we examined internal residues that may affect the mutually exclusive relationship between inactivation and closed-state blockade by 4-aminopyridine (4-AP) (Campbell, D.L., Y. Qu, R.L. Rasmussen, and H.C. Strauss. 1993. J. Gen. Physiol. 101:603–626; Shieh, C.-C., and G.E. Kirsch. 1994. Biophys. J. 67:2316–2325). A double mutation V[404,406]I in the distal section of the S6 region of the protein drastically slowed channel inactivation and deactivation, and significantly reduced the blockade by 4-AP. In addition, recovery from inactivation was slightly faster, but the pore properties were not significantly affected. Consistent with a more stable open state and disrupted closed state inactivation, V[404,406]I also caused hyperpolarizing and depolarizing shifts of the peak conductance–voltage curve (∼5 mV) and the prepulse inactivation curve (>10 mV), respectively. By contrast, the analogous mutations (V[556,558]I) in a K+ channel that undergoes N- and C-type inactivation (Kv1.4) did not affect macroscopic inactivation but dramatically slowed deactivation and recovery from inactivation, and eliminated open-channel blockade by 4-AP. Mutation of a Kv4-specifc residue in the S4–S5 loop (C322S) of Kv4.1 also altered gating and 4-AP sensitivity in a manner that closely resembles the effects of V[404,406]I. However, this mutant did not exhibit

  1. Modulation of Kv4.3 current by accessory subunits.

    PubMed

    Deschênes, Isabelle; Tomaselli, Gordon F

    2002-09-25

    Kv4.3 encodes the pore-forming subunit of the cardiac transient outward potassium current (I(to)). hKv4.3-encoded current does not fully replicate cardiac I(to), suggesting a functionally significant role for accessory subunits. KChIP2 associates with Kv4.3 and modifies hKv4.3-encoded currents but does not replicate native I(to). We examined the effect of several ancillary subunits expressed in the heart on hKv4.3-encoded currents. Remarkably, the ancillary subunits Kvbeta(3), minK, MiRP-1, the Na channel beta(1) and KChIP2 increased the density and modified the gating of hKv4.3 current. hKv4.3 promiscuously assembles with ancillary subunits in vitro, functionally modifying the encoded currents; however, the physiological significance is uncertain. PMID:12297301

  2. Retigabine, a Kv7.2/Kv7.3-Channel Opener, Attenuates Drug-Induced Seizures in Knock-In Mice Harboring Kcnq2 Mutations

    PubMed Central

    Ihara, Yukiko; Tomonoh, Yuko; Deshimaru, Masanobu; Zhang, Bo; Uchida, Taku; Ishii, Atsushi; Hirose, Shinichi

    2016-01-01

    The hetero-tetrameric voltage-gated potassium channel Kv7.2/Kv7.3, which is encoded by KCNQ2 and KCNQ3, plays an important role in limiting network excitability in the neonatal brain. Kv7.2/Kv7.3 dysfunction resulting from KCNQ2 mutations predominantly causes self-limited or benign epilepsy in neonates, but also causes early onset epileptic encephalopathy. Retigabine (RTG), a Kv7.2/ Kv7.3-channel opener, seems to be a rational antiepileptic drug for epilepsies caused by KCNQ2 mutations. We therefore evaluated the effects of RTG on seizures in two strains of knock-in mice harboring different Kcnq2 mutations, in comparison to the effects of phenobarbital (PB), which is the first-line antiepileptic drug for seizures in neonates. The subjects were heterozygous knock-in mice (Kcnq2Y284C/+ and Kcnq2A306T/+) bearing the Y284C or A306T Kcnq2 mutation, respectively, and their wild-type (WT) littermates, at 63–100 days of age. Seizures induced by intraperitoneal injection of kainic acid (KA, 12mg/kg) were recorded using a video-electroencephalography (EEG) monitoring system. Effects of RTG on KA-induced seizures of both strains of knock-in mice were assessed using seizure scores from a modified Racine’s scale and compared with those of PB. The number and total duration of spike bursts on EEG and behaviors monitored by video recording were also used to evaluate the effects of RTG and PB. Both Kcnq2Y284C/+ and Kcnq2A306T/+ mice showed significantly more KA-induced seizures than WT mice. RTG significantly attenuated KA-induced seizure activities in both Kcnq2Y284C/+ and Kcnq2A306T/+ mice, and more markedly than PB. This is the first reported evidence of RTG ameliorating KA-induced seizures in knock-in mice bearing mutations of Kcnq2, with more marked effects than those observed with PB. RTG or other Kv7.2-channel openers may be considered as first-line antiepileptic treatments for epilepsies resulting from KCNQ2 mutations. PMID:26910900

  3. Conceptual design of 275 kV class high-Tc superconducting cable

    NASA Astrophysics Data System (ADS)

    Mukoyama, S.; Yagi, M.; Fujiwara, N.; Ichikawa, H.

    2010-11-01

    High-temperature superconducting (HTS) cables are expected to be next generation transmission line because of the compact, lightweight, large capacity, and low loss features. Especially, since the YBa 2Cu 3O x (YBCO) tape has a high critical current, high magnetic-field property, low AC loss, and low cost, using YBCO tapes for a HTS cable seems to be one of the most promising ways to make the HTS cable attractive. Therefore, YBCO HTS cables have been studied extensively in Japan, the United States, Korea, and many other countries. We now believe that 275 kV class HTS cables will be used for future large capacity lines based on the needs of Japanese transmission networks for bulk transmission power in overhead transmission lines or gas insulated transmission lines (GIL). We started to develop the 275 kV class HTS cable for the new energy and industrial technology development organization (NEDO) project at 2008, and we have studied the applicability and the environmental and economic advantages of the 275 kV cable. This paper will introduce advantages and a conceptual design of the 275 kV HTS cable.

  4. Effect of tyrphostin AG879 on Kv4.2 and Kv4.3 potassium channels

    PubMed Central

    Yu, Haibo; Zou, Beiyan; Wang, Xiaoliang; Li, Min

    2015-01-01

    Background and Purpose A-type potassium channels (IA) are important proteins for modulating neuronal membrane excitability. The expression and activity of Kv4.2 channels are critical for neurological functions and pharmacological inhibitors of Kv4.2 channels may have therapeutic potential for Fragile X syndrome. While screening various compounds, we identified tyrphostin AG879, a tyrosine kinase inhibitor, as a Kv4.2 inhibitor from. In the present study we characterized the effect of AG879 on cloned Kv4.2/Kv channel-interacting protein 2 (KChIP2) channels. Experimental Approach To screen the library of pharmacologically active compounds, the thallium flux assay was performed on HEK-293 cells transiently-transfected with Kv4.2 cDNA using the Maxcyte transfection system. The effects of AG879 were further examined on CHO-K1 cells expressing Kv4.2/KChIP2 channels using a whole-cell patch-clamp technique. Key Results Tyrphostin AG879 selectively and dose-dependently inhibited Kv4.2 and Kv4.3 channels. In Kv4.2/KChIP2 channels, AG879 induced prominent acceleration of the inactivation rate, use-dependent block and slowed the recovery from inactivation. AG879 induced a hyperpolarizing shift in the voltage-dependence of the steady-state inactivation of Kv4.2 channels without apparent effect on the V1/2 of the voltage-dependent activation. The blocking effect of AG879 was enhanced as channel inactivation increased. Furthermore, AG879 significantly inhibited the A-type potassium currents in the cultured hippocampus neurons. Conclusion and Implications AG879 was identified as a selective and potent inhibitor the Kv4.2 channel. AG879 inhibited Kv4.2 channels by preferentially interacting with the open state and further accelerating their inactivation. PMID:25752739

  5. Modulation of Closed−State Inactivation in Kv2.1/Kv6.4 Heterotetramers as Mechanism for 4−AP Induced Potentiation

    PubMed Central

    Labro, Alain J.; Snyders, Dirk J.

    2015-01-01

    The voltage−gated K+ (Kv) channel subunits Kv2.1 and Kv2.2 are expressed in almost every tissue. The diversity of Kv2 current is increased by interacting with the electrically silent Kv (KvS) subunits Kv5−Kv6 and Kv8−Kv9, into functional heterotetrameric Kv2/KvS channels. These Kv2/KvS channels possess unique biophysical properties and display a more tissue-specific expression pattern, making them more desirable pharmacological and therapeutic targets. However, little is known about the pharmacological properties of these heterotetrameric complexes. We demonstrate that Kv5.1, Kv8.1 and Kv9.3 currents were inhibited differently by the channel blocker 4−aminopyridine (4−AP) compared to Kv2.1 homotetramers. In contrast, Kv6.4 currents were potentiated by 4−AP while displaying moderately increased affinities for the channel pore blockers quinidine and flecainide. We found that the 4−AP induced potentiation of Kv6.4 currents was caused by modulation of the Kv6.4−mediated closed−state inactivation: suppression by 4−AP of the Kv2.1/Kv6.4 closed−state inactivation recovered a population of Kv2.1/Kv6.4 channels that was inactivated at resting conditions, i.e. at a holding potential of −80 mV. This modulation also resulted in a slower initiation and faster recovery from closed−state inactivation. Using chimeric substitutions between Kv6.4 and Kv9.3 subunits, we demonstrated that the lower half of the S6 domain (S6c) plays a crucial role in the 4−AP induced potentiation. These results demonstrate that KvS subunits modify the pharmacological response of Kv2 subunits when assembled in heterotetramers and illustrate the potential of KvS subunits to provide unique pharmacological properties to the heterotetramers, as is the case for 4−AP on Kv2.1/Kv6.4 channels. PMID:26505474

  6. 220-kV combined optical transformer

    NASA Astrophysics Data System (ADS)

    Cui, Ying; Ye, Miaoyuan; Zhu, Yong; Xu, Yan; Luo, Sunan

    2000-10-01

    The magneto-optic current and electro-optic voltage sensing technologies were combined into a single phase unit which provides a new solution for combined current and voltage sensing and relaying applications. It senses voltage and current by making utilities of Pockels electro-optic effect of a BGO(Bi4Ge3O12) crystal and Faraday magneto-optic effect of a close-loop flint. The 220 kV high voltage is directly applied on the BGO crystal without capacitive dividers. This design permits true optical voltage measurement to be performed for the highest possible accuracy and stability. In the current sensor, a light beam circles around the current carrying conductor. The effect of the location movement of the conductor and external magnetic field are reduced. It has superior advantages of compact size, light- weight and better performance compared with conventional current and voltage transformers. Composite bushing consisting of a fiberglass tube support and silicone rubber that is filled with SF6 gas is employed for HV insulation. Connections between the Combined Optical Transformers in the substation and the metering and protecting unit in the control house are via optical fiber cable. The fundamental principal, system design and test result of this new 220 kV Combined Optical Transformer will be introduced in this paper.

  7. Dysfunction of the Heteromeric KV7.3/KV7.5 Potassium Channel is Associated with Autism Spectrum Disorders

    PubMed Central

    Gilling, Mette; Rasmussen, Hanne B.; Calloe, Kirstine; Sequeira, Ana F.; Baretto, Marta; Oliveira, Guiomar; Almeida, Joana; Lauritsen, Marlene B.; Ullmann, Reinhard; Boonen, Susanne E.; Brondum-Nielsen, Karen; Kalscheuer, Vera M.; Tümer, Zeynep; Vicente, Astrid M.; Schmitt, Nicole; Tommerup, Niels

    2012-01-01

    Heterozygous mutations in the KCNQ3 gene on chromosome 8q24 encoding the voltage-gated potassium channel KV7.3 subunit have previously been associated with rolandic epilepsy and idiopathic generalized epilepsy (IGE) including benign neonatal convulsions. We identified a de novo t(3;8) (q21;q24) translocation truncating KCNQ3 in a boy with childhood autism. In addition, we identified a c.1720C > T [p.P574S] nucleotide change in three unrelated individuals with childhood autism and no history of convulsions. This nucleotide change was previously reported in patients with rolandic epilepsy or IGE and has now been annotated as a very rare SNP (rs74582884) in dbSNP. The p.P574S KV7.3 variant significantly reduced potassium current amplitude in Xenopus laevis oocytes when co-expressed with KV7.5 but not with KV7.2 or KV7.4. The nucleotide change did not affect trafficking of heteromeric mutant KV7.3/2, KV7.3/4, or KV7.3/5 channels in HEK 293 cells or primary rat hippocampal neurons. Our results suggest that dysfunction of the heteromeric KV7.3/5 channel is implicated in the pathogenesis of some forms of autism spectrum disorders, epilepsy, and possibly other psychiatric disorders and therefore, KCNQ3 and KCNQ5 are suggested as candidate genes for these disorders. PMID:23596459

  8. Combined MV + kV inverse treatment planning for optimal kV dose incorporation in IGRT

    NASA Astrophysics Data System (ADS)

    Grelewicz, Zachary; Wiersma, Rodney D.

    2014-04-01

    Despite the existence of real-time kV intra-fractional tumor tracking strategies for many years, clinical adoption has been held back by concern over the excess kV imaging dose cost to the patient when imaging in continuous fluoroscopic mode. This work aims to solve this problem by investigating, for the first time, the use of convex optimization tools to optimally integrate this excess kV imaging dose into the MV therapeutic dose in order to make real-time kV tracking clinically feasible. Phase space files modeling both a 6 MV treatment beam and a kV on-board-imaging beam of a commercial LINAC were generated with BEAMnrc, and used to generate dose influence matrices in DOSXYZnrc for ten previously treated lung cancer patients. The dose optimization problem for IMRT, formulated as a quadratic problem, was modified to include additional constraints as required for real-time kV intra-fractional tracking. An interior point optimizer was used to solve the modified optimization problem. It was found that when using large kV imaging apertures during fluoroscopic tracking, combined MV + kV optimization lead to a 0.5%-5.17% reduction in the total number of monitor units assigned to the MV beam due to inclusion of the kV dose over the ten patients. This was accompanied by a reduction of up to 42% of the excess kV dose compared to standard MV IMRT with kV tracking. For all kV field sizes considered, combined MV + kV optimization provided prescription dose to the treatment volume coverage equal to the no-imaging case, yet superior to standard MV IMRT with non-optimized kV fluoroscopic tracking. With combined MV + kV optimization, it is possible to quantify in a patient specific way the dosimetric effect of real-time imaging on the patient. Such information is necessary when substantial kV imaging is performed.

  9. Tau-Dependent Kv4.2 Depletion and Dendritic Hyperexcitability in a Mouse Model of Alzheimer's Disease

    PubMed Central

    Hall, Alicia M.; Throesch, Benjamin T.; Buckingham, Susan C.; Markwardt, Sean J.; Peng, Yin; Wang, Qin

    2015-01-01

    Neuronal hyperexcitability occurs early in the pathogenesis of Alzheimer's disease (AD) and contributes to network dysfunction in AD patients. In other disorders with neuronal hyperexcitability, dysfunction in the dendrites often contributes, but dendritic excitability has not been directly examined in AD models. We used dendritic patch-clamp recordings to measure dendritic excitability in the CA1 region of the hippocampus. We found that dendrites, more so than somata, of hippocampal neurons were hyperexcitable in mice overexpressing Aβ. This dendritic hyperexcitability was associated with depletion of Kv4.2, a dendritic potassium channel important for regulating dendritic excitability and synaptic plasticity. The antiepileptic drug, levetiracetam, blocked Kv4.2 depletion. Tau was required, as crossing with tau knock-out mice also prevented both Kv4.2 depletion and dendritic hyperexcitability. Dendritic hyperexcitability induced by Kv4.2 deficiency exacerbated behavioral deficits and increased epileptiform activity in hAPP mice. We conclude that increased dendritic excitability, associated with changes in dendritic ion channels including Kv4.2, may contribute to neuronal dysfunction in early stages AD. PMID:25878292

  10. Tau-dependent Kv4.2 depletion and dendritic hyperexcitability in a mouse model of Alzheimer's disease.

    PubMed

    Hall, Alicia M; Throesch, Benjamin T; Buckingham, Susan C; Markwardt, Sean J; Peng, Yin; Wang, Qin; Hoffman, Dax A; Roberson, Erik D

    2015-04-15

    Neuronal hyperexcitability occurs early in the pathogenesis of Alzheimer's disease (AD) and contributes to network dysfunction in AD patients. In other disorders with neuronal hyperexcitability, dysfunction in the dendrites often contributes, but dendritic excitability has not been directly examined in AD models. We used dendritic patch-clamp recordings to measure dendritic excitability in the CA1 region of the hippocampus. We found that dendrites, more so than somata, of hippocampal neurons were hyperexcitable in mice overexpressing Aβ. This dendritic hyperexcitability was associated with depletion of Kv4.2, a dendritic potassium channel important for regulating dendritic excitability and synaptic plasticity. The antiepileptic drug, levetiracetam, blocked Kv4.2 depletion. Tau was required, as crossing with tau knock-out mice also prevented both Kv4.2 depletion and dendritic hyperexcitability. Dendritic hyperexcitability induced by Kv4.2 deficiency exacerbated behavioral deficits and increased epileptiform activity in hAPP mice. We conclude that increased dendritic excitability, associated with changes in dendritic ion channels including Kv4.2, may contribute to neuronal dysfunction in early stages AD. PMID:25878292

  11. Kv10.1 K(+) channel: from physiology to cancer.

    PubMed

    Ouadid-Ahidouch, Halima; Ahidouch, Ahmed; Pardo, Luis A

    2016-05-01

    K(+) ions play a major role in many cellular processes. The deregulation of K(+) signaling is associated with a variety of diseases including cancer. Ether-à-go-go-1 (Eag1, Kv10.1, KCNH1) is a member of the voltage-activated potassium channel family and was the first K(+) channel to be associated with oncogenesis and tumor development. Interestingly, in healthy tissue, Kv10.1 is only detected in the central nervous system, where it is involved in the regulation of excitability under repeated stimulation. Kv10.1 is in contrast robustly expressed in over 70 % human tumors, where its expression seems to be controlled by key regulators of proliferation and survival such as p53 and E2F1, often altered in cancer. Otherwise, Kv10.1 is involved in cell proliferation, survival, angiogenesis, migration, and invasion. This review aims to provide a comprehensive overview of the current status of research on the role of Kv10.1 channel in physiopathology. Focus is placed on biophysical and pharmacological properties of Kv10.1 channel, as well as its cycling, trafficking, and its role in the neuron and cancer. The possible mechanisms by which Kv10.1 channel affects tumor cell migration and survival in breast cancer and its regulation by extracellular proteins are discussed. PMID:26743871

  12. Solution of K-V envelope equations

    SciTech Connect

    Anderson, O.A.

    1995-04-01

    The envelope equations for a KV beam with space charge have been analyzed systematically by an e expansion followed by integrations. The focusing profile as a function of axial length is assumed to be symmetric but otherwise arbitrary. Given the bean current, emittance, and peak focusing field, we find the envelopes a(s) and b(s) and obtain , a{sub max}, {sigma}, and {sigma}{sub 0}. Explicit results are presented for various truncations of the expansion. The zeroth order results correspond to those from the well-known smooth approximation; the same convenient format is retained for the higher order cases. The first order results, involving single correction terms, give 3--10 times better accuracy and are good to {approximately}1% at {sigma}{sub 0} = 70{degree}. Third order gives a factor of 10--30 improvement over the smooth approximation and derived quantities accurate to {approximately}1% at {sigma}{sub 0} = 112 {degree}. The first order expressions are convenient design tools. They lend themselves to variable energy problems and have been applied to the design, construction, and testing of ESQ accelerators at LBL.

  13. [The story of K.V. Tjellesen].

    PubMed

    Clemmensen, Peter

    2005-01-01

    This is the story about a Danish pharmaceutical wholesaler. The story begins in 1909 in some basement rooms located in Vester Boulevard 42 (which was later to become the Boulevard of H.C. Andersen) in Copenhagen. The founder of the company, pharmacist Knud Valdemar Tjellesen, lived in the very same building. In the beginning, the company mainly sold chemicals and produced and sold chemical-technical products. In 1930, K.V. Tjellesen became the proprietor of the pharmacy Sct. Johannes Apotek situated in Fredensgade 5, Nørrebro in Copenhagen. The company then moved into some offices that were located just behind the pharmacy. Already in 1918, K.V. Tjellesen traded pharmaceutical specialties, which were imported, to a certain extent, from Germany through a purchasing office in Hamburg. In 1938, the son of Knud Valdemar Tjellesen, pharmacist Paul Tjellesen, joined the wholesale company charged with the primary task of intensifying the sale of the company's international agency products. At the same time, the general partnership K.V. Tjellesen was founded. In the time leading up to the Second World War and during this time, business was of course complicated by currency and import restrictions, and it was very difficult to make deliveries to the pharmacies. In Copenhagen, the articles were delivered by 10-12 bicycle delivery boys. In the beginning, the wholesale company mainly catered for Copenhagen and Zealand, but due to the excellent performance of Paul Tjellesen, more and more customers emerged in the rest of the country. In 1954, the company moved to Niels Ebbesens Vej 29 in Frederiksberg given the need for larger facilities. In 1963, an increased space requirement, once again, forced the company to buy one of the neighbouring buildings on Niels Ebbesens Vej and H.C. Orsteds Vej. For many years after, H.C. Orsteds Vej 22 was the official address of the company. After more than 45 years in the company, Paul Tjellesen agreed with his son-in-law Peter Sch

  14. Spironolactone and its main metabolite canrenoic acid block hKv1.5, Kv4.3 and Kv7.1+minK channels

    PubMed Central

    Gómez, Ricardo; Núñez, Lucía; Caballero, Ricardo; Vaquero, Miguel; Tamargo, Juan; Delpón, Eva

    2005-01-01

    Both spironolactone (SP) and its main metabolite, canrenoic acid (CA), prolong cardiac action potential duration and decrease the Kv11.1 (HERG) current. We examined the effects of SP and CA on cardiac hKv1.5, Kv4.3 and Kv7.1+minK channels that generate the human IKur, Ito1 and IKs, which contribute to the control of human cardiac action potential duration. hKv1.5 currents were recorded in stably transfected mouse fibroblasts and Kv4.3 and Kv7.1+minK in transiently transfected Chinese hamster ovary cells using the whole-cell patch clamp. SP (1 μM) and CA (1 nM) inhibited hKv1.5 currents by 23.2±3.2 and 18.9±2.7%, respectively, shifted the midpoint of the activation curve to more negative potentials and delayed the time course of tail deactivation. SP (1 μM) and CA (1 nM) inhibited the total charge crossing the membrane through Kv4.3 channels at +50 mV by 27.1±6.4 and 27.4±5.7%, respectively, and accelerated the time course of current decay. CA, but not SP, shifted the inactivation curve to more hyperpolarised potentials (Vh−37.0±1.8 vs −40.8±1.6 mV, n=10, P<0.05). SP (10 μM) and CA (1 nM) also inhibited Kv7.1+minK currents by 38.6±2.3 and 22.1±1.4%, respectively, without modifying the voltage dependence of channel activation. SP, but not CA, slowed the time course of tail current decay. CA (1 nM) inhibited the IKur (29.2±5.5%) and the Ito1 (16.1±3.9%) recorded in mouse ventricular myocytes and the IK (21.8±6.9%) recorded in guinea-pig ventricular myocytes. A mathematical model of human atrial action potentials demonstrated that K+ blocking effects of CA resulted in a lengthening of action potential duration, both in normal and atrial fibrillation simulated conditions. The results demonstrated that both SP and CA directly block hKv1.5, Kv4.3 and Kv7.1+minK channels, CA being more potent for these effects. Since peak free plasma concentrations of CA ranged between 3 and 16 nM, these results indicated that blockade of these human

  15. 61. THREE LINE WIRING DIAGRAM, 33 KV & NO. 1 ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    61. THREE LINE WIRING DIAGRAM, 33 KV & NO. 1 TRANS BANK, SANTA ANA NO. 1 HYDRO PLANT, OCTOBER 27, 1958. SCE drawing no. 428058-1. - Santa Ana River Hydroelectric System, SAR-1 Powerhouse, Redlands, San Bernardino County, CA

  16. Kv3.4 channel function and dysfunction in nociceptors

    PubMed Central

    Ritter, David M; Zemel, Benjamin M; Lepore, Angelo C; Covarrubias, Manuel

    2015-01-01

    Recently, we reported the isolation of the Kv3.4 current in dorsal root ganglion (DRG) neurons and described dysregulation of this current in a spinal cord injury (SCI) model of chronic pain. These studies strongly suggest that rat Kv3.4 channels are major regulators of excitability in DRG neurons from pups and adult females, where they help determine action potential (AP) repolarization and spiking properties. Here, we characterized the Kv3.4 current in rat DRG neurons from adult males and show that it transfers 40–70% of the total repolarizing charge during the AP across all ages and sexes. Following SCI, we also found remodeling of the repolarizing currents during the AP. In the light of these studies, homomeric Kv3.4 channels expressed in DRG nociceptors are emerging novel targets that may help develop new approaches to treat neuropathic pain. PMID:26039360

  17. Transient voltages in a tranformer connected to a 500 kV transmission line

    SciTech Connect

    Indulkar, C.S.; Kothari, D.P.; Saha, S.N.

    1985-09-01

    The objective of the study described in this paper was to formulate a simple way of calculating the distribution of transient voltages at different points of a cascaded system consisting of a transformer winding connected to a transmission line. A method of calculating the steady-state voltage distribution based on a simple eigenvalue technique for the analysis of identical ladder networks representing the line and the transformer winding is described which is then used in conjunction with the numerical inversion of the Fourier integral to obtain the transient voltage waveforms at various points in a 500 kV line and in the transformer winding.

  18. Caveolin interaction governs Kv1.3 lipid raft targeting

    PubMed Central

    Pérez-Verdaguer, Mireia; Capera, Jesusa; Martínez-Mármol, Ramón; Camps, Marta; Comes, Núria; Tamkun, Michael M.; Felipe, Antonio

    2016-01-01

    The spatial localization of ion channels at the cell surface is crucial for their functional role. Many channels localize in lipid raft microdomains, which are enriched in cholesterol and sphingolipids. Caveolae, specific lipid rafts which concentrate caveolins, harbor signaling molecules and their targets becoming signaling platforms crucial in cell physiology. However, the molecular mechanisms involved in such spatial localization are under debate. Kv1.3 localizes in lipid rafts and participates in the immunological response. We sought to elucidate the mechanisms of Kv1.3 surface targeting, which govern leukocyte physiology. Kv1 channels share a putative caveolin-binding domain located at the intracellular N-terminal of the channel. This motif, lying close to the S1 transmembrane segment, is situated near the T1 tetramerization domain and the determinants involved in the Kvβ subunit association. The highly hydrophobic domain (FQRQVWLLF) interacts with caveolin 1 targeting Kv1.3 to caveolar rafts. However, subtle variations of this cluster, putative ancillary associations and different structural conformations can impair the caveolin recognition, thereby altering channel’s spatial localization. Our results identify a caveolin-binding domain in Kv1 channels and highlight the mechanisms that govern the regulation of channel surface localization during cellular processes. PMID:26931497

  19. Caveolin interaction governs Kv1.3 lipid raft targeting.

    PubMed

    Pérez-Verdaguer, Mireia; Capera, Jesusa; Martínez-Mármol, Ramón; Camps, Marta; Comes, Núria; Tamkun, Michael M; Felipe, Antonio

    2016-01-01

    The spatial localization of ion channels at the cell surface is crucial for their functional role. Many channels localize in lipid raft microdomains, which are enriched in cholesterol and sphingolipids. Caveolae, specific lipid rafts which concentrate caveolins, harbor signaling molecules and their targets becoming signaling platforms crucial in cell physiology. However, the molecular mechanisms involved in such spatial localization are under debate. Kv1.3 localizes in lipid rafts and participates in the immunological response. We sought to elucidate the mechanisms of Kv1.3 surface targeting, which govern leukocyte physiology. Kv1 channels share a putative caveolin-binding domain located at the intracellular N-terminal of the channel. This motif, lying close to the S1 transmembrane segment, is situated near the T1 tetramerization domain and the determinants involved in the Kvβ subunit association. The highly hydrophobic domain (FQRQVWLLF) interacts with caveolin 1 targeting Kv1.3 to caveolar rafts. However, subtle variations of this cluster, putative ancillary associations and different structural conformations can impair the caveolin recognition, thereby altering channel's spatial localization. Our results identify a caveolin-binding domain in Kv1 channels and highlight the mechanisms that govern the regulation of channel surface localization during cellular processes. PMID:26931497

  20. Kv7.5 Potassium Channel Subunits Are the Primary Targets for PKA-Dependent Enhancement of Vascular Smooth Muscle Kv7 Currents.

    PubMed

    Mani, Bharath K; Robakowski, Christina; Brueggemann, Lyubov I; Cribbs, Leanne L; Tripathi, Abhishek; Majetschak, Matthias; Byron, Kenneth L

    2016-03-01

    Kv7 (KCNQ) channels, formed as homo- or heterotetramers of Kv7.4 and Kv7.5 α-subunits, are important regulators of vascular smooth muscle cell (VSMC) membrane voltage. Recent studies demonstrate that direct pharmacological modulation of VSMC Kv7 channel activity can influence blood vessel contractility and diameter. However, the physiologic regulation of Kv7 channel activity is still poorly understood. Here, we study the effect of cAMP/protein kinase A (PKA) activation on whole cell K(+) currents through endogenous Kv7.5 channels in A7r5 rat aortic smooth muscle cells or through Kv7.4/Kv7.5 heteromeric channels natively expressed in rat mesenteric artery smooth muscle cells. The contributions of specific α-subunits are further dissected using exogenously expressed human Kv7.4 and Kv7.5 homo- or heterotetrameric channels in A7r5 cells. Stimulation of Gαs-coupled β-adrenergic receptors with isoproterenol induced PKA-dependent activation of endogenous Kv7.5 currents in A7r5 cells. The receptor-mediated enhancement of Kv7.5 currents was mimicked by pharmacological agents that increase [cAMP] (forskolin, rolipram, 3-isobutyl-1-methylxanthine, and papaverine) or mimic cAMP (8-bromo-cAMP); the 2- to 4-fold PKA-dependent enhancement of currents was also observed with exogenously expressed Kv7.5 channels. In contrast, exogenously-expressed heterotetrameric Kv7.4/7.5 channels in A7r5 cells or native mesenteric artery smooth muscle Kv7.4/7.5 channels were only modestly enhanced, and homo-tetrameric Kv7.4 channels were insensitive to this regulatory pathway. Correspondingly, proximity ligation assays indicated that isoproterenol induced PKA-dependent phosphorylation of exogenously expressed Kv7.5 channel subunits, but not of Kv7.4 subunits. These results suggest that signal transduction-mediated responsiveness of vascular smooth muscle Kv7 channel subunits to cAMP/PKA activation follows the order of Kv7.5 > Kv7.4/Kv7.5 > Kv7.4. PMID:26700561

  1. Development of 66 kV/6.9 kV 2 MV A prototype HTS power transformer

    NASA Astrophysics Data System (ADS)

    Bohno, T.; Tomioka, A.; Imaizumi, M.; Sanuki, Y.; Yamamoto, T.; Yasukawa, Y.; Ono, H.; Yagi, Y.; Iwadate, K.

    2005-10-01

    We have developed the technology of the producing a HTS magnet for the power transformer. Three subjects have been mainly studied, high voltage technologies, large current and low AC loss technologies and sub-cooling system technologies to establish the technology of 66 kV/6.9 kV 10 MV A class HTS power transformer. In order to verify the validity of elemental technologies, such as high voltage technologies, large current and low AC loss technologies and sub-cooling system technologies, single-phase 2 MV A class 66 kV/6.9 kV prototype HTS transformer was manufactured and tested. In the load loss (AC loss) measurement, it was obtained that the measured value of 633 W was almost corresponding to the calculated value of 576 W at the rated operation of 2 MV A. Moreover, the breakdown was not found all voltage withstand test. These test results indicate that elemental technologies were established for the development of 66 kV/6.9 kV 10 MV A class HTS power transformer.

  2. MinK, MiRP1, and MiRP2 diversify Kv3.1 and Kv3.2 potassium channel gating.

    PubMed

    Lewis, Anthony; McCrossan, Zoe A; Abbott, Geoffrey W

    2004-02-27

    High frequency firing in mammalian neurons requires ultra-rapid delayed rectifier potassium currents generated by homomeric or heteromeric assemblies of Kv3.1 and Kv3.2 potassium channel alpha subunits. Kv3.1 alpha subunits can also form slower activating channels by coassembling with MinK-related peptide 2 (MiRP2), a single transmembrane domain potassium channel ancillary subunit. Here, using channel subunits cloned from rat and expressed in Chinese hamster ovary cells, we show that modulation by MinK, MiRP1, and MiRP2 is a general mechanism for slowing of Kv3.1 and Kv3.2 channel activation and deactivation and acceleration of inactivation, creating a functionally diverse range of channel complexes. MiRP1 also negatively shifts the voltage dependence of Kv3.1 and Kv3.2 channel activation. Furthermore, MinK, MiRP1, and MiRP2 each form channels with Kv3.1-Kv3.2 heteromers that are kinetically distinct from one another and from MiRP/homomeric Kv3 channels. The findings illustrate a mechanism for dynamic expansion of the functional repertoire of Kv3.1 and Kv3.2 potassium currents and suggest roles for these alpha subunits outside the scope of sustained rapid neuronal firing. PMID:14679187

  3. Kv3.3 potassium channels and spinocerebellar ataxia.

    PubMed

    Zhang, Yalan; Kaczmarek, Leonard K

    2016-08-15

    The voltage-dependent potassium channel subunit Kv3.3 is expressed at high levels in cerebellar Purkinje cells, in auditory brainstem nuclei and in many other neurons capable of firing at high rates. In the cerebellum, it helps to shape the very characteristic complex spike of Purkinje cells. Kv3.3 differs from other closely related channels in that human mutations in the gene encoding Kv3.3 (KCNC3) result in a unique neurodegenerative disease termed spinocerebellar ataxia type 13 (SCA13). This primarily affects the cerebellum, but also results in extracerebellar symptoms. Different mutations produce either early onset SCA13, associated with delayed motor and impaired cognitive skill acquisition, or late onset SCA13, which typically produces cerebellar degeneration in middle age. This review covers the localization and physiological function of Kv3.3 in the central nervous system and how the normal function of the channel is altered by the disease-causing mutations. It also describes experimental approaches that are being used to understand how Kv3.3 mutations are linked to neuronal survival, and to develop strategies for treatment. PMID:26442672

  4. Skeletal muscle Kv7 (KCNQ) channels in myoblast differentiation and proliferation

    SciTech Connect

    Roura-Ferrer, Meritxell; Sole, Laura; Martinez-Marmol, Ramon; Villalonga, Nuria; Felipe, Antonio

    2008-05-16

    Voltage-dependent K{sup +} channels (Kv) are involved in myocyte proliferation and differentiation by triggering changes in membrane potential and regulating cell volume. Since Kv7 channels may participate in these events, the purpose of this study was to investigate whether skeletal muscle Kv7.1 and Kv7.5 were involved during proliferation and myogenesis. Here we report that, while myotube formation did not regulate Kv7 channels, Kv7.5 was up-regulated during cell cycle progression. Although, Kv7.1 mRNA also increased during the G{sub 1}-phase, pharmacological evidence mainly involves Kv7.5 in myoblast growth. Our results indicate that the cell cycle-dependent expression of Kv7.5 is involved in skeletal muscle cell proliferation.

  5. Waltz Mill testing of 345-kV PPP cable

    SciTech Connect

    Burghardt, R.R. )

    1991-09-01

    A 345-kV PPP-insulated cable was subjected to a two-year accelerated life test program at the EPRI Waltz Mill Cable Test Facility. Testing started in November 1985 and was successfully completed in September 1988. The program included conductor temperatures ranging from 85{degrees}C to 105{degrees}C and line-to-line voltages from 362 kV to 474 kV. Cyclic testing was performed during 17 of the 24 months. Dissipation factor measurements were made throughout the program. The measurements indicated no deterioration of the cable or splices as a consequence of the high temperatures and voltages applied to them in this test program. 2 refs., 24 figs.

  6. Networks.

    ERIC Educational Resources Information Center

    Maughan, George R.; Petitto, Karen R.; McLaughlin, Don

    2001-01-01

    Describes the connectivity features and options of modern campus communication and information system networks, including signal transmission (wire-based and wireless), signal switching, convergence of networks, and network assessment variables, to enable campus leaders to make sound future-oriented decisions. (EV)

  7. The pore region of the Kv1.2alpha subunit is an important component of recombinant Kv1.2 channel oxygen sensitivity.

    PubMed

    Conforti, Laura; Takimoto, Koichi; Petrovic, Milan; Pongs, Olaf; Millhorn, David

    2003-06-27

    Oxygen-sensitive K(+) channels are important elements in the cellular response to hypoxia. Although much progress has been made in identifying their molecular composition, the structural components associated to their O(2)-sensitivity are not yet understood. Recombinant Kv1.2 currents expressed in Xenopus oocytes are inhibited by a decrease in O(2) availability. On the contrary, heterologous Kv2.1 channels are O(2)-insensitive. To elucidate the protein segment responsible for the O(2)-sensitivity of Kv1.2 channels, we analyzed the response to anoxia of Kv1.2/Kv2.1 chimeric channels. Expression of chimeric Kv2.1 channels each containing the S4, the S1-S3 or the S6-COOH segments of Kv1.2 polypeptide resulted in a K(+) current insensitive to anoxia. In contrast, transferring the S5-S6 segment of Kv1.2 into Kv2.1 produced an O(2)-sensitive K(+) current. Finally, mutating a redox-sensitive methionine residue (M380) of Kv1.2 polypeptide did not affect O(2)-sensitivity. Thus, the pore and its surrounding regions of Kv1.2 polypeptide confer its hypoxic inhibition. This response is independent on the redox modulation of methionine residues in this protein segment. PMID:12804584

  8. 230 kV operation of a substation designed for 115 kV by controlling voltage transients

    SciTech Connect

    Lannes, W.J.; Cheramie, W.J.; Priest, K.W.; Wilhelm, M.R.

    1984-09-01

    Arresters applied at the terminals of gas insulated substations have been used successfully for protection from switching and lightning surges. This arrester application for gas insulated substations permits the use of lower insulation levels and therefore more economical substations. In principle, the same technique can be applied to air insulated substations. This paper describes the analysis of transient voltages in a substation designed for 115 kV which is scheduled for operation at 230 kV. Both arrester rating and location are considered in this analysis. It is shown that complete substation protection is possible by addition of both arresters and capacitors.

  9. The use of polymer horizontal vee arms for 120kV to 230kV conversion

    SciTech Connect

    Doubley, D. )

    1991-01-01

    This paper reports on Detroit Edison which is using the horizontal-vee configuration to convert 120 kV tower lines to 230 kV. This approach is an improvement over previous methods of existing line conversion. The design provides reduced construction costs; both in labor and material. It also provides improvement in line structure strength as well. The all polymer horizontal-vees as replacement for crossarms on existing towers make a substantial improvement in productivity. They are lighter, easier to handle, and less likely to break than their porcelain equivalents.

  10. At 1200 Gallery, Block 63, looking west, showing 230 kv ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    At 1200 Gallery, Block 63, looking west, showing 230 kv cables. These lines were installed in the 1960s and are no longer used. - Columbia Basin Project, Grand Coulee Dam & Franklin D. Roosevelt Lake, Across Columbia River, Southeast of Town of Grand Coulee, Grand Coulee, Grant County, WA

  11. Kv1 channels and neural processing in vestibular calyx afferents.

    PubMed

    Meredith, Frances L; Kirk, Matthew E; Rennie, Katherine J

    2015-01-01

    Potassium-selective ion channels are important for accurate transmission of signals from auditory and vestibular sensory end organs to their targets in the central nervous system. During different gravity conditions, astronauts experience altered input signals from the peripheral vestibular system resulting in sensorimotor dysfunction. Adaptation to altered sensory input occurs, but it is not explicitly known whether this involves synaptic modifications within the vestibular epithelia. Future investigations of such potential plasticity require a better understanding of the electrophysiological mechanisms underlying the known heterogeneity of afferent discharge under normal conditions. This study advances this understanding by examining the role of the Kv1 potassium channel family in mediating action potentials in specialized vestibular afferent calyx endings in the gerbil crista and utricle. Pharmacological agents selective for different sub-types of Kv1 channels were tested on membrane responses in whole cell recordings in the crista. Kv1 channels sensitive to α-dendrotoxin and dendrotoxin-K were found to prevail in the central regions, whereas K(+) channels sensitive to margatoxin, which blocks Kv1.3 and 1.6 channels, were more prominent in peripheral regions. Margatoxin-sensitive currents showed voltage-dependent inactivation. Dendrotoxin-sensitive currents showed no inactivation and dampened excitability in calyces in central neuroepithelial regions. The differential distribution of Kv1 potassium channels in vestibular afferents supports their importance in accurately relaying gravitational and head movement signals through specialized lines to the central nervous system. Pharmacological modulation of specific groups of K(+) channels could help alleviate vestibular dysfunction on earth and in space. PMID:26082693

  12. Kv1 channels and neural processing in vestibular calyx afferents

    PubMed Central

    Meredith, Frances L.; Kirk, Matthew E.; Rennie, Katherine J.

    2015-01-01

    Potassium-selective ion channels are important for accurate transmission of signals from auditory and vestibular sensory end organs to their targets in the central nervous system. During different gravity conditions, astronauts experience altered input signals from the peripheral vestibular system resulting in sensorimotor dysfunction. Adaptation to altered sensory input occurs, but it is not explicitly known whether this involves synaptic modifications within the vestibular epithelia. Future investigations of such potential plasticity require a better understanding of the electrophysiological mechanisms underlying the known heterogeneity of afferent discharge under normal conditions. This study advances this understanding by examining the role of the Kv1 potassium channel family in mediating action potentials in specialized vestibular afferent calyx endings in the gerbil crista and utricle. Pharmacological agents selective for different sub-types of Kv1 channels were tested on membrane responses in whole cell recordings in the crista. Kv1 channels sensitive to α-dendrotoxin and dendrotoxin-K were found to prevail in the central regions, whereas K+ channels sensitive to margatoxin, which blocks Kv1.3 and 1.6 channels, were more prominent in peripheral regions. Margatoxin-sensitive currents showed voltage-dependent inactivation. Dendrotoxin-sensitive currents showed no inactivation and dampened excitability in calyces in central neuroepithelial regions. The differential distribution of Kv1 potassium channels in vestibular afferents supports their importance in accurately relaying gravitational and head movement signals through specialized lines to the central nervous system. Pharmacological modulation of specific groups of K+ channels could help alleviate vestibular dysfunction on earth and in space. PMID:26082693

  13. Networking.

    ERIC Educational Resources Information Center

    Duvall, Betty

    Networking is an information giving and receiving system, a support system, and a means whereby women can get ahead in careers--either in new jobs or in current positions. Networking information can create many opportunities: women can talk about how other women handle situations and tasks, and previously established contacts can be used in…

  14. Survey of Magnetic Fields Near BPA 230-kV and 500-kV Transmission Lines.

    SciTech Connect

    Perrin, Nancy; Aggarwal, Rajinder Pal; Bracken, T. Daniel

    1991-05-20

    The purpose of this study was to characterize typical levels and variability of 60Hz magnetic fields at the centerline and edge of right-of-way of Bonneville Power Administration (BPA) 230-kV and 500-kV transmission lines. This was accomplished by taking magnetic field measurements at over 800 spans in Oregon and Washington. The spans were sampled using a stratified random sampling procedure with region (East vs. West), voltage (230-kV vs 500-kV), and circuit configuration as strata. There were five different circuit configuration groups for each region/voltage category requiring a total of 200 strata. Magnetic field measurements were taken at 13 locations under each span using an EMDEX-C as a survey meter. Additional information recorded for each span included conductor height (at 10 locations), right-of-way width, longitudinal and lateral slope, time of day, vegetation, terrain, weather conditions, temperature, wind speed, span length and presence of other lines in the corridor. 9 refs., 17 figs., 26 tabs.

  15. Extracellular potassium inhibits Kv7.1 potassium channels by stabilizing an inactivated state.

    PubMed

    Larsen, Anders Peter; Steffensen, Annette Buur; Grunnet, Morten; Olesen, Søren-Peter

    2011-08-17

    Kv7.1 (KCNQ1) channels are regulators of several physiological processes including vasodilatation, repolarization of cardiomyocytes, and control of secretory processes. A number of Kv7.1 pore mutants are sensitive to extracellular potassium. We hypothesized that extracellular potassium also modulates wild-type Kv7.1 channels. The Kv7.1 currents were measured in Xenopus laevis oocytes at different concentrations of extracellular potassium (1-50 mM). As extracellular potassium was elevated, Kv7.1 currents were reduced significantly more than expected from theoretical calculations based on the Goldman-Hodgkin-Katz flux equation. Potassium inhibited the steady-state current with an IC(50) of 6.0 ± 0.2 mM. Analysis of tail-currents showed that potassium increased the fraction of channels in the inactivated state. Similarly, the recovery from inactivation was slowed by potassium, suggesting that extracellular potassium stabilizes an inactivated state in Kv7.1 channels. The effect of extracellular potassium was absent in noninactivating Kv7.1/KCNE1 and Kv7.1/KCNE3 channels, further supporting a stabilized inactivated state as the underlying mechanism. Interestingly, coexpression of Kv7.1 with KCNE2 did not attenuate the inhibition by potassium. In a number of other Kv channels, including Kv1.5, Kv4.3, and Kv7.2-5 channels, currents were only minimally reduced by an increase in extracellular potassium as expected. These results show that extracellular potassium modulates Kv7.1 channels and suggests that physiological changes in potassium concentrations may directly control the function of Kv7.1 channels. This may represent a novel regulatory mechanism of excitability and of potassium transport in tissues expressing Kv7.1 channels. PMID:21843472

  16. Proteomic analyses of native brain KV4.2 channel complexes

    PubMed Central

    Marionneau, Céline; LeDuc, Richard D.; Rohrs, Henry W.; Link, Andrew J.; Townsend, R. Reid; Nerbonne, Jeanne M.

    2010-01-01

    Somatodendritic A-type (IA) voltage-gated K+ (KV) channels are key regulators of neuronal excitability, functioning to control action potential waveforms, repetitive firing and the responses to synaptic inputs. Rapidly activating and inactivating somatodendritic IA channels are encoded by KV4 α subunits and accumulating evidence suggests that these channels function as components of macromolecular protein complexes. Mass spectrometry (MS)-based proteomic approaches were developed and exploited here to identify potential components and regulators of native brain KV4.2-encoded IA channel complexes. Using anti-KV4.2 specific antibodies, KV4.2 channel complexes were immunoprecipitated from adult wild type mouse brain. Parallel control experiments were performed on brain samples isolated from (KV4.2−/−) mice harboring a targeted disruption of the KCND2 (KV4.2) locus. Three proteomic strategies were employed: an in-gel approach, coupled to one-dimensional liquid chromatography-tandem MS (1D-LC-MS/MS), and two in-solution approaches, followed by 1D-or 2D-LC-MS/MS. The targeted in-gel 1D-LC-MS/MS analyses demonstrated the presence of the KV4 α subunits (KV4.2, KV4.3 and KV4.1) and the KV4 accessory, KChIP (KChIPI-4) and DPP (DPP6 and 10), proteins in native brain KV4.2 channel complexes. The more comprehensive, in-solution approach, coupled to 2D-LC-MS/MS, also called Multidimensional Protein Identification Technology (MudPIT), revealed that additional regulatory proteins, including the KV channel accessory subunit KVβ1, are also components of native brain KV4.2 channel complexes. Additional biochemical and functional approaches will be required to elucidate the physiological roles of these newly identified KV4 interacting proteins. PMID:19713751

  17. Kv4.2 Knockout Mice Have Hippocampal-Dependent Learning and Memory Deficits

    ERIC Educational Resources Information Center

    Lugo, Joaquin N.; Brewster, Amy L.; Spencer, Corinne M.; Anderson, Anne E.

    2012-01-01

    Kv4.2 channels contribute to the transient, outward K[superscript +] current (A-type current) in hippocampal dendrites, and modulation of this current substantially alters dendritic excitability. Using Kv4.2 knockout (KO) mice, we examined the role of Kv4.2 in hippocampal-dependent learning and memory. We found that Kv4.2 KO mice showed a deficit…

  18. IA Channels Encoded by Kv1.4 and Kv4.2 Regulate Circadian Period of PER2 Expression in the Suprachiasmatic Nucleus

    PubMed Central

    Granados-Fuentes, Daniel; Hermanstyne, Tracey O.; Carrasquillo, Yarimar; Nerbonne, Jeanne M.; Herzog, Erik D.

    2016-01-01

    Neurons in the suprachiasmatic nucleus (SCN), the master circadian pacemaker in mammals, display daily rhythms in electrical activity with more depolarized resting potentials and higher firing rates during the day than at night. Although these daily variations in the electrical properties of SCN neurons are required for circadian rhythms in physiology and behavior, the mechanisms linking changes in neuronal excitability to the molecular clock are not known. Recently, we reported that mice deficient for either Kcna4 (Kv1.4−/−) or Kcnd2 (Kv4.2−/−; but not Kcnd3, Kv4.3−/−), voltage-gated K+ (Kv) channel poreforming subunits that encode subthreshold, rapidly activating, and inactivating K+ currents (IA), have shortened (0.5 h) circadian periods in SCN firing and in locomotor activity compared with wild-type (WT) mice. In the experiments here, we used a mouse (Per2Luc) line engineered with a bioluminescent reporter construct, PERIOD2::LUCIFERASE (PER2::LUC), replacing the endogenous Per2 locus, to test the hypothesis that the loss of Kv1.4- or Kv4.2-encoded IA channels also modifies circadian rhythms in the expression of the clock protein PERIOD2 (PER2). We found that SCN explants from Kv1.4−/−Per2Luc and Kv4.2−/−Per2Luc, but not Kv4.3−/−Per2Luc, mice have significantly shorter (by approximately 0.5 h) circadian periods in PER2 rhythms, compared with explants from Per2Luc mice, revealing that the membrane properties of SCN neurons feedback to regulate clock (PER2) expression. The combined loss of both Kv1.4- and Kv4.2-encoded IA channels in Kv1.4−/−/Kv4.2−/−Per2Luc SCN explants did not result in any further alterations in PER2 rhythms. Interestingly, however, mice lacking both Kv1.4 and Kv4.2 show a striking (approximately 1.8 h) advance in their daily activity onset in a light cycle compared with WT mice, suggesting additional roles for Kv1.4- and Kv4.2-encoded IA channels in controlling the light-dependent responses of neurons within

  19. A Kv7.2 mutation associated with early onset epileptic encephalopathy with suppression-burst enhances Kv7/M channel activity.

    PubMed

    Devaux, Jérôme; Abidi, Affef; Roubertie, Agathe; Molinari, Florence; Becq, Hélène; Lacoste, Caroline; Villard, Laurent; Milh, Mathieu; Aniksztejn, Laurent

    2016-05-01

    Mutations in the KCNQ2 gene encoding the voltage-gated potassium channel subunit Kv7.2 cause early onset epileptic encephalopathy (EOEE). Most mutations have been shown to induce a loss of function or to affect the subcellular distribution of Kv7 channels in neurons. Herein, we investigated functional consequences and subcellular distribution of the p.V175L mutation of Kv7.2 (Kv7.2(V175L) ) found in a patient presenting EOEE. We observed that the mutation produced a 25-40 mV hyperpolarizing shift of the conductance-voltage relationship of both the homomeric Kv7.2(V175L) and heteromeric Kv7.2(V175L) /Kv7.3 channels compared to wild-type channels and a 10 mV hyperpolarizing shift of Kv7.2(V175L) /Kv7.2/Kv7.3 channels in a 1:1:2 ratio mimicking the patient situation. Mutant channels also displayed faster activation kinetics and an increased current density that was prevented by 1 μm linopirdine. The p.V175L mutation did not affect the protein expression of Kv7 channels and its localization at the axon initial segment. We conclude that p.V175L is a gain of function mutation. This confirms previous observations showing that mutations having opposite consequences on M channels can produce EOEE. These findings alert us that drugs aiming to increase Kv7 channel activity might have adverse effects in EOEE in the case of gain-of-function variants. PMID:27030113

  20. Symmetric Kv1.5 Blockers Discovered by Focused Screening

    PubMed Central

    2012-01-01

    Guided by computational methods, a set of 1920 compounds were selected from the AstraZeneca corporate collection and screened for Kv1.5 activity. To facilitate rapid generation of structure–activity relationships, special attention was given to selecting subsets of structurally similar molecules by using a maximum common substructure similarity-based procedure. The focused screen hit rate was relatively high (12%). More importantly, a structural series featured by the symmetric 1,2-diphenylethane-1,2-diamine substructure was identified as potent Kv.1.5 blockers. The property profile for the series is shown to meet stringent lead-optimization criteria, providing a springboard for the development of a new and safe treatment for atrial fibrillation. PMID:24900546

  1. A computer controlled 100 kV pulse generator

    SciTech Connect

    Kupferman, S.L.; Booker, S.R.; Meissner, H.J.

    1989-01-01

    A high voltage pulse generator used primarily for calibrating precision high voltage dividers has been redesigned and rebuilt for state of the art operation. It now has the capability to run a divider calibration, compare the results with historical data for the divider, and then print a calibration certificate, all under computer control. Thirteen component instruments are controlled via three IEEE 488 buses. A voice synthesizer is used to inform the operator of the progress of the calibration. Measurement precision is in the 0.1 % range. A heat exchanger is used to control the temperature of the oil bath to allow calibration of dividers between 13/degree/C (55/degree/F) and 21/degree/C (70/degree/F). Positive or negative pulse polarity may be selected and pulse magnitude is settable from less than 10 kV to greater than 300 kV. 1 ref., 8 figs., 1 tab.

  2. Injection into a circular machine with a KV distribution

    SciTech Connect

    Crosbie, E.; Symon, K.

    1995-12-31

    In order to achieve a maximum space charge limit in the IPNS-II synchrotron it is desirable to inject a Kapchinskij-Vladimirskij (KV) distribution (1). We rederive the KV distribution, first starting from a smoothed Hamiltonian and then for the full alternating gradient case. The microcanonical distribution can be generalized slightly so as to allow one to alter the aspect ratio of the beam ellipse. The KV distribution requires that the injected particles all have the same total transverse oscillation energy, and also that they are distributed uniformly throughout the entire energy shell. This requires painting the injected beam uniformly in the three independent dimensions of the energy shell. We have devised two scenarios for doing this, one involving a suitable variation of the x and y injected amplitudes during the injection process, and the second involving introducing a small coupling between the x and y motions. We have written a program to simulate the injection process which includes the turn-to-turn forces between the (500) injected turns. If we omit the turn-to-turn forces then the resulting space charge density distributions are indeed very nearly uniform within a circular beam cross section for either KV injection scenario, but are neither uniform nor circular for other plausible scenarios. With turn-to-turn forces included, the interturn scattering can be fairly important and the resulting density distributions tend to develop lower density halos. If we add a gradient bump to simulate magnetic quadrupole errors in the lattice, then the effects of half-integral resonances can be clearly seen. When the space charge forces between turns depress the tune to a resonance, beam growth keeps the tunes constant at the edge of the stop band, unless the resonance is crossed quickly.

  3. Kv1.3 potassium channel mediates macrophage migration in atherosclerosis by regulating ERK activity.

    PubMed

    Kan, Xiao-Hong; Gao, Hai-Qing; Ma, Zhi-Yong; Liu, Lin; Ling, Ming-Ying; Wang, Yuan-Yuan

    2016-02-01

    Ion channels expressed in macrophages have been tightly related to atherosclerosis by coupling cellular function. How the voltage-gated potassium channels (Kv) affect macrophage migration remain unknown. The aim of our study is to investigate whether Kv1.3-ERK signaling pathway plays an important role in the process. We explored the expression of Kv1.3 in coronary atherosclerotic heart disease and found Kv1.3 channel was increased in acute coronary syndrome patients. Treatment of RAW264.7 cells with Kv1.3 small interfering RNA, suppressed cell migration. The expression of phosphorylated ERK1/2 also decreased after knockdown of Kv1.3. On the other hand, overexpression of Kv1.3 channel promoted cell migration and ERK1/2 phosphorylation. U-0126, the mitogen-activated protein kinase inhibitors, could reverse macrophage migration induced by Kv1.3 channel overexpression. Downregulation of Kv1.3 channel by siRNA could not further inhibit cell migration when cells were treated with U-0126. It means that ERK is downstream signal of Kv1.3 channel. We concluded that Kv1.3 may stimulate macrophage migration through the activation of ERK. PMID:26748289

  4. Regulation of Kv1 channel trafficking by the mamba snake neurotoxin dendrotoxin K

    PubMed Central

    Vacher, Helene; Mohapatra, Durga P.; Misonou, Hiroaki; Trimmer, James S.

    2009-01-01

    Modulation of voltage-gated potassium (Kv) channel surface expression can profoundly affect neuronal excitability. Some but not all mammalian Shaker or Kv1 α subunits contain a dominant endoplasmic reticulum (ER) retention signal in their pore region, preventing surface expression of Kv1.1 homotetrameric channels, and of heteromeric Kv1 channels containing more than one Kv1.1 subunit. The critical amino acid residues within this ER pore-region retention signal are also critical for high-affinity binding of snake dendrotoxins (DTX). This suggests that ER retention may be mediated by an ER protein with a domain structurally similar to that of DTX. One facet of such a model is that expression of soluble DTX in the ER lumen should compete for binding to the retention protein and allow for surface expression of retained Kv1.1. Here we show that luminal DTX expression dramatically increased both the level of cell surface Kv1.1 immunofluorescence staining and the proportion of Kv1.1 with processed N-linked oligosaccharides. Electrophysiological analyses showed that luminal DTX expression led to significant increases in Kv1.1 currents. Together these data showed that luminal DTX expression increases surface expression of functional Kv1.1 homotetrameric channels, and support a model whereby a DTX-like ER protein regulates abundance of cell surface Kv1 channels. PMID:17185748

  5. Physical and functional interaction of KV10.1 with Rabaptin-5 impacts ion channel trafficking.

    PubMed

    Ninkovic, Milena; Mitkovski, Mišo; Kohl, Tobias; Stühmer, Walter; Pardo, Luis A

    2012-09-21

    K(V)10.1 is a potassium channel expressed in brain and implicated in tumor progression. We have searched for proteins interacting with K(V)10.1 and identified Rabaptin-5, an effector of the Rab5 GTPase. Both proteins co-localize on large early endosomes induced by Rab5 hyperactivity. Silencing of Rabaptin-5 induces down-regulation of recycling of K(V)10.1 channel in transfected cells and reduction of K(V)10.1 current density in cells natively expressing K(V)10.1, indicating a role of Rabaptin-5 in channel trafficking. K(V)10.1 co-localizes, but does not physically interact, with Rab7 and Rab11. Our data highlights the complex control of the amount of K(V)10.1 channels on the cell surface. PMID:22841712

  6. A Quantitative Analysis of Neurons with Kv3 Potassium Channel Subunits–Kv3.1b and Kv3.2–in Macaque Primary Visual Cortex

    PubMed Central

    Constantinople, Christine M.; Disney, Anita A; Maffie, Jonathan; Rudy, Bernardo; Hawken, Michael J

    2010-01-01

    Voltage-gated potassium channels that are composed of Kv3 subunits exhibit distinct electrophysiological properties: activation at more depolarized potentials than other voltage-gated K+ channels and fast kinetics. These channels have been shown to contribute to the high-frequency firing of fast-spiking (FS) GABAergic interneurons in the rat and mouse brain. In the rodent neocortex, there are distinct patterns of expression for the Kv3.1b and Kv3.2 channel subunits and of co-expression of these subunits with neurochemical markers, such as the calcium-binding proteins parvalbumin (PV) and calbindin D-28K (CB). The distribution of Kv3 channels and interrelationship with calcium-binding protein expression has not been investigated in primate cortex. We used immunoperoxidase and immunofluorescent labeling and stereological counting techniques to characterize the laminar and cell-type distributions of Kv3-ir neurons in macaque V1. We found that across the cortical layers ~25% of both Kv3.1b- and Kv3.2-ir neurons are non-GABAergic. In contrast all Kv3-ir neurons in rodent cortex are GABAergic (Chow et al., 1999). The putatively excitatory Kv3-ir neurons were mostly located in layers 2, 3 and 4b. Further, the proportion of Kv3-ir neurons that express PV or CB also differs between macaque V1 and rodent cortex. These data indicate that, within the population of cortical neurons, a broader population of neurons, encompassing cells of a wider range of morphological classes may be capable of sustaining high-frequency firing in macaque V1. PMID:19634181

  7. Inhibition of KV7 Channels Protects the Rat Heart against Myocardial Ischemia and Reperfusion Injury.

    PubMed

    Hedegaard, Elise R; Johnsen, Jacob; Povlsen, Jonas A; Jespersen, Nichlas R; Shanmuganathan, Jeffrey A; Laursen, Mia R; Kristiansen, Steen B; Simonsen, Ulf; Bøtker, Hans Erik

    2016-04-01

    The voltage-gated KV7 (KCNQ) potassium channels are activated by ischemia and involved in hypoxic vasodilatation. We investigated the effect of KV7 channel modulation on cardiac ischemia and reperfusion injury and its interaction with cardioprotection by ischemic preconditioning (IPC). Reverse-transcription polymerase chain reaction revealed expression of KV7.1, KV7.4, and KV7.5 in the left anterior descending rat coronary artery and all KV7 subtypes (KV7.1-KV7.5) in the left and right ventricles of the heart. Isolated hearts were subjected to no-flow global ischemia and reperfusion with and without IPC. Infarct size was quantified by 2,3,5-triphenyltetrazolium chloride staining. Two blockers of KV7 channels, XE991 [10,10-bis(4-pyridinylmethyl)-9(10H)-anthracenone] (10 µM) and linopirdine (10 µM), reduced infarct size and exerted additive infarct reduction to IPC. An opener of KV7 channels, flupirtine (10 µM) abolished infarct size reduction by IPC. Hemodynamics were measured using a catheter inserted in the left ventricle and postischemic left ventricular recovery improved in accordance with reduction of infarct size and deteriorated with increased infarct size. XE991 (10 µM) reduced coronary flow in the reperfusion phase and inhibited vasodilatation in isolated small branches of the left anterior descending coronary artery during both simulated ischemia and reoxygenation. KV7 channels are expressed in rat coronary arteries and myocardium. Inhibition of KV7 channels exerts cardioprotection and opening of KV7 channels abrogates cardioprotection by IPC. Although safety issues should be further addressed, our findings suggest a potential role for KV7 blockers in the treatment of ischemia-reperfusion injury. PMID:26869667

  8. The results of systems tests of the 500 kV busbar controllable shunting reactor in the Tavricheskaya substation

    SciTech Connect

    Gusev, S. I.; Karpov, V. N.; Kiselev, A. N.; Kochkin, V. I.

    2009-09-15

    The results of systems tests of the 500 kV busbar magnetization-controllable shunting reactor (CSR), set up in the Tavricheskaya substation, including measurements of the quality of the electric power, the harmonic composition of the network currents of the reactor for different values of the reactive power consumed, the determination of the regulating characteristics of the reactor, the speed of response of the shunting reactor in the current and voltage stabilization modes, and also the operation of the reactor under dynamic conditions for different perturbations, are presented. The results obtained are analyzed.

  9. Unconventional EGF-induced ERK1/2-mediated Kv1.3 endocytosis.

    PubMed

    Martínez-Mármol, Ramón; Comes, Núria; Styrczewska, Katarzyna; Pérez-Verdaguer, Mireia; Vicente, Rubén; Pujadas, Lluís; Soriano, Eduardo; Sorkin, Alexander; Felipe, Antonio

    2016-04-01

    The potassium channel Kv1.3 plays roles in immunity, neuronal development and sensory discrimination. Regulation of Kv1.3 by kinase signaling has been studied. In this context, EGF binds to specific receptors (EGFR) and triggers tyrosine kinase-dependent signaling, which down-regulates Kv1.3 currents. We show that Kv1.3 undergoes EGF-dependent endocytosis. This EGF-mediated mechanism is relevant because is involved in adult neural stem cell fate determination. We demonstrated that changes in Kv1.3 subcellular distribution upon EGFR activation were due to Kv1.3 clathrin-dependent endocytosis, which targets the Kv1.3 channels to the lysosomal degradative pathway. Interestingly, our results further revealed that relevant tyrosines and other interacting motifs, such as PDZ and SH3 domains, were not involved in the EGF-dependent Kv1.3 internalization. However, a new, and yet undescribed mechanism, of ERK1/2-mediated threonine phosphorylation is crucial for the EGF-mediated Kv1.3 endocytosis. Our results demonstrate that EGF triggers the down-regulation of Kv1.3 activity and its expression at the cell surface, which is important for the development and migration of adult neural progenitors. PMID:26542799

  10. The Role of KV7.3 in Regulating Osteoblast Maturation and Mineralization

    PubMed Central

    Yang, Ji Eun; Song, Min Seok; Shen, Yiming; Ryu, Pan Dong; Lee, So Yeong

    2016-01-01

    KCNQ (KV7) channels are voltage-gated potassium (KV) channels, and the function of KV7 channels in muscles, neurons, and sensory cells is well established. We confirmed that overall blockade of KV channels with tetraethylammonium augmented the mineralization of bone-marrow-derived human mesenchymal stem cells during osteogenic differentiation, and we determined that KV7.3 was expressed in MG-63 and Saos-2 cells at the mRNA and protein levels. In addition, functional KV7 currents were detected in MG-63 cells. Inhibition of KV7.3 by linopirdine or XE991 increased the matrix mineralization during osteoblast differentiation. This was confirmed by alkaline phosphatase, osteocalcin, and osterix in MG-63 cells, whereas the expression of Runx2 showed no significant change. The extracellular glutamate secreted by osteoblasts was also measured to investigate its effect on MG-63 osteoblast differentiation. Blockade of KV7.3 promoted the release of glutamate via the phosphorylation of extracellular signal-regulated kinase 1/2-mediated upregulation of synapsin, and induced the deposition of type 1 collagen. However, activation of KV7.3 by flupirtine did not produce notable changes in matrix mineralization during osteoblast differentiation. These results suggest that KV7.3 could be a novel regulator in osteoblast differentiation. PMID:26999128

  11. Unconventional EGF-induced ERK1/2-mediated Kv1.3 endocytosis

    PubMed Central

    Martínez-Mármol, Ramón; Comes, Núria; Styrczewska, Katarzyna; Pérez-Verdaguer, Mireia; Vicente, Rubén; Pujadas, Lluís; Soriano, Eduardo; Sorkin, Alexander

    2016-01-01

    The potassium channel Kv1.3 plays roles in immunity, neuronal development and sensory discrimination. Regulation of Kv1.3 by kinase signaling has been studied. In this context, EGF binds to specific receptors (EGFR) and triggers tyrosine kinase-dependent signaling, which down-regulates Kv1.3 currents. We show that Kv1.3 undergoes EGF-dependent endocytosis. This EGF-mediated mechanism is relevant because is involved in adult neural stem cell fate determination. We demonstrated that changes in Kv1.3 subcellular distribution upon EGFR activation were due to Kv1.3 clathrin-dependent endocytosis, which targets the Kv1.3 channels to the lysosomal degradative pathway. Interestingly, our results further revealed that relevant tyrosines and other interacting motifs, such as PDZ and SH3 domains, were not involved in the EGF-dependent Kv1.3 internalization. However, a new, and yet undescribed mechanism, of ERK1/2-mediated threonine phosphorylation is crucial for the EGF-mediated Kv1.3 endocytosis. Our results demonstrate that EGF triggers the down-regulation of Kv1.3 activity and its expression at the cell surface, which is important for the development and migration of adult neural progenitors. PMID:26542799

  12. The Role of KV7.3 in Regulating Osteoblast Maturation and Mineralization.

    PubMed

    Yang, Ji Eun; Song, Min Seok; Shen, Yiming; Ryu, Pan Dong; Lee, So Yeong

    2016-01-01

    KCNQ (KV7) channels are voltage-gated potassium (KV) channels, and the function of KV7 channels in muscles, neurons, and sensory cells is well established. We confirmed that overall blockade of KV channels with tetraethylammonium augmented the mineralization of bone-marrow-derived human mesenchymal stem cells during osteogenic differentiation, and we determined that KV7.3 was expressed in MG-63 and Saos-2 cells at the mRNA and protein levels. In addition, functional KV7 currents were detected in MG-63 cells. Inhibition of KV7.3 by linopirdine or XE991 increased the matrix mineralization during osteoblast differentiation. This was confirmed by alkaline phosphatase, osteocalcin, and osterix in MG-63 cells, whereas the expression of Runx2 showed no significant change. The extracellular glutamate secreted by osteoblasts was also measured to investigate its effect on MG-63 osteoblast differentiation. Blockade of KV7.3 promoted the release of glutamate via the phosphorylation of extracellular signal-regulated kinase 1/2-mediated upregulation of synapsin, and induced the deposition of type 1 collagen. However, activation of KV7.3 by flupirtine did not produce notable changes in matrix mineralization during osteoblast differentiation. These results suggest that KV7.3 could be a novel regulator in osteoblast differentiation. PMID:26999128

  13. Neuronal Voltage-Gated K+ (Kv) Channels Function in Macromolecular Complexes

    PubMed Central

    Norris, Aaron J.; Foeger, Nicholas C.; Nerbonne, Jeanne M.

    2010-01-01

    Considerable evidence indicates that native neuronal voltage-gated K+ (Kv) currents reflect the functioning of macromolecular Kv channel complexes, composed of pore-forming (α) subunits, cytosolic and transmembrane accessory subunits, together with regulatory and scaffolding proteins. The individual components of these macromolecular complexes appear to influence the stability, the trafficking, the localization and/or the biophysical properties of the channels. Recent studies suggest that Kv channel accessory subunits subserve multiple roles in the generation of native neuronal Kv channels. Additional recent findings suggest that Kv channel accessory subunits can respond to changes in intracellular Ca2+ or metabolism and thereby integrate signaling pathways to regulate Kv channel expression and properties. Although studies in heterologous cells have provided important insights into the effects of accessory subunits on Kv channel expression/properties, it has become increasingly clear that experiments in neurons are required to define the physiological roles of Kv channel accessory and associated proteins. A number of technological and experimental hurdles remain that must be overcome in the design, execution and interpretation of experiments aimed at detailing the functional roles of accessory subunits and associated proteins in the generation of native neuronal Kv channels. With the increasing association of altered Kv channel functioning with neurological disorders, the potential impact of these efforts is clear. PMID:20813163

  14. Developmental Expression of Kv Potassium Channels at the Axon Initial Segment of Cultured Hippocampal Neurons

    PubMed Central

    Sánchez-Ponce, Diana; DeFelipe, Javier; Garrido, Juan José; Muñoz, Alberto

    2012-01-01

    Axonal outgrowth and the formation of the axon initial segment (AIS) are early events in the acquisition of neuronal polarity. The AIS is characterized by a high concentration of voltage-dependent sodium and potassium channels. However, the specific ion channel subunits present and their precise localization in this axonal subdomain vary both during development and among the types of neurons, probably determining their firing characteristics in response to stimulation. Here, we characterize the developmental expression of different subfamilies of voltage-gated potassium channels in the AISs of cultured mouse hippocampal neurons, including subunits Kv1.2, Kv2.2 and Kv7.2. In contrast to the early appearance of voltage-gated sodium channels and the Kv7.2 subunit at the AIS, Kv1.2 and Kv2.2 subunits were tethered at the AIS only after 10 days in vitro. Interestingly, we observed different patterns of Kv1.2 and Kv2.2 subunit expression, with each confined to distinct neuronal populations. The accumulation of Kv1.2 and Kv2.2 subunits at the AIS was dependent on ankyrin G tethering, it was not affected by disruption of the actin cytoskeleton and it was resistant to detergent extraction, as described previously for other AIS proteins. This distribution of potassium channels in the AIS further emphasizes the heterogeneity of this structure in different neuronal populations, as proposed previously, and suggests corresponding differences in action potential regulation. PMID:23119056

  15. An engineered scorpion toxin analogue with improved Kv1.3 selectivity displays reduced conformational flexibility

    PubMed Central

    Bartok, Adam; Fehér, Krisztina; Bodor, Andrea; Rákosi, Kinga; Tóth, Gábor K.; Kövér, Katalin E.; Panyi, Gyorgy; Varga, Zoltan

    2015-01-01

    The voltage-gated Kv1.3 K+ channel plays a key role in the activation of T lymphocytes. Kv1.3 blockers selectively suppress immune responses mediated by effector memory T cells, which indicates the great potential of selective Kv1.3 inhibitors in the therapy of certain autoimmune diseases. Anuroctoxin (AnTx), a 35-amino-acid scorpion toxin is a high affinity blocker of Kv1.3, but also blocks Kv1.2 with similar potency. We designed and produced three AnTx variants: ([F32T]-AnTx, [N17A]-AnTx, [N17A/F32T]-AnTx) using solid-phase synthesis with the goal of improving the selectivity of the toxin for Kv1.3 over Kv1.2 while keeping the high affinity for Kv1.3. We used the patch-clamp technique to determine the blocking potency of the synthetic toxins on hKv1.3, mKv1.1, hKv1.2 and hKCa3.1 channels. Of the three variants [N17A/F32T]-AnTx maintained the high affinity of the natural peptide for Kv1.3 but became more than 16000-fold selective over Kv1.2. NMR data and molecular dynamics simulations suggest that the more rigid structure with restricted conformational space of the double substituted toxin compared to the flexible wild-type one is an important determinant of toxin selectivity. Our results provide the foundation for the possibility of the production and future therapeutic application of additional, even more selective toxins targeting various ion channels. PMID:26689143

  16. Kv2 dysfunction after peripheral axotomy enhances sensory neuron responsiveness to sustained input.

    PubMed

    Tsantoulas, Christoforos; Zhu, Lan; Yip, Ping; Grist, John; Michael, Gregory J; McMahon, Stephen B

    2014-01-01

    Peripheral nerve injuries caused by trauma are associated with increased sensory neuron excitability and debilitating chronic pain symptoms. Axotomy-induced alterations in the function of ion channels are thought to largely underlie the pathophysiology of these phenotypes. Here, we characterise the mRNA distribution of Kv2 family members in rat dorsal root ganglia (DRG) and describe a link between Kv2 function and modulation of sensory neuron excitability. Kv2.1 and Kv2.2 were amply expressed in cells of all sizes, being particularly abundant in medium-large neurons also immunoreactive for neurofilament-200. Peripheral axotomy led to a rapid, robust and long-lasting transcriptional Kv2 downregulation in the DRG, correlated with the onset of mechanical and thermal hypersensitivity. The consequences of Kv2 loss-of-function were subsequently investigated in myelinated neurons using intracellular recordings on ex vivo DRG preparations. In naïve neurons, pharmacological Kv2.1/Kv2.2 inhibition by stromatoxin-1 (ScTx) resulted in shortening of action potential (AP) after-hyperpolarization (AHP). In contrast, ScTx application on axotomized neurons did not alter AHP duration, consistent with the injury-induced Kv2 downregulation. In accordance with a shortened AHP, ScTx treatment also reduced the refractory period and improved AP conduction to the cell soma during high frequency stimulation. These results suggest that Kv2 downregulation following traumatic nerve lesion facilitates greater fidelity of repetitive firing during prolonged input and thus normal Kv2 function is postulated to limit neuronal excitability. In summary, we have profiled Kv2 expression in sensory neurons and provide evidence for the contribution of Kv2 dysfunction in the generation of hyperexcitable phenotypes encountered in chronic pain states. PMID:24252178

  17. Complementary functions of SK and Kv7/M potassium channels in excitability control and synaptic integration in rat hippocampal dentate granule cells

    PubMed Central

    Mateos-Aparicio, Pedro; Murphy, Ricardo; Storm, Johan F

    2014-01-01

    The dentate granule cells (DGCs) form the most numerous neuron population of the hippocampal memory system, and its gateway for cortical input. Yet, we have only limited knowledge of the intrinsic membrane properties that shape their responses. Since SK and Kv7/M potassium channels are key mechanisms of neuronal spiking and excitability control, afterhyperpolarizations (AHPs) and synaptic integration, we studied their functions in DGCs. The specific SK channel blockers apamin or scyllatoxin increased spike frequency (excitability), reduced early spike frequency adaptation, fully blocked the medium-duration AHP (mAHP) after a single spike or spike train, and increased postsynaptic EPSP summation after spiking, but had no effect on input resistance (Rinput) or spike threshold. In contrast, blockade of Kv7/M channels by XE991 increased Rinput, lowered the spike threshold, and increased excitability, postsynaptic EPSP summation, and EPSP–spike coupling, but only slightly reduced mAHP after spike trains (and not after single spikes). The SK and Kv7/M channel openers 1-EBIO and retigabine, respectively, had effects opposite to the blockers. Computational modelling reproduced many of these effects. We conclude that SK and Kv7/M channels have complementary roles in DGCs. These mechanisms may be important for the dentate network function, as CA3 neurons can be activated or inhibition recruited depending on DGC firing rate. PMID:24366266

  18. 30-kV proton injector for PIGMI

    SciTech Connect

    Hamm, R.W.; Mueller, D.W.; Sturgess, R.G.

    1981-01-01

    A 30-kV proton injector designed for matching a 31-mA proton beam into the radio-frequency quadrupole (RFQ) section of the PIGMI accelerator has been constructed and tested. This injector uses a small efficient duoplasmatron ion source and a single-gap extraction system for creating a convergent ion beam, and a three-element unipotential einzel lens for focusing the ion beam into the RFQ. A description of this prototype injector is presented, along with the experimental data obtained during the testing of this system.

  19. Pollution performance of 110 kV metal oxide arresters

    SciTech Connect

    Chrzan, K.; Pohl, Z.; Grzybowski, S.; Koehler, W.

    1997-04-01

    Pollution test results of single unit 110 kV metal oxide surge arresters with porcelain housing according to the solid layer and salt fog methods are presented. During 6 hours of testing, the internal and external charge and maximum temperature along the varistor column were measured. The formation of single stable dry bands on the housing was often observed, especially during salt fog tests. In such cases, the varistor temperature can reach about 70 C. The simple electrical model of the arrester enabling calculations of voltages and currents as a function of arrester and pollution parameters is shown.

  20. 500 kV shield wires; Sectionalize or ground everywhere

    SciTech Connect

    Tuominen, M.W. )

    1992-01-01

    The primary purpose of shield wires on transmission lines in lightning protection. Development of fiber-optic shied wires added communications capabilities. Schemes for adapting fiber-optic shield wires to sectionalized shield wire designs remain untested. It is a present policy to segment and insulate 500 kV shield wires. Computer modeling and supportive field measurements have revealed a characteristic distribution of grounded tower voltage versus distance for long lines. The intent of this paper is to report these results plus predict the outcome of grounding shield wires on transmission lines already in place. Shield wires are often called overhead ground wires and abbreviated OHGW.

  1. Mapping the Interaction Anatomy of BmP02 on Kv1.3 Channel.

    PubMed

    Wu, B; Wu, B F; Feng, Y J; Tao, J; Ji, Y H

    2016-01-01

    The potassium channel Kv 1.3 plays a vital part in the activation of T lymphocytes and is an attractive pharmacological target for autoimmune diseases. BmP02, a 28-residue peptide isolated from Chinese scorpion (Buthus martensi Karsch) venom, is a potent and selective Kv1.3 channel blocker. However, the mechanism through which BmP02 recognizes and inhibits the Kv1.3 channel is still unclear. In the present study, a complex molecular model of Kv1.3-BmP02 was developed by docking analysis and molecular dynamics simulations. From these simulations, it appears the large β-turn (residues 10-16) of BmP02 might be the binding interface with Kv 1.3. These results were confirmed by scanning alanine mutagenesis of BmP02, which identified His9, Lys11 and Lys13, which lie within BmP02's β-turn, as key residues for interacting with Kv1.3. Based on these results and molecular modeling, two negatively charged residues of Kv1.3, D421 and D422, located in turret region, were predicted to act as the binding site for BmP02. Mutation of these residues reduced sensitivity of Kv 1.3 to BmP02 inhibition, suggesting that electrostatic interactions play a crucial role in Kv1.3-BmP02 interaction. This study revealed the molecular basis of Kv 1.3 recognition by BmP02 venom, and provides a novel interaction model for Kv channel-specific blocker complex, which may help guide future drug-design for Kv1.3-related channelopathies. PMID:27403813

  2. First 765-kV SF/sub 6/ station proves successful

    SciTech Connect

    Wolff, R.F.

    1980-08-01

    The experience gained by American Electric Power Service Corp during the design, construction, and operation of the worlds first 765-kV gas-insulated substation proves the effectiveness of this design. Although the cost effectiveness of SF/sub 6/ insulated design increases as the voltage level increases, the combined cost of the 765-kV station and its associated 138-kV station was greater than the cost of equivalent air-insulated substations.

  3. Mapping the Interaction Anatomy of BmP02 on Kv1.3 Channel

    PubMed Central

    Wu, B.; Wu, B. F.; Feng, Y. J.; Tao, J.; Ji, Y. H.

    2016-01-01

    The potassium channel Kv 1.3 plays a vital part in the activation of T lymphocytes and is an attractive pharmacological target for autoimmune diseases. BmP02, a 28-residue peptide isolated from Chinese scorpion (Buthus martensi Karsch) venom, is a potent and selective Kv1.3 channel blocker. However, the mechanism through which BmP02 recognizes and inhibits the Kv1.3 channel is still unclear. In the present study, a complex molecular model of Kv1.3-BmP02 was developed by docking analysis and molecular dynamics simulations. From these simulations, it appears the large β-turn (residues 10–16) of BmP02 might be the binding interface with Kv 1.3. These results were confirmed by scanning alanine mutagenesis of BmP02, which identified His9, Lys11 and Lys13, which lie within BmP02’s β-turn, as key residues for interacting with Kv1.3. Based on these results and molecular modeling, two negatively charged residues of Kv1.3, D421 and D422, located in turret region, were predicted to act as the binding site for BmP02. Mutation of these residues reduced sensitivity of Kv 1.3 to BmP02 inhibition, suggesting that electrostatic interactions play a crucial role in Kv1.3-BmP02 interaction. This study revealed the molecular basis of Kv 1.3 recognition by BmP02 venom, and provides a novel interaction model for Kv channel-specific blocker complex, which may help guide future drug-design for Kv1.3-related channelopathies. PMID:27403813

  4. Expression and function of KV2-containing channels in human urinary bladder smooth muscle

    PubMed Central

    Hristov, Kiril L.; Chen, Muyan; Afeli, Serge A. Y.; Cheng, Qiuping; Rovner, Eric S.

    2012-01-01

    The functional role of the voltage-gated K+ (KV) channels in human detrusor smooth muscle (DSM) is largely unexplored. Here, we provide molecular, electrophysiological, and functional evidence for the expression of KV2.1, KV2.2, and the electrically silent KV9.3 subunits in human DSM. Stromatoxin-1 (ScTx1), a selective inhibitor of KV2.1, KV2.2, and KV4.2 homotetrameric channels and of KV2.1/9.3 heterotetrameric channels, was used to examine the role of these channels in human DSM function. Human DSM tissues were obtained during open bladder surgeries from patients without a history of overactive bladder. Freshly isolated human DSM cells were studied using RT-PCR, immunocytochemistry, live-cell Ca2+ imaging, and the perforated whole cell patch-clamp technique. Isometric DSM tension recordings of human DSM isolated strips were conducted using tissue baths. RT-PCR experiments showed mRNA expression of KV2.1, KV2.2, and KV9.3 (but not KV4.2) channel subunits in human isolated DSM cells. KV2.1 and KV2.2 protein expression was confirmed by Western blot analysis and immunocytochemistry. Perforated whole cell patch-clamp experiments revealed that ScTx1 (100 nM) inhibited the amplitude of the voltage step-induced KV current in freshly isolated human DSM cells. ScTx1 (100 nM) significantly increased the intracellular Ca2+ level in DSM cells. In human DSM isolated strips, ScTx1 (100 nM) increased the spontaneous phasic contraction amplitude and muscle force, and enhanced the amplitude of the electrical field stimulation-induced contractions within the range of 3.5–30 Hz stimulation frequencies. These findings reveal that ScTx1-sensitive KV2-containing channels are key regulators of human DSM excitability and contractility and may represent new targets for pharmacological or genetic intervention for bladder dysfunction. PMID:22422395

  5. Endogenous KCNE Subunits Govern Kv2.1 K+ Channel Activation Kinetics in Xenopus Oocyte Studies

    PubMed Central

    Gordon, Earl; Roepke, Torsten K.; Abbott, Geoffrey W.

    2006-01-01

    Kv2.1 is a voltage-gated potassium (Kv) channel that generates delayed rectifier currents in mammalian heart and brain. The biophysical properties of Kv2.1 and other ion channels have been characterized by functional expression in heterologous systems, and most commonly in Xenopus laevis oocytes. A number of previous oocyte-based studies of mammalian potassium channels have revealed expression-level-dependent changes in channel properties, leading to the suggestion that endogenous oocyte factors regulate channel gating. Here, we show that endogenous oocyte potassium channel KCNE ancillary subunits xMinK and xMiRP2 slow the activation of oocyte-expressed mammalian Kv2.1 channels two-to-fourfold. This produces a sigmoidal relationship between Kv2.1 current density and activation rate in oocyte-based two-electrode voltage clamp studies. The effect of endogenous xMiRP2 and xMinK on Kv2.1 activation is diluted at high Kv2.1 expression levels, or by RNAi knockdown of either endogenous subunit. RNAi knockdown of both xMiRP2 and xMinK eliminates the correlation between Kv2.1 expression level and activation kinetics. The data demonstrate a molecular basis for expression-level-dependent changes in Kv channel gating observed in heterologous expression studies. PMID:16326911

  6. Wild Horse 69-kV transmission line environmental assessment

    SciTech Connect

    1996-12-01

    Hill County Electric Cooperative Inc. (Hill County) proposes to construct and operate a 69-kV transmission line from its North Gildford Substation in Montana north to the Canadian border. A vicinity project area map is enclosed as a figure. TransCanada Power Corporation (TCP), a Canadian power-marketing company, will own and construct the connecting 69-kV line from the international border to Express Pipeline`s pump station at Wild Horse, Alberta. This Environmental Assessment is prepared for the Department of Energy (DOE) as lead federal agency to comply with the requirements of the National Environmental Policy Act (NEPA), as part of DOE`s review and approval process of the applications filed by Hill County for a DOE Presidential Permit and License to Export Electricity to a foreign country. The purpose of the proposed line is to supply electric energy to a crude oil pump station in Canada, owned by Express Pipeline Ltd. (Express). The pipeline would transport Canadian-produced oil from Hardisty, Alberta, Canada, to Caster, Wyoming. The Express Pipeline is scheduled to be constructed in 1996--97 and will supply crude oil to refineries in Wyoming and the midwest.

  7. High power thyristors with 5 kV blocking voltage. Volume 1: Development of high-voltage-thyristors (4.5 kV) with good dynamic properties

    NASA Technical Reports Server (NTRS)

    Lock, K.; Patalong, H.; Platzoeder, K.

    1979-01-01

    Using neutron irradiated silicon with considerably lower spread in resistivity as compared to conventionally doped silicon it was possible to produce power thyristors with breakdown voltages between 3.5 kV and 5.5 kV. The thyristor pellets have a diameter of 50 mm. Maximum average on-state currents of 600 to 800 A can be reached with these elements. The dynamic properties of the thryistors could be improved to allow standard applications up to maximum repetitive voltages of 4.5 kV.

  8. The Tetramerization Domain Potentiates Kv4 Channel Function by Suppressing Closed-State Inactivation

    PubMed Central

    Tang, Yi-Quan; Zhou, Jing-Heng; Yang, Fan; Zheng, Jie; Wang, KeWei

    2014-01-01

    A-type Kv4 potassium channels undergo a conformational change toward a nonconductive state at negative membrane potentials, a dynamic process known as pre-open closed states or closed-state inactivation (CSI). CSI causes inhibition of channel activity without the prerequisite of channel opening, thus providing a dynamic regulation of neuronal excitability, dendritic signal integration, and synaptic plasticity at resting. However, the structural determinants underlying Kv4 CSI remain largely unknown. We recently showed that the auxiliary KChIP4a subunit contains an N-terminal Kv4 inhibitory domain (KID) that directly interacts with Kv4.3 channels to enhance CSI. In this study, we utilized the KChIP4a KID to probe key structural elements underlying Kv4 CSI. Using fluorescence resonance energy transfer two-hybrid mapping and bimolecular fluorescence complementation-based screening combined with electrophysiology, we identified the intracellular tetramerization (T1) domain that functions to suppress CSI and serves as a receptor for the binding of KID. Disrupting the Kv4.3 T1-T1 interaction interface by mutating C110A within the C3H1 motif of T1 domain facilitated CSI and ablated the KID-mediated enhancement of CSI. Furthermore, replacing the Kv4.3 T1 domain with the T1 domain from Kv1.4 (without the C3H1 motif) or Kv2.1 (with the C3H1 motif) resulted in channels functioning with enhanced or suppressed CSI, respectively. Taken together, our findings reveal a novel (to our knowledge) role of the T1 domain in suppressing Kv4 CSI, and that KChIP4a KID directly interacts with the T1 domain to facilitate Kv4.3 CSI, thus leading to inhibition of channel function. PMID:25185545

  9. Vasorelaxant effects of novel Kv7.4 channel enhancers ML213 and NS15370

    PubMed Central

    Jepps, T A; Bentzen, B H; Stott, J B; Povstyan, O V; Sivaloganathan, K; Dalby-Brown, W; Greenwood, I A

    2014-01-01

    Background and Purpose The KCNQ-encoded voltage-gated potassium channel family (Kv7.1-Kv7.5) are established regulators of smooth muscle contractility, where Kv7.4 and Kv7.5 predominate. Various Kv7.2–7.5 channel enhancers have been developed that have been shown to cause a vasorelaxation in both rodent and human blood vessels. Recently, two novel Kv7 channel enhancers have been identified, ML213 and NS15370, that show increased potency, particularly on Kv7.4 channels. The aim of this study was to characterize the effects of these novel enhancers in different rat blood vessels and compare them with Kv7 enhancers (S-1, BMS204352, retigabine) described previously. We also sought to determine the binding sites of the new Kv7 enhancers. Key Results Both ML213 and NS15370 relaxed segments of rat thoracic aorta, renal artery and mesenteric artery in a concentration-dependent manner. In the mesenteric artery ML213 and NS15370 displayed EC50s that were far lower than other Kv7 enhancers tested. Current-clamp experiments revealed that both novel enhancers, at low concentrations, caused significant hyperpolarization in mesenteric artery smooth muscle cells. In addition, we determined that the stimulatory effect of these enhancers relied on a tryptophan residue located in the S5 domain, which is the same binding site for the other Kv7 enhancers tested in this study. Conclusions and Implications This study has identified and characterized ML213 and NS15370 as potent vasorelaxants in different blood vessels, thereby highlighting these new compounds as potential therapeutics for various smooth muscle disorders. PMID:24909207

  10. Stabilization of Kv1.5 channel protein by the inotropic agent olprinone.

    PubMed

    Endo, Ryo; Kurata, Yasutaka; Notsu, Tomomi; Li, Peili; Morikawa, Kumi; Kondo, Takehito; Ogura, Kazuyoshi; Miake, Junichiro; Yoshida, Akio; Shirayoshi, Yasuaki; Ninomiya, Haruaki; Higaki, Katsumi; Kuwabara, Masanari; Yamamoto, Kazuhiro; Inagaki, Yoshimi; Hisatome, Ichiro

    2015-10-15

    Olprinone is an inotropic agent that inhibits phosphodiesterase (PDE) III and causes vasodilation. Olprinone has been shown to be less proarrhythmic and possibly affect expression of functional Kv1.5 channels that confer the ultra-rapid delayed-rectifier K+ channel current (IKur) responsible for action potential repolarization. To reveal involvement of Kv1.5 channels in the less arrhythmic effect of olprinone, we examined effects of the agent on the stability of Kv1.5 channel proteins expressed in COS7 cells. Olprinone at 30-1000 nM increased the protein level of Kv1.5 channels in a concentration-dependent manner. Chase experiments showed that olprinone delayed degradation of Kv1.5 channels. Olprinone increased the immunofluorescent signal of Kv1.5 channels in the endoplasmic reticulum (ER) and Golgi apparatus as well as on the cell surface. Kv1.5-mediated membrane currents, measured as 4-aminopyridine-sensitive currents, were increased by olprinone without changes in their activation kinetics. A protein transporter inhibitor, colchicine, abolished the olprinone-induced increase of Kv.1.5-mediated currents. The action of olprinone was inhibited by 4-aminopyridine, and was not mimicked by the application of 8-Bromo-cAMP. Taken together, we conclude that olprinone stabilizes Kv1.5 proteins at the ER through an action as a chemical chaperone, and thereby increases the density of Kv1.5 channels on the cell membrane. The enhancement of Kv1.5 currents could underlie less arrhythmogenicity of olprinone. PMID:26368666

  11. Behavioural and functional characterization of Kv10.1 (Eag1) knockout mice.

    PubMed

    Ufartes, Roser; Schneider, Tomasz; Mortensen, Lena Sünke; de Juan Romero, Camino; Hentrich, Klaus; Knoetgen, Hendrik; Beilinson, Vadim; Moebius, Wiebke; Tarabykin, Victor; Alves, Frauke; Pardo, Luis A; Rawlins, J Nicholas P; Stuehmer, Walter

    2013-06-01

    Kv10.1 (Eag1), member of the Kv10 family of voltage-gated potassium channels, is preferentially expressed in adult brain. The aim of the present study was to unravel the functional role of Kv10.1 in the brain by generating knockout mice, where the voltage sensor and pore region of Kv10.1 were removed to render non-functional proteins through deletion of exon 7 of the KCNH1 gene using the '3 Lox P strategy'. Kv10.1-deficient mice show no obvious alterations during embryogenesis and develop normally to adulthood; cortex, hippocampus and cerebellum appear anatomically normal. Other tests, including general health screen, sensorimotor functioning and gating, anxiety, social behaviour, learning and memory did not show any functional aberrations in Kv10.1 null mice. Kv10.1 null mice display mild hyperactivity and longer-lasting haloperidol-induced catalepsy, but there was no difference between genotypes in amphetamine sensitization and withdrawal, reactivity to apomorphine and haloperidol in the prepulse inhibition tests or to antidepressants in the haloperidol-induced catalepsy. Furthermore, electrical properties of Kv10.1 in cerebellar Purkinje cells did not show any difference between genotypes. Bearing in mind that Kv10.1 is overexpressed in over 70% of all human tumours and that its inhibition leads to a reduced tumour cell proliferation, the fact that deletion of Kv10.1 does not show a marked phenotype is a prerequisite for utilizing Kv10.1 blocking and/or reduction techniques, such as siRNA, to treat cancer. PMID:23424202

  12. The short-circuit test results of 6.9 kV/2.3 kV 400 kVA-class YBCO model transformer

    NASA Astrophysics Data System (ADS)

    Tomioka, A.; Otonari, T.; Ogata, T.; Iwakuma, M.; Okamoto, H.; Hayashi, H.; Iijima, Y.; Saito, T.; Gosho, Y.; Tanabe, K.; Izumi, T.; Shiohara, Y.

    2011-11-01

    We are developing an elemental technology for 66 kV/6.9 kV 20 MVA-class power transformer with YBCO conductors. The protection of short-circuit technology is one of the elemental technologies for HTS transformer. Since short-circuit current is much higher than critical current of YBCO tape, there is a possibility that superconducting characteristics may be damaged during short-circuit period. We made a conductor to compose the YBCO tape with copper tape. We manufactured 6.9 kV/2.3 kV 400 kVA-class YBCO model transformer using this conductor and performed short-circuit current test. The short-circuit current of primary winding was 346 A which was about six times larger than the rated current. The I-V characteristics of the winding did not change before and after the test. We may consider this conductor withstands short-circuit current.

  13. Inter-Subunit Interactions across the Upper Voltage Sensing-Pore Domain Interface Contribute to the Concerted Pore Opening Transition of Kv Channels

    PubMed Central

    Shem-Ad, Tzilhav; Irit, Orr; Yifrach, Ofer

    2013-01-01

    The tight electro-mechanical coupling between the voltage-sensing and pore domains of Kv channels lies at the heart of their fundamental roles in electrical signaling. Structural data have identified two voltage sensor pore inter-domain interaction surfaces, thus providing a framework to explain the molecular basis for the tight coupling of these domains. While the contribution of the intra-subunit lower domain interface to the electro-mechanical coupling that underlies channel opening is relatively well understood, the contribution of the inter-subunit upper interface to channel gating is not yet clear. Relying on energy perturbation and thermodynamic coupling analyses of tandem-dimeric Shaker Kv channels, we show that mutation of upper interface residues from both sides of the voltage sensor-pore domain interface stabilizes the closed channel state. These mutations, however, do not affect slow inactivation gating. We, moreover, find that upper interface residues form a network of state-dependent interactions that stabilize the open channel state. Finally, we note that the observed residue interaction network does not change during slow inactivation gating. The upper voltage sensing-pore interaction surface thus only undergoes conformational rearrangements during channel activation gating. We suggest that inter-subunit interactions across the upper domain interface mediate allosteric communication between channel subunits that contributes to the concerted nature of the late pore opening transition of Kv channels. PMID:24340010

  14. Manipulating Kv4.2 identifies a specific component of hippocampal pyramidal neuron A-current that depends upon Kv4.2 expression

    PubMed Central

    Lauver, Aaron; Yuan, Li-Lian; Jeromin, Andreas; Nadin, Brian M.; Rodríguez, José J.; Davies, Heather A.; Stewart, Michael G.; Wu, Gang-Yi; Pfaffinger, Paul J.

    2012-01-01

    The somatodendritic A-current, ISA, in hippocampal CA1 pyramidal neurons regulates the processing of synaptic inputs and the amplitude of back propagating action potentials into the dendritic tree, as well as the action potential firing properties at the soma. In this study, we have used RNA interference and over-expression to show that expression of the Kv4.2 gene specifically regulates the ISA component of A-current in these neurons. In dissociated hippocampal pyramidal neuron cultures, or organotypic cultured CA1 pyramidal neurons, the expression level of Kv4.2 is such that the ISA channels are maintained in the population at a peak conductance of approximately 950 pS/pF. Suppression of Kv4.2 transcripts in hippocampal pyramidal neurons using an RNA interference vector suppresses ISA current by 60% in 2 days, similar to the effect of expressing dominant-negative Kv4 channel constructs. Increasing the expression of Kv4.2 in these neurons increases the level of ISA to 170% of the normal set point without altering the biophysical properties. Our results establish a specific role for native Kv4.2 transcripts in forming and maintaining ISA current at characteristic levels in hippocampal pyramidal neurons. PMID:17026528

  15. Application of intelligent optimal kV scanning technology (CARE kV) in dual-source computed tomography (DSCT) coronary angiography

    PubMed Central

    Zhang, Jun; Kang, Shaolei; Han, Dan; Xie, Xiaojie; Deng, Yaming

    2015-01-01

    This study aims to evaluate the applications and values of dual-source computed tomography (DSCT) intelligent optimal kV scanning technology (CARE kV) in coronary CT angiography (CCTA). 150 patients with normal body mass index were performed DSCT coronary angiography, then randomly divided into the “Semi”, 120,100 and 80 kV Group, and the 2 “on” groups, with 30 patients in each group. The first 5 groups used the reference voltage as 120 kV, and the reference current as 400 mAs, while the other group used the reference voltage as 100 kV, and the reference current as 400 mAs. The image quality, average CT value, image noise, signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR) and radiation dose were comparatively analyzed among the 5 groups. The image quality scores of the five groups showed no significant difference (P > 0.05); the average CT values and image noises had significance (P < 0.05), while SNR and CNR showed no significant difference (P > 0.05). The 80 kV group showed the biggest noise, with the CT value as 700 HU, while the radiation dose was the lowest, followed by the on group. As for the patients with normal body mass index (BMI), CARE kV-“on” could obtain high-quality images and lower radiation dose for CCTA, while the operation was simple and convenient. PMID:26770354

  16. Phosphatidic acid modulation of Kv channel voltage sensor function.

    PubMed

    Hite, Richard K; Butterwick, Joel A; MacKinnon, Roderick

    2014-01-01

    Membrane phospholipids can function as potent regulators of ion channel function. This study uncovers and investigates the effect of phosphatidic acid on Kv channel gating. Using the method of reconstitution into planar lipid bilayers, in which protein and lipid components are defined and controlled, we characterize two effects of phosphatidic acid. The first is a non-specific electrostatic influence on activation mediated by electric charge density on the extracellular and intracellular membrane surfaces. The second is specific to the presence of a primary phosphate group, acts only through the intracellular membrane leaflet and depends on the presence of a particular arginine residue in the voltage sensor. Intracellular phosphatidic acid accounts for a nearly 50 mV shift in the midpoint of the activation curve in a direction consistent with stabilization of the voltage sensor's closed conformation. These findings support a novel mechanism of voltage sensor regulation by the signaling lipid phosphatidic acid. PMID:25285449

  17. Developmental expression of Kv1 voltage-gated potassium channels in the avian hypothalamus.

    PubMed

    Doczi, Megan A; Vitzthum, Carl M; Forehand, Cynthia J

    2016-03-11

    Specialized hypothalamic neurons integrate the homeostatic balance between food intake and energy expenditure, processes that may become dysregulated during the development of diabetes, obesity, and other metabolic disorders. Shaker family voltage-gated potassium channels (Kv1) contribute to the maintenance of resting membrane potential, action potential characteristics, and neurotransmitter release in many populations of neurons, although hypothalamic Kv1 channel expression has been largely unexplored. Whole-cell patch clamp recordings from avian hypothalamic brain slices demonstrate a developmental shift in the electrophysiological properties of avian arcuate nucleus neurons, identifying an increase in outward ionic current that corresponds with action potential maturation. Additionally, RT-PCR experiments identified the early expression of Kv1.2, Kv1.3, and Kv1.5 mRNA in the embryonic avian hypothalamus, suggesting that these channels may underlie the electrophysiological changes observed in these neurons. Real-time quantitative PCR analysis on intact microdissections of embryonic hypothalamic tissue revealed a concomitant increase in Kv1.2 and Kv1.5 gene expression at key electrophysiological time points during development. This study is the first to demonstrate hypothalamic mRNA expression of Kv1 channels in developing avian embryos and may suggest a role for voltage-gated ion channel regulation in the physiological patterning of embryonic hypothalamic circuits governing energy homeostasis. PMID:26845562

  18. The Link between Ion Permeation and Inactivation Gating of Kv4 Potassium Channels

    PubMed Central

    Shahidullah, Mohammad; Covarrubias, Manuel

    2003-01-01

    Kv4 potassium channels undergo rapid inactivation but do not seem to exhibit the classical N-type and C-type mechanisms present in other Kv channels. We have previously hypothesized that Kv4 channels preferentially inactivate from the preopen closed state, which involves regions of the channel that contribute to the internal vestibule of the pore. To further test this hypothesis, we have examined the effects of permeant ions on gating of three Kv4 channels (Kv4.1, Kv4.2, and Kv4.3) expressed in Xenopus oocytes. Rb+ is an excellent tool for this purpose because its prolonged residency time in the pore delays K+ channel closing. The data showed that, only when Rb+ carried the current, both channel closing and the development of macroscopic inactivation are slowed (1.5- to 4-fold, relative to the K+ current). Furthermore, macroscopic Rb+ currents were larger than K+ currents (1.2- to 3-fold) as the result of a more stable open state, which increases the maximum open probability. These results demonstrate that pore occupancy can influence inactivation gating in a manner that depends on how channel closing impacts inactivation from the preopen closed state. By examining possible changes in ionic selectivity and the influence of elevating the external K+ concentration, additional experiments did not support the presence of C-type inactivation in Kv4 channels. PMID:12547775

  19. VizieR Online Data Catalog: KV Vel and TW Crv JHK light curves (Ribeiro+, 2011)

    NASA Astrophysics Data System (ADS)

    Ribeiro, T.; Baptista, R.

    2010-11-01

    Near-infrared time resolved photometric observations of KV Vel and TW Crv. KV Vel was observed quasi-simultaneously in JHK bands while TW Crv was observed on the H and K bands. Table contains the flux calibrated photometric measurements for each object on each band and consists of phase, flux and flux error. (1 data file).

  20. Context view of RyantoRainbow 100kv Transmission Line showing copper wire ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    Context view of Ryan-to-Rainbow 100kv Transmission Line showing copper wire conductors leaving roof of Ryan Powerhouse. View to east - Ryan Hydroelectric Facility, Ryan-to-Rainbow 100 kV Transmission Line, West bank of Missouri River, northeast of Great Falls, Great Falls, Cascade County, MT

  1. Kv4 Potassium Channels Modulate Hippocampal EPSP-Spike Potentiation and Spatial Memory in Rats

    ERIC Educational Resources Information Center

    Truchet, Bruno; Manrique, Christine; Sreng, Leam; Chaillan, Franck A.; Roman, Francois S.; Mourre, Christiane

    2012-01-01

    Kv4 channels regulate the backpropagation of action potentials (b-AP) and have been implicated in the modulation of long-term potentiation (LTP). Here we showed that blockade of Kv4 channels by the scorpion toxin AmmTX3 impaired reference memory in a radial maze task. In vivo, AmmTX3 intracerebroventricular (i.c.v.) infusion increased and…

  2. 75 FR 56051 - Bemidji to Grand Rapids Minnesota 230 kV Transmission Line Project

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-15

    ...; ] DEPARTMENT OF AGRICULTURE Rural Utilities Service Bemidji to Grand Rapids Minnesota 230 kV Transmission Line... Rapids, Minnesota 230 kV Transmission Line Project (``Project'') in Minnesota. The Final EIS was prepared... application of Minnkota Power Cooperative, Inc. for RUS financing to construct a 230 kilovolt...

  3. Solid-State Fault Current Limiter Development : Design and Testing Update of a 15kV SSCL Power Stack

    SciTech Connect

    Dr. Ram Adapa; Mr. Dante Piccone

    2012-04-30

    ABSTRACT The Solid-State Fault Current Limiter (SSCL) is a promising technology that can be applied to utility power delivery systems to address the problem of increasing fault currents associated with load growth. As demand continues to grow, more power is added to utility system either by increasing generator capacity or by adding distributed generators, resulting in higher available fault currents, often beyond the capabilities of the present infrastructure. The SSCL is power-electronics based equipment designed to work with the present utility system to address this problem. The SSCL monitors the line current and dynamically inserts additional impedance into the line in the event of a fault being detected. The SSCL is based on a modular design and can be configured for 5kV through 69kV systems at nominal current ratings of 1000A to 4000A. Results and Findings This report provides the final test results on the development of 15kV class SSCL single phase power stack. The scope of work included the design of the modular standard building block sub-assemblies, the design and manufacture of the power stack and the testing of the power stack for the key functional tests of continuous current capability and fault current limiting action. Challenges and Objectives Solid-State Current Limiter technology impacts a wide spectrum of utility engineering and operating personnel. It addresses the problems associated with load growth both at Transmission and Distribution class networks. The design concept is pioneering in terms of developing the most efficient and compact power electronics equipment for utility use. The initial test results of the standard building blocks are promising. The independent laboratory tests of the power stack are promising. However the complete 3 phase system needs rigorous testing for performance and reliability. Applications, Values, and Use The SSCL is an intelligent power-electronics device which is modular in design and can provide current

  4. From Pan-Reactive KV7 Channel Opener to Subtype Selective Opener/Inhibitor by Addition of a Methyl Group

    PubMed Central

    Blom, Sigrid Marie; Rottländer, Mario; Kehler, Jan; Bundgaard, Christoffer; Schmitt, Nicole; Jensen, Henrik Sindal

    2014-01-01

    The voltage-gated potassium channels of the KV7 family (KV7.1–5) play important roles in controlling neuronal excitability and are therefore attractive targets for treatment of CNS disorders linked to hyperexcitability. One of the main challenges in developing KV7 channel active drugs has been to identify compounds capable of discriminating between the neuronally expressed subtypes (KV7.2–5), aiding the identification of the subunit composition of KV7 currents in various tissues, and possessing better therapeutic potential for particular indications. By taking advantage of the structure-activity relationship of acrylamide KV7 channel openers and the effects of these compounds on mutant KV7 channels, we have designed and synthesized a novel KV7 channel modulator with a unique profile. The compound, named SMB-1, is an inhibitor of KV7.2 and an activator of KV7.4. SMB-1 inhibits KV7.2 by reducing the current amplitude and increasing the time constant for the slow component of the activation kinetics. The activation of KV7.4 is seen as an increase in the current amplitude and a slowing of the deactivation kinetics. Experiments studying mutant channels with a compromised binding site for the KV7.2–5 opener retigabine indicate that SMB-1 binds within the same pocket as retigabine for both inhibition of KV7.2 and activation of KV7.4. SMB-1 may serve as a valuable tool for KV7 channel research and may be used as a template for further design of better subtype selective KV7 channel modulators. A compound with this profile could hold novel therapeutic potential such as the treatment of both positive and cognitive symptoms in schizophrenia. PMID:24956197

  5. Kv7.2 regulates the function of peripheral sensory neurons

    PubMed Central

    King, Chih H.; Lancaster, Eric; Salomon, Daniela; Peles, Elior; Scherer, Steven S.

    2014-01-01

    The Kv7 (KCNQ) family of voltage-gated K+ channels regulates cellular excitability. The functional role of Kv7.2 has been hampered by the lack of a viable Kcnq2-null animal model. In this study, we generated homozygous Kcnq2-null sensory neurons using the Cre-Lox system; in these mice, Kv7.2 expression is absent in the peripheral sensory neurons, whereas the expression of other molecular components of nodes (including Kv7.3), paranodes, and juxtaparanodes is not altered. The conditional Kcnq2-null animals exhibit normal motor performance, but have increased thermal hyperalgesia and mechanical allodynia. Whole cell patch recording technique demonstrates that Kcnq2-null sensory neurons have increased excitability and reduced spike frequency adaptation. Taken together, our results suggest that the loss of Kv7.2 activity increases the excitability of primary sensory neurons. PMID:24687876

  6. Effects of dalfampridine and its metabolites on cloned human potassium channels Kv 1.1, Kv 1.2, and Kv 1.4 expressed in human embryonic kidney cells

    PubMed Central

    Caggiano, Anthony; Blight, Andrew; Parry, Tom J.

    2013-01-01

    Background Dalfampridine (4-aminopyridine; 4-AP) is a potassium channel blocker that has been available in the United States as a treatment to improve walking in patients with multiple sclerosis. 4-AP is well-characterized in vitro with regard to inhibition of neuronal potassium channels, but the potential contribution of its metabolites to clinical activity has not been determined. This study evaluated the concentration–response of 4-AP and its two primary metabolites, 3-hydroxy-4-aminopyridine and 3-hydroxy-4-aminopyridine sulfate, for inhibition of the potassium channels Kv 1.1, Kv 1.2, and Kv 1.4, which are considered candidates for mediating effects of 4-AP on action potential conduction because of their presence in axonal membranes. Methods Stable transfection of cDNA for Kv 1.1, Kv 1.2, and Kv 1.4 was performed into HEK293 cells, and colonies of cells containing each channel were selected and maintained under appropriate cell culture conditions. Electrophysiological measurements were performed using a patch-clamp technique in at least three cells for each concentration (50, 500, 5000, and 50,000 μM) of 4-AP and the two metabolites, with each cell acting as its own control. Concentration–response curves were constructed for 4-AP and each metabolite. Data were analyzed using nonlinear least-squares fit, and concentrations inhibiting the channels by 50% (IC50) were estimated. Results 4-AP induced similar concentration-dependent inhibition profiles of all three potassium channels, resulting in a narrow range of IC50 values across channels (242 µM to 399 µM). Across the three channels, the IC50 values of 3-hydroxy-4-aminopyridine and 3-hydroxy-4-aminopyridine sulfate were 1–2 orders of magnitude higher (less potent) than those of 4-AP. Conclusions 3-Hydroxy-4-aminopyridine and 3-hydroxy-4-aminopyridine sulfate demonstrated low in vitro potency for Kv 1.1, Kv 1.2, and Kv 1.4 inhibition, suggesting that these metabolites are unlikely to contribute to

  7. Modeling the binding modes of Kv1.5 potassium channel and blockers.

    PubMed

    Yang, Qian; Du, Lupei; Wang, Xiaojian; Li, Minyong; You, Qidong

    2008-09-01

    The ultra-rapid delayed rectifier potassium current (I(Kur)), encoded by Kv1.5 gene, is the critical determinant of Phase I repolarization of action potential duration (APD). The evidences that Kv1.5 gene expresses more extensively in human atrial myocytes than in ventricle and the I(Kur) currents has not been recorded in the human ventricle, suggest Kv1.5 potassium channel as a selective target for the treatment of atrial fibrillation (AF). Recent mutagenesis studies have provided us some evidences that are useful in designing Kv1.5 blockers. In order to further evaluate these molecular biological information, the homology model of Kv1.5 potassium channel was established based on the Kv1.2 crystal structure (PDB entry: 2A79) using MODELLER 9v2 program. After the molecular dynamics refinement, the optimized homology model was assessed as a reliable structure by PROCHECK, ERRAT, WHAT-IF, PROSA2003 and DOPE graph. The results of molecular docking studies on different Kv1.5 inhibitors are in agreement with the published mutagenesis data. Based on the docking conformations, a pharmacophore model was developed by HipHop algorithm in order to probe the common features of blockers. By analyzing the results, active site architecture, certain key residues and pharmacophore common-features that are responsible for substrate specificity were identified on the Kv1.5 potassium channel, which would be very helpful in understanding the blockade mechanism of Kv1.5 potassium channel and providing insights into rational design of novel Kv1.5 blockers. PMID:18485768

  8. Dysregulation of Kv3.4 channels in dorsal root ganglia following spinal cord injury.

    PubMed

    Ritter, David M; Zemel, Benjamin M; Hala, Tamara J; O'Leary, Michael E; Lepore, Angelo C; Covarrubias, Manuel

    2015-01-21

    Spinal cord injury (SCI) patients develop chronic pain involving poorly understood central and peripheral mechanisms. Because dysregulation of the voltage-gated Kv3.4 channel has been implicated in the hyperexcitable state of dorsal root ganglion (DRG) neurons following direct injury of sensory nerves, we asked whether such a dysregulation also plays a role in SCI. Kv3.4 channels are expressed in DRG neurons, where they help regulate action potential (AP) repolarization in a manner that depends on the modulation of inactivation by protein kinase C (PKC)-dependent phosphorylation of the channel's inactivation domain. Here, we report that, 2 weeks after cervical hemicontusion SCI, injured rats exhibit contralateral hypersensitivity to stimuli accompanied by accentuated repetitive spiking in putative DRG nociceptors. Also in these neurons at 1 week after laminectomy and SCI, Kv3.4 channel inactivation is impaired compared with naive nonsurgical controls. At 2-6 weeks after laminectomy, however, Kv3.4 channel inactivation returns to naive levels. Conversely, Kv3.4 currents at 2-6 weeks post-SCI are downregulated and remain slow-inactivating. Immunohistochemistry indicated that downregulation mainly resulted from decreased surface expression of the Kv3.4 channel, as whole-DRG-protein and single-cell mRNA transcript levels did not change. Furthermore, consistent with Kv3.4 channel dysregulation, PKC activation failed to shorten the AP duration of small-diameter DRG neurons. Finally, re-expressing synthetic Kv3.4 currents under dynamic clamp conditions dampened repetitive spiking in the neurons from SCI rats. These results suggest a novel peripheral mechanism of post-SCI pain sensitization implicating Kv3.4 channel dysregulation and potential Kv3.4-based therapeutic interventions. PMID:25609640

  9. Requisite Role of Kv1.5 Channels in Coronary Metabolic Dilation

    PubMed Central

    Bardakjian, Raffi; Kolz, Christopher; Enrick, Molly; Hakobyan, Tatevik; Kmetz, John; Bratz, Ian; Luli, Jordan; Nagane, Masaki; Khan, Nadeem; Hou, Huagang; Kuppusamy, Periannan; Graham, Jacqueline; Fu, Frances Kwan; Janota, Danielle; Oyewumi, Moses O.; Logan, Suzanna; Lindner, Jonathan R.; Chilian, William M.

    2016-01-01

    Rationale In the working heart coronary blood flow is linked to the production of metabolites, which modulate tone of smooth muscle in a redox-dependent manner. Voltage-gated potassium channels, which play a role in controlling membrane potential in vascular smooth muscle, have certain members that are redox sensitive. Objective To determine the role of redox-sensitive Kv1.5 channels in coronary metabolic flow regulation. Methods and Results In mice (wild type [WT], Kv1.5 null [Kv1.5−/−], and Kv1.5−/− and WT with inducible, smooth muscle specific expression of Kv1.5 channels) we measured mean arterial pressure (MAP), myocardial blood flow (MBF), myocardial tissue pO2, and ejection fraction (EF) before and after inducing cardiac stress with norepinephrine (NE). Cardiac work (CW) was estimated as the product of MAP and heart rate. Isolated arteries were studied to establish if genetic alterations modified vascular reactivity. Despite higher levels of CW in the Kv1.5−/− (versus WT at baseline and all doses of NE), MBF was lower in Kv1.5−/− than in WT. At high levels of CW, tissue pO2 dropped significantly along with EF. Expression of Kv1.5 channels in smooth muscle in the null background rescued this phenotype of impaired metabolic dilation. In isolated vessels from Kv1.5−/− mice, relaxation to H2O2 was impaired, but responses to adenosine and acetylcholine were normal compared to WT. Conclusions Kv1.5 channels in vascular smooth muscle play a critical role in coupling myocardial blood flow to cardiac metabolism. Absence of these channels disassociates metabolism from flow resulting in cardiac pump dysfunction and tissue hypoxia. PMID:26224794

  10. Nuclear Localization and Functional Characteristics of Voltage-gated Potassium Channel Kv1.3*

    PubMed Central

    Jang, Soo Hwa; Byun, Jun Kyu; Jeon, Won-Il; Choi, Seon Young; Park, Jin; Lee, Bo Hyung; Yang, Ji Eun; Park, Jin Bong; O'Grady, Scott M.; Kim, Dae-Yong; Ryu, Pan Dong; Joo, Sang-Woo; Lee, So Yeong

    2015-01-01

    It is widely known that ion channels are expressed in the plasma membrane. However, a few studies have suggested that several ion channels including voltage-gated K+ (Kv) channels also exist in intracellular organelles where they are involved in the biochemical events associated with cell signaling. In the present study, Western blot analysis using fractionated protein clearly indicates that Kv1.3 channels are expressed in the nuclei of MCF7, A549, and SNU-484 cancer cells and human brain tissues. In addition, Kv1.3 is located in the plasma membrane and the nucleus of Jurkat T cells. Nuclear membrane hyperpolarization after treatment with margatoxin (MgTX), a specific blocker of Kv1.3 channels, provides evidence for functional channels at the nuclear membrane of A549 cells. MgTX-induced hyperpolarization is abolished in the nuclei of Kv1.3 silenced cells, and the effects of MgTX are dependent on the magnitude of the K+ gradient across the nuclear membrane. Selective Kv1.3 blockers induce the phosphorylation of cAMP response element-binding protein (CREB) and c-Fos activation. Moreover, Kv1.3 is shown to form a complex with the upstream binding factor 1 in the nucleus. Chromatin immunoprecipitation assay reveals that Sp1 transcription factor is directly bound to the promoter region of the Kv1.3 gene, and the Sp1 regulates Kv1.3 expression in the nucleus of A549 cells. These results demonstrate that Kv1.3 channels are primarily localized in the nucleus of several types of cancer cells and human brain tissues where they are capable of regulating nuclear membrane potential and activation of transcription factors, such as phosphorylated CREB and c-Fos. PMID:25829491

  11. Nuclear localization and functional characteristics of voltage-gated potassium channel Kv1.3.

    PubMed

    Jang, Soo Hwa; Byun, Jun Kyu; Jeon, Won-Il; Choi, Seon Young; Park, Jin; Lee, Bo Hyung; Yang, Ji Eun; Park, Jin Bong; O'Grady, Scott M; Kim, Dae-Yong; Ryu, Pan Dong; Joo, Sang-Woo; Lee, So Yeong

    2015-05-15

    It is widely known that ion channels are expressed in the plasma membrane. However, a few studies have suggested that several ion channels including voltage-gated K(+) (Kv) channels also exist in intracellular organelles where they are involved in the biochemical events associated with cell signaling. In the present study, Western blot analysis using fractionated protein clearly indicates that Kv1.3 channels are expressed in the nuclei of MCF7, A549, and SNU-484 cancer cells and human brain tissues. In addition, Kv1.3 is located in the plasma membrane and the nucleus of Jurkat T cells. Nuclear membrane hyperpolarization after treatment with margatoxin (MgTX), a specific blocker of Kv1.3 channels, provides evidence for functional channels at the nuclear membrane of A549 cells. MgTX-induced hyperpolarization is abolished in the nuclei of Kv1.3 silenced cells, and the effects of MgTX are dependent on the magnitude of the K(+) gradient across the nuclear membrane. Selective Kv1.3 blockers induce the phosphorylation of cAMP response element-binding protein (CREB) and c-Fos activation. Moreover, Kv1.3 is shown to form a complex with the upstream binding factor 1 in the nucleus. Chromatin immunoprecipitation assay reveals that Sp1 transcription factor is directly bound to the promoter region of the Kv1.3 gene, and the Sp1 regulates Kv1.3 expression in the nucleus of A549 cells. These results demonstrate that Kv1.3 channels are primarily localized in the nucleus of several types of cancer cells and human brain tissues where they are capable of regulating nuclear membrane potential and activation of transcription factors, such as phosphorylated CREB and c-Fos. PMID:25829491

  12. Retigabine holds KV7 channels open and stabilizes the resting potential

    PubMed Central

    Corbin-Leftwich, Aaron; Mossadeq, Sayeed M.; Ha, Junghoon; Ruchala, Iwona; Le, Audrey Han Ngoc

    2016-01-01

    The anticonvulsant Retigabine is a KV7 channel agonist used to treat hyperexcitability disorders in humans. Retigabine shifts the voltage dependence for activation of the heteromeric KV7.2/KV7.3 channel to more negative potentials, thus facilitating activation. Although the molecular mechanism underlying Retigabine’s action remains unknown, previous studies have identified the pore region of KV7 channels as the drug’s target. This suggested that the Retigabine-induced shift in voltage dependence likely derives from the stabilization of the pore domain in an open (conducting) conformation. Testing this idea, we show that the heteromeric KV7.2/KV7.3 channel has at least two open states, which we named O1 and O2, with O2 being more stable. The O1 state was reached after short membrane depolarizations, whereas O2 was reached after prolonged depolarization or during steady state at the typical neuronal resting potentials. We also found that activation and deactivation seem to follow distinct pathways, suggesting that the KV7.2/KV7.3 channel activity displays hysteresis. As for the action of Retigabine, we discovered that this agonist discriminates between open states, preferentially acting on the O2 state and further stabilizing it. Based on these findings, we proposed a novel mechanism for the therapeutic effect of Retigabine whereby this drug reduces excitability by enhancing the resting potential open state stability of KV7.2/KV7.3 channels. To address this hypothesis, we used a model for action potential (AP) in Xenopus laevis oocytes and found that the resting membrane potential became more negative as a function of Retigabine concentration, whereas the threshold potential for AP firing remained unaltered. PMID:26880756

  13. Dysregulation of Kv3.4 Channels in Dorsal Root Ganglia Following Spinal Cord Injury

    PubMed Central

    Ritter, David M.; Zemel, Benjamin M.; Hala, Tamara J.; O'Leary, Michael E.; Lepore, Angelo C.

    2015-01-01

    Spinal cord injury (SCI) patients develop chronic pain involving poorly understood central and peripheral mechanisms. Because dysregulation of the voltage-gated Kv3.4 channel has been implicated in the hyperexcitable state of dorsal root ganglion (DRG) neurons following direct injury of sensory nerves, we asked whether such a dysregulation also plays a role in SCI. Kv3.4 channels are expressed in DRG neurons, where they help regulate action potential (AP) repolarization in a manner that depends on the modulation of inactivation by protein kinase C (PKC)-dependent phosphorylation of the channel's inactivation domain. Here, we report that, 2 weeks after cervical hemicontusion SCI, injured rats exhibit contralateral hypersensitivity to stimuli accompanied by accentuated repetitive spiking in putative DRG nociceptors. Also in these neurons at 1 week after laminectomy and SCI, Kv3.4 channel inactivation is impaired compared with naive nonsurgical controls. At 2–6 weeks after laminectomy, however, Kv3.4 channel inactivation returns to naive levels. Conversely, Kv3.4 currents at 2–6 weeks post-SCI are downregulated and remain slow-inactivating. Immunohistochemistry indicated that downregulation mainly resulted from decreased surface expression of the Kv3.4 channel, as whole-DRG-protein and single-cell mRNA transcript levels did not change. Furthermore, consistent with Kv3.4 channel dysregulation, PKC activation failed to shorten the AP duration of small-diameter DRG neurons. Finally, re-expressing synthetic Kv3.4 currents under dynamic clamp conditions dampened repetitive spiking in the neurons from SCI rats. These results suggest a novel peripheral mechanism of post-SCI pain sensitization implicating Kv3.4 channel dysregulation and potential Kv3.4-based therapeutic interventions. PMID:25609640

  14. Retigabine holds KV7 channels open and stabilizes the resting potential.

    PubMed

    Corbin-Leftwich, Aaron; Mossadeq, Sayeed M; Ha, Junghoon; Ruchala, Iwona; Le, Audrey Han Ngoc; Villalba-Galea, Carlos A

    2016-03-01

    The anticonvulsant Retigabine is a KV7 channel agonist used to treat hyperexcitability disorders in humans. Retigabine shifts the voltage dependence for activation of the heteromeric KV7.2/KV7.3 channel to more negative potentials, thus facilitating activation. Although the molecular mechanism underlying Retigabine's action remains unknown, previous studies have identified the pore region of KV7 channels as the drug's target. This suggested that the Retigabine-induced shift in voltage dependence likely derives from the stabilization of the pore domain in an open (conducting) conformation. Testing this idea, we show that the heteromeric KV7.2/KV7.3 channel has at least two open states, which we named O1 and O2, with O2 being more stable. The O1 state was reached after short membrane depolarizations, whereas O2 was reached after prolonged depolarization or during steady state at the typical neuronal resting potentials. We also found that activation and deactivation seem to follow distinct pathways, suggesting that the KV7.2/KV7.3 channel activity displays hysteresis. As for the action of Retigabine, we discovered that this agonist discriminates between open states, preferentially acting on the O2 state and further stabilizing it. Based on these findings, we proposed a novel mechanism for the therapeutic effect of Retigabine whereby this drug reduces excitability by enhancing the resting potential open state stability of KV7.2/KV7.3 channels. To address this hypothesis, we used a model for action potential (AP) in Xenopus laevis oocytes and found that the resting membrane potential became more negative as a function of Retigabine concentration, whereas the threshold potential for AP firing remained unaltered. PMID:26880756

  15. 14C SIRI samples at CNA: Measurements at 200 kV and 1000 kV

    NASA Astrophysics Data System (ADS)

    Santos Arévalo, Francisco-Javier; Gómez Martínez, Isabel; Agulló García, Lidia

    2015-10-01

    The Sixth International Radiocarbon Intercomparison (SIRI) exercise has taken place during late 2013 and 2014. 13 samples were distributed for AMS (Accelerator Mass Spectrometry) and 5 for radiometric laboratories, including one sample exclusively for radiometric laboratories. Being the first opportunity for our laboratory to participate actively in an intercomparison exercise, we have prepared and measured the samples in the two existing AMS dedicated facilities at the Centro Nacional de Aceleradores (CNA): SARA (Spanish Accelerator for Radionuclide Analysis), a 1 MV multielemental AMS system from HVEE, and Micadas, a 200 kV radiocarbon dating system designed by ETH. Results are presented for the two systems, together with a description of both the sample preparation and measurement procedures.

  16. Operational results of the spallation neutron source (SNS) polyphase converter-modulator for the 140 KV klystron RF system

    SciTech Connect

    Reass, W. A.; Doss, James D.; Gribble, R. F.; Lynch, M. T.; Rees, D. E.; Tallerico, P. J.; Borovina, D. L.

    2001-01-01

    This paper describes the first operational results of the 140 kV, 1 MW average, 11 MW peak, zero-voltageswitching, 20 kHz polyphase bridge, boost converter-modulator for klystron pulse application. The DC-DC converter derives the buss voltages from a standard 13.8 kV to 2100 Y substation cast-core transformer. Energy storage and filtering is provided by self-clearing metallized hazy polypropylene traction capacitors. Three 'H-Bridge' Insulated Gate Bipolar Transistor (IGBT) switching networks are used to generate the polyphase 20 kHz transformer primary drive waveforms. The 20 kHz drive waveforms are chirped the appropriate duration to generate the desired klystron pulse width. Pulse-Width Modulation (PWM) of the individual 20 kHz pulses is utilized to provide regulated output waveforms with adaptive feedforward and feedback techniques. The boost transformer design utilizes amorphous nanocrystalline material that provides the required low core loss at design flux levels and switching frequencies. Resonant shunt peaking is used on the transformer secondary to boost output voltage and resonate transformer leakage inductance. With the appropriate transformer leakage inductance and peaking capacitance, zero-voltage-switching of the IGBT's is attained, minimizing switching losses. Reviews of these design parameters and an examination of the first operational results will be performed.

  17. OPERATIONAL RESULTS OF THE SPALLATION NEUTRON SOURCE (SNS) POLYPHASE CONVERTER-MODULATOR FOR THE 140 KV KLYSTRON RF SYSTEMS

    SciTech Connect

    W.A. REASS; J.D. DOSS; ET AL

    2001-06-01

    This paper describes the first operational results of the 140 kV, 1 MW average, 11 MW peak, zero-voltage-switching, 20 kHz polyphase bridge, boost converter-modulator for klystron pulse application. The DC-DC converter derives the buss voltages from a standard 13.8 kV to 2100 Y substation cast-core transformer. Energy storage and filtering is provided by self-clearing metallized hazy polypropylene traction capacitors. Three ''H-Bridge'' Insulated Gate Bipolar Transistor (IGBT) switching networks are used to generate the polyphase 20 kHz transformer primary drive waveforms. The 20 kHz drive waveforms are chirped the appropriate duration to generate the desired klystron pulse width. Pulse-Width Modulation (PWM) of the individual 20 kHz pulses is utilized to provide regulated output waveforms with adaptive feedforward and feedback techniques. The boost transformer design utilizes amorphous nanocrystalline material that provides the required low core loss at design flux levels and switching frequencies. Resonant shunt peaking is used on the transformer secondary to boost output voltage and resonate transformer leakage inductance. With the appropriate transformer leakage inductance and peaking capacitance, zero-voltage-switching of the IGBT's is attained, minimizing switching losses. Reviews of these design parameters and an examination of the first operational results will be performed.

  18. Tungsten anode spectral model using interpolating cubic splines: Unfiltered x-ray spectra from 20 kV to 640 kV

    PubMed Central

    Hernandez, Andrew M.; Boone, John M.

    2014-01-01

    Purpose: Monte Carlo methods were used to generate lightly filtered high resolution x-ray spectra spanning from 20 kV to 640 kV. Methods: X-ray spectra were simulated for a conventional tungsten anode. The Monte Carlo N-Particle eXtended radiation transport code (MCNPX 2.6.0) was used to produce 35 spectra over the tube potential range from 20 kV to 640 kV, and cubic spline interpolation procedures were used to create piecewise polynomials characterizing the photon fluence per energy bin as a function of x-ray tube potential. Using these basis spectra and the cubic spline interpolation, 621 spectra were generated at 1 kV intervals from 20 to 640 kV. The tungsten anode spectral model using interpolating cubic splines (TASMICS) produces minimally filtered (0.8 mm Be) x-ray spectra with 1 keV energy resolution. The TASMICS spectra were compared mathematically with other, previously reported spectra. Results: Using paired t-test analyses, no statistically significant difference (i.e., p > 0.05) was observed between compared spectra over energy bins above 1% of peak bremsstrahlung fluence. For all energy bins, the correlation of determination (R2) demonstrated good correlation for all spectral comparisons. The mean overall difference (MOD) and mean absolute difference (MAD) were computed over energy bins (above 1% of peak bremsstrahlung fluence) and over all the kV permutations compared. MOD and MAD comparisons with previously reported spectra were 2.7% and 9.7%, respectively (TASMIP), 0.1% and 12.0%, respectively [R. Birch and M. Marshall, “Computation of bremsstrahlung x-ray spectra and comparison with spectra measured with a Ge(Li) detector,” Phys. Med. Biol. 24, 505–517 (1979)], 0.4% and 8.1%, respectively (Poludniowski), and 0.4% and 8.1%, respectively (AAPM TG 195). The effective energy of TASMICS spectra with 2.5 mm of added Al filtration ranged from 17 keV (at 20 kV) to 138 keV (at 640 kV); with 0.2 mm of added Cu filtration the effective energy was 9 ke

  19. Tungsten anode spectral model using interpolating cubic splines: Unfiltered x-ray spectra from 20 kV to 640 kV

    SciTech Connect

    Hernandez, Andrew M.; Boone, John M.

    2014-04-15

    Purpose: Monte Carlo methods were used to generate lightly filtered high resolution x-ray spectra spanning from 20 kV to 640 kV. Methods: X-ray spectra were simulated for a conventional tungsten anode. The Monte Carlo N-Particle eXtended radiation transport code (MCNPX 2.6.0) was used to produce 35 spectra over the tube potential range from 20 kV to 640 kV, and cubic spline interpolation procedures were used to create piecewise polynomials characterizing the photon fluence per energy bin as a function of x-ray tube potential. Using these basis spectra and the cubic spline interpolation, 621 spectra were generated at 1 kV intervals from 20 to 640 kV. The tungsten anode spectral model using interpolating cubic splines (TASMICS) produces minimally filtered (0.8 mm Be) x-ray spectra with 1 keV energy resolution. The TASMICS spectra were compared mathematically with other, previously reported spectra. Results: Using pairedt-test analyses, no statistically significant difference (i.e., p > 0.05) was observed between compared spectra over energy bins above 1% of peak bremsstrahlung fluence. For all energy bins, the correlation of determination (R{sup 2}) demonstrated good correlation for all spectral comparisons. The mean overall difference (MOD) and mean absolute difference (MAD) were computed over energy bins (above 1% of peak bremsstrahlung fluence) and over all the kV permutations compared. MOD and MAD comparisons with previously reported spectra were 2.7% and 9.7%, respectively (TASMIP), 0.1% and 12.0%, respectively [R. Birch and M. Marshall, “Computation of bremsstrahlung x-ray spectra and comparison with spectra measured with a Ge(Li) detector,” Phys. Med. Biol. 24, 505–517 (1979)], 0.4% and 8.1%, respectively (Poludniowski), and 0.4% and 8.1%, respectively (AAPM TG 195). The effective energy of TASMICS spectra with 2.5 mm of added Al filtration ranged from 17 keV (at 20 kV) to 138 keV (at 640 kV); with 0.2 mm of added Cu filtration the effective energy was 9

  20. Modeling-independent elucidation of inactivation pathways in recombinant and native A-type Kv channels.

    PubMed

    Fineberg, Jeffrey D; Ritter, David M; Covarrubias, Manuel

    2012-11-01

    A-type voltage-gated K(+) (Kv) channels self-regulate their activity by inactivating directly from the open state (open-state inactivation [OSI]) or by inactivating before they open (closed-state inactivation [CSI]). To determine the inactivation pathways, it is often necessary to apply several pulse protocols, pore blockers, single-channel recording, and kinetic modeling. However, intrinsic hurdles may preclude the standardized application of these methods. Here, we implemented a simple method inspired by earlier studies of Na(+) channels to analyze macroscopic inactivation and conclusively deduce the pathways of inactivation of recombinant and native A-type Kv channels. We investigated two distinct A-type Kv channels expressed heterologously (Kv3.4 and Kv4.2 with accessory subunits) and their native counterparts in dorsal root ganglion and cerebellar granule neurons. This approach applies two conventional pulse protocols to examine inactivation induced by (a) a simple step (single-pulse inactivation) and (b) a conditioning step (double-pulse inactivation). Consistent with OSI, the rate of Kv3.4 inactivation (i.e., the negative first derivative of double-pulse inactivation) precisely superimposes on the profile of the Kv3.4 current evoked by a single pulse because the channels must open to inactivate. In contrast, the rate of Kv4.2 inactivation is asynchronous, already changing at earlier times relative to the profile of the Kv4.2 current evoked by a single pulse. Thus, Kv4.2 inactivation occurs uncoupled from channel opening, indicating CSI. Furthermore, the inactivation time constant versus voltage relation of Kv3.4 decreases monotonically with depolarization and levels off, whereas that of Kv4.2 exhibits a J-shape profile. We also manipulated the inactivation phenotype by changing the subunit composition and show how CSI and CSI combined with OSI might affect spiking properties in a full computational model of the hippocampal CA1 neuron. This work unambiguously

  1. Mitochondrial Ultrastructure and Glucose Signaling Pathways Attributed to the Kv1.3 Ion Channel.

    PubMed

    Kovach, Christopher P; Al Koborssy, Dolly; Huang, Zhenbo; Chelette, Brandon M; Fadool, James M; Fadool, Debra A

    2016-01-01

    Gene-targeted deletion of the potassium channel Kv1.3 (Kv1.3(-∕-)) results in "Super-smeller" mice with a sensory phenotype that includes an increased olfactory ability linked to changes in olfactory circuitry, increased abundance of olfactory cilia, and increased expression of odorant receptors and the G-protein, Golf. Kv1.3(-∕-) mice also have a metabolic phenotype including lower body weight and decreased adiposity, increased total energy expenditure (TEE), increased locomotor activity, and resistance to both diet- and genetic-induced obesity. We explored two cellular aspects to elucidate the mechanism by which loss of Kv1.3 channel in the olfactory bulb (OB) may enhance glucose utilization and metabolic rate. First, using in situ hybridization we find that Kv1.3 and the insulin-dependent glucose transporter type 4 (GLUT4) are co-localized to the mitral cell layer of the OB. Disruption of Kv1.3 conduction via construction of a pore mutation (W386F Kv1.3) was sufficient to independently translocate GLUT4 to the plasma membrane in HEK 293 cells. Because olfactory sensory perception and the maintenance of action potential (AP) firing frequency by mitral cells of the OB is highly energy demanding and Kv1.3 is also expressed in mitochondria, we next explored the structure of this organelle in mitral cells. We challenged wildtype (WT) and Kv1.3(-∕-) male mice with a moderately high-fat diet (MHF, 31.8 % kcal fat) for 4 months and then examined OB ultrastructure using transmission electron microscopy. In WT mice, mitochondria were significantly enlarged following diet-induced obesity (DIO) and there were fewer mitochondria, likely due to mitophagy. Interestingly, mitochondria were significantly smaller in Kv1.3(-∕-) mice compared with that of WT mice. Similar to their metabolic resistance to DIO, the Kv1.3(-∕-) mice had unchanged mitochondria in terms of cross sectional area and abundance following a challenge with modified diet. We are very interested to

  2. Modeling-independent elucidation of inactivation pathways in recombinant and native A-type Kv channels

    PubMed Central

    Fineberg, Jeffrey D.; Ritter, David M.

    2012-01-01

    A-type voltage-gated K+ (Kv) channels self-regulate their activity by inactivating directly from the open state (open-state inactivation [OSI]) or by inactivating before they open (closed-state inactivation [CSI]). To determine the inactivation pathways, it is often necessary to apply several pulse protocols, pore blockers, single-channel recording, and kinetic modeling. However, intrinsic hurdles may preclude the standardized application of these methods. Here, we implemented a simple method inspired by earlier studies of Na+ channels to analyze macroscopic inactivation and conclusively deduce the pathways of inactivation of recombinant and native A-type Kv channels. We investigated two distinct A-type Kv channels expressed heterologously (Kv3.4 and Kv4.2 with accessory subunits) and their native counterparts in dorsal root ganglion and cerebellar granule neurons. This approach applies two conventional pulse protocols to examine inactivation induced by (a) a simple step (single-pulse inactivation) and (b) a conditioning step (double-pulse inactivation). Consistent with OSI, the rate of Kv3.4 inactivation (i.e., the negative first derivative of double-pulse inactivation) precisely superimposes on the profile of the Kv3.4 current evoked by a single pulse because the channels must open to inactivate. In contrast, the rate of Kv4.2 inactivation is asynchronous, already changing at earlier times relative to the profile of the Kv4.2 current evoked by a single pulse. Thus, Kv4.2 inactivation occurs uncoupled from channel opening, indicating CSI. Furthermore, the inactivation time constant versus voltage relation of Kv3.4 decreases monotonically with depolarization and levels off, whereas that of Kv4.2 exhibits a J-shape profile. We also manipulated the inactivation phenotype by changing the subunit composition and show how CSI and CSI combined with OSI might affect spiking properties in a full computational model of the hippocampal CA1 neuron. This work unambiguously

  3. KV7 channels are involved in hypoxia-induced vasodilatation of porcine coronary arteries

    PubMed Central

    Hedegaard, E R; Nielsen, B D; Kun, A; Hughes, A D; Krøigaard, C; Mogensen, S; Matchkov, V V; Fröbert, O; Simonsen, U

    2014-01-01

    BACKGROUND AND PURPOSE Hypoxia causes vasodilatation of coronary arteries, but the underlying mechanisms are poorly understood. We hypothesized that hypoxia reduces intracellular Ca2+ concentration ([Ca2+]i) by opening of K channels and release of H2S. EXPERIMENTAL APPROACH Porcine coronary arteries without endothelium were mounted for measurement of isometric tension and [Ca2+]i, and the expression of voltage-gated K channels KV7 channels (encoded by KCNQ genes) and large-conductance calcium-activated K channels (KCa1.1) was examined. Voltage clamp assessed the role of KV7 channels in hypoxia. KEY RESULTS Gradual reduction of oxygen concentration from 95 to 1% dilated the precontracted coronary arteries and this was associated with reduced [Ca2+]i in PGF2α (10 μM)-contracted arteries whereas no fall in [Ca2+]i was observed in 30 mM K-contracted arteries. Blockers of ATP-sensitive voltage-gated potassium channels and KCa1.1 inhibited hypoxia-induced dilatation in PGF2α-contracted arteries; this inhibition was more marked in the presence of the Kv7 channel blockers, XE991 and linopirdine, while a KV7.1 blocker, failed to change hypoxic vasodilatation. XE991 also inhibited H2S- and adenosine-induced vasodilatation. PCR revealed the expression of KV7.1, KV7.4, KV7.5 and KCa1.1 channels, and KCa1.1, KV7.4 and KV7.5 were also identified by immunoblotting. Voltage clamp studies showed the XE991-sensitive current was more marked in hypoxic conditions. CONCLUSION The KV7.4 and KV7.5 channels, which we identified in the coronary arteries, appear to have a major role in hypoxia-induced vasodilatation. The voltage clamp results further support the involvement of KV7 channels in this vasodilatation. Activation of these KV7 channels may be induced by H2S and adenosine. PMID:24111896

  4. SU-E-I-23: A General KV Constrained Optimization of CNR for CT Abdominal Imaging

    SciTech Connect

    Weir, V; Zhang, J

    2015-06-15

    Purpose: While Tube current modulation has been well accepted for CT dose reduction, kV adjusting in clinical settings is still at its early stage. This is mainly due to the limited kV options of most current CT scanners. kV adjusting can potentially reduce radiation dose and optimize image quality. This study is to optimize CT abdomen imaging acquisition based on the assumption of a continuous kV, with the goal to provide the best contrast to noise ratio (CNR). Methods: For a given dose (CTDIvol) level, the CNRs at different kV and pitches were measured with an ACR GAMMEX phantom. The phantom was scanned in a Siemens Sensation 64 scanner and a GE VCT 64 scanner. A constrained mathematical optimization was used to find the kV which led to the highest CNR for the anatomy and pitch setting. Parametric equations were obtained from polynomial fitting of plots of kVs vs CNRs. A suitable constraint region for optimization was chosen. Subsequent optimization yielded a peak CNR at a particular kV for different collimations and pitch setting. Results: The constrained mathematical optimization approach yields kV of 114.83 and 113.46, with CNRs of 1.27 and 1.11 at the pitch of 1.2 and 1.4, respectively, for the Siemens Sensation 64 scanner with the collimation of 32 x 0.625mm. An optimized kV of 134.25 and 1.51 CNR is obtained for a GE VCT 64 slice scanner with a collimation of 32 x 0.625mm and a pitch of 0.969. At 0.516 pitch and 32 x 0.625 mm an optimized kV of 133.75 and a CNR of 1.14 was found for the GE VCT 64 slice scanner. Conclusion: CNR in CT image acquisition can be further optimized with a continuous kV option instead of current discrete or fixed kV settings. A continuous kV option is a key for individualized CT protocols.

  5. Mitochondrial Ultrastructure and Glucose Signaling Pathways Attributed to the Kv1.3 Ion Channel

    PubMed Central

    Kovach, Christopher P.; Al Koborssy, Dolly; Huang, Zhenbo; Chelette, Brandon M.; Fadool, James M.; Fadool, Debra A.

    2016-01-01

    Gene-targeted deletion of the potassium channel Kv1.3 (Kv1.3−∕−) results in “Super-smeller” mice with a sensory phenotype that includes an increased olfactory ability linked to changes in olfactory circuitry, increased abundance of olfactory cilia, and increased expression of odorant receptors and the G-protein, Golf. Kv1.3−∕− mice also have a metabolic phenotype including lower body weight and decreased adiposity, increased total energy expenditure (TEE), increased locomotor activity, and resistance to both diet- and genetic-induced obesity. We explored two cellular aspects to elucidate the mechanism by which loss of Kv1.3 channel in the olfactory bulb (OB) may enhance glucose utilization and metabolic rate. First, using in situ hybridization we find that Kv1.3 and the insulin-dependent glucose transporter type 4 (GLUT4) are co-localized to the mitral cell layer of the OB. Disruption of Kv1.3 conduction via construction of a pore mutation (W386F Kv1.3) was sufficient to independently translocate GLUT4 to the plasma membrane in HEK 293 cells. Because olfactory sensory perception and the maintenance of action potential (AP) firing frequency by mitral cells of the OB is highly energy demanding and Kv1.3 is also expressed in mitochondria, we next explored the structure of this organelle in mitral cells. We challenged wildtype (WT) and Kv1.3−∕− male mice with a moderately high-fat diet (MHF, 31.8 % kcal fat) for 4 months and then examined OB ultrastructure using transmission electron microscopy. In WT mice, mitochondria were significantly enlarged following diet-induced obesity (DIO) and there were fewer mitochondria, likely due to mitophagy. Interestingly, mitochondria were significantly smaller in Kv1.3−∕− mice compared with that of WT mice. Similar to their metabolic resistance to DIO, the Kv1.3−∕− mice had unchanged mitochondria in terms of cross sectional area and abundance following a challenge with modified diet. We are very

  6. Transient Hippocampal Down-Regulation of Kv1.1 Subunit mRNA during Associative Learning in Rats

    ERIC Educational Resources Information Center

    Kourrich, Said; Manrique, Christine; Salin, Pascal; Mourre, Christiane

    2005-01-01

    Voltage-gated potassium channels (Kv) are critically involved in learning and memory processes. It is not known, however, whether the expression of the Kv1.1 subunit, constituting Kv1 channels, can be specifically regulated in brain areas important for learning and memory processing. Radioactive in situ hybridization was used to evaluate the…

  7. Effect of methamphetamine on the microglial damage: role of potassium channel Kv1.3.

    PubMed

    Wang, Jun; Qian, Wenyi; Liu, Jingli; Zhao, Jingjing; Yu, Pan; Jiang, Lei; Zhou, Jing; Gao, Rong; Xiao, Hang

    2014-01-01

    Methamphetamine (Meth) abusing represents a major public health problem worldwide. Meth has long been known to induce neurotoxicity. However, the mechanism is still remained poorly understood. Growing evidences indicated that the voltage-gated potassium channels (Kv) were participated in neuronal damage and microglia function. With the whole cell patch clamp, we found that Meth significantly increased the outward K⁺ currents, therefore, we explored whether Kv1.3, one of the major K⁺ channels expressed in microglia, was involved in Meth-induced microglia damage. Our study showed that Meth significantly increased the cell viability in a dose dependent manner, while the Kv blocker, tetraethylamine (TEA), 4-Aminopyridine (4-AP) and Kv1.3 specific antagonist margatoxin (MgTx), prevented against the damage mediated by Meth. Interestingly, treatment of cells with Meth resulted in increasing expression of Kv1.3 rather than Kv1.5, at both mRNA and protein level, which is partially blocked by MgTx. Furthermore, Meth also stimulated a significant increased expression of IL-6 and TNF-α at protein level, which was significantly inhibited by MgTx. Taken together, these results demonstrated that Kv1.3 was involved in Meth-mediated microglial damage, providing the potential target for the development of therapeutic strategies for Meth abuse. PMID:24533129

  8. Contribution of Kv7 channels to natriuretic peptide mediated vasodilation in normal and hypertensive rats.

    PubMed

    Stott, Jennifer B; Barrese, Vincenzo; Jepps, Thomas A; Leighton, Emma V; Greenwood, Iain A

    2015-03-01

    The Kv7 family of voltage-gated potassium channels are expressed within the vasculature where they are key regulators of vascular tone and mediate cAMP-linked endogenous vasodilator responses, a pathway that is compromised in hypertension. However, the role of Kv7 channels in non-cAMP-linked vasodilator pathways has not been investigated. Natriuretic peptides are potent vasodilators, which operate primarily through the activation of a cGMP-dependent signaling pathway. This study investigated the putative role of Kv7 channels in natriuretic peptide-dependent relaxations in the vasculature of normal and hypertensive animals. Relaxant responses of rat aorta to both atrial and C-type natriuretic peptides and the nitric oxide donor sodium nitroprusside were impaired by the Kv7 blocker linopirdine (10 μmol/L) but not by the Kv7.1-specific blocker HMR1556 (10 μmol/L) and other K(+) channel blockers. In contrast, only the atrial natriuretic peptide response was sensitive to linopirdine in the renal artery. These Kv7-mediated responses were attenuated in arteries from hypertensive rats. Quantitative polymerase chain reaction showed that A- and B-type natriuretic peptide receptors were expressed at high levels in the aorta and renal artery from normal and spontaneously hypertensive rats. This study provides the first evidence that natriuretic peptide responses are impaired in hypertension and that recruitment of Kv7 channels is a key component of natriuretic peptide-dependent vasodilations. PMID:25547342

  9. Suppression of KV7/KCNQ potassium channel enhances neuronal differentiation of PC12 cells.

    PubMed

    Zhou, Najing; Huang, Sha; Li, Li; Huang, Dongyang; Yan, Yunli; Du, Xiaona; Zhang, Hailin

    2016-10-01

    Membrane potential shift driven by electrical activity is critical in determining the cell fate of proliferation or differentiation. As such, the ion channels that underlie the membrane electrical activity play an important role in cell proliferation/differentiation. KV7/KCNQ potassium channels are critical in determining the resting membrane potentials in many neuronal cells. However, the role of these channels in cell differentiation is not well studied. In the present study, we used PC12 cells as well as primary cultured rat cortical neurons to study the role and mechanism of KV7/KCNQ in neuronal differentiation. NGF induced PC12 cell differentiation into neuron-like cells with growth of neurites showing typical growth cone-like extensions. The Kv7/KCNQ blocker XE991 promoted NGF-induced neurite outgrowth, whereas Kv7/KCNQ opener retigabine (RTG) inhibited outgrowth. M-type Kv7 channels are likely involved in regulating neurite growth because overexpression of KCNQ2/Q3 inhibited neurite growth whereas suppression of KCNQ2/Q3 with shRNA promoted neurite growth. Membrane depolarization possibly underpins enhanced neurite growth induced by the suppression of Kv7/KCNQ. Additionally, high extracellular K(+) likely induced membrane depolarization and also promoted neurite growth. Finally, T-type Ca(2+) channels may be involved in membrane-depolarization-induced neurite growth. This study provides a new perspective for understanding neuronal differentiation as well as KV7/KCNQ channel function. PMID:27450567

  10. Regulation of Human Kv1.4 Channel Activity by the Antidepressant Metergoline.

    PubMed

    Yeom, Hye Duck; Lee, Jun-Ho

    2016-01-01

    Metergoline is an ergot-derived psychoactive drug that is a ligand for various serotonin and dopamine receptors. Little is known about the effect of metergoline on different types of receptors and ion channels. Potassium channels are the most diverse group of ion channels. Kv1.4, a shaker family K channel alpha subunit, is one of a family of voltage gated K channels that mediates transient and rapid inactivating A-type currents and N-type inactivation. We demonstrated previously that metergoline inhibited the activity of neuronal voltage-dependent Na(+) channels in Xenopus laevis oocytes (Acta Pharmacol. Sin., 35, 2014, Lee et al.). In this study, we sought to elucidate the regulatory effects underlying metergoline-induced human Kv1.4 channel inhibition. We used the two electrode voltage-clamp (TEVC) technique to investigate the effect of metergoline on human Kv1.4 channel currents in Xenopus laevis oocytes expressing human Kv1.4 alpha subunits. Interestingly, metergoline treatment also induced inhibition of peak currents in human Kv1.4 channels in a concentration-dependent manner. The IC50 of peak currents of hKv1.4 currents was 3.6±0.6 µM. These results indicate that metergoline might regulate the human Kv1.4 channel activity that is expressed in X. laevis oocytes. Further, this regulation of potassium currents by metergoline might be one of the pharmacological actions of metergoline-mediated psychoactivity. PMID:27251511

  11. CEBAF 200 kV Inverted Electron Gun

    SciTech Connect

    Grames, J M; Clark, J; Hansknecht, J; Poelker, M; Stutzman, M L; Suleiman, R; Surles-Law, K.E.L.; BastaniNejad, M; McCarter, J J

    2011-03-01

    Two DC high volt­age GaAs pho­to­guns have been built at Jef­fer­son Lab based on a com­pact in­vert­ed in­su­la­tor de­sign. One pho­to­gun pro­vides the po­lar­ized elec­tron beam at CEBAF and op­er­ates at 130 kV bias volt­age. The other gun is used for high av­er­age cur­rent life­time stud­ies at a ded­i­cat­ed test fa­cil­i­ty and has been op­er­at­ed at bias volt­age up to 225 kV. The ad­van­tages of high­er DC volt­age for CEBAF in­clude re­duced space-charge emit­tance growth and the po­ten­tial for pro­longed pho­to­cath­ode life­time. How­ev­er, a con­se­quence of op­er­at­ing at high­er volt­ages is the in­creased like­li­hood of field emis­sion or break­down, both of which are un­ac­cept­able. High­lights of the R&D stud­ies lead­ing to­ward a pro­duc­tion 200keV GaAs pho­to­gun for CEBAF will be pre­sent­ed.

  12. Kv4.2 knockout mice have hippocampal-dependent learning and memory deficits.

    PubMed

    Lugo, Joaquin N; Brewster, Amy L; Spencer, Corinne M; Anderson, Anne E

    2012-05-01

    Kv4.2 channels contribute to the transient, outward K(+) current (A-type current) in hippocampal dendrites, and modulation of this current substantially alters dendritic excitability. Using Kv4.2 knockout (KO) mice, we examined the role of Kv4.2 in hippocampal-dependent learning and memory. We found that Kv4.2 KO mice showed a deficit in the learning phase of the Morris water maze (MWM) and significant impairment in the probe trial compared with wild type (WT). Kv4.2 KO mice also demonstrated a specific deficit in contextual learning in the fear-conditioning test, without impairment in the conditioned stimulus or new context condition. Kv4.2 KO mice had normal activity, anxiety levels, and prepulse inhibition compared with WT mice. A compensatory increase in tonic inhibition has been previously described in hippocampal slice recordings from Kv4.2 KO mice. In an attempt to decipher whether increased tonic inhibition contributed to the learning and memory deficits in Kv4.2 KO mice, we administered picrotoxin to block GABA(A) receptors (GABA(A)R), and thereby tonic inhibition. This manipulation had no effect on behavior in the WT or KO mice. Furthermore, total protein levels of the α5 or δ GABA(A)R subunits, which contribute to tonic inhibition, were unchanged in hippocampus. Overall, our findings add to the growing body of evidence, suggesting an important role for Kv4.2 channels in hippocampal-dependent learning and memory. PMID:22505720

  13. Voltage-dependent gating and gating charge measurements in the Kv1.2 potassium channel

    PubMed Central

    Ishida, Itzel G.; Rangel-Yescas, Gisela E.; Carrasco-Zanini, Julia

    2015-01-01

    Much has been learned about the voltage sensors of ion channels since the x-ray structure of the mammalian voltage-gated potassium channel Kv1.2 was published in 2005. High resolution structural data of a Kv channel enabled the structural interpretation of numerous electrophysiological findings collected in various ion channels, most notably Shaker, and permitted the development of meticulous computational simulations of the activation mechanism. The fundamental premise for the structural interpretation of functional measurements from Shaker is that this channel and Kv1.2 have the same characteristics, such that correlation of data from both channels would be a trivial task. We tested these assumptions by measuring Kv1.2 voltage-dependent gating and charge per channel. We found that the Kv1.2 gating charge is near 10 elementary charges (eo), ∼25% less than the well-established 13–14 eo in Shaker. Next, we neutralized positive residues in the Kv1.2 S4 transmembrane segment to investigate the cause of the reduction of the gating charge and found that, whereas replacing R1 with glutamine decreased voltage sensitivity to ∼50% of the wild-type channel value, mutation of the subsequent arginines had a much smaller effect. These data are in marked contrast to the effects of charge neutralization in Shaker, where removal of the first four basic residues reduces the gating charge by roughly the same amount. In light of these differences, we propose that the voltage-sensing domains (VSDs) of Kv1.2 and Shaker might undergo the same physical movement, but the septum that separates the aqueous crevices in the VSD of Kv1.2 might be thicker than Shaker’s, accounting for the smaller Kv1.2 gating charge. PMID:25779871

  14. Effect of mosapride on Kv4.3 potassium channels expressed in CHO cells.

    PubMed

    Sung, Ki-Wug; Hahn, Sang June

    2013-10-01

    Mosapride and cisapride are gastroprokinetic agents with 5-hydroxytryptamine4 receptor agonist activity and have been widely used in the treatment of a variety of gastrointestinal disorders. The effects of mosapride and cisapride on cloned Kv4.3 channels stably expressed in Chinese hamster ovary cells were investigated using the whole-cell patch-clamp technique. Mosapride and cisapride inhibited Kv4.3 in a concentration-dependent manner with IC50 values of 15.2 and 9.8 μM, respectively. Mosapride accelerated the rate of inactivation and activation of Kv4.3 in a concentration-dependent manner and thereby decreased the time to peak. The rate constants of association (k +1) and dissociation (k -1) for mosapride were 9.9 μM(-1) s(-1) and 151.3 s(-1), respectively. The K D (k -1/k +1) was 16.2 μM, similar to the IC50 value calculated from the concentration-response curve. Voltage-dependent inhibition by mosapride was observed in the voltage range for channel opening but was not observed over a voltage range in which all Kv4.3 channels were open. Both the steady-state activation and inactivation curves of Kv4.3 were shifted in the hyperpolarizing direction in the presence of mosapride. Mosapride also caused a substantial acceleration in closed-state inactivation of Kv4.3. Mosapride produced use-dependent inhibition, which was consistent with a slow recovery from inactivation of Kv4.3. M1 and norcisapride, the major metabolites of mosapride and cisapride, respectively, had little or no effect on Kv4.3. These results indicate that mosapride inhibits Kv4.3 by both preferential binding to the open state of the channels during depolarization and acceleration of the closed-state inactivation at subthreshold potentials. PMID:23793103

  15. Voltage-dependent gating and gating charge measurements in the Kv1.2 potassium channel.

    PubMed

    Ishida, Itzel G; Rangel-Yescas, Gisela E; Carrasco-Zanini, Julia; Islas, León D

    2015-04-01

    Much has been learned about the voltage sensors of ion channels since the x-ray structure of the mammalian voltage-gated potassium channel Kv1.2 was published in 2005. High resolution structural data of a Kv channel enabled the structural interpretation of numerous electrophysiological findings collected in various ion channels, most notably Shaker, and permitted the development of meticulous computational simulations of the activation mechanism. The fundamental premise for the structural interpretation of functional measurements from Shaker is that this channel and Kv1.2 have the same characteristics, such that correlation of data from both channels would be a trivial task. We tested these assumptions by measuring Kv1.2 voltage-dependent gating and charge per channel. We found that the Kv1.2 gating charge is near 10 elementary charges (eo), ∼25% less than the well-established 13-14 eo in Shaker. Next, we neutralized positive residues in the Kv1.2 S4 transmembrane segment to investigate the cause of the reduction of the gating charge and found that, whereas replacing R1 with glutamine decreased voltage sensitivity to ∼50% of the wild-type channel value, mutation of the subsequent arginines had a much smaller effect. These data are in marked contrast to the effects of charge neutralization in Shaker, where removal of the first four basic residues reduces the gating charge by roughly the same amount. In light of these differences, we propose that the voltage-sensing domains (VSDs) of Kv1.2 and Shaker might undergo the same physical movement, but the septum that separates the aqueous crevices in the VSD of Kv1.2 might be thicker than Shaker's, accounting for the smaller Kv1.2 gating charge. PMID:25779871

  16. Design, construction, and operational results of an 800-A, 10-kV hot deck amplifier

    SciTech Connect

    Reass, W.A.

    1984-06-01

    This paper describes the electrical design, implementation, and operational results of a high fidelity (feedback regulated) 800 A, 10-kV hard tube, hot deck amplifier. The amplifier can produce any linear waveform to 800-A for 30 ms and beyond (depending on main energy storage). The present use is to drive the vertical field (VF) control windings on ZT-40M, a toroidal reversed field pinch plasma physics experiment. Although our application requires only 10 kV (8 MW) of switching, anode voltage may be as high as 40 kV (32 MW).

  17. Development of 600 kV triple resonance pulse transformer.

    PubMed

    Li, Mingjia; Zhang, Faqiang; Liang, Chuan; Xu, Zhou

    2015-06-01

    In this paper, a triple-resonance pulse transformer based on an air-core transformer is introduced. The voltage across the high-voltage winding of the air-core transformer is significantly less than the output voltage; instead, the full output voltage appears across the tuning inductor. The maximum ratio of peak load voltage to peak transformer voltage is 2.77 in theory. By analyzing pulse transformer's lossless circuit, the analytical expression for the output voltage and the characteristic equation of the triple-resonance circuit are presented. Design method for the triple-resonance pulse transformer (iterated simulation method) is presented, and a triple-resonance pulse transformer is developed based on the existing air-core transformer. The experimental results indicate that the maximum ratio of peak voltage across the load to peak voltage across the high-voltage winding of the air-core transformer is approximately 2.0 and the peak output voltage of the triple-resonance pulse transformer is approximately 600 kV. PMID:26133858

  18. Phosphatidic acid modulation of Kv channel voltage sensor function

    PubMed Central

    Hite, Richard K; Butterwick, Joel A; MacKinnon, Roderick

    2014-01-01

    Membrane phospholipids can function as potent regulators of ion channel function. This study uncovers and investigates the effect of phosphatidic acid on Kv channel gating. Using the method of reconstitution into planar lipid bilayers, in which protein and lipid components are defined and controlled, we characterize two effects of phosphatidic acid. The first is a non-specific electrostatic influence on activation mediated by electric charge density on the extracellular and intracellular membrane surfaces. The second is specific to the presence of a primary phosphate group, acts only through the intracellular membrane leaflet and depends on the presence of a particular arginine residue in the voltage sensor. Intracellular phosphatidic acid accounts for a nearly 50 mV shift in the midpoint of the activation curve in a direction consistent with stabilization of the voltage sensor's closed conformation. These findings support a novel mechanism of voltage sensor regulation by the signaling lipid phosphatidic acid. DOI: http://dx.doi.org/10.7554/eLife.04366.001 PMID:25285449

  19. Modeling of a 22.9 kV 50 MVA superconducting power cable based on PSCAD/EMTDC for application to the Icheon substation in Korea

    NASA Astrophysics Data System (ADS)

    Lee, S.; Yoon, J.; Lee, B.; Yang, B.

    2011-11-01

    Two projects for high temperature superconducting (HTS) power cable have been carried out in Korea since 2001. One of them is a HTS cable project in DAPAS (Development of Advanced Power system by Applied Superconductivity technologies) program funded by the Ministry of Education, Science and Technology. In this project, LS Cable Ltd. (LSC) and Korea Electrotechnology Research Institute (KERI) jointly developed a 22.9 kV, 50 MVA, 3 phase, 100 m HTS cable using first generation (1G) HTS wire in 2006. The HTS cable system has been tested in a power test center of Korea Electric Power Corporation (KEPCO). LSC and KEPCO have been developing a 22.9 kV, 50 MVA, 3 phase, 500 m HTS cable system using second generation (2G) HTS wire since 2008, based on the technology of the DAPAS project. This project is called as GENI (Green Superconducting Electric Power Network at the Icheon Substation) project. The target of GENI project is to install and operate the HTS cable system in the Icheon substation located in near Seoul. In order to analyze the Icheon substation power system employing the HTS cable, an analysis model of the HTS cable is necessary. This paper describes the development of an analysis model for the 22.9 kV, 50 MVA HTS cable that will be applied to the Icheon substation in Korea.

  20. Determination Of X-Ray Tube Potential (kV) Waveform By A Noninvasive Evaluation Of Radiation Output (NERO)

    NASA Astrophysics Data System (ADS)

    Simon, William E.; Richards, Doug

    1981-07-01

    A method has been developed to noninvasively measure the instantaneous potential (kV) applied to an x-ray tube. The method uses differentially filtered x-ray detectors whose outputs during the exposure are converted to digital signals and stored in a memory array. The conversions are made every 125 microseconds. After the exposure, a microprocessor calculates the ratios of the detector outputs; computes the kV waveform from stored calibration data; and digitally displays the kV value. A resolution of +/-0.5 kV at 110 kV has been achieved.

  1. Kv4.2 and accessory dipeptidyl peptidase-like protein 10 (DPP10) subunit preferentially form a 4:2 (Kv4.2:DPP10) channel complex.

    PubMed

    Kitazawa, Masahiro; Kubo, Yoshihiro; Nakajo, Koichi

    2015-09-11

    Kv4 is a member of the voltage-gated K(+) channel family and forms a complex with various accessory subunits. Dipeptidyl aminopeptidase-like protein (DPP) is one of the auxiliary subunits for the Kv4 channel. Although DPP has been well characterized and is known to increase the current amplitude and accelerate the inactivation and recovery from inactivation of Kv4 current, it remains to be determined how many DPPs bind to one Kv4 channel. To examine whether the expression level of DPP changes the biophysical properties of Kv4, we expressed Kv4.2 and DPP10 in different ratios in Xenopus oocytes and analyzed the currents under two-electrode voltage clamp. The current amplitude and the speed of recovery from inactivation of Kv4.2 changed depending on the co-expression level of DPP10. This raised the possibility that the stoichiometry of the Kv4.2-DPP10 complex is variable and affects the biophysical properties of Kv4.2. We next determined the stoichiometry of DPP10 alone by subunit counting using single-molecule imaging. Approximately 70% of the DPP10 formed dimers in the plasma membrane, and the rest existed as monomers in the absence of Kv4.2. We next determined the stoichiometry of the Kv4.2-DPP10 complex; Kv4.2-mCherry and mEGFP-DPP10 were co-expressed in different ratios and the stoichiometries of Kv4.2-DPP10 complexes were evaluated by the subunit counting method. The stoichiometry of the Kv4.2-DPP10 complex was variable depending on the relative expression level of each subunit, with a preference for 4:2 stoichiometry. This preference may come from the bulky dimeric structure of the extracellular domain of DPP10. PMID:26209633

  2. Toxins Targeting the KV1.3 Channel: Potential Immunomodulators for Autoimmune Diseases

    PubMed Central

    Zhao, Yipeng; Huang, Jie; Yuan, Xiaolu; Peng, Biwen; Liu, Wanhong; Han, Song; He, Xiaohua

    2015-01-01

    Autoimmune diseases are usually accompanied by tissue injury caused by autoantigen-specific T-cells. KV1.3 channels participate in modulating calcium signaling to induce T-cell proliferation, immune activation and cytokine production. Effector memory T (TEM)-cells, which play major roles in many autoimmune diseases, are controlled by blocking KV1.3 channels on the membrane. Toxins derived from animal venoms have been found to selectively target a variety of ion channels, including KV1.3. By blocking the KV1.3 channel, these toxins are able to suppress the activation and proliferation of TEM cells and may improve TEM cell-mediated autoimmune diseases, such as multiple sclerosis and type I diabetes mellitus. PMID:25996605

  3. Status of research and development for 1200-kV transmission and substation equipment

    SciTech Connect

    Walldorf, S.P.; Gnadt, P.A.

    1985-01-01

    In 1975 the US Department of Energy (DOE), Division of Electric Energy Systems (EES), initiated a program for the research and development (R and D) of 1200-kV transmission and substation equipment with a goal of developing the technology capable of three-phase power transfer of 10 GW. This R and D program has encompassed all the major and essential transmission cable, substation apparatus, and specialized instrumentation for a 1200-kV system; and it has produced a number of full-scale 1200-kV components that have successfully undergone or are still undergoing accelerated field testing. This paper presents a review of the overall DOE/EES 1200-kV R and D program, describes the major components developed, highlights their key design aspects, indicates the present status of development and testing, and presents a discussion of future commercial implications.

  4. In silico identification of PAP-1 binding sites in the Kv1.2 potassium channel.

    PubMed

    Jorgensen, Christian; Darré, Leonardo; Vanommeslaeghe, Kenno; Omoto, Kiyoyuki; Pryde, David; Domene, Carmen

    2015-04-01

    Voltage-gated potassium channels of the Kv1 family play a crucial role in the generation and transmission of electrical signals in excitable cells affecting neuronal and cardiac activities. Small-molecule blockage of these channels has been proposed to occur via a cooperative mechanism involving two main blocking sites: the inner-pore site located below the selectivity filter, and a side-pocket cavity located between the pore and the voltage sensor. Using 0.5 μs molecular dynamics simulation trajectories complemented by docking calculations, the potential binding sites of the PAP-1 (5-(4-phenoxybutoxy)psoralen) blocker to the crystal structure of Kv1.2 channel have been studied. The presence of both mentioned blocking sites at Kv1.2 is confirmed, adding evidence in favor of a cooperative channel blockage mechanism. These observations provide insight into drug modulation that will guide further developments of Kv inhibitors. PMID:25734225

  5. PKC and AMPK regulation of Kv1.5 potassium channels

    PubMed Central

    Andersen, Martin Nybo; Skibsbye, Lasse; Tang, Chuyi; Petersen, Frederic; MacAulay, Nanna; Rasmussen, Hanne Borger; Jespersen, Thomas

    2015-01-01

    The voltage-gated Kv1.5 potassium channel, conducting the ultra-rapid rectifier K+ current (IKur), is regulated through several pathways. Here we investigate if Kv1.5 surface expression is controlled by the 2 kinases PKC and AMPK, using Xenopus oocytes, MDCK cells and atrial derived HL-1 cells. By confocal microscopy combined with electrophysiology we demonstrate that PKC activation reduces Kv1.5 current, through a decrease in membrane expressed channels. AMPK activation was found to decrease the membrane expression in MDCK cells, but not in HL-1 cells and was furthermore shown to be dependent on co-expression of Nedd4–2 in Xenopus oocytes. These results indicate that Kv1.5 channels are regulated by both kinases, although through different molecular mechanisms in different cell systems. PMID:26043299

  6. PKC and AMPK regulation of Kv1.5 potassium channels.

    PubMed

    Andersen, Martin Nybo; Skibsbye, Lasse; Tang, Chuyi; Petersen, Frederic; MacAulay, Nanna; Rasmussen, Hanne Borger; Jespersen, Thomas

    2015-01-01

    The voltage-gated Kv1.5 potassium channel, conducting the ultra-rapid rectifier K(+) current (IKur), is regulated through several pathways. Here we investigate if Kv1.5 surface expression is controlled by the 2 kinases PKC and AMPK, using Xenopus oocytes, MDCK cells and atrial derived HL-1 cells. By confocal microscopy combined with electrophysiology we demonstrate that PKC activation reduces Kv1.5 current, through a decrease in membrane expressed channels. AMPK activation was found to decrease the membrane expression in MDCK cells, but not in HL-1 cells and was furthermore shown to be dependent on co-expression of Nedd4-2 in Xenopus oocytes. These results indicate that Kv1.5 channels are regulated by both kinases, although through different molecular mechanisms in different cell systems. PMID:26043299

  7. O2-sensitive K+ channels: role of the Kv1.2 -subunit in mediating the hypoxic response.

    PubMed

    Conforti, L; Bodi, I; Nisbet, J W; Millhorn, D E

    2000-05-01

    One of the early events in O2 chemoreception is inhibition of O2-sensitive K+ (KO2) channels. Characterization of the molecular composition of the native KO2 channels in chemosensitive cells is important to understand the mechanism(s) that couple O2 to the KO2 channels. The rat phaeochromocytoma PC12 clonal cell line expresses an O2-sensitive voltage-dependent K+ channel similar to that recorded in other chemosensitive cells. Here we examine the possibility that the Kv1.2 alpha-subunit comprises the KO2 channel in PC12 cells. Whole-cell voltage-clamp experiments showed that the KO2 current in PC12 cells is inhibited by charybdotoxin, a blocker of Kv1.2 channels. PC12 cells express the Kv1.2 alpha-subunit of K+ channels: Western blot analysis with affinity-purified anti-Kv1.2 antibody revealed a band at approximately 80 kDa. Specificity of this antibody was established in Western blot and immunohystochemical studies. Anti-Kv1.2 antibody selectively blocked Kv1.2 current expressed in the Xenopus oocyte, but had no effect on Kv2.1 current. Anti-Kv1.2 antibody dialysed through the patch pipette completely blocked the KO2 current, while the anti-Kv2.1 and irrelevant antibodies had no effect. The O2 sensitivity of recombinant Kv1.2 and Kv2.1 channels was studied in Xenopus oocytes. Hypoxia inhibited the Kv1.2 current only. These findings show that the KO2 channel in PC12 cells belongs to the Kv1 subfamily of K+ channels and that the Kv1.2 alpha-subunit is important in conferring O2 sensitivity to this channel. PMID:10790158

  8. O2-sensitive K+ channels: role of the Kv1.2 α-subunit in mediating the hypoxic response

    PubMed Central

    Conforti, Laura; Bodi, Ilona; Nisbet, John W; Millhorn, David E

    2000-01-01

    One of the early events in O2 chemoreception is inhibition of O2-sensitive K+ (KO2) channels. Characterization of the molecular composition of the native KO2 channels in chemosensitive cells is important to understand the mechanism(s) that couple O2 to the KO2 channels. The rat phaeochromocytoma PC12 clonal cell line expresses an O2-sensitive voltage-dependent K+ channel similar to that recorded in other chemosensitive cells. Here we examine the possibility that the Kv1.2 α-subunit comprises the KO2 channel in PC12 cells. Whole-cell voltage-clamp experiments showed that the KO2 current in PC12 cells is inhibited by charybdotoxin, a blocker of Kv1.2 channels. PC12 cells express the Kv1.2 α-subunit of K+ channels: Western blot analysis with affinity-purified anti-Kv1.2 antibody revealed a band at ≈80 kDa. Specificity of this antibody was established in Western blot and immunohystochemical studies. Anti-Kv1.2 antibody selectively blocked Kv1.2 current expressed in the Xenopus oocyte, but had no effect on Kv2.1 current. Anti-Kv1.2 antibody dialysed through the patch pipette completely blocked the KO2 current, while the anti-Kv2.1 and irrelevant antibodies had no effect. The O2 sensitivity of recombinant Kv1.2 and Kv2.1 channels was studied in Xenopus oocytes. Hypoxia inhibited the Kv1.2 current only. These findings show that the KO2 channel in PC12 cells belongs to the Kv1 subfamily of K+ channels and that the Kv1.2 α-subunit is important in conferring O2 sensitivity to this channel. PMID:10790158

  9. Overexpression of Tau Downregulated the mRNA Levels of Kv Channels and Improved Proliferation in N2A Cells

    PubMed Central

    Li, Xiantao; Hu, Ximu; Li, Xiaoqing; Hao, Xuran

    2015-01-01

    Microtubule binding protein tau has a crucial function in promoting the assembly and stabilization of microtubule. Besides tuning the action potentials, voltage-gated K+ channels (Kv) are important for cell proliferation and appear to play a role in the development of cancer. However, little is known about the possible interaction of tau with Kv channels in various tissues. In the present study, tau plasmids were transiently transfected into mouse neuroblastoma N2A cells to explore the possible linkages between tau and Kv channels. This treatment led to a downregulation of mRNA levels of several Kv channels, including Kv2.1, Kv3.1, Kv4.1, Kv9.2, and KCNH4, but no significant alteration was observed for Kv5.1 and KCNQ4. Furthermore, the macroscopic currents through Kv channels were reduced by 36.5% at +60 mV in tau-tranfected N2A cells. The proliferation rates of N2A cells were also improved by the induction of tau expression and the incubation of TEA (tetraethylammonium) for 48 h by 120.9% and 149.3%, respectively. Following the cotransfection with tau in HEK293 cells, the mRNA levels and corresponding currents of Kv2.1 were significantly declined compared with single Kv2.1 transfection. Our data indicated that overexpression of tau declined the mRNA levels of Kv channels and related currents. The effects of tau overexpression on Kv channels provided an alternative explanation for low sensitivity to anti-cancer chemicals in some specific cancer tissues. PMID:25590133

  10. E3 ligase CHIP and Hsc70 regulate Kv1.5 protein expression and function in mammalian cells.

    PubMed

    Li, Peili; Kurata, Yasutaka; Maharani, Nani; Mahati, Endang; Higaki, Katsumi; Hasegawa, Akira; Shirayoshi, Yasuaki; Yoshida, Akio; Kondo, Tatehito; Kurozawa, Youichi; Yamamoto, Kazuhiro; Ninomiya, Haruaki; Hisatome, Ichiro

    2015-09-01

    Kv1.5 confers ultra-rapid delayed-rectifier potassium channel current (IKur) which contributes to repolarization of the atrial action potential. Kv1.5 proteins, degraded via the ubiquitin-proteasome pathway, decreased in some atrial fibrillation patients. Carboxyl-terminus heat shock cognate 70-interacting protein (CHIP), an E3 ubiquitin ligase, is known to ubiquitinate short-lived proteins. Here, we investigated the roles of CHIP in Kv1.5 degradation to provide insights into the mechanisms of Kv1.5 decreases and treatments targeting Kv1.5 for atrial fibrillation. Coexpression of CHIP with Kv1.5 in HEK293 cells increased Kv1.5 protein ubiquitination and decreased the protein level. Immunofluorescence revealed decreases of Kv1.5 proteins in the endoplasmic reticulum and on the cell membrane. A siRNA against CHIP suppressed Kv1.5 protein ubiquitination and increased its protein level. CHIP mutants, lacking either the N-terminal tetratricopeptide region domain or the C-terminal U-box domain, failed to exert these effects on Kv1.5 proteins. Immunoprecipitation showed that CHIP formed complexes with Kv1.5 proteins and heat shock cognate protein 70 (Hsc70). Effects of Hsc70 on Kv1.5 were similar to CHIP by altering interaction of CHIP with Kv1.5 protein. Coexpression of CHIP and Hsc70 with Kv1.5 additionally enhanced Kv1.5 ubiquitination. Kv1.5 currents were decreased by overexpression of CHIP or Hsc70 but were increased by knockdown of CHIP or Hsc70 in HEK 293 cells stably expressing Kv1.5. These effects of CHIP and Hsc70 were also observed on endogenous Kv1.5 in HL-1 mouse cardiomyocytes, decreasing IKur and prolonging action potential duration. These results indicate that CHIP decreases the Kv1.5 protein level and functional channel by facilitating its degradation in concert with chaperone Hsc70. PMID:26232501

  11. Different KChIPs Compete for Heteromultimeric Assembly with Pore-Forming Kv4 Subunits

    PubMed Central

    Zhou, Jingheng; Tang, Yiquan; Zheng, Qin; Li, Meng; Yuan, Tianyi; Chen, Liangyi; Huang, Zhuo; Wang, KeWei

    2015-01-01

    Auxiliary Kv channel-interacting proteins 1–4 (KChIPs1–4) coassemble with pore-forming Kv4 α-subunits to form channel complexes underlying somatodendritic subthreshold A-type current that regulates neuronal excitability. It has been hypothesized that different KChIPs can competitively bind to Kv4 α-subunit to form variable channel complexes that can exhibit distinct biophysical properties for modulation of neural function. In this study, we use single-molecule subunit counting by total internal reflection fluorescence microscopy in combinations with electrophysiology and biochemistry to investigate whether different isoforms of auxiliary KChIPs, KChIP4a, and KChIP4bl, can compete for binding of Kv4.3 to coassemble heteromultimeric channel complexes for modulation of channel function. To count the number of photobleaching steps solely from cell membrane, we take advantage of a membrane tethered k-ras-CAAX peptide that anchors cytosolic KChIP4 proteins to the surface for reduction of background noise. Single-molecule subunit counting reveals that the number of KChIP4 isoforms in Kv4.3-KChIP4 complexes can vary depending on the KChIP4 expression level. Increasing the amount of KChIP4bl gradually reduces bleaching steps of KChIP4a isoform proteins, and vice versa. Further analysis of channel gating kinetics from different Kv4-KChIP4 subunit compositions confirms that both KChIP4a and KChIP4bl can modulate the channel complex function upon coassembly. Taken together, our findings show that auxiliary KChIPs can heteroassemble with Kv4 in a competitive manner to form heteromultimeric Kv4-KChIP4 channel complexes that are biophysically distinct and regulated under physiological or pathological conditions. PMID:26039167

  12. A Dipeptidyl Aminopeptidase–like Protein Remodels Gating Charge Dynamics in Kv4.2 Channels

    PubMed Central

    Dougherty, Kevin; Covarrubias, Manuel

    2006-01-01

    Dipeptidyl aminopeptidase–like proteins (DPLPs) interact with Kv4 channels and thereby induce a profound remodeling of activation and inactivation gating. DPLPs are constitutive components of the neuronal Kv4 channel complex, and recent observations have suggested the critical functional role of the single transmembrane segment of these proteins (Zagha, E., A. Ozaita, S.Y. Chang, M.S. Nadal, U. Lin, M.J. Saganich, T. McCormack, K.O. Akinsanya, S.Y. Qi, and B. Rudy. 2005. J. Biol. Chem. 280:18853–18861). However, the underlying mechanism of action is unknown. We hypothesized that a unique interaction between the Kv4.2 channel and a DPLP found in brain (DPPX-S) may remodel the channel's voltage-sensing domain. To test this hypothesis, we implemented a robust experimental system to measure Kv4.2 gating currents and study gating charge dynamics in the absence and presence of DPPX-S. The results demonstrated that coexpression of Kv4.2 and DPPX-S causes a −26 mV parallel shift in the gating charge-voltage (Q-V) relationship. This shift is associated with faster outward movements of the gating charge over a broad range of relevant membrane potentials and accelerated gating charge return upon repolarization. In sharp contrast, DPPX-S had no effect on gating charge movements of the Shaker B Kv channel. We propose that DPPX-S destabilizes resting and intermediate states in the voltage-dependent activation pathway, which promotes the outward gating charge movement. The remodeling of gating charge dynamics may involve specific protein–protein interactions of the DPPX-S's transmembrane segment with the voltage-sensing and pore domains of the Kv4.2 channel. This mechanism may determine the characteristic fast operation of neuronal Kv4 channels in the subthreshold range of membrane potentials. PMID:17130523

  13. Cortactin Is Required for N-cadherin Regulation of Kv1.5 Channel Function*

    PubMed Central

    Cheng, Lan; Yung, Aaron; Covarrubias, Manuel; Radice, Glenn L.

    2011-01-01

    The intercalated disc serves as an organizing center for various cell surface components at the termini of the cardiomyocyte, thus ensuring proper mechanoelectrical coupling throughout the myocardium. The cell adhesion molecule, N-cadherin, is an essential component of the intercalated disc. Cardiac-specific deletion of N-cadherin leads to abnormal electrical conduction and sudden arrhythmic death in mice. The mechanisms linking the loss of N-cadherin in the heart and spontaneous malignant ventricular arrhythmias are poorly understood. To investigate whether ion channel remodeling contributes to arrhythmogenesis in N-cadherin conditional knock-out (N-cad CKO) mice, cardiac myocyte excitability and voltage-gated potassium channel (Kv), as well as inwardly rectifying K+ channel remodeling, were investigated in N-cad CKO cardiomyocytes by whole cell patch clamp recordings. Action potential duration was prolonged in N-cad CKO ventricle myocytes compared with wild type. Relative to wild type, IK,slow density was significantly reduced consistent with decreased expression of Kv1.5 and Kv accessory protein, Kcne2, in the N-cad CKO myocytes. The decreased Kv1.5/Kcne2 expression correlated with disruption of the actin cytoskeleton and reduced cortactin at the sarcolemma. Biochemical experiments revealed that cortactin co-immunoprecipitates with Kv1.5. Finally, cortactin was required for N-cadherin-mediated enhancement of Kv1.5 channel activity in a heterologous expression system. Our results demonstrate a novel mechanistic link among the cell adhesion molecule, N-cadherin, the actin-binding scaffold protein, cortactin, and Kv channel remodeling in the heart. These data suggest that in addition to gap junction remodeling, aberrant Kv1.5 channel function contributes to the arrhythmogenic phenotype in N-cad CKO mice. PMID:21507952

  14. Characterization of a fluoroscopic imaging system for kV and MV radiography.

    PubMed

    Drake, D G; Jaffray, D A; Wong, J W

    2000-05-01

    An on-line kilovoltage (kV) imaging system has been implemented on a medical linear accelerator to verify radiotherapy field placement. A kV x-ray tube is mounted on the accelerator at 90 degrees to the megavoltage (MV) source and shares the same isocenter. Nearly identical CCD-based fluoroscopic imagers are mounted opposite the two x-ray sources. These systems are being used in a clinical study of patient setup error that examines the advantage of kV imaging for on-line localization. In the investigation reported here, the imaging performance of the kV and MV systems are characterized to provide support to the conclusions of the studies of setup error. A spatial-frequency-dependent linear systems model is used to predict the detective quantum efficiencies (DQEs) of the two systems. Each is divided into a series of gain and spreading stages. The parameters of each stage are either measured or obtained from the literature. The model predicts the system gain to within 7% of the measured gain for the MV system and to within 10% for the kV system. The systems' noise power spectra (NPSs) and modulation transfer functions (MTFs) are measured to construct the measured DQEs. X-ray fluences are calculated using modeled polyenergetic spectra. Measured DQEs agree well with those predicted by the model. The model reveals that the MV system is well optimized, and is x-ray quantum noise limited at low spatial frequencies. The kV system is suboptimal, but for purposes of patient positioning yields images superior to those produced by the MV system. This is attributed to the kV system's higher DQE and to the inherently higher contrasts present at kV energies. PMID:10841392

  15. Role of Kv7 channels in responses of the pulmonary circulation to hypoxia.

    PubMed

    Sedivy, Vojtech; Joshi, Shreena; Ghaly, Youssef; Mizera, Roman; Zaloudikova, Marie; Brennan, Sean; Novotna, Jana; Herget, Jan; Gurney, Alison M

    2015-01-01

    Hypoxic pulmonary vasoconstriction (HPV) is a beneficial mechanism that diverts blood from hypoxic alveoli to better ventilated areas of the lung, but breathing hypoxic air causes the pulmonary circulation to become hypertensive. Responses to airway hypoxia are associated with depolarization of smooth muscle cells in the pulmonary arteries and reduced activity of K(+) channels. As Kv7 channels have been proposed to play a key role in regulating the smooth muscle membrane potential, we investigated their involvement in the development of HPV and hypoxia-induced pulmonary hypertension. Vascular effects of the selective Kv7 blocker, linopirdine, and Kv7 activator, flupirtine, were investigated in isolated, saline-perfused lungs from rats maintained for 3-5 days in an isobaric hypoxic chamber (FiO2 = 0.1) or room air. Linopirdine increased vascular resistance in lungs from normoxic, but not hypoxic rats. This effect was associated with reduced mRNA expression of the Kv7.4 channel α-subunit in hypoxic arteries, whereas Kv7.1 and Kv7.5 were unaffected. Flupirtine had no effect in normoxic lungs but reduced vascular resistance in hypoxic lungs. Moreover, oral dosing with flupirtine (30 mg/kg/day) prevented short-term in vivo hypoxia from increasing pulmonary vascular resistance and sensitizing the arteries to acute hypoxia. These findings suggest a protective role for Kv7.4 channels in the pulmonary circulation, limiting its reactivity to pressor agents and preventing hypoxia-induced pulmonary hypertension. They also provide further support for the therapeutic potential of Kv7 activators in pulmonary vascular disease. PMID:25361569

  16. Role of Kv7 channels in responses of the pulmonary circulation to hypoxia

    PubMed Central

    Joshi, Shreena; Ghaly, Youssef; Mizera, Roman; Zaloudikova, Marie; Brennan, Sean; Novotna, Jana; Herget, Jan; Gurney, Alison M.

    2014-01-01

    Hypoxic pulmonary vasoconstriction (HPV) is a beneficial mechanism that diverts blood from hypoxic alveoli to better ventilated areas of the lung, but breathing hypoxic air causes the pulmonary circulation to become hypertensive. Responses to airway hypoxia are associated with depolarization of smooth muscle cells in the pulmonary arteries and reduced activity of K+ channels. As Kv7 channels have been proposed to play a key role in regulating the smooth muscle membrane potential, we investigated their involvement in the development of HPV and hypoxia-induced pulmonary hypertension. Vascular effects of the selective Kv7 blocker, linopirdine, and Kv7 activator, flupirtine, were investigated in isolated, saline-perfused lungs from rats maintained for 3–5 days in an isobaric hypoxic chamber (FiO2 = 0.1) or room air. Linopirdine increased vascular resistance in lungs from normoxic, but not hypoxic rats. This effect was associated with reduced mRNA expression of the Kv7.4 channel α-subunit in hypoxic arteries, whereas Kv7.1 and Kv7.5 were unaffected. Flupirtine had no effect in normoxic lungs but reduced vascular resistance in hypoxic lungs. Moreover, oral dosing with flupirtine (30 mg/kg/day) prevented short-term in vivo hypoxia from increasing pulmonary vascular resistance and sensitizing the arteries to acute hypoxia. These findings suggest a protective role for Kv7.4 channels in the pulmonary circulation, limiting its reactivity to pressor agents and preventing hypoxia-induced pulmonary hypertension. They also provide further support for the therapeutic potential of Kv7 activators in pulmonary vascular disease. PMID:25361569

  17. First structure on MoronytoRainbow 100kV Transmission Line below Morony Dam ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    First structure on Morony-to-Rainbow 100kV Transmission Line below Morony Dam and Power House. Three-pole H-frame structure with historic porcelain suspension insulators, jumper supports insulators, overhead ground wires, and pole stubs. View to east-northeast - Morony Hydroelectric Facility, Morony-to-Rainbow 100 kV Transmission Line, West bank of the Missouri River, Great Falls, Cascade County, MT

  18. Thalamic Kv7 channels: pharmacological properties and activity control during noxious signal processing

    PubMed Central

    Cerina, Manuela; Szkudlarek, Hanna J; Coulon, Philippe; Meuth, Patrick; Kanyshkova, Tatyana; Nguyen, Xuan Vinh; Göbel, Kerstin; Seidenbecher, Thomas; Meuth, Sven G; Pape, Hans-Christian; Budde, Thomas

    2015-01-01

    Background and Purpose The existence of functional Kv7 channels in thalamocortical (TC) relay neurons and the effects of the K+-current termed M-current (IM) on thalamic signal processing have long been debated. Immunocytochemical evidence suggests their presence in this brain region. Therefore, we aimed to verify their existence, pharmacological properties and function in regulating activity in neurons of the ventrobasal thalamus (VB). Experimental Approach Characterization of Kv7 channels was performed by combining in vitro, in vivo and in silico techniques with a pharmacological approach. Retigabine (30 μM) and XE991 (20 μM), a specific Kv7 channel enhancer and blocker, respectively, were applied in acute brain slices during electrophysiological recordings. The effects of intrathalamic injection of retigabine (3 mM, 300 nL) and/or XE991 (2 mM, 300 nL) were investigated in freely moving animals during hot-plate tests by recording behaviour and neuronal activity. Key Results Kv7.2 and Kv7.3 subunits were found to be abundantly expressed in TC neurons of mouse VB. A slow K+-current with properties of IM was activated by retigabine and inhibited by XE991. Kv7 channel activation evoked membrane hyperpolarization, a reduction in tonic action potential firing, and increased burst firing in vitro and in computational models. Single-unit recordings and pharmacological intervention demonstrated a specific burst-firing increase upon IM activation in vivo. A Kv7 channel-mediated increase in pain threshold was associated with fewer VB units responding to noxious stimuli, and increased burst firing in responsive neurons. Conclusions and Implications Kv7 channel enhancement alters somatosensory activity and may reflect an anti-nociceptive mechanism during acute pain processing. PMID:25684311

  19. 29 CFR 1926.1409 - Power line safety (over 350 kV).

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... professional engineer who is a qualified person with respect to electrical power transmission and distribution. ... 29 Labor 8 2014-07-01 2014-07-01 false Power line safety (over 350 kV). 1926.1409 Section 1926... Construction § 1926.1409 Power line safety (over 350 kV). The requirements of § 1926.1407 and § 1926.1408...

  20. 29 CFR 1926.1409 - Power line safety (over 350 kV).

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... professional engineer who is a qualified person with respect to electrical power transmission and distribution. ... 29 Labor 8 2013-07-01 2013-07-01 false Power line safety (over 350 kV). 1926.1409 Section 1926... Construction § 1926.1409 Power line safety (over 350 kV). The requirements of § 1926.1407 and § 1926.1408...

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

    PubMed

    Bilet, Arne; Bauer, Christiane K

    2012-01-01

    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. PMID:23226420

  2. Distinct Cell- and Layer-Specific Expression Patterns and Independent Regulation of Kv2 Channel Subtypes in Cortical Pyramidal Neurons

    PubMed Central

    Bishop, Hannah I.; Guan, Dongxu; Bocksteins, Elke; Parajuli, Laxmi Kumar; Murray, Karl D.; Cobb, Melanie M.; Misonou, Hiroaki; Zito, Karen; Foehring, Robert C.

    2015-01-01

    The Kv2 family of voltage-gated potassium channel α subunits, comprising Kv2.1 and Kv2.2, mediate the bulk of the neuronal delayed rectifier K+ current in many mammalian central neurons. Kv2.1 exhibits robust expression across many neuron types and is unique in its conditional role in modulating intrinsic excitability through changes in its phosphorylation state, which affect Kv2.1 expression, localization, and function. Much less is known of the highly related Kv2.2 subunit, especially in forebrain neurons. Here, through combined use of cortical layer markers and transgenic mouse lines, we show that Kv2.1 and Kv2.2 are localized to functionally distinct cortical cell types. Kv2.1 expression is consistently high throughout all cortical layers, especially in layer (L) 5b pyramidal neurons, whereas Kv2.2 expression is primarily limited to neurons in L2 and L5a. In addition, L4 of primary somatosensory cortex is strikingly devoid of Kv2.2 immunolabeling. The restricted pattern of Kv2.2 expression persists in Kv2.1-KO mice, suggesting distinct cell- and layer-specific functions for these two highly related Kv2 subunits. Analyses of endogenous Kv2.2 in cortical neurons in situ and recombinant Kv2.2 expressed in heterologous cells reveal that Kv2.2 is largely refractory to stimuli that trigger robust, phosphorylation-dependent changes in Kv2.1 clustering and function. Immunocytochemistry and voltage-clamp recordings from outside-out macropatches reveal distinct cellular expression patterns for Kv2.1 and Kv2.2 in intratelencephalic and pyramidal tract neurons of L5, indicating circuit-specific requirements for these Kv2 paralogs. Together, these results support distinct roles for these two Kv2 channel family members in mammalian cortex. SIGNIFICANCE STATEMENT Neurons within the neocortex are arranged in a laminar architecture and contribute to the input, processing, and/or output of sensory and motor signals in a cell- and layer-specific manner. Neurons of different

  3. Kv3.1 uses a timely resurgent K+ current to secure action potential repolarization

    PubMed Central

    Labro, Alain J.; Priest, Michael F.; Lacroix, Jérôme J.; Snyders, Dirk J.; Bezanilla, Francisco

    2015-01-01

    High-frequency action potential (AP) transmission is essential for rapid information processing in the central nervous system. Voltage-dependent Kv3 channels play an important role in this process thanks to their high activation threshold and fast closure kinetics, which reduce the neuron's refractory period. However, premature Kv3 channel closure leads to incomplete membrane repolarization, preventing sustainable AP propagation. Here, we demonstrate that Kv3.1b channels solve this problem by producing resurgent K+ currents during repolarization, thus ensuring enough repolarizing power to terminate each AP. Unlike previously described resurgent Na+ and K+ currents, Kv3.1b's resurgent current does not originate from recovery of channel block or inactivation but results from a unique combination of steep voltage-dependent gating kinetics and ultra-fast voltage-sensor relaxation. These distinct properties are readily transferrable onto an orthologue Kv channel by transplanting the voltage-sensor's S3–S4 loop, providing molecular insights into the mechanism by which Kv3 channels contribute to high-frequency AP transmission. PMID:26673941

  4. Elm Creek: NSP establishes new 345 kV substation in Twin Cities area

    SciTech Connect

    Ray, B.; Spaulding, P.; Olson, D.

    1996-11-01

    Northern States Power Company (NSP) has proposed to expand the existing Elm Creek substation located in the City of Maple Grove for 345 kV development. The planned expansion will not require acquisition of any new 345 kV transmission line right-of-way. The northwestern metropolitan area of Twin Cities (Minneapolis/St. Paul) has been experiencing significant growth in the residential, commercial and industrial sectors. This necessitates substantial transmission and distribution system improvements in the near future involving millions of dollars of capital investment, to ensure adequate load serving capability and maintain the desired reliability. Relaxing thermal transmission limitations in this area will be achieved by strategic establishment of a new 345 kV source at the existing Elm Creek site in year 2000. The central location of the station in relation to the area experiencing rapid load development makes it an excellent choice for the desired 345 kV source establishment. One 115 kV circuit from Elm Creek to Champlin substation is being constructed in 1997 which will act as an additional transmission outlet from the 345 kV source. Recently completed Long-Range Geographic Planning Study results strongly reinforce the need for such prudent improvements.

  5. Induction of stable ER–plasma-membrane junctions by Kv2.1 potassium channels

    PubMed Central

    Fox, Philip D.; Haberkorn, Christopher J.; Akin, Elizabeth J.; Seel, Peter J.; Krapf, Diego; Tamkun, Michael M.

    2015-01-01

    ABSTRACT Junctions between cortical endoplasmic reticulum (cER) and the plasma membrane are a subtle but ubiquitous feature in mammalian cells; however, very little is known about the functions and molecular interactions that are associated with neuronal ER–plasma-membrane junctions. Here, we report that Kv2.1 (also known as KCNB1), the primary delayed-rectifier K+ channel in the mammalian brain, induces the formation of ER–plasma-membrane junctions. Kv2.1 localizes to dense, cell-surface clusters that contain non-conducting channels, indicating that they have a function that is unrelated to membrane-potential regulation. Accordingly, Kv2.1 clusters function as membrane-trafficking hubs, providing platforms for delivery and retrieval of multiple membrane proteins. Using both total internal reflection fluorescence and electron microscopy we demonstrate that the clustered Kv2.1 plays a direct structural role in the induction of stable ER–plasma-membrane junctions in both transfected HEK 293 cells and cultured hippocampal neurons. Glutamate exposure results in a loss of Kv2.1 clusters in neurons and subsequent retraction of the cER from the plasma membrane. We propose Kv2.1-induced ER–plasma-membrane junctions represent a new macromolecular plasma-membrane complex that is sensitive to excitotoxic insult and functions as a scaffolding site for both membrane trafficking and Ca2+ signaling. PMID:25908859

  6. The Role of Kv1.2 Channel in Electrotaxis Cell Migration.

    PubMed

    Zhang, Gaofeng; Edmundson, Mathew; Telezhkin, Vsevolod; Gu, Yu; Wei, Xiaoqing; Kemp, Paul J; Song, Bing

    2016-06-01

    Voltage-gated potassium Kv1.2 channels play pivotal role in maintaining of resting membrane potential and, consequently, regulation of cellular excitability of neurons. Endogenously generated electric field (EF) have been proven as an important regulator for cell migration and tissue repair. The mechanisms of ion channel involvement in EF-induced cell responses are extensively studied but largely are poorly understood. In this study we generated three COS-7 clones with different expression levels of Kv1.2 channel, and confirmed their functional variations with patch clamp analysis. Time-lapse imaging analysis showed that EF-induced cell migration response was Kv1.2 channel expression level depended. Inhibition of Kv1.2 channels with charybdotoxin (ChTX) constrained the sensitivity of COS-7 cells to EF stimulation more than their motility. Immunocytochemistry and pull-down analyses demonstrated association of Kv1.2 channels with actin-binding protein cortactin and its re-localization to the cathode-facing membrane at EF stimulation, which confirms the mechanism of EF-induced directional migration. This study displays that Kv1.2 channels represent an important physiological link in EF-induced cell migration. The described mechanism suggests a potential application of EF which may improve therapeutic performance in curing injuries of neuronal and/or cardiac tissue repair, post operational therapy, and various degenerative syndromes. PMID:26580832

  7. Physiological modulators of Kv3.1 channels adjust firing patterns of auditory brain stem neurons.

    PubMed

    Brown, Maile R; El-Hassar, Lynda; Zhang, Yalan; Alvaro, Giuseppe; Large, Charles H; Kaczmarek, Leonard K

    2016-07-01

    Many rapidly firing neurons, including those in the medial nucleus of the trapezoid body (MNTB) in the auditory brain stem, express "high threshold" voltage-gated Kv3.1 potassium channels that activate only at positive potentials and are required for stimuli to generate rapid trains of actions potentials. We now describe the actions of two imidazolidinedione derivatives, AUT1 and AUT2, which modulate Kv3.1 channels. Using Chinese hamster ovary cells stably expressing rat Kv3.1 channels, we found that lower concentrations of these compounds shift the voltage of activation of Kv3.1 currents toward negative potentials, increasing currents evoked by depolarization from typical neuronal resting potentials. Single-channel recordings also showed that AUT1 shifted the open probability of Kv3.1 to more negative potentials. Higher concentrations of AUT2 also shifted inactivation to negative potentials. The effects of lower and higher concentrations could be mimicked in numerical simulations by increasing rates of activation and inactivation respectively, with no change in intrinsic voltage dependence. In brain slice recordings of mouse MNTB neurons, both AUT1 and AUT2 modulated firing rate at high rates of stimulation, a result predicted by numerical simulations. Our results suggest that pharmaceutical modulation of Kv3.1 currents represents a novel avenue for manipulation of neuronal excitability and has the potential for therapeutic benefit in the treatment of hearing disorders. PMID:27052580

  8. 26Al measurements below 500 kV in charge state 2+

    NASA Astrophysics Data System (ADS)

    Müller, Arnold Milenko; Christl, Marcus; Lachner, Johannes; Synal, Hans-Arno; Vockenhuber, Christof; Zanella, Claudia

    2015-10-01

    The use of helium as stripper gas improved the measurement efficiency of compact AMS systems for many radionuclides significantly because of a higher mean charge state and reduced scattering losses compared with other conventional gases. Recent tests at the ETH 500 kV AMS facility (Tandy) with aluminum have demonstrated that a transmission of more than 50% is achievable in the charge state 2+ at terminal voltages between 300 and 500 kV. On the other hand the m/q interference of 13C1+ entering the detector at very high intensity has to be suppressed. Based on first positive results with a very simple absorber cell a more elaborate absorber detector configuration was designed and built in order to eliminate the carbon interference. The suppression of carbon with the new detector-absorber design has been studied extensively at 300 kV (950 keV) and 500 kV (1550 keV) and the results are compared with simulated data. With the new configuration an overall transmission for 26Al of more than 42% at 500 kV and about 30% at 300 kV terminal voltage is achieved, while 26Al/27Al blank ratios of aluminum targets in the range of 5-14·10-15 are measured.

  9. Kv3.1 uses a timely resurgent K(+) current to secure action potential repolarization.

    PubMed

    Labro, Alain J; Priest, Michael F; Lacroix, Jérôme J; Snyders, Dirk J; Bezanilla, Francisco

    2015-01-01

    High-frequency action potential (AP) transmission is essential for rapid information processing in the central nervous system. Voltage-dependent Kv3 channels play an important role in this process thanks to their high activation threshold and fast closure kinetics, which reduce the neuron's refractory period. However, premature Kv3 channel closure leads to incomplete membrane repolarization, preventing sustainable AP propagation. Here, we demonstrate that Kv3.1b channels solve this problem by producing resurgent K(+) currents during repolarization, thus ensuring enough repolarizing power to terminate each AP. Unlike previously described resurgent Na(+) and K(+) currents, Kv3.1b's resurgent current does not originate from recovery of channel block or inactivation but results from a unique combination of steep voltage-dependent gating kinetics and ultra-fast voltage-sensor relaxation. These distinct properties are readily transferrable onto an orthologue Kv channel by transplanting the voltage-sensor's S3-S4 loop, providing molecular insights into the mechanism by which Kv3 channels contribute to high-frequency AP transmission. PMID:26673941

  10. Induction of stable ER-plasma-membrane junctions by Kv2.1 potassium channels.

    PubMed

    Fox, Philip D; Haberkorn, Christopher J; Akin, Elizabeth J; Seel, Peter J; Krapf, Diego; Tamkun, Michael M

    2015-06-01

    Junctions between cortical endoplasmic reticulum (cER) and the plasma membrane are a subtle but ubiquitous feature in mammalian cells; however, very little is known about the functions and molecular interactions that are associated with neuronal ER-plasma-membrane junctions. Here, we report that Kv2.1 (also known as KCNB1), the primary delayed-rectifier K(+) channel in the mammalian brain, induces the formation of ER-plasma-membrane junctions. Kv2.1 localizes to dense, cell-surface clusters that contain non-conducting channels, indicating that they have a function that is unrelated to membrane-potential regulation. Accordingly, Kv2.1 clusters function as membrane-trafficking hubs, providing platforms for delivery and retrieval of multiple membrane proteins. Using both total internal reflection fluorescence and electron microscopy we demonstrate that the clustered Kv2.1 plays a direct structural role in the induction of stable ER-plasma-membrane junctions in both transfected HEK 293 cells and cultured hippocampal neurons. Glutamate exposure results in a loss of Kv2.1 clusters in neurons and subsequent retraction of the cER from the plasma membrane. We propose Kv2.1-induced ER-plasma-membrane junctions represent a new macromolecular plasma-membrane complex that is sensitive to excitotoxic insult and functions as a scaffolding site for both membrane trafficking and Ca(2+) signaling. PMID:25908859

  11. Kv1.1 channels act as mechanical brake in the senses of touch and pain.

    PubMed

    Hao, Jizhe; Padilla, Françoise; Dandonneau, Mathieu; Lavebratt, Catharina; Lesage, Florian; Noël, Jacques; Delmas, Patrick

    2013-03-01

    Molecular determinants of threshold sensitivity of mammalian mechanoreceptors are unknown. Here, we identify a mechanosensitive (MS) K(+) current (IKmech) that governs mechanical threshold and adaptation of distinct populations of mechanoreceptors. Toxin profiling and transgenic mouse studies indicate that IKmech is carried by Kv1.1-Kv1.2 heteromers. Mechanosensitivity is attributed to Kv1.1 subunits, through facilitation of voltage-dependent open probability. IKmech is expressed in high-threshold C-mechano-nociceptors (C-HTMRs) and Aβ-mechanoreceptors, but not in low-threshold C-mechanoreceptors. IKmech opposes depolarization induced by slow/ultraslow MS cation currents in C-HTMRs, thereby shifting mechanical threshold for firing to higher values. However, due to kinetics mismatch with rapidly-adapting MS cation currents, IKmech tunes firing adaptation but not mechanical threshold in Aβ-mechanoreceptors. Expression of Kv1.1 dominant negative or inhibition of Kv1.1/IKmech caused severe mechanical allodynia but not heat hyperalgesia. By balancing the activity of excitatory mechanotransducers, Kv1.1 acts as a mechanosensitive brake that regulates mechanical sensitivity of fibers associated with mechanical perception. PMID:23473320

  12. Molecular identification of Kvα subunits that contribute to the oxygen-sensitive K+ current of chemoreceptor cells of the rabbit carotid body

    PubMed Central

    Sanchez, Diego; López-López, Jose R; Pérez-García, M Teresa; Sanz-Alfayate, Gloria; Obeso, Ana; Ganfornina, Maria D; Gonzalez, Constancio

    2002-01-01

    Rabbit carotid body (CB) chemoreceptor cells possess a fast-inactivating K+ current that is specifically inhibited by hypoxia. We have studied the expression of Kvα subunits, which might be responsible for this current. RT-PCR experiments identified the expression of Kv1.4, Kv3.4, Kv4.1 and Kv4.3 mRNAs in the rabbit CB. There was no expression of Kv3.3 or Kv4.2 transcripts. Immunocytochemistry with antibodies to tyrosine hydroxylase (anti-TH) and to specific Kv subunits revealed the expression of Kv3.4 and Kv4.3 in chemoreceptor cells, while Kv1.4 was only found in nerve fibres. Kv4.1 mRNA was also found in chemoreceptor cells following in situ hybridization combined with anti-TH antibody labelling. Kv4.1 and Kv4.3 appeared to be present in all chemoreceptor cells, but Kv3.4 was only expressed in a population of them. Electrophysiological experiments applying specific toxins or antibodies demonstrated that both Kv3.4 and Kv4.3 participate in the oxygen-sensitive K+ current of chemoreceptor cells. However, toxin application experiments confirmed a larger contribution of members of the Kv4 subfamily. [Ca2+]i measurements under hypoxic conditions and immunocytochemistry experiments in dispersed CB cells demonstrated the expression of Kv3.4 and Kv4.3 in oxygen-sensitive cells; the presence of Kv3.4 in the chemoreceptor cell membrane was not required for the response to low PO2. In summary, three Kv subunits (Kv3.4, Kv4.1 and Kv4.3) may be involved in the fast-inactivating outward K+ current of rabbit CB chemoreceptor cells. The homogeneous distribution of the Kv4 subunits in chemoreceptor cells, along with their electrophysiological properties, suggest that Kv4.1, Kv4.3, or their heteromultimers, are the molecular correlate of the oxygen-sensitive K+ channel. PMID:12122138

  13. KV7 channels regulate muscle tone and nonadrenergic noncholinergic relaxation of the rat gastric fundus.

    PubMed

    Ipavec, V; Martire, M; Barrese, V; Taglialatela, M; Currò, D

    2011-10-01

    Voltage-dependent type 7 K+ (KV7) channels play important physiological roles in neurons and muscle cells. The aims of the present study were to investigate the motor effects of KV7 channel modulators in the rat gastric fundus and the expression of KV7 channels in this tissue. Muscle tone and electrical field stimulation (EFS)-evoked relaxations of precontracted longitudinal muscle strips of the rat gastric fundus were investigated under nonadrenergic noncholinergic conditions by organ bath studies. Gene expression was studied by real-time PCR and tissue localization of channels was investigated by immunohistochemistry. The KV7 channel blocker XE-991 induced concentration-dependent contractions, with mean pD2 and Emax of 5.4 and 48% of the maximal U46619-induced contraction, respectively. The KV7 channel activators retigabine and flupirtine concentration-dependently relaxed U46619-precontracted strips, with pD2s of 4.7 and 4.4 and Emax of 93% and 91% of the maximal relaxation induced by papaverine, respectively. XE-991 concentration-dependently inhibited retigabine-induced relaxation with a pIC50 of 6.2. XE-991 and DMP-543, another KV7 channel blocker, increased by 13-25% or reduced by 11-21% the relaxations evoked by low- or high-frequency EFS, respectively. XE-991 also reduced the relaxation induced by vasoactive intestinal polypeptide (VIP) by 33% of controls. Transcripts encoded by all KV7 genes were detected in the fundus, with 7.4 and 7.5 showing the highest expression levels. KV7.4 and 7.5 channels were visualized by confocal immunofluorescence in both circular and longitudinal muscle layers. In conclusion, in the rat proximal stomach, KV7 channels appear to contribute to the resting muscle tone and to VIP- and high-frequency EFS-induced relaxation. KV7 channel activators could be useful relaxant agents of the gastric smooth muscle. PMID:21740972

  14. Kv8.1, a new neuronal potassium channel subunit with specific inhibitory properties towards Shab and Shaw channels.

    PubMed Central

    Hugnot, J P; Salinas, M; Lesage, F; Guillemare, E; de Weille, J; Heurteaux, C; Mattéi, M G; Lazdunski, M

    1996-01-01

    Outward rectifier K+ channels have a characteristic structure with six transmembrane segments and one pore region. A new member of this family of transmembrane proteins has been cloned and called Kv8.1. Kv8.1 is essentially present in the brain where it is located mainly in layers II, IV and VI of the cerebral cortex, in hippocampus, in CA1-CA4 pyramidal cell layer as well in granule cells of the dentate gyrus, in the granule cell layer and in the Purkinje cell layer of the cerebellum. The Kv8.1 gene is in the 8q22.3-8q24.1 region of the human genome. Although Kv8.1 has the hallmarks of functional subunits of outward rectifier K+ channels, injection of its cRNA in Xenopus oocytes does not produce K+ currents. However Kv8.1 abolishes the functional expression of members of the Kv2 and Kv3 subfamilies, suggesting that the functional role of Kv8.1 might be to inhibit the function of a particular class of outward rectifier K+ channel types. Immunoprecipitation studies have demonstrated that inhibition occurs by formation of heteropolymeric channels, and results obtained with Kv8.1 chimeras have indicated that association of Kv8.1 with other types of subunits is via its N-terminal domain. Images PMID:8670833

  15. Affinity and selectivity of ShK toxin for the Kv1 potassium channels from free energy simulations.

    PubMed

    Rashid, M Harunur; Kuyucak, Serdar

    2012-04-26

    The voltage-gated potassium channel Kv1.3 is an attractive target for treatment of autoimmune diseases. ShK toxin from sea anemone is one of the most potent blockers of Kv1.3, and therefore ShK and its analogues have been proposed as therapeutic leads for such diseases. Increasing the selectivity of the proposed leads for Kv1.3 over other Kv1 channels is a major issue in this endeavor. Here we study binding of ShK toxin to Kv1 channels using free energy simulation methods. Homology models for Kv1.1 and Kv1.3 channels are constructed using the crystal structure of Kv1.2. The initial poses for the Kv1.x-ShK complexes are obtained using HADDOCK, which are then refined via molecular dynamics simulations. The binding mode in each complex is characterized by identifying the strongly interacting residues, which compare well with available mutagenesis studies. For each complex, the potential of mean force is calculated from umbrella sampling simulations, and the corresponding absolute binding free energy is determined. The computed binding free energies are in good agreement with the experimental data, which increases the confidence on the model complexes. The insights gained on Kv1.x-ShK binding modes will be valuable in the development of new ShK analogues with better selectivity properties. PMID:22480371

  16. Kv4 Channels Underlie the Subthreshold-Operating A-type K+-current in Nociceptive Dorsal Root Ganglion Neurons

    PubMed Central

    Phuket, Thanawath Ratanadilok Na; Covarrubias, Manuel

    2009-01-01

    The dorsal root ganglion (DRG) contains heterogeneous populations of sensory neurons including primary nociceptive neurons and C-fibers implicated in pain signaling. Recent studies have demonstrated DRG hyperexcitability associated with downregulation of A-type K+ channels; however, the molecular correlate of the corresponding A-type K+ current (IA) has remained hypothetical. Kv4 channels may underlie the IA in DRG neurons. We combined electrophysiology, molecular biology (Whole-Tissue and Single-Cell RT-PCR) and immunohistochemistry to investigate the molecular basis of the IA in acutely dissociated DRG neurons from 7- to 8-day-old rats. Whole-cell recordings demonstrate a robust tetraethylammonium-resistant (20 mM) and 4-aminopyridine-sensitive (5 mM) IA. Matching Kv4 channel properties, activation and inactivation of this IA occur in the subthreshold range of membrane potentials and the rate of recovery from inactivation is rapid and voltage-dependent. Among Kv4 transcripts, the DRG expresses significant levels of Kv4.1 and Kv4.3 mRNAs. Also, single small-medium diameter DRG neurons (∼30 μm) exhibit correlated frequent expression of mRNAs encoding Kv4.1 and Nav1.8, a known nociceptor marker. In contrast, the expressions of Kv1.4 and Kv4.2 mRNAs at the whole-tissue and single-cell levels are relatively low and infrequent. Kv4 protein expression in nociceptive DRG neurons was confirmed by immunohistochemistry, which demonstrates colocalization of Kv4.3 and Nav1.8, and negligible expression of Kv4.2. Furthermore, specific dominant-negative suppression and overexpression strategies confirmed the contribution of Kv4 channels to IA in DRG neurons. Contrasting the expression patterns of Kv4 channels in the central and peripheral nervous systems, we discuss possible functional roles of these channels in primary sensory neurons. PMID:19668710

  17. Functional and molecular evidence for Kv7 channel subtypes in human detrusor from patients with and without bladder outflow obstruction.

    PubMed

    Svalø, Julie; Sheykhzade, Majid; Nordling, Jørgen; Matras, Christina; Bouchelouche, Pierre

    2015-01-01

    The aim of the study was to investigate whether Kv7 channels and their ancillary β-subunits, KCNE, are functionally expressed in the human urinary bladder. Kv7 channels were examined at the molecular level and by functional studies using RT-qPCR and myography, respectively. We found mRNA expression of KCNQ1, KCNQ3-KCNQ5 and KCNE1-5 in the human urinary bladder from patients with normal bladder function (n = 7) and in patients with bladder outflow obstruction (n = 3). Interestingly, a 3.4-fold up-regulation of KCNQ1 was observed in the latter. The Kv7 channel subtype selective modulators, ML277 (activator of Kv7.1 channels, 10 μM) and ML213 (activator of Kv7.2, Kv7.4, Kv7.4/7.5 and Kv7.5 channels, 10 μM), reduced the tone of 1 μM carbachol pre-constricted bladder strips. XE991 (blocker of Kv7.1-7.5 channels, 10 μM) had opposing effects as it increased contractions achieved with 20 mM KPSS. Furthermore, we investigated if there is interplay between Kv7 channels and β-adrenoceptors. Using cumulative additions of isoprenaline (β-adrenoceptor agonist) and forskolin (adenylyl cyclase activator) in combination with the Kv7 channel activator and blocker, retigabine and XE991, we did not find interplay between Kv7 channels and β-adrenoceptors in the human urinary bladder. The performed gene expression analysis combined with the organ bath studies imply that compounds that activate Kv7 channels could be useful for treatment of overactive bladder syndrome. PMID:25692982

  18. High reliability low jitter 80 kV pulse generator.

    SciTech Connect

    Savage, Mark Edward; Stoltzfus, Brian Scott

    2009-06-01

    Switching can be considered to be the essence of pulsed power. Time accurate switch/trigger systems with low inductance are useful in many applications. This article describes a unique switch geometry coupled with a low-inductance capacitive energy store. The system provides a fast-rising high voltage pulse into a low impedance load. It can be challenging to generate high voltage (more than 50 kilovolts) into impedances less than 10 {Omega}, from a low voltage control signal with a fast rise time and high temporal accuracy. The required power amplification is large, and is usually accomplished with multiple stages. The multiple stages can adversely affect the temporal accuracy and the reliability of the system. In the present application, a highly reliable and low jitter trigger generator was required for the Z pulsed-power facility [M. E. Savage, L. F. Bennett, D. E. Bliss, W. T. Clark, R. S. Coats,J. M. Elizondo, K. R. LeChien, H. C. Harjes, J. M. Lehr, J. E. Maenchen, D. H. McDaniel, M. F. Pasik, T. D. Pointon, A. C. Owen, D. B. Seidel, D. L. Smith, B. S. Stoltzfus, K.W. Struve, W.A. Stygar, L.K. Warne, and J. R. Woodworth, 2007 IEEE Pulsed Power Conference, Albuquerque, NM (IEEE, Piscataway, NJ, 2007), p. 979]. The large investment in each Z experiment demands low prefire probability and low jitter simultaneously. The system described here is based on a 100 kV DC-charged high-pressure spark gap, triggered with an ultraviolet laser. The system uses a single optical path for simultaneously triggering two parallel switches, allowing lower inductance and electrode erosion with a simple optical system. Performance of the system includes 6 ns output rise time into 5.6 {Omega}, 550 ps one-sigma jitter measured from the 5 V trigger to the high voltage output, and misfire probability less than 10{sup -4}. The design of the system and some key measurements will be shown in the paper. We will discuss the design goals related to high reliability and low jitter. While

  19. Effects of dapoxetine on cloned Kv1.5 channels expressed in CHO cells.

    PubMed

    Jeong, Imju; Yoon, Shin Hee; Hahn, Sang June

    2012-07-01

    The effects of dapoxetine were examined on cloned Kv1.5 channels stably expressed in Chinese hamster ovary cells using the whole-cell patch clamp technique. Dapoxetine decreased the peak amplitude of Kv1.5 currents and accelerated the decay rate of current inactivation in a concentration-dependent manner with an IC ( 50 ) of 11.6 μM. Kinetic analysis of the time-dependent effects of dapoxetine on Kv1.5 current decay yielded the apparent association (k (+1 )) and dissociation (k (-1 )) rate constants of 2.8 μM(-1) s(-1) and 34.2 s(-1), respectively. The theoretical K ( D ) value, derived by k (-1 )/k (+1 ), yielded 12.3 μM, which was reasonably similar to the IC ( 50 ) value obtained from the concentration-response curve. Dapoxetine decreased the tail current amplitude and slowed the deactivation process of Kv1.5, which resulted in a tail crossover phenomenon. The block by dapoxetine is voltage-dependent and steeply increased at potentials between -10 and +10 mV, which correspond to the voltage range of channel activation. At more depolarized potentials, a weaker voltage dependence was observed (δ=0.31). Dapoxetine had no effect on the steady-state activation of Kv1.5 but shifted the steady-state inactivation curves in a hyperpolarizing direction. Dapoxetine produced a use-dependent block of Kv1.5 at frequencies of 1 and 2 Hz and slowed the time course for recovery of inactivation. These effects were reversible after washout of the drug. Our results indicate that dapoxetine blocks Kv1.5 currents by interacting with the channel in both the open and inactivated states of the channel. PMID:22538641

  20. Blockade of Kv1.5 by paroxetine, an antidepressant drug

    PubMed Central

    Lee, Hyang Mi; Hahn, Sang June

    2016-01-01

    Paroxetine, a selective serotonin reuptake inhibitor (SSRI), has been reported to have an effect on several ion channels including human ether-a-go-go-related gene in a SSRI-independent manner. These results suggest that paroxetine may cause side effects on cardiac system. In this study, we investigated the effect of paroxetine on Kv1.5, which is one of cardiac ion channels. The action of paroxetine on the cloned neuronal rat Kv1.5 channels stably expressed in Chinese hamster ovary cells was investigated using the whole-cell patch-clamp technique. Paroxetine reduced Kv1.5 whole-cell currents in a reversible concentration-dependent manner, with an IC50 value and a Hill coefficient of 4.11 µM and 0.98, respectively. Paroxetine accelerated the decay rate of inactivation of Kv1.5 currents without modifying the kinetics of current activation. The inhibition increased steeply between -30 and 0 mV, which corresponded with the voltage range for channel opening. In the voltage range positive to 0 mV, inhibition displayed a weak voltage dependence, consistent with an electrical distance δ of 0.32. The binding (k+1) and unbinding (k-1) rate constants for paroxetine-induced block of Kv1.5 were 4.9 µM-1s-1 and 16.1 s-1, respectively. The theoretical KD value derived by k-1/k+1 yielded 3.3 µM. Paroxetine slowed the deactivation time course, resulting in a tail crossover phenomenon when the tail currents, recorded in the presence and absence of paroxetine, were superimposed. Inhibition of Kv1.5 by paroxetine was use-dependent. The present results suggest that paroxetine acts on Kv1.5 currents as an open-channel blocker. PMID:26807026

  1. Determinants of frequency-dependent regulation of Kv1.2-containing potassium channels

    PubMed Central

    Baronas, Victoria A.; Yang, Runying; Vilin, Yury Y.; Kurata, Harley T.

    2016-01-01

    ABSTRACT Voltage-gated potassium channels are important regulators of electrical excitation in many tissues, with Kv1.2 standing out as an essential contributor in the CNS. Genetic deletion of Kv1.2 invariably leads to early lethality in mice. In humans, mutations affecting Kv1.2 function are linked to epileptic encephalopathy and movement disorders. We have demonstrated that Kv1.2 is subject to a unique regulatory mechanism in which repetitive stimulation leads to dramatic potentiation of current. In this study, we explore the properties and molecular determinants of this use-dependent potentiation/activation. First, we examine how alterations in duty cycle (depolarization and repolarization/recovery times) affect the onset and extent of use-dependent activation. Also, we use trains of repetitive depolarizations to test the effects of a variety of Thr252 (S2-S3 linker) mutations on use-dependent activation. Substitutions of Thr with some sterically similar amino acids (Ser, Val, and Met, but not Cys) retain use-dependent activation, while bulky or charged amino acid substitutions eliminate use-dependence. Introduction of Thr at the equivalent position in other Kv1 channels (1.1, 1.3, 1.4), was not sufficient to transfer the phenotype. We hypothesize that use-dependent activation of Kv1.2 channels is mediated by an extrinsic regulator that binds preferentially to the channel closed state, with Thr252 being necessary but not sufficient for this interaction to alter channel function. These findings extend the conclusions of our recent demonstration of use-dependent activation of Kv1.2-containing channels in hippocampal neurons, by adding new details about the molecular mechanism underlying this effect. PMID:26646078

  2. DPP6 Localization in Brain Supports Function as a Kv4 Channel Associated Protein

    PubMed Central

    Clark, Brian D.; Kwon, Elaine; Maffie, Jon; Jeong, Hyo-Young; Nadal, Marcela; Strop, Pavel; Rudy, Bernardo

    2008-01-01

    The gene encoding the dipeptidyl peptidase-like protein DPP6 (also known as DPPX) has been associated with human neural disease. However, until recently no function had been found for this protein. It has been proposed that DPP6 is an auxiliary subunit of neuronal Kv4 K+ channels, the ion channels responsible for the somato-dendritic A-type K+ current, an ionic current with crucial roles in the regulation of firing frequency, dendritic integration and synaptic plasticity. This view has been supported mainly by studies showing that DPP6 is necessary to generate channels with biophysical properties resembling the native channels in some neurons. However, independent evidence that DPP6 is a component of neuronal Kv4 channels in the brain, and whether this protein has other functions in the CNS is still lacking. We generated antibodies to DPP6 proteins to compare their distribution in brain with that of the Kv4 pore-forming subunits. DPP6 proteins were prominently expressed in neuronal populations expressing Kv4.2 proteins and both types of protein were enriched in the dendrites of these cells, strongly supporting the hypothesis that DPP6 is an associated protein of Kv4 channels in brain neurons. The observed similarity in the cellular and subcellular patterns of expression of both proteins suggests that this is the main function of DPP6 in brain. However, we also found that DPP6 antibodies intensely labeled the hippocampal mossy fiber axons, which lack Kv4 proteins, suggesting that DPP6 proteins may have additional, Kv4-unrelated functions. PMID:18978958

  3. Marys Lake 69/115-kV transmission line upgrade and substation expansion projects

    SciTech Connect

    1996-05-01

    Western Area Power Administration (Western) and the Platte River Power Authority (Platte River) propose to upgrade portions of the existing electric transmission and substation system that serves the Town of Estes Park, Colorado. The existing transmission lines between the Estes Power Plant Switchyard and the Marys Lake Substation include a 115,000 volt (115-kV) line and 69,000 volt (69-kV) line. Approximately one mile is a double-circuit 115/69-kV line on steel lattice structures, and approximately two miles consists of separate single-circuit 115-kV and a 69-kV lines, constructed on wood H-Frame structures. Both lines were constructed in 1951 by the US Bureau of Reclamation. The existing transmission lines are on rights-of-way (ROW) that vary from 75 feet to 120 feet and are owned by Western. There are 48 landowners adjacent to the existing ROW. All of the houses were built adjacent to the existing ROW after the transmission lines were constructed. Upgrading the existing 69-kV transmission line between the Marys Lake Substation and the Estes Power Plant Switchyard to 115-kV and expanding the Marys Lake Substation was identified as the most effective way in which to improve electric service to Estes Park. The primary purpose and need of the proposed project is to improve the reliability of electric service to the Town of Estes Park. Lack of reliability has been a historical concern, and reliability will always be less than desired until physical improvements are made to the electrical facilities serving Estes Park.

  4. PIST (GOPC) modulates the oncogenic voltage-gated potassium channel KV10.1.

    PubMed

    Herrmann, Solveig; Ninkovic, Milena; Kohl, Tobias; Pardo, Luis A

    2013-01-01

    Although crucial for their correct function, the mechanisms controlling surface expression of ion channels are poorly understood. In the case of the voltage-gated potassium channel KV10.1, this is determinant not only for its physiological function in brain, but also for its pathophysiology in tumors and possible use as a therapeutic target. The Golgi resident protein PIST binds several membrane proteins, thereby modulating their expression. Here we describe a PDZ domain-mediated interaction of KV10.1 and PIST, which enhances surface levels of KV10.1. The functional, but not the physical interaction of both proteins is dependent on the coiled-coil and PDZ domains of PIST; insertion of eight amino acids in the coiled-coil domain to render the neural form of PIST (nPIST) and the corresponding short isoform in an as-of-yet unknown form abolishes the effect. In addition, two new isoforms of PIST (sPIST and nsPIST) lacking nearly the complete PDZ domain were cloned and shown to be ubiquitously expressed. PIST and KV10.1 co-precipitate from native and expression systems. nPIST also showed interaction, but did not alter the functional expression of the channel. We could not document physical interaction between KV10.1 and sPIST, but it reduced KV10.1 functional expression in a dominant-negative manner. nsPIST showed weak physical interaction and no functional effect on KV10.1. We propose these isoforms to work as modulators of PIST function via regulating the binding on interaction partners. PMID:23966943

  5. S-glutathionylation of an auxiliary subunit confers redox sensitivity to Kv4 channel inactivation.

    PubMed

    Jerng, Henry H; Pfaffinger, Paul J

    2014-01-01

    Reactive oxygen species (ROS) regulate ion channels, modulate neuronal excitability, and contribute to the etiology of neurodegenerative disorders. ROS differentially suppress fast "ball-and-chain" N-type inactivation of cloned Kv1 and Kv3 potassium channels but not of Kv4 channels, likely due to a lack of reactive cysteines in Kv4 N-termini. Recently, we discovered that N-type inactivation of Kv4 channel complexes can be independently conferred by certain N-terminal variants of Kv4 auxiliary subunits (DPP6a, DPP10a). Here, we report that both DPP6a and DPP10a, like Kv subunits with redox-sensitive N-type inactivation, contain a highly conserved cysteine in their N-termini (Cys-13). To test if N-type inactivation mediated by DPP6a or DPP10a is redox sensitive, Xenopus oocyte recordings were performed to examine the effects of two common oxidants, tert-butyl hydroperoxide (tBHP) and diamide. Both oxidants markedly modulate DPP6a- or DPP10a-conferred N-type inactivation of Kv4 channels, slowing the overall inactivation and increasing the peak current. These functional effects are fully reversed by the reducing agent dithiothreitol (DTT) and appear to be due to a selective modulation of the N-type inactivation mediated by these auxiliary subunits. Mutation of DPP6a Cys-13 to serine eliminated the tBHP or diamide effects, confirming the importance of Cys-13 to the oxidative regulation. Biochemical studies designed to elucidate the underlying molecular mechanism show no evidence of protein-protein disulfide linkage formation following cysteine oxidation. Instead, using a biotinylated glutathione (BioGEE) reagent, we discovered that oxidation by tBHP or diamide leads to S-glutathionylation of Cys-13, suggesting that S-glutathionylation underlies the regulation of fast N-type inactivation by redox. In conclusion, our studies suggest that Kv4-based A-type current in neurons may show differential redox sensitivity depending on whether DPP6a or DPP10a is highly expressed

  6. S-Glutathionylation of an Auxiliary Subunit Confers Redox Sensitivity to Kv4 Channel Inactivation

    PubMed Central

    Jerng, Henry H.; Pfaffinger, Paul J.

    2014-01-01

    Reactive oxygen species (ROS) regulate ion channels, modulate neuronal excitability, and contribute to the etiology of neurodegenerative disorders. ROS differentially suppress fast “ball-and-chain” N-type inactivation of cloned Kv1 and Kv3 potassium channels but not of Kv4 channels, likely due to a lack of reactive cysteines in Kv4 N-termini. Recently, we discovered that N-type inactivation of Kv4 channel complexes can be independently conferred by certain N-terminal variants of Kv4 auxiliary subunits (DPP6a, DPP10a). Here, we report that both DPP6a and DPP10a, like Kv subunits with redox-sensitive N-type inactivation, contain a highly conserved cysteine in their N-termini (Cys-13). To test if N-type inactivation mediated by DPP6a or DPP10a is redox sensitive, Xenopus oocyte recordings were performed to examine the effects of two common oxidants, tert-butyl hydroperoxide (tBHP) and diamide. Both oxidants markedly modulate DPP6a- or DPP10a-conferred N-type inactivation of Kv4 channels, slowing the overall inactivation and increasing the peak current. These functional effects are fully reversed by the reducing agent dithiothreitol (DTT) and appear to be due to a selective modulation of the N-type inactivation mediated by these auxiliary subunits. Mutation of DPP6a Cys-13 to serine eliminated the tBHP or diamide effects, confirming the importance of Cys-13 to the oxidative regulation. Biochemical studies designed to elucidate the underlying molecular mechanism show no evidence of protein-protein disulfide linkage formation following cysteine oxidation. Instead, using a biotinylated glutathione (BioGEE) reagent, we discovered that oxidation by tBHP or diamide leads to S-glutathionylation of Cys-13, suggesting that S-glutathionylation underlies the regulation of fast N-type inactivation by redox. In conclusion, our studies suggest that Kv4-based A-type current in neurons may show differential redox sensitivity depending on whether DPP6a or DPP10a is highly expressed

  7. Positive Allosteric Modulation of Kv Channels by Sevoflurane: Insights into the Structural Basis of Inhaled Anesthetic Action.

    PubMed

    Liang, Qiansheng; Anderson, Warren D; Jones, Shelly T; Souza, Caio S; Hosoume, Juliana M; Treptow, Werner; Covarrubias, Manuel

    2015-01-01

    Inhalational general anesthesia results from the poorly understood interactions of haloethers with multiple protein targets, which prominently includes ion channels in the nervous system. Previously, we reported that the commonly used inhaled anesthetic sevoflurane potentiates the activity of voltage-gated K+ (Kv) channels, specifically, several mammalian Kv1 channels and the Drosophila K-Shaw2 channel. Also, previous work suggested that the S4-S5 linker of K-Shaw2 plays a role in the inhibition of this Kv channel by n-alcohols and inhaled anesthetics. Here, we hypothesized that the S4-S5 linker is also a determinant of the potentiation of Kv1.2 and K-Shaw2 by sevoflurane. Following functional expression of these Kv channels in Xenopus oocytes, we found that converse mutations in Kv1.2 (G329T) and K-Shaw2 (T330G) dramatically enhance and inhibit the potentiation of the corresponding conductances by sevoflurane, respectively. Additionally, Kv1.2-G329T impairs voltage-dependent gating, which suggests that Kv1.2 modulation by sevoflurane is tied to gating in a state-dependent manner. Toward creating a minimal Kv1.2 structural model displaying the putative sevoflurane binding sites, we also found that the positive modulations of Kv1.2 and Kv1.2-G329T by sevoflurane and other general anesthetics are T1-independent. In contrast, the positive sevoflurane modulation of K-Shaw2 is T1-dependent. In silico docking and molecular dynamics-based free-energy calculations suggest that sevoflurane occupies distinct sites near the S4-S5 linker, the pore domain and around the external selectivity filter. We conclude that the positive allosteric modulation of the Kv channels by sevoflurane involves separable processes and multiple sites within regions intimately involved in channel gating. PMID:26599217

  8. Elimination of fast inactivation in Kv4 A-type potassium channels by an auxiliary subunit domain

    PubMed Central

    Holmqvist, Mats H.; Cao, Jie; Hernandez-Pineda, Ricardo; Jacobson, Michael D.; Carroll, Karen I.; Sung, M. Amy; Betty, Maria; Ge, Pei; Gilbride, Kevin J.; Brown, Melissa E.; Jurman, Mark E.; Lawson, Deborah; Silos-Santiago, Inmaculada; Xie, Yu; Covarrubias, Manuel; Rhodes, Kenneth J.; Distefano, Peter S.; An, W. Frank

    2002-01-01

    The Kv4 A-type potassium currents contribute to controlling the frequency of slow repetitive firing and back-propagation of action potentials in neurons and shape the action potential in heart. Kv4 currents exhibit rapid activation and inactivation and are specifically modulated by K-channel interacting proteins (KChIPs). Here we report the discovery and functional characterization of a modular K-channel inactivation suppressor (KIS) domain located in the first 34 aa of an additional KChIP (KChIP4a). Coexpression of KChIP4a with Kv4 α-subunits abolishes fast inactivation of the Kv4 currents in various cell types, including cerebellar granule neurons. Kinetic analysis shows that the KIS domain delays Kv4.3 opening, but once the channel is open, it disrupts rapid inactivation and slows Kv4.3 closing. Accordingly, KChIP4a increases the open probability of single Kv4.3 channels. The net effects of KChIP4a and KChIP1–3 on Kv4 gating are quite different. When both KChIP4a and KChIP1 are present, the Kv4.3 current shows mixed inactivation profiles dependent on KChIP4a/KChIP1 ratios. The KIS domain effectively converts the A-type Kv4 current to a slowly inactivating delayed rectifier-type potassium current. This conversion is opposite to that mediated by the Kv1-specific “ball” domain of the Kvβ1 subunit. Together, these results demonstrate that specific auxiliary subunits with distinct functions actively modulate gating of potassium channels that govern membrane excitability. PMID:11805342

  9. Positive Allosteric Modulation of Kv Channels by Sevoflurane: Insights into the Structural Basis of Inhaled Anesthetic Action

    PubMed Central

    Liang, Qiansheng; Anderson, Warren D.; Jones, Shelly T.; Souza, Caio S.; Hosoume, Juliana M.; Treptow, Werner; Covarrubias, Manuel

    2015-01-01

    Inhalational general anesthesia results from the poorly understood interactions of haloethers with multiple protein targets, which prominently includes ion channels in the nervous system. Previously, we reported that the commonly used inhaled anesthetic sevoflurane potentiates the activity of voltage-gated K+ (Kv) channels, specifically, several mammalian Kv1 channels and the Drosophila K-Shaw2 channel. Also, previous work suggested that the S4-S5 linker of K-Shaw2 plays a role in the inhibition of this Kv channel by n-alcohols and inhaled anesthetics. Here, we hypothesized that the S4-S5 linker is also a determinant of the potentiation of Kv1.2 and K-Shaw2 by sevoflurane. Following functional expression of these Kv channels in Xenopus oocytes, we found that converse mutations in Kv1.2 (G329T) and K-Shaw2 (T330G) dramatically enhance and inhibit the potentiation of the corresponding conductances by sevoflurane, respectively. Additionally, Kv1.2-G329T impairs voltage-dependent gating, which suggests that Kv1.2 modulation by sevoflurane is tied to gating in a state-dependent manner. Toward creating a minimal Kv1.2 structural model displaying the putative sevoflurane binding sites, we also found that the positive modulations of Kv1.2 and Kv1.2-G329T by sevoflurane and other general anesthetics are T1-independent. In contrast, the positive sevoflurane modulation of K-Shaw2 is T1-dependent. In silico docking and molecular dynamics-based free-energy calculations suggest that sevoflurane occupies distinct sites near the S4-S5 linker, the pore domain and around the external selectivity filter. We conclude that the positive allosteric modulation of the Kv channels by sevoflurane involves separable processes and multiple sites within regions intimately involved in channel gating. PMID:26599217

  10. Tracking single Kv2.1 channels in live cells reveals anomalous subdiffusion and ergodicity breaking

    NASA Astrophysics Data System (ADS)

    Weigel, Aubrey; Simon, Blair; Tamkun, Michael; Krapf, Diego

    2011-03-01

    The dynamic organization of the plasma membrane is responsible for essential cellular processes, such as receptor trafficking and signaling. By studying the dynamics of transmembrane proteins a greater understanding of these processes as a whole can be achieved. It is broadly observed that the diffusion pattern of membrane protein displays anomalous subdiffusion. However, the mechanisms responsible for this behavior are not yet established. We explore the dynamics of the voltage gated potassium channel Kv2.1 by using single-particle tracking. We analyze Kv2.1 channel trajectories in terms of the time and ensemble distributions of square displacements. Our results reveal that all Kv2.1 channels experience anomalous subdiffusion and we observe that the Kv2.1 diffusion pattern is non-ergodic. We further investigated the role of the actin cytoskeleton in these channel dynamics by applying actin depolymerizing drugs. It is seen that with the breakdown of the actin cytoskeleton the Kv2.1 channel trajectories recover ergodicity.

  11. Role of Kv1.3 mitochondrial potassium channel in apoptotic signalling in lymphocytes.

    PubMed

    Gulbins, Erich; Sassi, Nicola; Grassmè, Heike; Zoratti, Mario; Szabò, Ildikò

    2010-01-01

    Mitochondria have been shown to play a pivotal role in apoptotic signalling in various cell types. We have recently reported that in lymphocytes the voltage-gated potassium channel Kv1.3, known to reside in the plasma membrane, is active also in the inner mitochondrial membrane. Upon induction of apoptosis, outer-membrane inserted Bax binds to and inhibits Kv1.3 resulting in hyperpolarization, an increase in reactive oxygen species production and cytochrome c release. In cells lacking Kv1.3 these events do not take place. Here, we present new data which further corroborates an important role of this channel in the sequence of events leading to Bax-induced cytochrome c release. Recombinant Kv1.3, when pre-incubated with Bax, prevents the actions of Bax at the level of mitochondria. Furthermore, we report the presence of Kv1.3 protein in mitochondria from PC3 and MCF-7 cancer cells, suggesting that this channel might play a role in the apoptotic signalling not only in lymphocytes but also in other cells. PMID:20114030

  12. Fe2O3 nanoparticles suppress Kv1.3 channels via affecting the redox activity of Kvβ2 subunit in Jurkat T cells

    NASA Astrophysics Data System (ADS)

    Yan, Li; Liu, Xiao; Liu, Wei-Xia; Tan, Xiao-Qiu; Xiong, Fei; Gu, Ning; Hao, Wei; Gao, Xue; Cao, Ji-Min

    2015-12-01

    Superparamagnetic iron oxide nanoparticles (SPIONs) are promising nanomaterials in medical practice due to their special magnetic characteristics and nanoscale size. However, their potential impacts on immune cells are not well documented. This study aims to investigate the effects of Fe2O3 nanoparticles (Fe2O3-NPs) on the electrophysiology of Kv1.3 channels in Jurkat T cells. Using the whole-cell patch-clamp technique, we demonstrate that incubation of Jurkat cells with Fe2O3-NPs dose- and time-dependently decreased the current density and shifted the steady-state inactivation curve and the recovery curve of Kv1.3 channels to a rightward direction. Fe2O3-NPs increased the NADP level but decreased the NADPH level of Jurkat cells. Direct induction of NADPH into the cytosole of Jurkat cells via the pipette abolished the rightward shift of the inactivation curve. In addition, transmission electron microscopy showed that Fe2O3-NPs could be endocytosed by Jurkat cells with relatively low speed and capacity. Fe2O3-NPs did not significantly affect the viability of Jurkat cells, but suppressed the expressions of certain cytokines (TNFα, IFNγ and IL-2) and interferon responsive genes (IRF-1 and PIM-1), and the time courses of Fe2O3-NPs endocytosis and effects on the expressions of cytokines and interferon responsive genes were compatible. We conclude that Fe2O3-NPs can be endocytosed by Jurkat cells and act intracellularly. Fe2O3-NPs decrease the current density and delay the inactivation and recovery kinetics of Kv1.3 channels in Jurkat cells by oxidizing NADPH and therefore disrupting the redox activity of the Kvβ2 auxiliary subunit, and as a result, lead to changes of the Kv1.3 channel function. These results suggest that iron oxide nanoparticles may affect T cell function by disturbing the activity of Kv1.3 channels. Further, the suppressing effects of Fe2O3-NPs on the expressions of certain inflammatory cytokines and interferon responsive genes suggest that iron

  13. Zinc Oxide Surge Arresters and HVDC 125kV-upgrade 500kV Converter Stations

    NASA Astrophysics Data System (ADS)

    Shirakawa, Shingo; Kobayashi, Takayuki; Matsushita, Yoshinao; Sakai, Takehisa; Suzuki, Hironori; Ozaki, Yuzo

    Gapless Metal (Zinc) Oxide Surge Arresters for a.c. systems contribute to the insulation co-ordination based on the suppression of lightning surges and switching surges. These gapless metal oxide surge arresters using ZnO elements are effective to HVDC systems. This paper describes basic characteristics of ZnO (zinc oxide) elements for d.c. systems and applications of gapless surge arresters to HVDC 125kV frequency converters, HVDC 250kV, upgrade HVDC 500kV converter stations, and HVDC 500kV cables of Japan through the experience of developments and applications of gapless metal oxide surge arresters.

  14. Selective down-regulation of KV2.1 function contributes to enhanced arterial tone during diabetes.

    PubMed

    Nieves-Cintrón, Madeline; Nystoriak, Matthew A; Prada, Maria Paz; Johnson, Kenneth; Fayer, William; Dell'Acqua, Mark L; Scott, John D; Navedo, Manuel F

    2015-03-20

    Enhanced arterial tone is a leading cause of vascular complications during diabetes. Voltage-gated K(+) (KV) channels are key regulators of vascular smooth muscle cells (VSMCs) contractility and arterial tone. Whether impaired KV channel function contributes to enhance arterial tone during diabetes is unclear. Here, we demonstrate a reduction in KV-mediated currents (IKv) in VSMCs from a high fat diet (HFD) mouse model of type 2 diabetes. In particular, IKv sensitive to stromatoxin (ScTx), a potent KV2 blocker, were selectively reduced in diabetic VSMCs. This was associated with decreased KV2-mediated regulation of arterial tone and suppression of the KV2.1 subunit mRNA and protein in VSMCs/arteries isolated from HFD mice. We identified protein kinase A anchoring protein 150 (AKAP150), via targeting of the phosphatase calcineurin (CaN), and the transcription factor nuclear factor of activated T-cells c3 (NFATc3) as required determinants of KV2.1 suppression during diabetes. Interestingly, substantial reduction in transcript levels for KV2.1 preceded down-regulation of large conductance Ca(2+)-activated K(+) (BKCa) channel β1 subunits, which are ultimately suppressed in chronic hyperglycemia to a similar extent. Together, our study supports the concept that transcriptional suppression of KV2.1 by activation of the AKAP150-CaN/NFATc3 signaling axis contributes to enhanced arterial tone during diabetes. PMID:25670860

  15. Voltage-Gated K+ Channel, Kv3.3 Is Involved in Hemin-Induced K562 Differentiation

    PubMed Central

    Song, Min Seok; Choi, Seon Young; Ryu, Pan Dong; Lee, So Yeong

    2016-01-01

    Voltage-gated K+ (Kv) channels are well known to be involved in cell proliferation. However, even though cell proliferation is closely related to cell differentiation, the relationship between Kv channels and cell differentiation remains poorly investigated. This study demonstrates that Kv3.3 is involved in K562 cell erythroid differentiation. Down-regulation of Kv3.3 using siRNA-Kv3.3 increased hemin-induced K562 erythroid differentiation through decreased activation of signal molecules such as p38, cAMP response element-binding protein, and c-fos. Down-regulation of Kv3.3 also enhanced cell adhesion by increasing integrin β3 and this effect was amplified when the cells were cultured with fibronectin. The Kv channels, or at least Kv3.3, appear to be associated with cell differentiation; therefore, understanding the mechanisms of Kv channel regulation of cell differentiation would provide important information regarding vital cellular processes. PMID:26849432

  16. Molecular basis for the toxin insensitivity of scorpion voltage-gated potassium channel MmKv1.

    PubMed

    Zhang, Chuangeng; Xie, Zili; Li, Xinxin; Chen, Jing; Feng, Jing; Lang, Yange; Yang, Weishan; Li, Wenxin; Chen, Zongyun; Yao, Jing; Cao, Zhijian; Wu, Yingliang

    2016-05-01

    Scorpions are insensitive to their own venoms, which contain various neurotoxins specific for mammalian or insect ion channels, whose molecular mechanism remains unsolved. Using MmKv1, a potassium channel identified from the genome of the scorpion Mesobuthus martensii, channel kinetic experiments showed that MmKv1 was a classical voltage-gated potassium channel with a voltage-dependent fast activation and slow inactivation. Compared with the human Kv1.3 channel (hKv1.3), the MmKv1 channel exhibited a remarkable insensitivity to both scorpion venom and toxin. The chimaeric channels of MmKv1 and hKv1.3 revealed that both turret and filter regions of the MmKv1 channel were critical for the toxin insensitivity of MmKv1. Furthermore, mutagenesis of the chimaeric channel indicated that two basic residues (Arg(399) and Lys(403)) in the MmKv1 turret region and Arg(425) in the MmKv1 filter region significantly affected its toxin insensitivity. Moreover, when these three basic residues of MmKv1 were simultaneously substituted with the corresponding residues from hKv1.3, the MmKv1-R399T/K403S/R425H mutant channels exhibited similar sensitivity to both scorpion venom and toxin to hKv1.3, which revealed the determining role of these three basic residues in the toxin insensitivity of the MmKv1 channel. More strikingly, a similar triad sequence structure is present in all Shaker-like channels from venomous invertebrates, which suggested a possible convergent functional evolution of these channels to enable them to resist their own venoms. Together, these findings first illustrate the mechanism by which scorpions are insensitive to their own venoms at the ion channel receptor level and enrich our knowledge of the insensitivity of scorpions and other venomous animals to their own venoms. PMID:26951716

  17. Structure of the KvAP voltage-dependent K+ channel and its dependence on the lipid membrane

    SciTech Connect

    Lee,S.; Lee, A.; Chen, J.; MacKinnon, R.; Chin, W.

    2005-01-01

    Voltage-dependent ion channels gate open in response to changes in cell membrane voltage. This form of gating permits the propagation of action potentials. We present two structures of the voltage-dependent K{sup +} channel KvAP, in complex with monoclonal Fv fragments (3.9 Angstroms) and without antibody fragments (8 Angstroms). We also studied KvAP with disulfide cross-bridges in lipid membranes. Analyzing these data in the context of the crystal structure of Kv1.2 and EPR data on KvAP we reach the following conclusions: (i) KvAP is similar in structure to Kv1.2 with a very modest difference in the orientation of its voltage sensor; (ii) mAb fragments are not the source of non-native conformations of KvAP in crystal structures; (iii) because KvAP contains separate loosely adherent domains, a lipid membrane is required to maintain their correct relative orientations, and (iv) the model of KvAP is consistent with the proposal of voltage sensing through the movement of an arginine-containing helix-turn-helix element at the protein-lipid interface.

  18. Structure of the KvAP voltage-dependent K+ channel and its dependence on the lipid membrane

    PubMed Central

    Lee, Seok-Yong; Lee, Alice; Chen, Jiayun; MacKinnon, Roderick

    2005-01-01

    Voltage-dependent ion channels gate open in response to changes in cell membrane voltage. This form of gating permits the propagation of action potentials. We present two structures of the voltage-dependent K+ channel KvAP, in complex with monoclonal Fv fragments (3.9 Å) and without antibody fragments (8 Å). We also studied KvAP with disulfide cross-bridges in lipid membranes. Analyzing these data in the context of the crystal structure of Kv1.2 and EPR data on KvAP we reach the following conclusions: (i) KvAP is similar in structure to Kv1.2 with a very modest difference in the orientation of its voltage sensor; (ii) mAb fragments are not the source of non-native conformations of KvAP in crystal structures; (iii) because KvAP contains separate loosely adherent domains, a lipid membrane is required to maintain their correct relative orientations, and (iv) the model of KvAP is consistent with the proposal of voltage sensing through the movement of an arginine-containing helix-turn-helix element at the protein-lipid interface. PMID:16223877

  19. Kv1.3 deletion biases T cells toward an immunoregulatory phenotype and renders mice resistant to autoimmune encephalomyelitis.

    PubMed

    Gocke, Anne R; Lebson, Lori A; Grishkan, Inna V; Hu, Lina; Nguyen, Hai M; Whartenby, Katharine A; Chandy, K George; Calabresi, Peter A

    2012-06-15

    Increasing evidence suggests ion channels have critical functions in the differentiation and plasticity of T cells. Kv1.3, a voltage-gated K(+) channel, is a functional marker and a pharmacological target for activated effector memory T cells. Selective Kv1.3 blockers have been shown to inhibit proliferation and cytokine production by human and rat effector memory T cells. We used Kv1.3 knockout (KO) mice to investigate the mechanism by which Kv1.3 blockade affects CD4(+) T cell differentiation during an inflammatory immune-mediated disease. Kv1.3 KO animals displayed significantly lower incidence and severity of myelin oligodendrocyte glycoprotein (MOG) peptide-induced experimental autoimmune encephalomyelitis. Kv1.3 was the only K(V) channel expressed in MOG 35-55-specific CD4(+) T cell blasts, and no K(V) current was present in MOG-specific CD4(+) T cell-blasts from Kv1.3 KO mice. Fewer CD4(+) T cells migrated to the CNS in Kv1.3 KO mice following disease induction, and Ag-specific proliferation of CD4(+) T cells from these mice was impaired with a corresponding cell-cycle delay. Kv1.3 was required for optimal expression of IFN-γ and IL-17, whereas its absence led to increased IL-10 production. Dendritic cells from Kv1.3 KO mice fully activated wild-type CD4(+) T cells, indicating a T cell-intrinsic defect in Kv1.3 KO mice. The loss of Kv1.3 led to a suppressive phenotype, which may contribute to the mechanism by which deletion of Kv1.3 produces an immunotherapeutic effect. Skewing of CD4(+) T cell differentiation toward Ag-specific regulatory T cells by pharmacological blockade or genetic suppression of Kv1.3 might be beneficial for therapy of immune-mediated diseases such as multiple sclerosis. PMID:22581856

  20. Diagnostic quality of CT pulmonary angiography in pulmonary thromboembolism: A comparison of three different kV values

    PubMed Central

    Yilmaz, Ömer; Üstün, Esma Dilek; Kayan, Mustafa; Kayan, Fatmanur; Aktaş, Aykut Recep; Unlu, Elif Nisa; Degirmenci, Bumin; Cetin, Meltem

    2013-01-01

    Background Our purpose was to evaluate the effectiveness of different kilovolt (kV) uses in computed tomography pulmonary angiography (CTPA) in the diagnosis of pulmonary thromboembolism (PTE). We also aimed to establish the optimal kV value and investigate the possibility of obtaining appropriate imaging quality with minimal radiation dose. Material/Methods We compared 120, 100, and 80 kV CTPA for 90 patients in whom PTE was clinically considered. The examinations were carried out using a 128 multislice CT device (Definition AS, Siemens Medical Solutions, Forchheim, Germany). Each kV value was used on 30 patients in 3 groups. Patients in all groups were compared with respect to the mean radiation dose they received, pulmonary arterial attenuation values, image quality, and motion artefacts. Results With respect to pulmonary arterial attenuation values, imaging with 80 kV yielded significantly higher values (p<0.05). However, no difference was found between 120 kV, 100 kV, and 80 kV with respect to image quality. Similarly, no significant difference was detected between the groups with respect to pulmonary artery contrasting and motion artefacts. Statistically significant differences were present in DLP values and effective dose among all 3 groups (p<0.001). Conclusions Using 80 kV as the low value in CTPA imaging for patients pre-diagnosed with PTE will increase the density of pulmonary arteries and decrease the amount of radiation received. PMID:24169688

  1. Identification of a key residue in Kv7.1 potassium channel essential for sensing external potassium ions

    PubMed Central

    Wang, Wenying; Flores, Maria Cristina Perez; Sihn, Choong-Ryoul; Kim, Hyo Jeong; Zhang, Yinuo; Doyle, Karen J.; Chiamvimonvat, Nipavan

    2015-01-01

    Kv7.1 voltage-gated K+ (Kv) channels are present in the apical membranes of marginal cells of the stria vascularis of the inner ear, where they mediate K+ efflux into the scala media (cochlear duct) of the cochlea. As such, they are exposed to the K+-rich (∼150 mM of external K+ (K+e)) environment of the endolymph. Previous studies have shown that Kv7.1 currents are substantially suppressed by high K+e (independent of the effects of altering the electrochemical gradient). However, the molecular basis for this inhibition, which is believed to involve stabilization of an inactivated state, remains unclear. Using sequence alignment of S5-pore linkers of several Kv channels, we identified a key residue, E290, found in only a few Kv channels including Kv7.1. We used substituted cysteine accessibility methods and patch-clamp analysis to provide evidence that the ability of Kv7.1 to sense K+e depends on E290, and that the charge at this position is essential for Kv7.1’s K+e sensitivity. We propose that Kv7.1 may use this feedback mechanism to maintain the magnitude of the endocochlear potential, which boosts the driving force to generate the receptor potential of hair cells. The implications of our findings transcend the auditory system; mutations at this position also result in long QT syndrome in the heart. PMID:25712016

  2. Waltz Mill testing of 765-kV paper-polypropylene-paper (PPP) cable. Final report

    SciTech Connect

    Burghardt, R.R.

    1992-06-01

    A 765-kV PPP-insulated cable was subjected to a 27-month accelerated life test program at the EPRI Waltz Mill Cable Test Facility. Testing started in August 1981 and was successfully completed in January 1985. The program included conductor temperatures ranging from 85{degree}C to 105{degree}C and line-to-line voltages from 800 kV to 1050 kV. Cyclic testing was performed during 20 of the 27 months. Dissipation factor measurements were made throughout the program. The measurements indicated no deterioration of the cable or splices as a consequence of the high temperatures and voltages applied to them in this test program.

  3. Waltz Mill testing of 765-kV paper-polypropylene-paper (PPP) cable

    SciTech Connect

    Burghardt, R.R. )

    1992-06-01

    A 765-kV PPP-insulated cable was subjected to a 27-month accelerated life test program at the EPRI Waltz Mill Cable Test Facility. Testing started in August 1981 and was successfully completed in January 1985. The program included conductor temperatures ranging from 85{degree}C to 105{degree}C and line-to-line voltages from 800 kV to 1050 kV. Cyclic testing was performed during 20 of the 27 months. Dissipation factor measurements were made throughout the program. The measurements indicated no deterioration of the cable or splices as a consequence of the high temperatures and voltages applied to them in this test program.

  4. Electrical Remodeling of Preoptic GABAergic Neurons Involves the Kv1.5 Subunit

    PubMed Central

    Tabarean, Iustin V.

    2014-01-01

    The electrogenic machinery of an excitable cell can adapt in response to changes in input, genetic deficit or in pathological conditions, however the underlying molecular mechanisms are not understood. In cases of genetic deletion it is commonly observed that a channel subunit from the same family replaces the missing one. We have previously reported that Kv4.2−/− preoptic GABAergic neurons display identical firing characteristics to those of wild-type neurons despite having reduced A-type currents, and that, surprisingly, they present a robust upregulation of a delayed rectifier current, the nature of which is unknown. Here, using pharmacology, qPCR and Western blots we report that, although the wild-type neurons express several Kv subunits, the upregulated current is conducted by the Kv1.5 subunit exclusively. Thus, this study reveals the molecular nature of a novel mechanism of electrical remodeling in central neurons. PMID:24797243

  5. Electrical remodeling of preoptic GABAergic neurons involves the Kv1.5 subunit.

    PubMed

    Tabarean, Iustin V

    2014-01-01

    The electrogenic machinery of an excitable cell can adapt in response to changes in input, genetic deficit or in pathological conditions, however the underlying molecular mechanisms are not understood. In cases of genetic deletion it is commonly observed that a channel subunit from the same family replaces the missing one. We have previously reported that Kv4.2-/- preoptic GABAergic neurons display identical firing characteristics to those of wild-type neurons despite having reduced A-type currents, and that, surprisingly, they present a robust upregulation of a delayed rectifier current, the nature of which is unknown. Here, using pharmacology, qPCR and Western blots we report that, although the wild-type neurons express several Kv subunits, the upregulated current is conducted by the Kv1.5 subunit exclusively. Thus, this study reveals the molecular nature of a novel mechanism of electrical remodeling in central neurons. PMID:24797243

  6. Status of 275 kV REBCO HTS Cable Development in the NEDO Project

    NASA Astrophysics Data System (ADS)

    Mukoyama, Shinichi; Yagi, Masashi; Okuma, Takeshi; Maruyama, Osamu; Shiohara, Yuu; Hayakawa, Naoki; Mizutani, Teruyoshi

    A 275 kV 3 kA high temperature superconducting cable (HTS cable), which could be used as a backbone power line in the future, was developed in the NEDO project called M-PACC. One of the most important developments of a high voltage HTS cable was the high voltage insulation technology. A design guideline and a test specification that was necessary to design, product and demonstrate of a 275 kV, 3 kA HTS cable have been studied by obtaining the various experimental data such as AC withstand voltage, impulse withstand voltage, partial discharge inception stress, and the V-t characteristics of the insulation, on the basis of the Japan Electrical Standards (JEC) and the International Electrotechnical Commission (IEC). Moreover, the 275 kV, 3 kA HTS cable with a length of 30 m was demonstrated under a long-term voltage and current loading test.

  7. Field Demonstration of a 24-kV Superconducting Cable at Detroit Edison

    SciTech Connect

    Kelley, Nathan; Corsaro, Pietro

    2004-12-01

    Customer acceptance of high temperature superconducting (HTS) cable technology requires a substantial field demonstration illustrating both the system's technical capabilities and its suitability for installation and operation within the utility environment. In this project, the world's first underground installation of an HTS cable using existing ductwork, a 120 meter demonstration cable circuit was designed and installed between the 24 kV bus distribution bus and a 120 kV-24 kV transformer at Detroit Edison's Frisbie substation. The system incorporated cables, accessories, a refrigeration system, and control instrumentation. Although the system was never put in operation because of problems with leaks in the cryostat, the project significantly advanced the state-of-the-art in the design and implementation of Warm Dielectric cable systems in substation applications. Lessons learned in this project are already being incorporated in several ongoing demonstration projects.

  8. Study on Preserving Public Security of Installing 22 kV Distribution Equipment on Pavements

    NASA Astrophysics Data System (ADS)

    Murata, Koichi; Oka, Keisuke; Uemura, Satoshi; Ariga, Yasuo

    To cope with restructuring of electricity market, application of 22kV distribution systems to areas with high load density, is promising. To expand them, it is important to install 22kV distribution equipment on pavements and lots without fences. On the other hand, the neutral grounding method is expected for the 22kV distribution equipment in terms of rationalization of insulation. In this case, one line-to-ground fault current will be large. Therefore, to make safety of people, it is essential to decrease the contact voltage. In this paper, we will perform the simulation of current distribution in the case of one line-to-ground fault and study the security of human from the viewpoint of both step voltage and contact voltage. We will also clarify that we can lower the contact voltage by grounding mutually insulated inner box and outer box separately. Then we will propose the improved grounding method.

  9. Hypoxia induces voltage-gated K+ (Kv) channel expression in pulmonary arterial smooth muscle cells through hypoxia-inducible factor-1 (HIF-1)

    PubMed Central

    Dong, Qian; Zhao, Ning; Xia, Cheng-kun; Du, Li-li; Fu, Xiao-xing; Du, Yi-mei

    2012-01-01

    Hypoxia-inducible factor-1 (HIF-1) regulates the expression of hypoxia-inducible genes by binding erythropoietin (EPO) enhancer fragments. Of these genes, HIF-1 upregulates voltage-gated K+1.2 channels (Kv1.2) in rat PC12 cells. Whether HIF-1 regulates hypoxia-induced Kv channel expression in cultured pulmonary artery smooth muscle cells (PASMCs), however, has not been determined. In this study, we investigated the effects of hypoxia on the expression of Kv1.2 Kv1.5, Kv2.1, and Kv9.3 channels in PASMCs and examined the direct role of HIF-1 by transfecting either wild type or mutant EPO enhancer fragments. Our results showed that 18 h exposure to hypoxia significantly increased the expression of Kv1.2, Kv1.5, Kv2.1, and Kv9.3; and this hypoxia-induced upregulation was completely inhibited after transfection with the wild type but not mutant EPO enhancer fragment. These results indicate that HIF-1 regulates hypoxia-stimulated induction of Kv1.2, Kv1.5, Kv2.1, and Kv9.3 channels in cultured PASMCs. PMID:22938542

  10. Redistribution of Kv2.1 ion channels on spinal motoneurons following peripheral nerve injury.

    PubMed

    Romer, Shannon H; Dominguez, Kathleen M; Gelpi, Marc W; Deardorff, Adam S; Tracy, Robert C; Fyffe, Robert E W

    2014-02-14

    Pathophysiological responses to peripheral nerve injury include alterations in the activity, intrinsic membrane properties and excitability of spinal neurons. The intrinsic excitability of α-motoneurons is controlled in part by the expression, regulation, and distribution of membrane-bound ion channels. Ion channels, such as Kv2.1 and SK, which underlie delayed rectifier potassium currents and afterhyperpolarization respectively, are localized in high-density clusters at specific postsynaptic sites (Deardorff et al., 2013; Muennich and Fyffe, 2004). Previous work has indicated that Kv2.1 channel clustering and kinetics are regulated by a variety of stimuli including ischemia, hypoxia, neuromodulator action and increased activity. Regulation occurs via channel dephosphorylation leading to both declustering and alterations in channel kinetics, thus normalizing activity (Misonou et al., 2004; Misonou et al., 2005; Misonou et al., 2008; Mohapatra et al., 2009; Park et al., 2006). Here we demonstrate using immunohistochemistry that peripheral nerve injury is also sufficient to alter the surface distribution of Kv2.1 channels on motoneurons. The dynamic changes in channel localization include a rapid progressive decline in cluster size, beginning immediately after axotomy, and reaching maximum within one week. With reinnervation, the organization and size of Kv2.1 clusters do not fully recover. However, in the absence of reinnervation Kv2.1 cluster sizes fully recover. Moreover, unilateral peripheral nerve injury evokes parallel, but smaller effects bilaterally. These results suggest that homeostatic regulation of motoneuron Kv2.1 membrane distribution after axon injury is largely independent of axon reinnervation. PMID:24355600

  11. Phosphoinositide 3-kinase dependent regulation of Kv channels in dendritic cells.

    PubMed

    Shumilina, Ekaterina; Zahir, Naima; Xuan, Nguyen Thi; Lang, Florian

    2007-01-01

    The phosphoinositide 3 (PI3) kinase plays a pivotal role in the regulation of dendritic cells (DCs), antigen-presenting cells that are able to initiate primary immune responses and to establish immunological memory. PI3 kinase is an endogenous suppressor of interleukin 12 (IL-12) production in DCs that is triggered by Toll-like receptor signaling. Inhibition of IL-12 production limits T helper 1 (Th1) polarization. On the other hand, PI3 kinase is an important regulator of various ion channels. The present study aimed to explore whether ion channels in DCs are regulated by PI3 kinase and whether they are important for DC function. To this end, DCs were isolated from murine bone marrow and ion channel activity was determined by patch clamp. As a result, DCs express voltage-gated K(+) channels (Kv), which are blocked by Stichodactyla helianthus toxin (ShK, 2.5 nM). A significant upregulation of Kv currents was observed upon maturation of DCs as induced by stimulation of the cells with lipopolysaccharide (LPS, 0.1 microg/ml, 48 h). A dramatic increase of Kv current amplitude was observed following preincubation of the cells with LY294002 (100 nM), a specific inhibitor of PI3 kinase. PI3 kinase inhibitor wortmannin (100 nM) similarly increased Kv current. LY294002 treatment was further followed by a significant increase of IL-12 production. ShK (100 nM) significantly blunted the stimulation of IL-12 release by LPS but not when the cells were first pretreated with LY294002. The observations point to Kv channel sensitive and Kv channel insensitive regulation of DC function. PMID:17982262

  12. The Role of Kv1.2 Channel in Electrotaxis Cell Migration

    PubMed Central

    Zhang, Gaofeng; Edmundson, Mathew; Telezhkin, Vsevolod; Gu, Yu; Wei, Xiaoqing; Kemp, Paul J.

    2015-01-01

    Voltage‐gated potassium Kv1.2 channels play pivotal role in maintaining of resting membrane potential and, consequently, regulation of cellular excitability of neurons. Endogenously generated electric field (EF) have been proven as an important regulator for cell migration and tissue repair. The mechanisms of ion channel involvement in EF‐induced cell responses are extensively studied but largely are poorly understood. In this study we generated three COS‐7 clones with different expression levels of Kv1.2 channel, and confirmed their functional variations with patch clamp analysis. Time‐lapse imaging analysis showed that EF‐induced cell migration response was Kv1.2 channel expression level depended. Inhibition of Kv1.2 channels with charybdotoxin (ChTX) constrained the sensitivity of COS‐7 cells to EF stimulation more than their motility. Immunocytochemistry and pull‐down analyses demonstrated association of Kv1.2 channels with actin‐binding protein cortactin and its re‐localization to the cathode‐facing membrane at EF stimulation, which confirms the mechanism of EF‐induced directional migration. This study displays that Kv1.2 channels represent an important physiological link in EF‐induced cell migration. The described mechanism suggests a potential application of EF which may improve therapeutic performance in curing injuries of neuronal and/or cardiac tissue repair, post operational therapy, and various degenerative syndromes. J. Cell. Physiol. 231: 1375–1384, 2016. © 2015 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc. PMID:26580832

  13. Kv1.3 channel blockade enhances the phagocytic function of RAW264.7 macrophages.

    PubMed

    Zhu, Hong; Yan, Li; Gu, JingLi; Hao, Wei; Cao, JiMin

    2015-09-01

    This study aimed to comprehend the largely unknown role of voltage-gated potassium channel 1.3 (Kv1.3) in the phagocytic function of macrophages. We found that blocking of the Kv1.3 channel with 100 pmol L(-1) Stichodactyla helianthus neurotoxin (ShK) enhanced the phagocytic capacities of both resting and lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages in the chicken erythrocyte system. In the fluorescein isothiocyanate (FITC)-labeled Escherichia coli k-12 system, ShK increased the phagocytic capacities of resting RAW264.7 cells, but not of the LPS-stimulated cells, as LPS alone stimulated almost saturated phagocytosis of the macrophages. ShK increased the nitric oxide (NO) production in LPS-activated cells, but not in resting RAW264.7 cells. There was no effect of ShK alone on the cytokine secretions in resting RAW264.7 cells, but it suppressed IL-1β secretion in LPS-stimulated RAW264.7 cells. At a concentration of 100 pmol L(-1), ShK did not affect the viability of the tested cells. Kv1.3 was expressed in RAW264.7 cells; this expression was downregulated by LPS, but significantly upregulated by disrupting caveolin-dependent endocytosis with filipin III. In addition, cytochalasin D, an inhibitor of actin polymerization, did not affect the Kv1.3 expression. Thus, blocking of the Kv1.3 channel enhances the phagocytic capacity and NO production of this cell line. Our results suggest that Kv1.3 channel serves as a negative regulator of phagocytosis in macrophages and can therefore be a potential target in the treatment of macrophage dysfunction. PMID:26354506

  14. Blockade of Kv1.5 channels by the antidepressant drug sertraline

    PubMed Central

    Lee, Hyang Mi; Hahn, Sang June

    2016-01-01

    Sertraline, a selective serotonin reuptake inhibitor (SSRI), has been reported to lead to cardiac toxicity even at therapeutic doses including sudden cardiac death and ventricular arrhythmia. And in a SSRI-independent manner, sertraline has been known to inhibit various voltage-dependent channels, which play an important role in regulation of cardiovascular system. In the present study, we investigated the action of sertraline on Kv1.5, which is one of cardiac ion channels. The eff ect of sertraline on the cloned neuronal rat Kv1.5 channels stably expressed in Chinese hamster ovary cells was investigated using the whole-cell patch-clamp technique. Sertraline reduced Kv1.5 whole-cell currents in a reversible concentration-dependent manner, with an IC50 value and a Hill coefficient of 0.71 µM and 1.29, respectively. Sertraline accelerated the decay rate of inactivation of Kv1.5 currents without modifying the kinetics of current activation. The inhibition increased steeply between –20 and 0 mV, which corresponded with the voltage range for channel opening. In the voltage range positive to +10 mV, inhibition displayed a weak voltage dependence, consistent with an electrical distance δ of 0.16. Sertraline slowed the deactivation time course, resulting in a tail crossover phenomenon when the tail currents, recorded in the presence and absence of sertraline, were superimposed. Inhibition of Kv1.5 by sertraline was use-dependent. The present results suggest that sertraline acts on Kv1.5 currents as an open-channel blocker. PMID:26937216

  15. Blockade of Kv1.5 channels by the antidepressant drug sertraline.

    PubMed

    Lee, Hyang Mi; Hahn, Sang June; Choi, Bok Hee

    2016-03-01

    Sertraline, a selective serotonin reuptake inhibitor (SSRI), has been reported to lead to cardiac toxicity even at therapeutic doses including sudden cardiac death and ventricular arrhythmia. And in a SSRI-independent manner, sertraline has been known to inhibit various voltage-dependent channels, which play an important role in regulation of cardiovascular system. In the present study, we investigated the action of sertraline on Kv1.5, which is one of cardiac ion channels. The eff ect of sertraline on the cloned neuronal rat Kv1.5 channels stably expressed in Chinese hamster ovary cells was investigated using the whole-cell patch-clamp technique. Sertraline reduced Kv1.5 whole-cell currents in a reversible concentration-dependent manner, with an IC 50 value and a Hill coefficient of 0.71 µM and 1.29, respectively. Sertraline accelerated the decay rate of inactivation of Kv1.5 currents without modifying the kinetics of current activation. The inhibition increased steeply between -20 and 0 mV, which corresponded with the voltage range for channel opening. In the voltage range positive to +10 mV, inhibition displayed a weak voltage dependence, consistent with an electrical distance δ of 0.16. Sertraline slowed the deactivation time course, resulting in a tail crossover phenomenon when the tail currents, recorded in the presence and absence of sertraline, were superimposed. Inhibition of Kv1.5 by sertraline was use-dependent. The present results suggest that sertraline acts on Kv1.5 currents as an open-channel blocker. PMID:26937216

  16. Proteomic Analysis Highlights the Molecular Complexities of Native Kv4 Channel Macromolecular Complexes

    PubMed Central

    Marionneau, Céline; Townsend, R Reid; Nerbonne, Jeanne M

    2010-01-01

    Voltage-gated K+ (Kv) channels are key determinants of membrane excitability in the nervous and cardiovascular systems, functioning to control resting membrane potentials, shape action potential waveforms and influence the responses to neurotransmitters and neurohormones. Consistent with this functional diversity, multiple types of Kv currents, with distinct biophysical properties and cellular/subcellular distributions, have been identified. Rapidly activating and inactivating Kv currents, typically referred to as IA (A-type) in neurons, for example, regulate repetitive firing rates, action potential back-propagation (into dendrites) and modulate synaptic responses. Currents with similar properties, referred to as Ito,f (fast transient outward), expressed in cardiomyocytes, control the early phase of myocardial action potential repolarization. A number of studies have demonstrated critical roles for pore-forming (α) subunits of the Kv4 subfamily in the generation of native neuronal IA and cardiac Ito,f channels. Studies in heterologous cells have also suggested important roles for a number of Kv channel accessory and regulatory proteins in the generation of functional IA and Ito,f channels. Quantitative mass spectrometry-based proteomic analysis is increasingly recognized as a rapid and, importantly, unbiased, approach to identify the components of native macromolecular protein complexes. The recent application of proteomic approaches to identify the components of native neuronal (and cardiac) Kv4 channel complexes has revealed even greater complexity than anticipated. The continued emphasis on development of improved biochemical and analytical proteomics methods seems certain to accelerate progress and to provide important new insights into the molecular determinants of native ion channel protein complexes. PMID:20959143

  17. Development of low-loss 765-kV pipe-type cable. Final report

    SciTech Connect

    Allam, E.M.

    1982-01-01

    The successful laboratory development of a 765-kV low-loss high-pressure pipe-type transmission cable and splice, employing an oil-impregnated laminate insulation is described. The laminate is PPP (a polypropylene film sandwiched between two layers of cellulose paper) and the impregnating fluid is hydrogenated polybutene oil. Test results on this cable demonstrated excellent ac and transient electric strength, low dielectric loss and acceptable bending performance. This system appears economically attractive compared to cellulose paper cables at 765 kV.

  18. Gating Charge Immobilization in Kv4.2 Channels: The Basis of Closed-State Inactivation

    PubMed Central

    Dougherty, Kevin; De Santiago-Castillo, Jose A.; Covarrubias, Manuel

    2008-01-01

    Kv4 channels mediate the somatodendritic A-type K+ current (ISA) in neurons. The availability of functional Kv4 channels is dynamically regulated by the membrane potential such that subthreshold depolarizations render Kv4 channels unavailable. The underlying process involves inactivation from closed states along the main activation pathway. Although classical inactivation mechanisms such as N- and P/C-type inactivation have been excluded, a clear understanding of closed-state inactivation in Kv4 channels has remained elusive. This is in part due to the lack of crucial information about the interactions between gating charge (Q) movement, activation, and inactivation. To overcome this limitation, we engineered a charybdotoxin (CTX)-sensitive Kv4.2 channel, which enabled us to obtain the first measurements of Kv4.2 gating currents after blocking K+ conduction with CTX (Dougherty and Covarrubias. 2006J. Gen. Physiol. 128:745–753). Here, we exploited this approach further to investigate the mechanism that links closed-state inactivation to slow Q-immobilization in Kv4 channels. The main observations revealed profound Q-immobilization at steady-state over a range of hyperpolarized voltages (−110 to −75 mV). Depolarization in this range moves <5% of the observable Q associated with activation and is insufficient to open the channels significantly. The kinetics and voltage dependence of Q-immobilization and ionic current inactivation between −153 and −47 mV are similar and independent of the channel's proximal N-terminal region (residues 2–40). A coupled state diagram of closed-state inactivation with a quasi-absorbing inactivated state explained the results from ionic and gating current experiments globally. We conclude that Q-immobilization and closed-state inactivation at hyperpolarized voltages are two manifestations of the same process in Kv4.2 channels, and propose that inactivation in the absence of N- and P/C-type mechanisms involves desensitization to

  19. TVA's 500-kV electric and magnetic fields: measurements and analyses

    SciTech Connect

    Not Available

    1983-09-01

    This report summarizes the results of the 500-kV electric and magnetic fields data project, performed by the Electrical Systems Group of the Tennessee Valley Authority (TVA). The objectives of the project were to measure, record, and analyze electric and magnetic fields in the vicintiy of TVA's 500-kV transmission lines. Also the effects of transmission line fields on the growth and development of selected agricultural and forest plant species were to be investigated. This report contains only the categorization and the statistical analysis of the fields data.

  20. The 80 kV electrostatic wire septum for AmPS

    NASA Astrophysics Data System (ADS)

    Vanderlinden, A.; Bijleveld, J. H. M.; Rookhuizen, H. Boer; Bruinsma, P. J. T.; Heine, E.; Lassing, P.; Prins, E.

    The characteristics of the wire septum for the Amsterdam Pulse Stretcher (AmPS) are summarized. In the extraction process of the AmPS the extracted beam is intercepted from the circulating beam by the 1 m long electrostatic wire septum. For a bending angle of 4.4 mrad, the maximum anode voltage is 80 kV. The system developed consists of a wire spacing of 0.65 mm between tungsten wires of 50 micrometers diameter. Stainless steel spring wires, bent in a half cylindrical carrier, stretch the septum wires two by two. Prototype tests were successful up to an anode voltage of 120 kV.

  1. K(V)7 potassium channels: a new therapeutic target in smooth muscle disorders.

    PubMed

    Stott, Jennifer B; Jepps, Thomas A; Greenwood, Iain A

    2014-04-01

    Potassium channels are key regulators of smooth muscle tone, with increases in activity resulting in hyperpolarisation of the cell membrane, which acts to oppose vasoconstriction. Several potassium channels exist within smooth muscle, but the KV7 family of voltage-gated potassium channels have been identified as being crucial mediators of this process in a variety of smooth muscle. Recently, KV7 channels have been shown to be involved in the pathogenesis of hypertension, as well as being implicated in other smooth muscle disorders, providing a new and inviting target for smooth muscle disorders. PMID:24333708

  2. Planning and installation of the 138 kV South Padre Island submarine cable

    SciTech Connect

    Cooper, J.H. ); Polasek, M.J. )

    1993-10-01

    This paper describes the planning, design and installation phases of a 138 kV submarine cable project which was recently completed by Central Power and Light (CPL) to improve the service reliability of South Padre Island and the town of Port Isabel. The project presented unique installation problems due to the shallow water depths combined with the necessity to minimize the environmental impact to sea grasses during the cable installation. This project resulted in the longest 138 kV extruded dielectric submarine cable circuit in the US.

  3. Concerted Trafficking Regulation of Kv2.1 and KATP Channels by Leptin in Pancreatic β-Cells.

    PubMed

    Wu, Yi; Shyng, Show-Ling; Chen, Pei-Chun

    2015-12-11

    In pancreatic β-cells, voltage-gated potassium 2.1 (Kv2.1) channels are the dominant delayed rectifier potassium channels responsible for action potential repolarization. Here, we report that leptin, a hormone secreted by adipocytes known to inhibit insulin secretion, causes a transient increase in surface expression of Kv2.1 channels in rodent and human β-cells. The effect of leptin on Kv2.1 surface expression is mediated by the AMP-activated protein kinase (AMPK). Activation of AMPK mimics whereas inhibition of AMPK occludes the effect of leptin. Inhibition of Ca(2+)/calmodulin-dependent protein kinase kinase β, a known upstream kinase of AMPK, also blocks the effect of leptin. In addition, the cAMP-dependent protein kinase (PKA) is involved in Kv2.1 channel trafficking regulation. Inhibition of PKA prevents leptin or AMPK activators from increasing Kv2.1 channel density, whereas stimulation of PKA is sufficient to promote Kv2.1 channel surface expression. The increased Kv2.1 surface expression by leptin is dependent on actin depolymerization, and pharmacologically induced actin depolymerization is sufficient to enhance Kv2.1 surface expression. The signaling and cellular mechanisms underlying Kv2.1 channel trafficking regulation by leptin mirror those reported recently for ATP-sensitive potassium (KATP) channels, which are critical for coupling glucose stimulation with membrane depolarization. We show that the leptin-induced increase in surface KATP channels results in more hyperpolarized membrane potentials than control cells at stimulating glucose concentrations, and the increase in Kv2.1 channels leads to a more rapid repolarization of membrane potential in cells firing action potentials. This study supports a model in which leptin exerts concerted trafficking regulation of KATP and Kv2.1 channels to coordinately inhibit insulin secretion. PMID:26453299

  4. KV7 channels contribute to paracrine, but not metabolic or ischemic, regulation of coronary vascular reactivity in swine.

    PubMed

    Goodwill, Adam G; Fu, Lijuan; Noblet, Jillian N; Casalini, Eli D; Sassoon, Daniel; Berwick, Zachary C; Kassab, Ghassan S; Tune, Johnathan D; Dick, Gregory M

    2016-03-15

    Hydrogen peroxide (H2O2) and voltage-dependent K(+) (KV) channels play key roles in regulating coronary blood flow in response to metabolic, ischemic, and paracrine stimuli. The KV channels responsible have not been identified, but KV7 channels are possible candidates. Existing data regarding KV7 channel function in the coronary circulation (limited to ex vivo assessments) are mixed. Thus we examined the hypothesis that KV7 channels are present in cells of the coronary vascular wall and regulate vasodilation in swine. We performed a variety of molecular, biochemical, and functional (in vivo and ex vivo) studies. Coronary arteries expressed KCNQ genes (quantitative PCR) and KV7.4 protein (Western blot). Immunostaining demonstrated KV7.4 expression in conduit and resistance vessels, perhaps most prominently in the endothelial and adventitial layers. Flupirtine, a KV7 opener, relaxed coronary artery rings, and this was attenuated by linopirdine, a KV7 blocker. Endothelial denudation inhibited the flupirtine-induced and linopirdine-sensitive relaxation of coronary artery rings. Moreover, linopirdine diminished bradykinin-induced endothelial-dependent relaxation of coronary artery rings. There was no effect of intracoronary flupirtine or linopirdine on coronary blood flow at the resting heart rate in vivo. Linopirdine had no effect on coronary vasodilation in vivo elicited by ischemia, H2O2, or tachycardia. However, bradykinin increased coronary blood flow in vivo, and this was attenuated by linopirdine. These data indicate that KV7 channels are expressed in some coronary cell type(s) and influence endothelial function. Other physiological functions of coronary vascular KV7 channels remain unclear, but they do appear to contribute to endothelium-dependent responses to paracrine stimuli. PMID:26825518

  5. Regulation of the Kv2.1 potassium channel by MinK and MiRP1

    PubMed Central

    McCrossan, Zoe A.; Roepke, Torsten K.; Lewis, Anthony; Panaghie, Gianina; Abbott, Geoffrey W.

    2010-01-01

    Kv2.1 is a voltage-gated potassium (Kv) channel α subunit expressed in mammalian heart and brain. MinK-related peptides (MiRPs), encoded by KCNE genes, are single transmembrane domain ancillary subunits that form complexes with Kv channel α subunits to modify their function. Mutations in human MinK (KCNE1) and MiRP1 (KCNE2) are associated with inherited and acquired forms of long QT syndrome (LQTS). Here, co-immunoprecipitations from rat heart tissue suggested that both MinK and MiRP1 form native cardiac complexes with Kv2.1. In whole-cell voltage clamp studies of subunits expressed in CHO cells, rat MinK and MiRP1 reduced Kv2.1 current density 3- and 2-fold respectively, slowed Kv2.1 activation (at +60 mV) 2- and 3-fold respectively, and each slowed Kv2.1 deactivation <2-fold. Human MinK slowed Kv2.1 activation 25%, while human MiRP1 slowed Kv2.1 activation and deactivation 2-fold. Inherited mutations in human MinK and MiRP1, previously associated with LQTS, were also evaluated. D76N-MinK and S74L-MinK reduced Kv2.1 current density (3-fold and 40%, respectively) and slowed deactivation (60 and 80%, respectively). Compared to wild-type human MiRP1-Kv2.1 complexes, channels formed with M54T- or I57T-MiRP1 showed greatly slowed activation (10-fold and 5-fold, respectively). The data broaden the potential roles of MinK and MiRP1 in cardiac physiology and support the possibility that inherited mutations in either subunit could contribute to cardiac arrhythmia by multiple mechanisms. PMID:19219384

  6. Modulation of KvAP Unitary Conductance and Gating by 1-Alkanols and Other Surface Active Agents

    PubMed Central

    Finol-Urdaneta, Rocio K.; McArthur, Jeffrey R.; Juranka, Peter F.; French, Robert J.; Morris, Catherine E.

    2010-01-01

    Abstract The actions of alcohols and anesthetics on ion channels are poorly understood. Controversy continues about whether bilayer restructuring is relevant to the modulatory effects of these surface active agents (SAAs). Some voltage-gated K channels (Kv), but not KvAP, have putative low affinity alcohol-binding sites, and because KvAP structures have been determined in bilayers, KvAP could offer insights into the contribution of bilayer mechanics to SAA actions. We monitored KvAP unitary conductance and macroscopic activation and inactivation kinetics in PE:PG/decane bilayers with and without exposure to classic SAAs (short-chain 1-alkanols, cholesterol, and selected anesthetics: halothane, isoflurane, chloroform). At levels that did not measurably alter membrane specific capacitance, alkanols caused functional changes in KvAP behavior including lowered unitary conductance, modified kinetics, and shifted voltage dependence for activation. A simple explanation is that the site of SAA action on KvAP is its entire lateral interface with the PE:PG/decane bilayer, with SAA-induced changes in surface tension and bilayer packing order combining to modulate the shape and stability of various conformations. The KvAP structural adjustment to diverse bilayer pressure profiles has implications for understanding desirable and undesirable actions of SAA-like drugs and, broadly, predicts that channel gating, conductance and pharmacology may differ when membrane packing order differs, as in raft versus nonraft domains. PMID:20197029

  7. 76 FR 19744 - Final Tropic to Hatch 138 kV Transmission Line Project Environmental Impact Statement and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-08

    ... Forest Service Final Tropic to Hatch 138 kV Transmission Line Project Environmental Impact Statement and... Impact Statement (FEIS) for the Tropic to Hatch 138 kV Transmission Line Project and a Proposed... Agency publishes this Notice of Availability in the Federal Register. ADDRESSES: Copies of the Tropic...

  8. Physiological Role of Kv1.3 Channel in T Lymphocyte Cell Investigated Quantitatively by Kinetic Modeling

    PubMed Central

    Feng, Jing; Wang, Wei; Wu, Yingliang; Ding, Jiuping

    2014-01-01

    Kv1.3 channel is a delayed rectifier channel abundant in human T lymphocytes. Chronic inflammatory and autoimmune disorders lead to the over-expression of Kv1.3 in T cells. To quantitatively study the regulatory mechanism and physiological function of Kv1.3 in T cells, it is necessary to have a precise kinetic model of Kv1.3. In this study, we firstly established a kinetic model capable to precisely replicate all the kinetic features for Kv1.3 channels, and then constructed a T-cell model composed of ion channels including Ca2+-release activated calcium (CRAC) channel, intermediate K+ (IK) channel, TASK channel and Kv1.3 channel for quantitatively simulating the changes in membrane potentials and local Ca2+ signaling messengers during activation of T cells. Based on the experimental data from current-clamp recordings, we successfully demonstrated that Kv1.3 dominated the membrane potential of T cells to manipulate the Ca2+ influx via CRAC channel. Our results revealed that the deficient expression of Kv1.3 channel would cause the less Ca2+ signal, leading to the less efficiency in secretion. This was the first successful attempt to simulate membrane potential in non-excitable cells, which laid a solid basis for quantitatively studying the regulatory mechanism and physiological role of channels in non-excitable cells. PMID:24594979

  9. The Dipeptidyl-Peptidase-Like Protein DPP6 Determines the Unitary Conductance of Neuronal Kv4.2 Channels

    PubMed Central

    De Santiago-Castillo, José A.; Rocha, Carmen A.; Nadal, Marcela S.; Rudy, Bernardo; Covarrubias, Manuel

    2009-01-01

    The neuronal subthreshold-operating A-type K+ current regulates electrical excitability, spike timing, and synaptic integration and plasticity. The Kv4 channels underlying this current have been implicated in epilepsy, regulation of dopamine release, and pain plasticity. However, the unitary conductance (γ) of neuronal somatodendritic A-type K+ channels composed of Kv4 pore-forming subunits is larger (∼7.5 pS) than that of Kv4 channels expressed singly in heterologous cells (∼4 pS). Here, we examined the putative novel contribution of the dipeptidyl-peptidase-like protein-6 DPP6-S to the γ of native [cerebellar granule neuron (CGN)] and reconstituted Kv4.2 channels. Coexpression of Kv4.2 proteins with DPP6-S was sufficient to match the γ of native CGN channels; and CGN Kv4 channels from dpp6 knock-out mice yielded a γ indistinguishable from that of Kv4.2 channels expressed singly. Moreover, suggesting electrostatic interactions, charge neutralization mutations of two N-terminal acidic residues in DPP6-S eliminated the increase in γ. Therefore, DPP6-S, as a membrane protein extrinsic to the pore domain, is necessary and sufficient to explain a fundamental difference between native and recombinant Kv4 channels. These observations may help to understand the molecular basis of neurological disorders correlated with recently identified human mutations in the dpp6 gene. PMID:19279261

  10. Silencing of Kv4.1 potassium channels inhibits cell proliferation of tumorigenic human mammary epithelial cells

    SciTech Connect

    Jang, Soo Hwa; Choi, Changsun; Hong, Seong-Geun; Yarishkin, Oleg V.; Bae, Young Min; Kim, Jae Gon; O'Grady, Scott M.; Kang, Kyung-Sun; Ryu, Pan Dong; Lee, So Yeong

    2009-06-26

    Potassium channel activity has been shown to facilitate cell proliferation in cancer cells. In the present study, the role of Kv4.1 channels in immortal and tumorigenic human mammary epithelial cells was investigated. Kv4.1 protein expression was positively correlated with tumorigenicity. Moreover, transfection with siRNAs targeting Kv4.1 mRNA suppressed proliferation of tumorigenic mammary epithelial cells. Experiments using mRNA isolated from human breast cancer tissues revealed that the level of Kv4.1 mRNA expression varied depending on the stage of the tumor. Kv4.1 protein expression increased during stages T2 and T3 compared to normal tissue. These results demonstrated that Kv4.1 plays a role in proliferation of tumorigenic human mammary epithelial cells. In addition, elevated Kv4.1 expression may be useful as a diagnostic marker for staging mammary tumors and selective blockers of Kv4.1 may serve to suppress tumor cell proliferation.

  11. Overview of MoronytoRainbow Line (background) in context with RyantoRainbow 100kV ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    Overview of Morony-to-Rainbow Line (background) in context with Ryan-to-Rainbow 100kV Transmission Lines 1 and 2 (center and foreground) about three miles southwest of Morony Dam and Powerhouse. View to north - Morony Hydroelectric Facility, Morony-to-Rainbow 100 kV Transmission Line, West bank of the Missouri River, Great Falls, Cascade County, MT

  12. Molecular Expression and Pharmacological Evidence for a Functional Role of Kv7 Channel Subtypes in Guinea Pig Urinary Bladder Smooth Muscle

    PubMed Central

    Afeli, Serge A. Y.; Malysz, John; Petkov, Georgi V.

    2013-01-01

    Voltage-gated Kv7 (KCNQ) channels are emerging as essential regulators of smooth muscle excitability and contractility. However, their physiological role in detrusor smooth muscle (DSM) remains to be elucidated. Here, we explored the molecular expression and function of Kv7 channel subtypes in guinea pig DSM by RT-PCR, qRT-PCR, immunohistochemistry, electrophysiology, and isometric tension recordings. In whole DSM tissue, mRNAs for all Kv7 channel subtypes were detected in a rank order: Kv7.1~Kv7.2Kv7.3~Kv7.5Kv7.4. In contrast, freshly-isolated DSM cells showed mRNA expression of: Kv7.1~Kv7.2Kv7.5Kv7.3~Kv7.4. Immunohistochemical confocal microscopy analyses of DSM, conducted by using co-labeling of Kv7 channel subtype-specific antibodies and α-smooth muscle actin, detected protein expression for all Kv7 channel subtypes, except for the Kv7.4, in DSM cells. L-364373 (R-L3), a Kv7.1 channel activator, and retigabine, a Kv7.2-7.5 channel activator, inhibited spontaneous phasic contractions and the 10-Hz electrical field stimulation (EFS)-induced contractions of DSM isolated strips. Linopiridine and XE991, two pan-Kv7 (effective at Kv7.1-Kv7.5 subtypes) channel inhibitors, had opposite effects increasing DSM spontaneous phasic and 10 Hz EFS-induced contractions. EFS-induced DSM contractions generated by a wide range of stimulation frequencies were decreased by L-364373 (10 µM) or retigabine (10 µM), and increased by XE991 (10 µM). Retigabine (10 µM) induced hyperpolarization and inhibited spontaneous action potentials in freshly-isolated DSM cells. In summary, Kv7 channel subtypes are expressed at mRNA and protein levels in guinea pig DSM cells. Their pharmacological modulation can control DSM contractility and excitability; therefore, Kv7 channel subtypes provide potential novel therapeutic targets for urinary bladder dysfunction. PMID:24073284

  13. Approaches targeting K(V)10.1 open a novel window for cancer diagnosis and therapy.

    PubMed

    Pardo, L A; Gómez-Varela, D; Major, F; Sansuk, K; Leurs, R; Downie, B R; Tietze, L F; Stühmer, W

    2012-01-01

    K(V)10.1 has recently become generally accepted as a promising cancer target, as it is ectopically expressed in the majority of solid tumors. Due to its cell-surface accessibility, K(V)10.1 has a strong potential for tumor treatment and diagnosis. Given that its mode of action is likely independent of conventional cancer pathways such as tyrosine kinases, K(V)10.1 opens a novel window for treating cancer. In this review we will give an overview of the current status of data linking K(V)10.1 to cancer, and propose techniques that could exploit K(V)10.1's properties for the management of cancer. PMID:22204340

  14. Complete Genome Sequence of Murine Pneumotropic Virus (Polyomaviridae) Clone pKV(37-1)

    PubMed Central

    Libbey, Jane E.

    2016-01-01

    The murine pneumotropic virus genome encoded by the pKV(37-1) clone was sequenced to completion. The regulatory region harbored a mutation not previously reported. The protein coding regions (large and small T antigens, viral proteins 1 to 3) showed multiple regions of high amino acid identity to the human, simian, and bovine polyomaviruses. PMID:27198030

  15. Complete Genome Sequence of Murine Pneumotropic Virus (Polyomaviridae) Clone pKV(37-1).

    PubMed

    Libbey, Jane E; Fujinami, Robert S

    2016-01-01

    The murine pneumotropic virus genome encoded by the pKV(37-1) clone was sequenced to completion. The regulatory region harbored a mutation not previously reported. The protein coding regions (large and small T antigens, viral proteins 1 to 3) showed multiple regions of high amino acid identity to the human, simian, and bovine polyomaviruses. PMID:27198030

  16. Kv4.2 Mediates Histamine Modulation of Preoptic Neuron Activity and Body Temperature

    PubMed Central

    Sethi, Jasmine; Sanchez-Alavez, Manuel; Tabarean, Iustin V.

    2011-01-01

    Histamine regulates arousal, circadian rhythms, and thermoregulation. Activation of H3 histamine receptors expressed by preoptic GABAergic neurons results in a decrease of their firing rate and hyperthermia. Here we report that an increase in the A-type K+ current in preoptic GABAergic neurons in response to activation of H3 histamine receptors results in decreased firing rate and hyperthermia in mice. The Kv4.2 subunit is required for these actions in spite of the fact that Kv4.2−/− preoptic GABAergic neurons display A-type currents and firing characteristics similar to those of wild-type neurons. This electrical remodeling is achieved by robust upregulation of the expression of the Kv4.1 subunit and of a delayed rectifier current. Dynamic clamp experiments indicate that enhancement of the A-type current by a similar amount to that induced by histamine is sufficient to mimic its robust effect on firing rates. These data indicate a central role played by the Kv4.2 subunit in histamine regulation of body temperature and its interaction with pERK1/2 downstream of the H3 receptor. We also reveal that this pathway provides a mechanism for selective modulation of body temperature at the beginning of the active phase of the circadian cycle. PMID:22220205

  17. Kv4.2 mediates histamine modulation of preoptic neuron activity and body temperature.

    PubMed

    Sethi, Jasmine; Sanchez-Alavez, Manuel; Tabarean, Iustin V

    2011-01-01

    Histamine regulates arousal, circadian rhythms, and thermoregulation. Activation of H3 histamine receptors expressed by preoptic GABAergic neurons results in a decrease of their firing rate and hyperthermia. Here we report that an increase in the A-type K⁺ current in preoptic GABAergic neurons in response to activation of H3 histamine receptors results in decreased firing rate and hyperthermia in mice. The Kv4.2 subunit is required for these actions in spite of the fact that Kv4.2⁻/⁻ preoptic GABAergic neurons display A-type currents and firing characteristics similar to those of wild-type neurons. This electrical remodeling is achieved by robust upregulation of the expression of the Kv4.1 subunit and of a delayed rectifier current. Dynamic clamp experiments indicate that enhancement of the A-type current by a similar amount to that induced by histamine is sufficient to mimic its robust effect on firing rates. These data indicate a central role played by the Kv4.2 subunit in histamine regulation of body temperature and its interaction with pERK1/2 downstream of the H3 receptor. We also reveal that this pathway provides a mechanism for selective modulation of body temperature at the beginning of the active phase of the circadian cycle. PMID:22220205

  18. A kernel-based method for markerless tumor tracking in kV fluoroscopic images.

    PubMed

    Zhang, Xiaoyong; Homma, Noriyasu; Ichiji, Kei; Abe, Makoto; Sugita, Norihiro; Takai, Yoshihiro; Narita, Yuichiro; Yoshizawa, Makoto

    2014-09-01

    Markerless tracking of respiration-induced tumor motion in kilo-voltage (kV) fluoroscopic image sequence is still a challenging task in real time image-guided radiation therapy (IGRT). Most of existing markerless tracking methods are based on a template matching technique or its extensions that are frequently sensitive to non-rigid tumor deformation and involve expensive computation. This paper presents a kernel-based method that is capable of tracking tumor motion in kV fluoroscopic image sequence with robust performance and low computational cost. The proposed tracking system consists of the following three steps. To enhance the contrast of kV fluoroscopic image, we firstly utilize a histogram equalization to transform the intensities of original images to a wider dynamical intensity range. A tumor target in the first frame is then represented by using a histogram-based feature vector. Subsequently, the target tracking is then formulated by maximizing a Bhattacharyya coefficient that measures the similarity between the tumor target and its candidates in the subsequent frames. The numerical solution for maximizing the Bhattacharyya coefficient is performed by a mean-shift algorithm. The proposed method was evaluated by using four clinical kV fluoroscopic image sequences. For comparison, we also implement four conventional template matching-based methods and compare their performance with our proposed method in terms of the tracking accuracy and computational cost. Experimental results demonstrated that the proposed method is superior to conventional template matching-based methods. PMID:25098382

  19. A kernel-based method for markerless tumor tracking in kV fluoroscopic images

    NASA Astrophysics Data System (ADS)

    Zhang, Xiaoyong; Homma, Noriyasu; Ichiji, Kei; Abe, Makoto; Sugita, Norihiro; Takai, Yoshihiro; Narita, Yuichiro; Yoshizawa, Makoto

    2014-09-01

    Markerless tracking of respiration-induced tumor motion in kilo-voltage (kV) fluoroscopic image sequence is still a challenging task in real time image-guided radiation therapy (IGRT). Most of existing markerless tracking methods are based on a template matching technique or its extensions that are frequently sensitive to non-rigid tumor deformation and involve expensive computation. This paper presents a kernel-based method that is capable of tracking tumor motion in kV fluoroscopic image sequence with robust performance and low computational cost. The proposed tracking system consists of the following three steps. To enhance the contrast of kV fluoroscopic image, we firstly utilize a histogram equalization to transform the intensities of original images to a wider dynamical intensity range. A tumor target in the first frame is then represented by using a histogram-based feature vector. Subsequently, the target tracking is then formulated by maximizing a Bhattacharyya coefficient that measures the similarity between the tumor target and its candidates in the subsequent frames. The numerical solution for maximizing the Bhattacharyya coefficient is performed by a mean-shift algorithm. The proposed method was evaluated by using four clinical kV fluoroscopic image sequences. For comparison, we also implement four conventional template matching-based methods and compare their performance with our proposed method in terms of the tracking accuracy and computational cost. Experimental results demonstrated that the proposed method is superior to conventional template matching-based methods.

  20. Compact 180-kV Marx generator triggered in atmospheric air by femtosecond laser filaments

    NASA Astrophysics Data System (ADS)

    Arantchouk, L.; Point, G.; Brelet, Y.; Larour, J.; Carbonnel, J.; André, Y.-B.; Mysyrowicz, A.; Houard, A.

    2014-03-01

    We developed a compact Marx generator triggered in atmospheric air by a single femtosecond laser beam undergoing filamentation. Voltage pulses of 180 kV could be generated with a subnanosecond jitter. The same laser beam was also used to initiate simultaneously guided discharges up to 21 cm long at the output of the generator.

  1. Design and development of a 5 kV isolated solid state switch

    NASA Technical Reports Server (NTRS)

    Holbrook, R. J.; Scapple, R. Y.; Keister, F. Z.; Gooder, S. T.

    1975-01-01

    A unique microcircuit intended for use as a shorting switch for large extraterrestrial solar cell arrays is described. The packaging design for the 5 kV isolated hybrid switch is different from most hybrid microcircuits in that it utilizes a compartmentalized plastic case (a portion of which is encapsulated), is not hermetic, and is designed for high voltage operation.

  2. Pivoting between Calmodulin Lobes Triggered by Calcium in the Kv7.2/Calmodulin Complex

    PubMed Central

    Alaimo, Alessandro; Alberdi, Araitz; Gomis-Perez, Carolina; Fernández-Orth, Juncal; Bernardo-Seisdedos, Ganeko; Malo, Covadonga; Millet, Oscar; Areso, Pilar; Villarroel, Alvaro

    2014-01-01

    Kv7.2 (KCNQ2) is the principal molecular component of the slow voltage gated M-channel, which strongly influences neuronal excitability. Calmodulin (CaM) binds to two intracellular C-terminal segments of Kv7.2 channels, helices A and B, and it is required for exit from the endoplasmic reticulum. However, the molecular mechanisms by which CaM controls channel trafficking are currently unknown. Here we used two complementary approaches to explore the molecular events underlying the association between CaM and Kv7.2 and their regulation by Ca2+. First, we performed a fluorometric assay using dansylated calmodulin (D-CaM) to characterize the interaction of its individual lobes to the Kv7.2 CaM binding site (Q2AB). Second, we explored the association of Q2AB with CaM by NMR spectroscopy, using 15N-labeled CaM as a reporter. The combined data highlight the interdependency of the N- and C-lobes of CaM in the interaction with Q2AB, suggesting that when CaM binds Ca2+ the binding interface pivots between the N-lobe whose interactions are dominated by helix B and the C-lobe where the predominant interaction is with helix A. In addition, Ca2+ makes CaM binding to Q2AB more difficult and, reciprocally, the channel weakens the association of CaM with Ca2+. PMID:24489773

  3. ANTIARRHYTHMIC DRUG-INDUCED INTERNALIZATION OF THE ATRIAL SPECIFIC K+ CHANNEL, Kv1.5

    PubMed Central

    Schumacher, Sarah M.; McEwen, Dyke P.; Zhang, Lian; Arendt, Kristin L.; Van Genderen, Kristin M.; Martens, Jeffrey R.

    2009-01-01

    Conventional antiarrhythmic drugs target the ion permeability of channels, but increasing evidence suggests that functional ion channel density can also be modified pharmacologically. Kv1.5 mediates the ultrarapid potassium current (IKur) that controls atrial action potential duration. Given the atrial specific expression of Kv1.5 and its alterations in human atrial fibrillation, significant effort has been made to identify novel channel blockers. In this study, treatment of HL-1 atrial myocytes expressing Kv1.5-GFP with the class I antiarrhythmic agent quinidine, resulted in a dose-, and temperature-dependent internalization of Kv1.5, concomitant with channel block. This quinidine-induced channel internalization was confirmed in acutely dissociated neonatal myocytes. Channel internalization was subunit-dependent, activity-independent, stereospecific, and blocked by pharmacologic disruption of the endocytic machinery. Pore block and channel internalization partially overlap in the structural requirements for drug binding. Surprisingly, quinidine-induced endocytosis was calcium-dependent and therefore unrecognized by previous biophysical studies focused on isolating channel-drug interactions. Importantly, while acute quinidine-induced internalization was reversible, chronic treatment led to channel degradation. Together, these data reveal a novel mechanism of antiarrhythmic drug action and highlight the possibility for new agents that selectively modulate the stability of channel protein in the membrane as an approach for treating cardiac arrhythmias. PMID:19443837

  4. Kv4 channel blockade reduces motor and neuropsychiatric symptoms in rodent models of Parkinson's disease.

    PubMed

    Aidi-Knani, Sabrine; Regaya, Imed; Amalric, Marianne; Mourre, Christiane

    2015-02-01

    The striatum, a major input structure of basal ganglia, integrates glutamatergic cortical and thalamic inputs to control psychomotor behaviors. Nigrostriatal dopamine (DA) neurodegeneration in Parkinson's disease causes a loss of spinal and glutamatergic synapses in the striatal medium spiny neurons (MSNs). Adaptive responses, a form of homeostatic plasticity, to these changes are caused by a decrease in a potassium Kv4 channel-dependent inactivating A-type potassium (KIA) current that increases the intrinsic excitability of MSNs. Nevertheless, the functional outcome of these compensatory mechanisms does not allow adequate behavioral recovery in vivo. We thus addressed the question of whether further blockade of Kv4 activity could enhance the striatal responsiveness of MSNs to DA depletion and restore normal function in vivo. To test this hypothesis, we examined the effects of a selective blocker of Kv4 channels, AmmTX3, on the motor, cognitive, and emotional symptoms produced by 6-hydroxydopamine lesions of the nigrostriatal DA pathway in rats. Striatal infusion of AmmTX3 (0.2-0.4 μg) reduced motor deficits, decreased anxiety, and restored short-term social and spatial memories. These results underlie the importance of Kv4 channels as players in the homeostatic responses, and, more importantly, provide a potential target for adjunctive therapies for Parkinson's disease. PMID:25356731

  5. Analysis on heat loss characteristics of a 10 kV HTS power substation

    NASA Astrophysics Data System (ADS)

    Teng, Yuping; Dai, Shaotao; Song, Naihao; Zhang, Jingye; Gao, Zhiyuan; Zhu, Zhiqin; Zhou, Weiwei; Wei, Zhourong; Lin, Liangzhen; Xiao, Liye

    2014-09-01

    A 10 kV High Temperature Superconducting power substation (10 kV HTS substation), supported by Chinese State 863 projects, was developed and has been running to supply power for several factories for more than two years at an industrial park of Baiyin, Gansu province in Northwest China. The system of the 10 kV HTS substation compositions, including a HTS cable, a HTS transformer, a SFCL, and a SMES, are introduced. The SMES works at liquid helium temperature and the other three apparatus operates under liquid nitrogen condition. There are mainly four types of heat losses existing in each HTS apparatus of the 10 kV HTS substation, including AC loss, Joule heat loss, conductive heat, and leak-in heat from cryostat. A small quantity of AC loss still exists due to the harmonic component of the current when it carries DC for HTS apparatus. The principle and basis for analysis of the heat losses are introduced and the total heat loss of each apparatus are calculated or estimated, which agree well with the test result. The analysis and result presented are of importance for the design of the refrigeration system.

  6. Design and performance of a 30 KV electron gun with ten independent cathodes & a magnetic lens.

    SciTech Connect

    Rudys, Joseph Matthew; Reed, Kim Warren

    2006-08-01

    Measurements on a 30 kV electron gun with ten independent cathodes, operating in a 6.5 Tesla (T) magnetic field are presented. An earlier paper covered the design of this electron gun [1]. Experimental results are compared to model predictions. Beam current is compared to theoretical space charge limited flow.

  7. Field evaluation of 69-kV outdoor Polysil insulators. Final report

    SciTech Connect

    Richenbacher, A.G.

    1985-03-01

    After three years of exposure to widely varying climates and environments, Polysil (polymer concrete) 69-kV post-type insulators are still performing satisfactorily. In all test situations, Polysil insulators performed as well as - sometimes even surpassed - their porcelain counterparts. They also demonstrated potential for substantially reducing insulator costs.

  8. Hayden-Blue River 345-kV transmission line project, Colorado

    SciTech Connect

    Not Available

    1982-09-01

    Tri-State Generation and Transmission Association, Inc., Colorado-Ute Electric Association, Inc., Platte River Power Authority, and Western Area Power Administration propose to construct and operate approximately 90 miles of 345-kilovolt (kV) transmission line between Hayden and the Blue River Valley in Colorado. The project would involve expansion of existing substation facilities at Hayden and construction of two new substations. The line would be operated at 230 kV initially. Estimated cost of the project is $37.8 million. The new line and substation facilities would provide a backup transmission path, satisfy the long-term needs in meeting the energy requirements, improve system reliability, improve system stability for the Craig and Hayden generating stations, and leave the existing 115-kV and 138-kV lines in operation to provide additional transmission capacity that would function as backup transmission during an outage on another line. Minute amounts of lands would be displaced. Construction activities would disturb critical ranges for elk and mule deer, elk calving areas, and the mating and nesting areas of greater sandhill cranes, great blue herons, sage grouse, golden eagles, and prairie falcons. Management of timberland would damage natural vegetation. The line would traverse 3.1 miles of flood-prone area, and as many as three transmission towers would lie within the floodplain of the Colorado River. The visual quality of land crossed by the line would be degraded somewhat.

  9. 29 CFR 1926.1409 - Power line safety (over 350 kV).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 29 Labor 8 2011-07-01 2011-07-01 false Power line safety (over 350 kV). 1926.1409 Section 1926.1409 Labor Regulations Relating to Labor (Continued) OCCUPATIONAL SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR (CONTINUED) SAFETY AND HEALTH REGULATIONS FOR CONSTRUCTION Cranes and Derricks...

  10. 29 CFR 1926.1409 - Power line safety (over 350 kV).

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 29 Labor 8 2012-07-01 2012-07-01 false Power line safety (over 350 kV). 1926.1409 Section 1926.1409 Labor Regulations Relating to Labor (Continued) OCCUPATIONAL SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR (CONTINUED) SAFETY AND HEALTH REGULATIONS FOR CONSTRUCTION Cranes and Derricks...

  11. 8. VIEW OF 100 kV SWITCHYARD. TELEPHONE BOOTH AND THREESTALL ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    8. VIEW OF 100 kV SWITCHYARD. TELEPHONE BOOTH AND THREE-STALL GARAGE. VIEW TO SOUTHWEST. - Rainbow Hydroelectric Facility, On north bank of Missouri River 2 miles Northeast of Great Falls, & end of Rainbow Dam Road, Great Falls, Cascade County, MT

  12. Block of cloned Kv4.3 potassium channels by dapoxetine.

    PubMed

    Jeong, Imju; Kim, Sae Woong; Yoon, Shin Hee; Hahn, Sang June

    2012-06-01

    Dapoxetine, a short-acting selective serotonin reuptake inhibitor, is widely prescribed for the treatment of patients with premature ejaculation. The effects of dapoxetine were examined on cloned Kv4.3 channels stably expressed in Chinese hamster ovary cells using the whole-cell patch-clamp technique. Dapoxetine not only reduced the peak amplitude of Kv4.3 currents but also accelerated the decay rate of current inactivation in a concentration-dependent manner. Thus, the concentration-dependent reduction in Kv4.3 was measured from the integral of the current during the depolarizing pulse. Dapoxetine decreased the integral of the Kv4.3 currents over the duration of a depolarizing pulse with an IC(50) of 5.3 μM. Analysis of the time dependence of the block gave estimates of an association rate constant (k(+1)) of 3.9 μM(-1)s(-1) and a dissociation rate constant (k(-1)) of 25.6s(-1). The K(D) (k(-1)/k(+1)) was 6.5 μM, similar to the IC(50) value calculated from the concentration-response curve. The block of Kv4.3 by dapoxetine was highly voltage-dependent at a membrane potential coinciding with the activation of the channels. The additional block by dapoxetine displayed a shallow voltage dependence (δ=0.21) in the full activation voltage range. The steady-state inactivation curves were shifted in the hyperpolarizing direction in the presence of dapoxetine. Dapoxetine also caused a substantial acceleration in closed-state inactivation. Dapoxetine produced a significant use-dependent block, which was accompanied by a delayed recovery from inactivation of Kv4.3 currents. These results indicated that dapoxetine potently blocks Kv4.3 currents by both preferentially binding to the open state of the channels and accelerating the closed-state inactivation. These data could provide insight into the mechanism underlying some of the therapeutic actions of this drug. PMID:22192593

  13. Diet-induced obesity resistance of Kv1.3−/− mice is olfactory bulb dependent

    PubMed Central

    Tucker, Kristal; Overton, J. Michael; Fadool, Debra Ann

    2012-01-01

    Gene-targeted deletion of the voltage-gated potassium channel, Kv1.3 (Kv1.3−/−), increases olfactory sensitivity and discriminatory ability, and causes resistance to diet-induced obesity (DIO) in mice. The objective of the present study was to determine if the enhanced olfactory ability of the Kv1.3−/− mouse contributes to the resistance to DIO. Kv1.3+/+ and Kv1.3−/− mice were subject to bilateral olfactory bulbectomy (OBX) or sham surgery at 9 weeks of age and placed on either a control chow diet (CF) or a 32% moderately high-fat diet (MHF). Caloric and water intake, locomotor activity, and oxygen consumption were monitored after 5 weeks of diet treatment. At the end of 26 weeks of diet treatment, fat pad weight and blood chemistry were evaluated. Kv1.3+/+ mice exhibited a significant increase in weight, adiposity, fasting glucose and fasting leptin in response to the MHF-diet, with or without OBX. When treated with a MHF-diet, Kv1.3−/− mice gained significantly less weight than Kv1.3 +/+ mice and exhibited a significant increase in light phase metabolism. OBX of Kv1.3−/− mice prevented the resistance to DIO and concomitant upregulation of light phase metabolism while decreasing dark phase metabolism and total energy expenditure. These findings suggest that pathways activated in Kv1.3 −/− that increased energy expenditure and led to resistance to DIO are olfactory bulb dependent. Thus, these findings add to a growing body of evidence suggesting that the olfactory system can modulate pathways involved in the regulation of energy balance. PMID:22435906

  14. Contribution of Kv1.2 Voltage-gated Potassium Channel to D2 Autoreceptor Regulation of Axonal Dopamine Overflow*

    PubMed Central

    Fulton, Stephanie; Thibault, Dominic; Mendez, Jose A.; Lahaie, Nicolas; Tirotta, Emanuele; Borrelli, Emiliana; Bouvier, Michel; Tempel, Bruce L.; Trudeau, Louis-Eric

    2011-01-01

    Impairments in axonal dopamine release are associated with neurological disorders such as schizophrenia and attention deficit hyperactivity disorder and pathophysiological conditions promoting drug abuse and obesity. The D2 dopamine autoreceptor (D2-AR) exerts tight regulatory control of axonal dopamine (DA) release through a mechanism suggested to involve K+ channels. To evaluate the contribution of Kv1 voltage-gated potassium channels of the Shaker gene family to the regulation of axonal DA release by the D2-AR, the present study employed expression analyses, real time measurements of striatal DA overflow, K+ current measurements and immunoprecipitation assays. Kv1.1, -1.2, -1.3, and -1.6 mRNA and protein were detected in midbrain DA neurons purified by fluorescence-activated cell sorting and in primary DA neuron cultures. In addition, Kv1.1, -1.2, and -1.6 were localized to DA axonal processes in the dorsal striatum. By means of fast scan cyclic voltammetry in striatal slice preparations, we found that the inhibition of stimulation-evoked DA overflow by a D2 agonist was attenuated by Kv1.1, -1.2, and -1.6 toxin blockers. A particular role for the Kv1.2 subunit in the process whereby axonal D2-AR inhibits DA overflow was established with the use of a selective Kv1.2 blocker and Kv1.2 knock-out mice. Moreover, we demonstrate the ability of D2-AR activation to increase Kv1.2 currents in co-transfected cells and its reliance on Gβγ subunit signaling along with the physical coupling of D2-AR and Kv1.2-containing channels in striatal tissue. These findings underline the contribution of Kv1.2 in the regulation of nigrostriatal DA release by the D2-AR and thereby offer a novel mechanism by which DA release is regulated. PMID:21233214

  15. A phenomenological kV beam model for cone-beam imaging

    NASA Astrophysics Data System (ADS)

    Bhagwat, Mandar S.; Blessing, Manuel; Lyatskaya, Yulia; Zygmanski, Piotr

    2010-10-01

    A phenomenological kV beam model was developed to address attenuation and scatter in radiographic images for the purpose of cone-beam imaging. Characterization of a kV beam in terms of the minimal number of parameters and calculation of attenuation and scatter in radiographs of scanned objects are the main applications of this model. Model parameters are derived from radiographs of homogeneous solid water phantoms for various depths and field sizes. The response of the cone-beam detector to kV beams is factorized into different contributions such as output factor, tissue-air ratio and off-axis ratio, with each contribution having an analytical representation. The formulas which are used to characterize the beam model in uniform phantoms are then extended to arbitrary objects using the concept of the water-equivalent pathlength. A weighted sum of three Gaussians in each direction models the dose deposition kernel. Detector response arising from the first Gaussian term can be interpreted as the primary signal while the second and third Gaussians constitute short- and long-range scatter. The model is then applied to predict the primary and scatter signals for arbitrary objects. A technique of scatter removal from the measured radiographs is investigated. The model accurately predicts detector response of varying-thickness phantoms such as multi-step and cylindrical phantoms. The scatter contributes over 90% to the total signal for 20 cm thick phantoms. The calculated scatter-to-primary ratio as a function of spatial coordinates agrees with Monte Carlo studies reported in the literature. Water-equivalent thickness related to primary and scatter contributions calculated from an analysis of radiographs results in an improved calibration technique suitable for CB-CT reconstruction. The kV beam model and the associated theoretical formulations can be utilized to characterize any kV beam line; however, for the specific study the OBI™ system (Varian) was used to obtain

  16. Investigation on X-Radiation for 126 kV Vacuum Interrupters

    NASA Astrophysics Data System (ADS)

    Yan, Jing; Liu, Zhiyuan; Geng, Yingsan; Zhang, Sheng; Zhang, Yingyao

    2016-05-01

    When subjected to high voltages between opened contacts, vacuum interrupters may emit X-rays. In order to ensure that these are of an acceptable level, vacuum interrupters should comply with the limits for X-ray emission and the test procedures to be carried out to verify this based on relevant standards and specifications. In this paper, a comprehensive experimental study has been performed for 126 kV vacuum interrupters used in a transmission system to understand the X-radiation level and its influence by three main parameters, namely applied power-frequency voltage, contact gap and power-frequency voltage conditioning. The radiation instrument is an FJ347 radiometer and the X-radiation dose was measured at the power-frequency test voltage. These tests demonstrated that the X-radiation emission level for a 126 kV vacuum interrupter did not exceed the following: 5 μSv per hour at a rated voltage of 126 kV and 150 μSv per hour at a power-frequency voltage of 230 kV at 1 m distance. The X-radiation dose increased with the applied power-frequency voltage increasing and decreased with the contact gap increasing. The X-radiation dose for 126 kV vacuum interrupters decreased by 57% after the conditioning procedure with a certain power-frequency voltage. During the conditioning procedure, the average value of the X-radiation dose was 4.49 mSv, which means if a professional conditions 180 interrupters per year, it will be safe at the 6.4 m distance. supported by National Key Basic Research Program of China (973 Program) (No. 2015CB251002)

  17. Blockade of Kv1.3 channels ameliorates radiation-induced brain injury

    PubMed Central

    Peng, Ying; Lu, Kui; Li, Zichen; Zhao, Yaodong; Wang, Yiping; Hu, Bin; Xu, Pengfei; Shi, Xiaolei; Zhou, Bin; Pennington, Michael; Chandy, K. George; Tang, Yamei

    2014-01-01

    Background Tumors affecting the head, neck, and brain account for significant morbidity and mortality. The curative efficacy of radiotherapy for these tumors is well established, but radiation carries a significant risk of neurologic injury. So far, neuroprotective therapies for radiation-induced brain injury are still limited. In this study we demonstrate that Stichodactyla helianthus (ShK)–170, a specific inhibitor of the voltage-gated potassium (Kv)1.3 channel, protected mice from radiation-induced brain injury. Methods Mice were treated with ShK-170 for 3 days immediately after brain irradiation. Radiation-induced brain injury was assessed by MRI scans and a Morris water maze. Pathophysiological change of the brain was measured by immunofluorescence. Gene and protein expressions of Kv1.3 and inflammatory factors were measured by quantitative real-time PCR, reverse transcription PCR, ELISA assay, and western blot analyses. Kv currents were recorded in the whole-cell configuration of the patch-clamp technique. Results Radiation increased Kv1.3 mRNA and protein expression in microglia. Genetic silencing of Kv1.3 by specific short interference RNAs or pharmacological blockade with ShK-170 suppressed radiation-induced production of the proinflammatory factors interleukin-6, cyclooxygenase-2, and tumor necrosis factor–α by microglia. ShK-170 also inhibited neurotoxicity mediated by radiation-activated microglia and promoted neurogenesis by increasing the proliferation of neural progenitor cells. Conclusions The therapeutic effect of ShK-170 is mediated by suppression of microglial activation and microglia-mediated neurotoxicity and enhanced neurorestoration by promoting proliferation of neural progenitor cells. PMID:24305723

  18. Combined kV and MV imaging for real-time tracking of implanted fiducial markers

    SciTech Connect

    Wiersma, R. D.; Mao Weihua; Xing, L.

    2008-04-15

    In the presence of intrafraction organ motion, target localization uncertainty can greatly hamper the advantage of highly conformal dose techniques such as intensity modulated radiation therapy (IMRT). To minimize the adverse dosimetric effect caused by tumor motion, a real-time knowledge of the tumor position is required throughout the beam delivery process. The recent integration of onboard kV diagnostic imaging together with MV electronic portal imaging devices on linear accelerators can allow for real-time three-dimensional (3D) tumor position monitoring during a treatment delivery. The aim of this study is to demonstrate a near real-time 3D internal fiducial tracking system based on the combined use of kV and MV imaging. A commercially available radiotherapy system equipped with both kV and MV imaging systems was used in this work. A hardware video frame grabber was used to capture both kV and MV video streams simultaneously through independent video channels at 30 frames per second. The fiducial locations were extracted from the kV and MV images using a software tool. The geometric tracking capabilities of the system were evaluated using a pelvic phantom with embedded fiducials placed on a moveable stage. The maximum tracking speed of the kV/MV system is approximately 9 Hz, which is primarily limited by the frame rate of the MV imager. The geometric accuracy of the system is found to be on the order of less than 1 mm in all three spatial dimensions. The technique requires minimal hardware modification and is potentially useful for image-guided radiation therapy systems.

  19. Critical contribution of KV1 channels to the regulation of coronary blood flow.

    PubMed

    Goodwill, Adam G; Noblet, Jillian N; Sassoon, Daniel; Fu, Lijuan; Kassab, Ghassan S; Schepers, Luke; Herring, B Paul; Rottgen, Trey S; Tune, Johnathan D; Dick, Gregory M

    2016-09-01

    Ion channels in smooth muscle control coronary vascular tone, but the identity of the potassium channels involved requires further investigation. The purpose of this study was to evaluate the functional role of KV1 channels on porcine coronary blood flow using the selective antagonist correolide. KV1 channel gene transcripts were found in porcine coronary arteries, with KCNA5 (encoding KV1.5) being most abundant (P < 0.001). Immunohistochemical staining demonstrated KV1.5 protein in the vascular smooth muscle layer of both porcine and human coronary arteries, including microvessels. Whole-cell patch-clamp experiments demonstrated significant correolide-sensitive (1-10 µM) current in coronary smooth muscle. In vivo studies included direct intracoronary infusion of vehicle or correolide into a pressure-clamped left anterior descending artery of healthy swine (n = 5 in each group) with simultaneous measurement of coronary blood flow. Intracoronary correolide (~0.3-3 µM targeted plasma concentration) had no effect on heart rate or systemic pressure, but reduced coronary blood flow in a dose-dependent manner (P < 0.05). Dobutamine (0.3-10 µg/kg/min) elicited coronary metabolic vasodilation and intracoronary correolide (3 µM) significantly reduced coronary blood flow at any given level of myocardial oxygen consumption (P < 0.001). Coronary artery occlusions (15 s) elicited reactive hyperemia and correolide (3 µM) reduced the flow volume repayment by approximately 30 % (P < 0.05). Taken together, these data support a major role for KV1 channels in modulating baseline coronary vascular tone and, perhaps, vasodilation in response to increased metabolism and transient ischemia. PMID:27496159

  20. Verapamil- and state-dependent effect of 2-aminoethylmethanethiosulphonate (MTSEA) on hKv1.3 channels

    PubMed Central

    Nikouee, Azadeh; Janbein, Malika; Grissmer, Stephan

    2012-01-01

    BACKGROUND AND PURPOSE T-cells usually express voltage-gated Kv1.3 channels. These channels are distinguished by their typical C-type inactivation. Therefore, to be able to rationally design drugs specific for the C-type inactivation state that may have therapeutic value in autoimmune disease therapy, it is necessary to identify those amino acids that are accessible for drug binding in C-type inactivated channels. EXPERIMENTAL APPROACH The influence of 2-aminoethylmethanethiosulphonate (MTSEA) on currents through wild-type human Kv1.3 (hKv1.3) and three mutant channels, hKv1.3_L418C, hKv1.3_T419C and hKv1.3_I420C, in the closed, open and inactivated states was investigated by the patch-clamp technique. KEY RESULTS Currents through hKv1.3_L418C and hKv1.3_T419C channels were irreversibly reduced after the external application of MTSEA in the open state but not in the inactivated and closed states. Currents through hKv1.3_I420C channels were irreversibly reduced in the open and inactivated states but not in the closed state. In the presence of verapamil, the MTSEA modification of hKv1.3_T419C and hKv1.3_I420C channels was prevented, while the MTSEA modification of hKv1.3_L418C channels was unaffected. CONCLUSION AND IMPLICATIONS From our experiments, we conclude that the activation gate of all mutant channels must be open for modification by MTSEA and must also be open during inactivation. In addition, the relative movement of the S6 segments that occur during C-type inactivation includes a movement of the side chains of the amino acids at positions 418 and 419 away from the pore lining. Furthermore, the overlapping binding site for MTSEA and verapamil does not include position 418 in hKv1.3 channels. PMID:22748056

  1. Decrease of a Current Mediated by Kv1.3 Channels Causes Striatal Cholinergic Interneuron Hyperexcitability in Experimental Parkinsonism.

    PubMed

    Tubert, Cecilia; Taravini, Irene R E; Flores-Barrera, Eden; Sánchez, Gonzalo M; Prost, María Alejandra; Avale, María Elena; Tseng, Kuei Y; Rela, Lorena; Murer, Mario Gustavo

    2016-09-01

    The mechanism underlying a hypercholinergic state in Parkinson's disease (PD) remains uncertain. Here, we show that disruption of the Kv1 channel-mediated function causes hyperexcitability of striatal cholinergic interneurons in a mouse model of PD. Specifically, our data reveal that Kv1 channels containing Kv1.3 subunits contribute significantly to the orphan potassium current known as IsAHP in striatal cholinergic interneurons. Typically, this Kv1 current provides negative feedback to depolarization that limits burst firing and slows the tonic activity of cholinergic interneurons. However, such inhibitory control of cholinergic interneuron excitability by Kv1.3-mediated current is markedly diminished in the parkinsonian striatum, suggesting that targeting Kv1.3 subunits and their regulatory pathways may have therapeutic potential in PD therapy. These studies reveal unexpected roles of Kv1.3 subunit-containing channels in the regulation of firing patterns of striatal cholinergic interneurons, which were thought to be largely dependent on KCa channels. PMID:27568555

  2. Cortactin controls surface expression of the voltage-gated potassium channel K(V)10.1.

    PubMed

    Herrmann, Solveig; Ninkovic, Milena; Kohl, Tobias; Lörinczi, Éva; Pardo, Luis A

    2012-12-28

    K(V)10.1 is a voltage-gated potassium channel aberrantly expressed in many cases of cancer, and participates in cancer initiation and tumor progression. Its action as an oncoprotein can be inhibited by a functional monoclonal antibody, indicating a role for channels located at the plasma membrane, accessible to the antibody. Cortactin is an actin-interacting protein implicated in cytoskeletal architecture and often amplified in several types of cancer. In this study, we describe a physical and functional interaction between cortactin and K(V)10.1. Binding of these two proteins occurs between the C terminus of K(V)10.1 and the proline-rich domain of cortactin, regions targeted by many post-translational modifications. This interaction is specific for K(V)10.1 and does not occur with K(V)10.2. Cortactin controls the abundance of K(V)10.1 at the plasma membrane and is required for functional expression of K(V)10.1 channels. PMID:23144454

  3. KV10.1 K(+)-channel plasma membrane discrete domain partitioning and its functional correlation in neurons.

    PubMed

    Jiménez-Garduño, Aura M; Mitkovski, Miso; Alexopoulos, Ioannis K; Sánchez, Araceli; Stühmer, Walter; Pardo, Luis A; Ortega, Alicia

    2014-03-01

    KV10.1 potassium channels are implicated in a variety of cellular processes including cell proliferation and tumour progression. Their expression in over 70% of human tumours makes them an attractive diagnostic and therapeutic target. Although their physiological role in the central nervous system is not yet fully understood, advances in their precise cell localization will contribute to the understanding of their interactions and function. We have determined the plasma membrane (PM) distribution of the KV10.1 protein in an enriched mouse brain PM fraction and its association with cholesterol- and sphingolipid-rich domains. We show that the KV10.1 channel has two different populations in a 3:2 ratio, one associated to and another excluded from Detergent Resistant Membranes (DRMs). This distribution of KV10.1 in isolated PM is cholesterol- and cytoskeleton-dependent since alteration of those factors changes the relationship to 1:4. In transfected HEK-293 cells with a mutant unable to bind Ca(2+)/CaM to KV10.1 protein, Kv10.1 distribution in DRM/non-DRM is 1:4. Mean current density was doubled in the cholesterol-depleted cells, without any noticeable effects on other parameters. These results demonstrate that recruitment of the KV10.1 channel to the DRM fractions involves its functional regulation. PMID:24269539

  4. Heteromeric Kv7.2/7.3 channels differentially regulate action potential initiation and conduction in neocortical myelinated axons.

    PubMed

    Battefeld, Arne; Tran, Baouyen T; Gavrilis, Jason; Cooper, Edward C; Kole, Maarten H P

    2014-03-01

    Rapid energy-efficient signaling along vertebrate axons is achieved through intricate subcellular arrangements of voltage-gated ion channels and myelination. One recently appreciated example is the tight colocalization of K(v)7 potassium channels and voltage-gated sodium (Na(v)) channels in the axonal initial segment and nodes of Ranvier. The local biophysical properties of these K(v)7 channels and the functional impact of colocalization with Na(v) channels remain poorly understood. Here, we quantitatively examined K(v)7 channels in myelinated axons of rat neocortical pyramidal neurons using high-resolution confocal imaging and patch-clamp recording. K(v)7.2 and 7.3 immunoreactivity steeply increased within the distal two-thirds of the axon initial segment and was mirrored by the conductance density estimates, which increased from ~12 (proximal) to 150 pS μm(-2) (distal). The axonal initial segment and nodal M-currents were similar in voltage dependence and kinetics, carried by K(v)7.2/7.3 heterotetramers, 4% activated at the resting membrane potential and rapidly activated with single-exponential time constants (~15 ms at 28 mV). Experiments and computational modeling showed that while somatodendritic K(v)7 channels are strongly activated by the backpropagating action potential to attenuate the afterdepolarization and repetitive firing, axonal K(v)7 channels are minimally recruited by the forward-propagating action potential. Instead, in nodal domains K(v)7.2/7.3 channels were found to increase Na(v) channel availability and action potential amplitude by stabilizing the resting membrane potential. Thus, K(v)7 clustering near axonal Na(v) channels serves specific and context-dependent roles, both restraining initiation and enhancing conduction of the action potential. PMID:24599470

  5. Heteromeric Kv7.2/7.3 Channels Differentially Regulate Action Potential Initiation and Conduction in Neocortical Myelinated Axons

    PubMed Central

    Battefeld, Arne; Tran, Baouyen T.; Gavrilis, Jason; Cooper, Edward C.

    2014-01-01

    Rapid energy-efficient signaling along vertebrate axons is achieved through intricate subcellular arrangements of voltage-gated ion channels and myelination. One recently appreciated example is the tight colocalization of Kv7 potassium channels and voltage-gated sodium (Nav) channels in the axonal initial segment and nodes of Ranvier. The local biophysical properties of these Kv7 channels and the functional impact of colocalization with Nav channels remain poorly understood. Here, we quantitatively examined Kv7 channels in myelinated axons of rat neocortical pyramidal neurons using high-resolution confocal imaging and patch-clamp recording. Kv7.2 and 7.3 immunoreactivity steeply increased within the distal two-thirds of the axon initial segment and was mirrored by the conductance density estimates, which increased from ∼12 (proximal) to 150 pS μm−2 (distal). The axonal initial segment and nodal M-currents were similar in voltage dependence and kinetics, carried by Kv7.2/7.3 heterotetramers, 4% activated at the resting membrane potential and rapidly activated with single-exponential time constants (∼15 ms at 28 mV). Experiments and computational modeling showed that while somatodendritic Kv7 channels are strongly activated by the backpropagating action potential to attenuate the afterdepolarization and repetitive firing, axonal Kv7 channels are minimally recruited by the forward-propagating action potential. Instead, in nodal domains Kv7.2/7.3 channels were found to increase Nav channel availability and action potential amplitude by stabilizing the resting membrane potential. Thus, Kv7 clustering near axonal Nav channels serves specific and context-dependent roles, both restraining initiation and enhancing conduction of the action potential. PMID:24599470

  6. Kv7 potassium channels in airway smooth muscle cells: signal transduction intermediates and pharmacological targets for bronchodilator therapy.

    PubMed

    Brueggemann, Lioubov I; Kakad, Priyanka P; Love, Robert B; Solway, Julian; Dowell, Maria L; Cribbs, Leanne L; Byron, Kenneth L

    2012-01-01

    Expression and function of Kv7 (KCNQ) voltage-activated potassium channels in guinea pig and human airway smooth muscle cells (ASMCs) were investigated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), patch-clamp electrophysiology, and precision-cut lung slices. qRT-PCR revealed expression of multiple KCNQ genes in both guinea pig and human ASMCs. Currents with electrophysiological and pharmacological characteristics of Kv7 currents were measured in freshly isolated guinea pig and human ASMCs. In guinea pig ASMCs, Kv7 currents were significantly suppressed by application of the bronchoconstrictor agonists methacholine (100 nM) or histamine (30 μM), but current amplitudes were restored by addition of a Kv7 channel activator, flupirtine (10 μM). Kv7 currents in guinea pig ASMCs were also significantly enhanced by another Kv7.2-7.5 channel activator, retigabine, and by celecoxib and 2,5-dimethyl celecoxib. In precision-cut human lung slices, constriction of airways by histamine was significantly reduced in the presence of flupirtine. Kv7 currents in both guinea pig and human ASMCs were inhibited by the Kv7 channel blocker XE991. In human lung slices, XE991 induced robust airway constriction, which was completely reversed by addition of the calcium channel blocker verapamil. These findings suggest that Kv7 channels in ASMCs play an essential role in the regulation of airway diameter and may be targeted pharmacologically to relieve airway hyperconstriction induced by elevated concentrations of bronchoconstrictor agonists. PMID:21964407

  7. Kv7 potassium channels in airway smooth muscle cells: signal transduction intermediates and pharmacological targets for bronchodilator therapy

    PubMed Central

    Brueggemann, Lioubov I.; Kakad, Priyanka P.; Love, Robert B.; Solway, Julian; Dowell, Maria L.; Cribbs, Leanne L.

    2012-01-01

    Expression and function of Kv7 (KCNQ) voltage-activated potassium channels in guinea pig and human airway smooth muscle cells (ASMCs) were investigated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), patch-clamp electrophysiology, and precision-cut lung slices. qRT-PCR revealed expression of multiple KCNQ genes in both guinea pig and human ASMCs. Currents with electrophysiological and pharmacological characteristics of Kv7 currents were measured in freshly isolated guinea pig and human ASMCs. In guinea pig ASMCs, Kv7 currents were significantly suppressed by application of the bronchoconstrictor agonists methacholine (100 nM) or histamine (30 μM), but current amplitudes were restored by addition of a Kv7 channel activator, flupirtine (10 μM). Kv7 currents in guinea pig ASMCs were also significantly enhanced by another Kv7.2–7.5 channel activator, retigabine, and by celecoxib and 2,5-dimethyl celecoxib. In precision-cut human lung slices, constriction of airways by histamine was significantly reduced in the presence of flupirtine. Kv7 currents in both guinea pig and human ASMCs were inhibited by the Kv7 channel blocker XE991. In human lung slices, XE991 induced robust airway constriction, which was completely reversed by addition of the calcium channel blocker verapamil. These findings suggest that Kv7 channels in ASMCs play an essential role in the regulation of airway diameter and may be targeted pharmacologically to relieve airway hyperconstriction induced by elevated concentrations of bronchoconstrictor agonists. PMID:21964407

  8. Inhibitory actions of the phosphatidylinositol 3-kinase inhibitor LY294002 on the human Kv1.5 channel

    PubMed Central

    Wu, J; Ding, W-G; Matsuura, H; Tsuji, K; Zang, W-J; Horie, M

    2009-01-01

    Background and purpose: Kv1.5 channels conduct the ultra-rapid delayed rectifier potassium current (IKur), and in humans, Kv1.5 channels are highly expressed in cardiac atria but are scarce in ventricles. Pharmacological blockade of human Kv1.5 (hKv1.5) has been regarded as effective for prevention and treatment of re-entry-based atrial tachyarrhythmias. Here we examined blockade of hKv1.5 channels by LY294002, a well-known inhibitor of phosphatidylinositol 3-kinase (PI3K). Experimental approach: hKv1.5 channels were heterologously expressed in Chinese hamster ovary cells. Effects of LY294002 on wild-type and mutant (T462C, H463C, T480A, R487V, A501V, I502A, I508A, L510A and V516A) hKv1.5 channels were examined by using the whole-cell patch-clamp method. Key results: LY294002 rapidly and reversibly inhibited hKv1.5 current in a concentration-dependent manner (IC50 of 7.9 µmol·L−1). In contrast, wortmannin, a structurally distinct inhibitor of PI3K, had little inhibitory effect on hKv1.5 current. LY294002 block of hKv1.5 current developed with time during depolarizing voltage-clamp steps, and this blockade was also voltage-dependent with a steep increase over the voltage range for channel openings. The apparent binding (k+1) and unbinding (k−1) rate constants were calculated to be 1.6 µmol·L−1−1·s−1 and 5.7 s−1 respectively. Inhibition by LY294002 was significantly reduced in several hKv1.5 mutant channels: T480A, R487V, I502A, I508A, L510A and V516A. Conclusions and implications: LY294002 acts directly on hKv1.5 currents as an open channel blocker, independently of its effects on PI3K activity. Amino acid residues located in the pore region (Thr480, Arg487) and the S6 segment (Ile502, Ile508, Leu510, Val516) appear to constitute potential binding sites for LY294002. PMID:19154427

  9. Measurement of backscattered x-ray spectra at the water surface in the energy range 60 kV to 120 kV.

    PubMed

    Aoki, Kiyoshi; Koyama, Masaki

    2002-04-01

    Backscattered x-ray spectra at the water surface have been measured by using a small silicon diode detector. The measurements have been made at tube voltages 60 kV to 120 kV (HVL 2.4-6.1 mm Al) and field sizes 5 x 5 cm2 to 30 x 30 cm2. The measured spectra are corrected for detector distortion and for the angular dependence of detector efficiency. The obtained backscattered spectrum has a lower mean energy and a narrower shape than the primary spectrum. The ratio of the mean energy of the backscattered spectrum to that of the primary spectrum is between 0.83 and 0.94. The ratio of the spectrum width at 10% of the continuous spectrum maximum is between 0.65 and 0.78. The change of spectral shape due to the field size is slight. In the high-voltage spectra, the peak due to the Compton scattering of tungsten Kalpha x-rays is observed. The backscatter factors (BSFs) calculated from the obtained spectra show a satisfactory agreement with other studies. The difference between the BSF defined as the ratio of air kerma and the BSF defined as the ratio of water kerma is also calculated; the maximum difference is 0.43%. The empirical equation showing the relation between the two BSFs is presented. PMID:11996064

  10. Note: Design and tests of a 13 kA-6.5 kV thyristor switch for a pulsed inductive vacuum ultraviolet source

    NASA Astrophysics Data System (ADS)

    Teske, C.; Lee, B.-J.; Jacoby, J.; Schweizer, W.; Sun, J. Chao

    2010-04-01

    In this paper, the design, construction, and test procedure of a closing switch prototype based on thyristors is described. In particular, details are given about the design criteria and about the triggering board architecture, which is a high side biased, self supplied unit using the electrical energy derived from a local snubber network for the gate drive. The structure guarantees a hard firing gate pulse for the required high dI /dt application. Further, the results of the prototype tests are presented and discussed. The stack assembly has a holding voltage of 6.5 kV and is used for switching a series resonant circuit with a ringing frequency of 12 kHz for a pulsed inductive vacuum ultraviolet source. Maximum current amplitudes of 13 kA and pulse energies of more than 600 J were switched during the test procedure.

  11. Materials science applications of a 120 kV FEG TEM/STEM: Triskaidekaphilia

    SciTech Connect

    Bentley, J.; Fisher, A.T.; Kenik, E.A.; Wang, Z.L.

    1991-01-01

    The introduction by several manufacturers of 200kV transmission electron microscopes (TEM) equipped with field emission guns affords the opportunity to assess their potential impact on materials science by examining applications of similar 100-120kV instruments that have been in use for more than a decade. This summary is based on results from a Philips EM400T/FEG configured as an analytical electron microscope (AEM) with a 6,585 scanning transmission (STEM) unit, EDAX 9100/70 or 9900 energy dispersive X-ray spectroscopy (EDS) systems, and Gatan 607 serial- or 666 parallel-detection electron energy-loss spectrometers (EELS). Examples in four areas that illustrate applications that are impossible or so difficult as to be impracticable with conventional thermionic electron guns are described.

  12. An electrically triggered 200 kV rail-gap switch for wide aperture excimer lasers

    NASA Astrophysics Data System (ADS)

    Endoh, A.; Watanabe, S.; Watanabe, M.

    1984-03-01

    A wide aperture (7 x 7 sq cm), high output energy (5 J in KrF and 13.8 J in XeCl), UV preionized excimer laser is described. A self-breakdown rail gap was employed as an output switch with the maximum voltage and current up to 230 kV and 300 kA, respectively. To solve the switching jitter problem associated with the self-breakdown, an electrical triggering was investigated. The measured minimum switching time delay and gap closing time were 40 and 10 ns, respectively. The number of channels up to 50 was observed with a uniform distribution over the 80-cm electrode length. The triggering jitter was measured to be less than a nanosecond. The maximum operation voltage of the triggered rail gap was 200 kV. The successful trigger operation was obtained in the range 30-98 percent of the self-breakdown voltage.

  13. Design and Overview of 100 kV Bushing for the DNB Injector of ITER

    NASA Astrophysics Data System (ADS)

    Shah, Sejal; Rajesh, S.; Nishad, S.; Srusti, B.; Bandyopadhyay, M.; Rotti, C.; Singh, M. J.; Roopesh, G.; Chakraborty, A. K.; Schunke, B.; Hemsworth, R.; Chareyre, J.; Svensson, L.

    2011-09-01

    The 100 kV bushing is one of the most important and technologically challenging Safety Important Class (SIC) components of the Diagnostic Neutral Beam (DNB) injector of ITER. It forms interface between gas insulated electrical transmission line and torus primary vacuum and acts as a vacuum feedthrough of ITER. Design optimization has been carried out to meet the electric and structural requirements based on its classification. Unlike HNB bushing, single stage bushing is designed to provide 100 kV isolation. Finite Element Analysis (FEA) based optimization has been carried out for electrostatic and structural analysis. Manufacturing assembly sequence is studied and presented in this paper. However validation of the same is foreseen from manufacturer.

  14. Design and Overview of 100 kV Bushing for the DNB Injector of ITER

    SciTech Connect

    Shah, Sejal; Bandyopadhyay, M.; Rotti, C.; Singh, M. J.; Roopesh, G.; Chakraborty, A. K.; Rajesh, S.; Nishad, S.; Srusti, B.; Schunke, B.; Hemsworth, R.; Chareyre, J.; Svensson, L.

    2011-09-26

    The 100 kV bushing is one of the most important and technologically challenging Safety Important Class (SIC) components of the Diagnostic Neutral Beam (DNB) injector of ITER. It forms interface between gas insulated electrical transmission line and torus primary vacuum and acts as a vacuum feedthrough of ITER. Design optimization has been carried out to meet the electric and structural requirements based on its classification. Unlike HNB bushing, single stage bushing is designed to provide 100 kV isolation. Finite Element Analysis (FEA) based optimization has been carried out for electrostatic and structural analysis. Manufacturing assembly sequence is studied and presented in this paper. However validation of the same is foreseen from manufacturer.

  15. Power frequency electric and magnetic fields from a 230 kV gas-insulated substation

    SciTech Connect

    Wong, P.S.; Rind, T.M. ); Harvey, S.M.; Scheer, R.R. . Research Div.)

    1994-07-01

    Gas-insulated substations (GIS), owing to their compact nature, offer an attractive alternative to conventional substations where space is limited, such as in urban areas. Consequently, it is important to address the issue of environmental conditions in and around the GIS. This paper presents the results of a survey of power-frequency electric and magnetic fields in and around a 230 kV/28 kV GIS. The survey was designed to cover the electric and magnetic fields from the substation equipment and from the power lines and cables surrounding the substation. It also includes a determination of the shielding effect of the GIS bus sheath. The information provided should allow the prediction of electric and magnetic field levels from other GIS of similar design.

  16. Schottky versus bipolar 3.3 kV SiC diodes

    NASA Astrophysics Data System (ADS)

    Pérez-Tomás, A.; Brosselard, P.; Hassan, J.; Jordà, X.; Godignon, P.; Placidi, M.; Constant, A.; Millán, J.; Bergman, J. P.

    2008-12-01

    A comparative study of the electrical characteristics of 3.3 kV SiC Schottky barrier (SBD), junction bipolar Schottky (JBS) and PiN diodes is presented. 3.3 kV class 4H-SiC SBD, JBS and PiN diodes have been fabricated with an analogous technology process on similar epi wafers. Diodes have been characterized in forward, reverse and switching mode in the 25 °C-300 °C temperature range. The optimum performance of the diodes depends on the adequate use of the unipolar or bipolar advantages and is established by the final application specifications. In this respect, a reverse recovery charge versus on-resistance diagram for different current densities is also presented. DC stress tests have been performed to investigate the forward voltage drift, related to the formation of stacking faults, during the bipolar mode of operation.

  17. Evaluation of Characteristics of Lightning Faults on 275kV Transmission Lines

    NASA Astrophysics Data System (ADS)

    Kawamura, Hironao; Itamoto, Naoki; Shinjo, Kazuo; Ishii, Masaru

    It is effective to design the lightning protection of transmission lines by employing simulation to be able to reproduce conditions of experienced lightning faults. As a result of the lightning faults on 275kV transmission lines in Hokuriku area, the frequency of only one-ground fault of upper or middle lines is higher. Especially, the frequency of one-ground faults of the middle lines in summer is the highest. It is thought that many one-ground faults in summer are caused by direct lightning strokes to phase conductors. Moreover, multi-ground faults caused by lightning strokes to tower tops or overhead ground wires also include many ground faults of the middle lines. In this paper, the experienced lightning faults on the 275kV transmission lines are reproduced by EMTP calculations and the characteristics of the ground-fault lines are examined.

  18. Development of 500 kV DC PPLP-insulated oil-filled submarine cable

    SciTech Connect

    Fujimori, A.; Tanaka, T.; Takashima, H.; Imajo, T.; Hata, R.; Tanabe, T.; Yoshida, S.; Kakihana, T.

    1996-01-01

    This paper outlines the development of a 500 kV DC oil-filled submarine cable capable of transmitting 2,800 MW with {+-} 500 kV 2800A bipole system. Polypropylene Laminated Paper (PPL) was employed as the insulation material, which is the worlds first application to DC cables. The conductor size is 3,000 mm{sup 2}, which is the largest size for submarine cables ever put into practical use. Through various fundamental and prototype tests, the cable proved to have excellent electrical characteristics for DC voltage as well as transient overvoltage. The cable and accessories are currently undergoing a long-term accelerated aging test as the final confirmation of their reliability and stability.

  19. Backside optimization for improving avalanche breakdown behavior of 4.5 kV IGBT

    NASA Astrophysics Data System (ADS)

    Xiaoli, Tian; Jiang, Lu; Yuan, Teng; Wenliang, Zhang; Shuojin, Lu; Yangjun, Zhu

    2015-03-01

    The static avalanche breakdown behavior of 4.5 kV high-voltage IGBT is studied by theory analysis and experiment. The avalanche breakdown behaviors of the 4.5 kV IGBTs with different backside structures are investigated and compared by using the curve tracer. The results show that the snap back behavior of the breakdown waveform is related to the bipolar PNP gain, which leads to the deterioration of the breakdown voltage. There are two ways to optimize the backside structure, one is increasing the implant dose of the N+ buffer layer, the other is decreasing the implant dose of the P+ collector layer. It is found that the optimized structure is effective in suppressing the snap back behavior and improving the breakdown characteristic of high voltage IGBT. Project supported by the National Major Science and Technology Special Project of China (No. 2011ZX02503-003).

  20. Targeting Kv1.3 channels to reduce white matter pathology after traumatic brain injury.

    PubMed

    Reeves, Thomas M; Trimmer, Patricia A; Colley, Beverly S; Phillips, Linda L

    2016-09-01

    Axonal injury is present in essentially all clinically significant cases of traumatic brain injury (TBI). While no effective treatment has been identified to date, experimental TBI models have shown promising axonal protection using immunosuppressants FK506 and Cyclosporine-A, with treatment benefits attributed to calcineurin inhibition or protection of mitochondrial function. However, growing evidence suggests neuroprotective efficacy of these compounds may also involve direct modulation of ion channels, and in particular Kv1.3. The present study tested whether blockade of Kv1.3 channels, using Clofazimine (CFZ), would alleviate TBI-induced white matter pathology in rodents. Postinjury CFZ administration prevented suppression of compound action potential (CAP) amplitude in the corpus callosum of adult rats following midline fluid percussion TBI, with injury and treatment effects primarily expressed in unmyelinated CAPs. Kv1.3 protein levels in callosal tissue extracts were significantly reduced postinjury, but this loss was prevented by CFZ treatment. In parallel, CFZ also attenuated the injury-induced elevation in pro-inflammatory cytokine IL1-β. The effects of CFZ on glial function were further studied using mixed microglia/astrocyte cell cultures derived from P3-5 mouse corpus callosum. Cultures of callosal glia challenged with lipopolysaccharide exhibited a dramatic increase in IL1-β levels, accompanied by reactive morphological changes in microglia, both of which were attenuated by CFZ treatment. These results support a cell specific role for Kv1.3 signaling in white matter pathology after TBI, and suggest a treatment approach based on the blockade of these channels. This therapeutic strategy may be especially efficacious for normalizing neuro-glial interactions affecting unmyelinated axons after TBI. PMID:27302680

  1. Environmental studies for a 1100-kV power line in Oregon. A special report

    SciTech Connect

    Rogers, L.; Hinds, R.

    1983-01-01

    In the first research project to monitor the effects of 1100-kV transmission on a natural ecosystem, researchers at Battelle, Pacific Northwest Laboratories have studied responses of wildlife, native plants, honeybees, and agricultural crops and cattle. No adverse effects were detected on wildlife, crops or cattle. Effects were observed on trees growing within the defined right of way and on honeybee colonies placed beneath the line.

  2. The varistor protected series capacitors at the 500 KV broadview substation

    SciTech Connect

    Barcus, J.M. ); Miske, S.A. Jr. ); Vitols, A.P. ); Maynard, H.M. ); Peterson, W.G. )

    1988-10-01

    Two 235 Mvar series capacitors have been successfully applied at the 500 kV Broadview substation of the Colstrip Transmission Project. The series capacitors have an overvoltage protection system based on metal oxide varistors. The system provides instantaneous bypass and reinsertion of the capacitors following external line section faults. The excellent performance of the equipment was demonstrated during staged fault tests performed in October, 1985. The banks have been in successful operation since January, 1986.

  3. [Mortality of people residing near electric power supply line with voltage of 500 kV].

    PubMed

    Gurvich, E B; Novokhatskaia, E A; Rubtsova, N B

    1996-01-01

    The epidemiologic study covered causes and levels of mortality in the settlement situated near electric power supply line (voltage is 500 kV). The work used retrospective cohort method adjusted for evaluation of mortality in general population. The study revealed no higher mortality risk with all the causes totally and with leading causal groups under influence of high frequency electromagnetic fields. However, higher relative mortality risk with leukemia and suicide appeared statistically insignificant. PMID:9019327

  4. Pseudosaccharin amines as potent and selective KV1.5 blockers.

    PubMed

    Lloyd, John; Finlay, Heather J; Kover, Alexander; Johnson, James; Pi, Zulan; Jiang, Ji; Neels, James; Cavallaro, Cullen; Wexler, Ruth; Conder, Mary Lee; Shi, Hong; Li, Danshi; Sun, Huabin; Chimalakonda, Anjaneya; Huang, Christine; Salvati, Mark; Levesque, Paul

    2015-11-01

    Phenethyl aminoheterocycles like compound 1 were known to be potent I(Kur) blockers although they lacked potency in vivo. Modification of the heterocycle led to the design and synthesis of pseudosaccharin amines. Compounds such as 14, 17d and 21c were found to be potent K(V)1.5 blockers and selective over other cardiac ion channels. These compounds had potent pharmacodynamic activity, however, they also showed off-target activities such as hemodynamic effects. PMID:25801931

  5. The contribution of Kv7 channels to pregnant mouse and human myometrial contractility.

    PubMed

    McCallum, Laura A; Pierce, Stephanie L; England, Sarah K; Greenwood, Iain A; Tribe, Rachel M

    2011-03-01

    Premature birth accounts for approximately 75% of neonatal mortality and morbidity in the developed world. Despite this, methods for identifying and treating women at risk of preterm labour are limited and many women still present in preterm labour requiring tocolytic therapy to suppress uterine contractility. The aim of this study was to assess the utility of Kv7 channel activators as potential uterine smooth muscle (myometrium) relaxants in tissues from pregnant mice and women. Myometrium was obtained from early and late pregnant mice and from lipopolysaccharide (LPS)-injected mice (day 15 of gestation; model of infection in pregnancy). Human myometrium was obtained at the time of Caesarean section from women at term (38-41 weeks). RT-PCR/qRT-PCR detected KCNQ and KCNE expression in mouse and human myometrium. In mice, there was a global suppression of all KCNQ isoforms, except KCNQ3, in early pregnancy (n= 6, P < 0.001 versus late pregnant); expression subsequently increased in late pregnancy (n= 6). KCNE isoforms were also gestationally regulated (P < 0.05). KCNQ and KCNE isoform expression was slightly down-regulated in myometrium from LPS-treated-mice versus controls (P < 0.05, n= 3-4). XE991 (10 μM, Kv7 inhibitor) significantly increased spontaneous myometrial contractions in vitro in both human and mouse myometrial tissues (P < 0.05) and retigabine/flupirtine (20 μM, Kv7 channel activators) caused profound myometrial relaxation (P < 0.05). In summary, Kv7 activators suppressed myometrial contraction and KCNQ gene expression was sustained throughout gestation, particularly at term. Consequently, activation of the encoded channels represents a novel mechanism for treatment of preterm labour. PMID:20132415

  6. On the relaxation of semi-Gaussian and K-V beams to thermal equilibrium

    SciTech Connect

    Lund, S.M.; Barnard, J.J.; Miller, J.M.

    1995-05-02

    A beam propagating in a continuous, linear focusing channel tends to relax to a thermal equilibrium state. We employ nonlinear conservation constraints to theoretically analyze changes in quantities that characterize both an initial semi-Gaussian beam with a matched rms beam envelope and a K-V beam under a relaxation to thermal equilibrium. Results from particle-in-cell simulations are compared to the theoretical predictions.

  7. 7. VIEW OF 100 kV SWITCHYARD WITH MISSOURI RIVER IN ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    7. VIEW OF 100 kV SWITCHYARD WITH MISSOURI RIVER IN THE BACKGROUND. ALONG THE LEFT SIDE FROM THE FOREGROUND ARE THE U.S. GOVERNMENT STORAGE SHED, TOOL HOUSE, THREE-STALL GARAGE, AND PARTIAL VIEW OF POWERHOUSE ADDITION. VIEW TO NORTHEAST. - Rainbow Hydroelectric Facility, On north bank of Missouri River 2 miles Northeast of Great Falls, & end of Rainbow Dam Road, Great Falls, Cascade County, MT

  8. Direct atomic resolution imaging of dislocation core structures in a 300 kV stem

    SciTech Connect

    McGibbon, A.J.; Pennycook, S.J.

    1995-05-01

    By employing the incoherent imaging technique of Z-contrast imaging in a 300kV STEM, the authors show that it is possible to provide directly interpretable, atomic resolution images of the sublattice in compound semiconductors. Using this approach, analysis of dislocations at an interface in the CdTe(001)/GaAs(001) system reveals unexpected core structures at Lomer dislocations.

  9. A 200 kV fast rise time, low jitter, trigger system with magnetic pulse sharpener

    SciTech Connect

    Jaitly, N.C.; Coleman, M.D.; Ramrus, A.; Earley, L.M.; Downing, J.N.; Reisch, H.H.; Caudill, L.D.; Eversol, S.A.

    1992-09-01

    The DARHT Facility is being designed at Los Alamos national Laboratory to produce high resolution flash radiographs of hydrodynamic experiments. Two linear induction accelerators (LIA), each in the range of 16 to 20 MeV, will be used to produce intense bremsstrahlung X-ray pulses of short duration (60 ns flat top). Each LIA will produce a 3 kA, high brightness, electron beam using a 4 MeV injector and a series of 250 kV induction cells. Technology demonstration of key accelerator subsystems is under progress at the DARHT Integrated Test Stand (ITS). The eight inductions cells present in the ITS are driven by a Maxwell prototype Induction Cell Pulsed Power supply (ICPPS) which provides 250 kV, 70 ns pulses via four Blumieins. Each Blumiein drives two cells and is triggered using independently controlled trigger units. This turnkey DARHT Trigger System, consisting of four separate trigger units, provides 200 kV trigger pulses with low jitter and fast rise time to each of the four Blumiein coaxial spark gaps. Details of the trigger system design and results obtained during extensive testing at Maxwell are described.

  10. Usefulness of targeting lymphocyte Kv1.3-channels in the treatment of respiratory diseases.

    PubMed

    Kazama, Itsuro; Tamada, Tsutomu; Tachi, Masahiro

    2015-10-01

    T lymphocytes predominantly express delayed rectifier K(+)-channels (Kv1.3) in their plasma membranes. Patch-clamp studies revealed that the channels play crucial roles in facilitating the calcium influx necessary to trigger lymphocyte activation and proliferation. Using selective channel inhibitors in experimental animal models, in vivo studies further revealed the clinically relevant relationship between the channel expression and the development of chronic respiratory diseases, in which chronic inflammation or the overstimulation of cellular immunity in the airways is responsible for the pathogenesis. In chronic respiratory diseases, such as chronic obstructive pulmonary disease, asthma, diffuse panbronchiolitis and cystic fibrosis, in addition to the supportive management for the symptoms, the anti-inflammatory effects of macrolide antibiotics were shown to be effective against the over-activation or proliferation of T lymphocytes. Recently, we provided physiological and pharmacological evidence that macrolide antibiotics, together with calcium channel blockers, HMG-CoA reductase inhibitors, and nonsteroidal anti-inflammatory drugs, effectively suppress the Kv1.3-channel currents in lymphocytes, and thus exert anti-inflammatory or immunomodulatory effects. In this review article, based on the findings obtained from recent in vivo and in vitro studies, we address the novel therapeutic implications of targeting the lymphocyte Kv1.3-channels for the treatment of chronic or acute respiratory diseases. PMID:26206235

  11. A 200 kV fast rise time, low jitter, trigger system with magnetic pulse sharpener

    SciTech Connect

    Jaitly, N.C.; Ramrus, A.; Coleman, M.D.; Earley, L.M.; Downing, J.N.; Reisch, H.H.; Caudill, L.D.; Eversol, S.A.

    1993-12-31

    The DARHT Facility is being designed at Los Alamos National Laboratory to produce high resolution flash radiographs of hydrodynamic experiments. Two linear induction accelerators (LIA), each in the range of 16 to 20 MeV, will be used to produce intense bremsstrahlung X-ray pulses of short duration (60 ns flat top). Each LIA will produce a 3 kA, high brightness, electron beam using a 4 MeV injector and a series of 250 kV induction cells. Technology demonstration of key accelerator subsystems is under progress at the DARHT Integrated Test Stand (ITS). The eight inductions cells present in the ITS are driven by a Maxwell prototype Induction Cell Pulsed Power Supply (ICPPS) which provides 250 kV, 70ns pulses via four Blumleins. Each Blumlein drives two cells and is triggered using independently controlled trigger units. This turnkey DARHT Trigger System, consisting of four separate trigger units, provides 200 kV trigger pulses with low jitter and fast rise time to each of the four Blumlein coaxial spark gaps. Details of the trigger system design and results obtained during extensive testing at Maxwell are described.

  12. Fiber optic voltage sensor for 420 kV electric power systems

    NASA Astrophysics Data System (ADS)

    Bohnert, Klaus M.; Kostovic, Jadran; Pequignot, P.

    2000-11-01

    We present an optical fiber voltage sensor for 420 kV electric power lines. The sensor exploits the converse piezoelectric effect of quartz and measures the voltage by a line integration of the electric field. The alternating voltage is partitioned to a series of four cylinder-shaped quartz crystals, which are embedded in polyurethane resin within a 3.2-m long insulator tube of fiber reinforced epoxy. The alternating piezoelectric deformations of the crystals are sensed by a common elliptical-core dual-mode fiber, which is wound onto the circumferential crystal surfaces. The fiber is interrogated using low coherence interferometry. We determine the dielectric design of the sensor from a numerical analysis of the electric field distribution within and in the vicinity of the sensor. We experimentally verify the dielectric reliability under ac overvoltages up to 520 kV root mean square and lightning and switching impulse voltages up to 1425 and 1050 kV, respectively. Further, we investigate the sensor performance including accuracy, dynamic range, bandwidth, and temperature dependence.

  13. Polyunsaturated fatty acids inhibit Kv1.4 by interacting with positively charged extracellular pore residues.

    PubMed

    Farag, N E; Jeong, D; Claydon, T; Warwicker, J; Boyett, M R

    2016-08-01

    Polyunsaturated fatty acids (PUFAs) modulate voltage-gated K(+) channel inactivation by an unknown site and mechanism. The effects of ω-6 and ω-3 PUFAs were investigated on the heterologously expressed Kv1.4 channel. PUFAs inhibited wild-type Kv1.4 during repetitive pulsing as a result of slowing of recovery from inactivation. In a mutant Kv1.4 channel lacking N-type inactivation, PUFAs reversibly enhanced C-type inactivation (Kd, 15-43 μM). C-type inactivation was affected by extracellular H(+) and K(+) as well as PUFAs and there was an interaction among the three: the effect of PUFAs was reversed during acidosis and abolished on raising K(+) Replacement of two positively charged residues in the extracellular pore (H508 and K532) abolished the effects of the PUFAs (and extracellular H(+) and K(+)) on C-type inactivation but had no effect on the lipoelectric modulation of voltage sensor activation, suggesting two separable interaction sites/mechanisms of action of PUFAs. Charge calculations suggest that the acidic head group of the PUFAs raises the pKa of H508 and this reduces the K(+) occupancy of the selectivity filter, stabilizing the C-type inactivated state. PMID:27281482

  14. A 6 kV arbitrary waveform generator for the Tevatron Electron Lens

    SciTech Connect

    Pfeffer, H.; Saewert, G.

    2011-11-09

    This paper reports on a 6 kV modulator built and installed at Fermilab to drive the electron gun anode for the Tevatron Electron Lens (TEL). The TEL was built with the intention of shifting the individual (anti)proton bunch tunes to even out the tune spread among all 36 bunches with the desire of improving Tevatron integrated luminosity. This modulator is essentially a 6 kV arbitrary waveform generator that enables the TEL to define the electron beam intensity on a bunch-by-bunch basis. A voltage waveform is constructed having a 7 μs duration that corresponds to the tune shift requirements of a 12-bunch (anti)proton beam pulse train. This waveform is played out for any one or all three bunch trains in the Tevatron. The programmed waveform voltages transition to different levels at time intervals corresponding to the 395 ns bunch spacing. In addition, complex voltage waveforms can be played out at a sustained rate of 143 kHz over the full 6 kV output range. This paper describes the novel design of the inductive adder topology employing five transformers. It describes the design aspects that minimize switching losses for this multi-kilovolt, high repetition rate and high duty factor application.

  15. A functional Kv1.2-hERG chimaeric channel expressed in Pichia pastoris

    NASA Astrophysics Data System (ADS)

    Dhillon, Mandeep S.; Cockcroft, Christopher J.; Munsey, Tim; Smith, Kathrine J.; Powell, Andrew J.; Carter, Paul; Wrighton, David C.; Rong, Hong-Lin; Yusaf, Shahnaz P.; Sivaprasadarao, Asipu

    2014-02-01

    Members of the six-transmembrane segment family of ion channels share a common structural design. However, there are sequence differences between the members that confer distinct biophysical properties on individual channels. Currently, we do not have 3D structures for all members of the family to help explain the molecular basis for the differences in their biophysical properties and pharmacology. This is due to low-level expression of many members in native or heterologous systems. One exception is rat Kv1.2 which has been overexpressed in Pichia pastoris and crystallised. Here, we tested chimaeras of rat Kv1.2 with the hERG channel for function in Xenopus oocytes and for overexpression in Pichia. Chimaera containing the S1-S6 transmembrane region of HERG showed functional and pharmacological properties similar to hERG and could be overexpressed and purified from Pichia. Our results demonstrate that rat Kv1.2 could serve as a surrogate to express difficult-to-overexpress members of the six-transmembrane segment channel family.

  16. A 6 kV arbitrary waveform generator for the Tevatron Electron Lens

    DOE PAGESBeta

    Pfeffer, H.; Saewert, G.

    2011-11-09

    This paper reports on a 6 kV modulator built and installed at Fermilab to drive the electron gun anode for the Tevatron Electron Lens (TEL). The TEL was built with the intention of shifting the individual (anti)proton bunch tunes to even out the tune spread among all 36 bunches with the desire of improving Tevatron integrated luminosity. This modulator is essentially a 6 kV arbitrary waveform generator that enables the TEL to define the electron beam intensity on a bunch-by-bunch basis. A voltage waveform is constructed having a 7 μs duration that corresponds to the tune shift requirements of amore » 12-bunch (anti)proton beam pulse train. This waveform is played out for any one or all three bunch trains in the Tevatron. The programmed waveform voltages transition to different levels at time intervals corresponding to the 395 ns bunch spacing. In addition, complex voltage waveforms can be played out at a sustained rate of 143 kHz over the full 6 kV output range. This paper describes the novel design of the inductive adder topology employing five transformers. It describes the design aspects that minimize switching losses for this multi-kilovolt, high repetition rate and high duty factor application.« less

  17. A Study of Electromagnetic Radiation of Corona Discharge Near 500-Kv Electric Installations

    SciTech Connect

    Korzhov, A. V.; Okrainskaya, I. S.; Sidorov, A. I.; Kufel'd, V. D.

    2004-01-15

    Data on the spectral composition and intensity of electromagnetic radiation of corona discharge are obtained in an experimental study performed on the outdoor switchgear of the Shagol 500-kV substation of the Chelyabinsk Enterprise of Trunk Transmission Grids and under a 500-kV Shagol - Kozyrevo overhead transmission line. The electromagnetic environment on the territory of the 500-kV outdoor switchgear is shown to be determined by narrow-band radiations (harmonics of the frequency of electric supply) and wide-band radiations due to corona discharges of high-voltage sources. This means that the personnel experience the action of a commercial-frequency electric field and electromagnetic radiation of a quite wide range, which is not allowed for by the existing guidelines. It is recommended to continue the study in cooperation with medical institutions in order to create guidelines that would allow for the joint action of commercial-frequency electric field and electromagnetic radiation and for the voltage in the line, the current load, the meteorological situation, and other factors.

  18. A functional Kv1.2-hERG chimaeric channel expressed in Pichia pastoris

    PubMed Central

    Dhillon, Mandeep S.; Cockcroft, Christopher J.; Munsey, Tim; Smith, Kathrine J.; Powell, Andrew J.; Carter, Paul; Wrighton, David C.; Rong, Hong-lin; Yusaf, Shahnaz P.; Sivaprasadarao, Asipu

    2014-01-01

    Members of the six-transmembrane segment family of ion channels share a common structural design. However, there are sequence differences between the members that confer distinct biophysical properties on individual channels. Currently, we do not have 3D structures for all members of the family to help explain the molecular basis for the differences in their biophysical properties and pharmacology. This is due to low-level expression of many members in native or heterologous systems. One exception is rat Kv1.2 which has been overexpressed in Pichia pastoris and crystallised. Here, we tested chimaeras of rat Kv1.2 with the hERG channel for function in Xenopus oocytes and for overexpression in Pichia. Chimaera containing the S1–S6 transmembrane region of HERG showed functional and pharmacological properties similar to hERG and could be overexpressed and purified from Pichia. Our results demonstrate that rat Kv1.2 could serve as a surrogate to express difficult-to-overexpress members of the six-transmembrane segment channel family. PMID:24569544

  19. Poster — Thur Eve — 10: Partial kV CBCT, complete kV CBCT and EPID in breast treatment: a dose comparison study for skin, breasts, heart and lungs

    SciTech Connect

    Roussin, E; Archambault, L K; Wierzbicki, W

    2014-08-15

    The advantages of kilovoltage cone beam CT (kV CBCT) imaging over electronic portal imaging device (EPID) such as accurate 3D anatomy, soft tissue visualization, fast rigid registration and enhanced precision on patient positioning has lead to its increasing use in clinics. The benefits of this imaging technique are at the cost of increasing the dose to healthy surrounding organs. Our center has moved toward the use of daily partial rotation kV CBCT to restrict the dose to healthy tissues. This study aims to better quantify radiation doses from different image-guidance techniques such as tangential EPID, complete and partial kV CBCT for breast treatments. Cross-calibrated ionization chambers and kV calibrated Gafchromic films were used to measure the dose to the heart, lungs, breasts and skin. It was found that performing partial kV CBCT decreases the heart dose by about 36%, the lungs dose by 31%, the contralateral breast dose by 41% and the ipsilateral breast dose by 43% when compared to a full rotation CBCT. The skin dose measured for a full rotation CBCT was about 0.8 cGy for the contralateral breast and about 0.3 cGy for the ipsilateral breast. The study is still ongoing and results on skin doses for partial rotation kV CBCT as well as for tangential EPID images are upcoming.

  20. IEEE 342 Node Low Voltage Networked Test System

    SciTech Connect

    Schneider, Kevin P.; Phanivong, Phillippe K.; Lacroix, Jean-Sebastian

    2014-07-31

    The IEEE Distribution Test Feeders provide a benchmark for new algorithms to the distribution analyses community. The low voltage network test feeder represents a moderate size urban system that is unbalanced and highly networked. This is the first distribution test feeder developed by the IEEE that contains unbalanced networked components. The 342 node Low Voltage Networked Test System includes many elements that may be found in a networked system: multiple 13.2kV primary feeders, network protectors, a 120/208V grid network, and multiple 277/480V spot networks. This paper presents a brief review of the history of low voltage networks and how they evolved into the modern systems. This paper will then present a description of the 342 Node IEEE Low Voltage Network Test System and power flow results.

  1. Kv1.3 potassium channels are localized in the immunological synapse formed between cytotoxic and target cells

    PubMed Central

    Panyi, G.; Vámosi, G.; Bacsó, Z.; Bagdány, M.; Bodnár, A.; Varga, Z.; Gáspár, R.; Mátyus, L.; Damjanovich, S.

    2004-01-01

    Membrane proteins of cytotoxic T cells specifically reorganize to form an immunological synapse (IS) on interaction with their specific target. In this paper, we investigated the redistribution of Kv1.3 channels, which are the dominant voltage-gated potassium channels, in the plasma membrane of allogen-activated human cytotoxic T lymphocytes (CTLs) on interacting with their specific target cells. Kv1.3 channels bearing a FLAG epitope were expressed in the CTLs and the cell-surface distribution of fluorescently labeled ion channels was determined from confocal laser-scanning microscopy images. FLAG epitope-tagged Kv1.3 channels showed a patchy distribution in CTLs not engaged with target cells, whereas the channels were accumulated in the IS formed between CTLs and specific target lymphocytes. Localization of Kv1.3 channels in the IS might open an unrevealed possibility in the regulation of ion channel activity by signaling molecules accumulated in the IS. PMID:14745040

  2. 250 kV 6 mA compact Cockcroft-Walton high-voltage power supply.

    PubMed

    Ma, Zhan-Wen; Su, Xiao-Dong; Lu, Xiao-Long; Wei, Zhen; Wang, Jun-Run; Huang, Zhi-Wu; Miao, Tian-You; Su, Tong-Ling; Yao, Ze-En

    2016-08-01

    A compact power supply system for a compact neutron generator has been developed. A 4-stage symmetrical Cockcroft-Walton circuit is adopted to produce 250 kV direct current high-voltage. A 2-stage 280 kV isolation transformer system is used to drive the ion source power supply. For a compact structure, safety, and reliability during the operation, the Cockcroft-Walton circuit and the isolation transformer system are enclosed in an epoxy vessel containing the transformer oil whose size is about ∅350 mm × 766 mm. Test results indicate that the maximum output voltage of the power supply is 282 kV, and the stability of the output voltage is better than 0.63% when the high voltage power supply is operated at 250 kV, 6.9 mA with the input voltage varying ±10%. PMID:27587170

  3. SU-E-I-29: Care KV: Dose It Influence Radiation Dose in Non-Contrast Examination of CT Abdomen/pelvis?

    SciTech Connect

    Zhang, J; Ganesh, H; Weir, V

    2015-06-15

    Purpose: CARE kV is a tool that automatically recommends optimal kV setting for individual patient for specific CT examination. The use of CARE kV depends on topogram and the user-selected contrast behavior. CARE kV is expected to reduce radiation dose while improving image quality. However, this may work only for certain groups of patients and/or certain CT examinations. This study is to investigate the effects of CARE kV on radiation dose of non-contrast examination of CT abdomen/pelvis. Methods: Radiation dose (CTDIvol and DLP) from patients who underwent abdomen/pelvis non-contrast examination with and without CARE kV were retrospectively reviewed. All patients were scanned in the same scanner (Siemens Somatom AS64). To mitigate any possible influences due to technologists’ unfamiliarity with the CARE kV, the data with CARE kV were retrieved 1.5 years after the start of CARE kV usage. T-test was used for significant difference in radiation dose. Results: Volume CTDIs and DLPs from 18 patients before and 24 patients after the use of CARE kV were obtained in a duration of one month. There is a slight increase in both average CTDIvol and average DLP with CARE kV compared to those without CARE kV (25.52 mGy vs. 22.65 mGy for CTDIvol; 1265.81 mGy-cm vs. 1199.19 mGy-cm). Statistically there was no significant difference. Without CARE kV, 140 kV was used in 9 of 18 patients, while with CARE KV, 140 kV was used in 15 of 24 patients. 80kV was not used in either group. Conclusion: The use of CARE kV may save time for protocol optimization and minimize variability among technologists. Radiation dose reduction was not observed in non-contrast examinations of CT abdomen/pelvis. This was partially because our CT protocols were tailored according to patient size before CARE kV and partially because of large size patients.

  4. SU-E-J-22: A Feasibility Study On KV-Based Whole Breast Radiation Patient Setup

    SciTech Connect

    Huang, Q; Zhang, M; Yue, N; Chen, T

    2015-06-15

    Purpose: In room kilovoltage x-ray (kV) imaging provides higher contrast than Megavoltage (MV) imaging with faster acquisition time compared with on-board cone-beam computed tomography (CBCT), thus improving patient setup accuracy and efficiency. In this study we evaluated the clinical feasibility of utilizing kV imaging for whole breast radiation patient setup. Methods: For six breast cancer patients with whole breast treatment plans using two opposed tangential fields, MV-based patient setup was conducted by aligning patient markers with in room lasers and MV portal images. Beam-eye viewed kV images were acquired using Varian OBI system after the set up process. In house software was developed to transfer MLC blocks information overlaying onto kV images to demonstrate the field shape for verification. KV-based patient digital shift was derived by performing rigid registration between kV image and the digitally reconstructed radiography (DRR) to align the bony structure. This digital shift between kV-based and MV-based setup was defined as setup deviation. Results: Six sets of kV images were acquired for breast patients. The mean setup deviation was 2.3mm, 2.2mm and 1.8mm for anterior-posterior, superior-inferior and left-right direction respectively. The average setup deviation magnitude was 4.3±1.7mm for six patients. Patient with large breast had a larger setup deviation (4.4–6.2mm). There was no strong correlation between MV-based shift and setup deviation. Conclusion: A preliminary clinical workflow for kV-based whole breast radiation setup was established and tested. We observed setup deviation of the magnitude below than 5mm. With the benefit of providing higher contrast and MLC block overlaid on the images for treatment field verification, it is feasible to use kV imaging for breast patient setup.

  5. Catalytic mechanism and substrate specificity of the β-subunit of the voltage-gated potassium (Kv) channel

    PubMed Central

    Tipparaju, Srinivas M.; Barski, Oleg A.; Srivastava, Sanjay; Bhatnagar, Aruni

    2008-01-01

    The β-subunits of voltage-gated potassium (Kv) channels are members of aldo-keto reductase (AKR) superfamily. These proteins regulate inactivation and membrane localization of Kv1 and Kv4 channels. The Kvβ proteins bind to pyridine nucleotides with high affinity; however, their catalytic properties remain unclear. Here we report that recombinant rat Kvβ2 catalyzes the reduction of a wide range of aldehydes and ketones. The rate of catalysis was slower (0.06 to 0.2 min−1) than that of other AKRs, but displayed the expected hyperbolic dependence on substrate concentration, with no evidence of allosteric cooperativity. Catalysis was prevented by site-directed substitution of Tyr-90 with phenylalanine, indicating that the acid-base catalytic residue, identified in other AKRs, has a conserved function in Kvβ2. The protein catalyzed the reduction of a broad range of carbonyls including aromatic carbonyls, electrophilic aldehydes and prostaglandins, phospholipid and sugar aldehydes. Little or no activity was detected with carbonyl steroids. Initial velocity profiles were consistent with an ordered bi-bi rapid-equilibrium mechanism in which NADPH binding precedes carbonyl binding. Significant primary kinetic isotope effects (2.0 – 3.1) were observed under single and multiple turnover conditions, indicating that the bond-breaking chemical step is rate-limiting. Structure-activity relationships with a series of para-substituted benzaldehydes indicated that the electronic interactions predominate during substrate binding and that no significant charge develops during the transition state. These data strengthen the view that Kvβ proteins are catalytically-active AKRs that impart redox-sensitivity to Kv channels. PMID:18672894

  6. Serum albumin attenuates the open-channel blocking effects of propofol on the human Kv1.5 channel.

    PubMed

    Kojima, Akiko; Bai, Jia-Yu; Ito, Yuki; Ding, Wei-Guang; Kitagawa, Hirotoshi; Matsuura, Hiroshi

    2016-07-15

    The intravenous anesthetic propofol modulates various ion channel functions. It is generally accepted that approximately 98% of propofol binds to blood constituents and that the free (unbound) drug preferentially affects target proteins including ion channels. However, modulatory effects of propofol on ion channels have not been previously explored in the presence of serum albumin. This study was designed to investigate the effects of serum albumin on the blocking action of propofol on the human Kv1.5 (hKv1.5) current. Whole-cell patch-clamp method was used to record the hKv1.5 channel current, heterologously expressed in Chinese hamster ovary cells, in the absence and presence of bovine serum albumin (BSA). Propofol induced a time-dependent decline of the hKv1.5 current during depolarizing steps and slowed the time course of tail current decay upon repolarization, supporting that propofol acts as an open-channel blocker. This blocking effect was reversible and concentration-dependent with an IC50 of 62.9±3.1μM (n = 6). Bath application of 1% BSA markedly reduced the blocking potency of propofol on hKv1.5 current (IC50 of 1116.0±491.4μM; n = 6). However, in the presence of BSA, the propofol-induced inhibition of hKv1.5 current was also accompanied by a gradual decline of activated current during depolarization and deceleration of deactivating tail current upon repolarization. The presence of BSA greatly attenuated the blocking potency of propofol on hKv1.5 channel without affecting the mode of action of propofol on the channel. Serum albumin thus appears to bind to propofol and thereby reducing effective concentrations of the drug for inhibition of hKv1.5 channel. PMID:27164421

  7. Increased aldosterone-dependent Kv1.5 recycling predisposes to pacing-induced atrial fibrillation in Kcne3-/- mice.

    PubMed

    Lisewski, Ulrike; Koehncke, Clemens; Wilck, Nicola; Buschmeyer, Bastian; Pieske, Burkert; Roepke, Torsten K

    2016-07-01

    Hyperaldosteronism is associated with an increased prevalence of atrial fibrillation (AF). Mutations in KCNE3 have been associated with AF, and Kcne3(-/-) mice exhibit hyperaldosteronism. In this study, we used recently developed Kcne3(-/-) mice to study atrial electrophysiology with respect to development of aldosterone-dependent AF. In invasive electrophysiology studies, Kcne3(-/-) mice displayed a reduced atrial effective refractory period (AERP) and inducible episodes of paroxysmal AF. The cellular arrhythmogenic correlate for AF predisposition was a significant increase in atrial Kv currents generated by the micromolar 4-aminopyridine-sensitive Kv current encoded by Kv1.5. Electrophysiological alterations in Kcne3(-/-) mice were aldosterone dependent and were associated with increased Rab4, -5, and -9-dependent recycling of Kv1.5 channels to the Z-disc/T-tubulus region and lateral membrane via activation of the Akt/AS160 pathway. Treatment with spironolactone inhibited Akt/AS160 phosphorylation, reduced Rab-dependent Kv1.5 recycling, normalized AERP and atrial Kv currents to the wild-type level, and reduced arrhythmia induction in Kcne3(-/-) mice. Kcne3 deletion in mice predisposes to AF by a heretofore unrecognized mechanism-namely, increased aldosterone-dependent Kv1.5 recycling via Rab GTPases. The findings uncover detailed molecular mechanisms underpinning a channelopathy-linked form of AF and emphasize the inevitability of considering extracardiac mechanisms in genetic arrhythmia syndromes.-Lisewski, U., Koehncke, C., Wilck, N., Buschmeyer, B., Pieske, B., Roepke, T. K. Increased aldosterone-dependent Kv1.5 recycling predisposes to pacing-induced atrial fibrillation in Kcne3(-/-) mice. PMID:26985008

  8. Kv1.3 channel-blocking immunomodulatory peptides from parasitic worms: implications for autoimmune diseases

    PubMed Central

    Chhabra, Sandeep; Chang, Shih Chieh; Nguyen, Hai M.; Huq, Redwan; Tanner, Mark R.; Londono, Luz M.; Estrada, Rosendo; Dhawan, Vikas; Chauhan, Satendra; Upadhyay, Sanjeev K.; Gindin, Mariel; Hotez, Peter J.; Valenzuela, Jesus G.; Mohanty, Biswaranjan; Swarbrick, James D.; Wulff, Heike; Iadonato, Shawn P.; Gutman, George A.; Beeton, Christine; Pennington, Michael W.; Norton, Raymond S.; Chandy, K. George

    2014-01-01

    The voltage-gated potassium (Kv) 1.3 channel is widely regarded as a therapeutic target for immunomodulation in autoimmune diseases. ShK-186, a selective inhibitor of Kv1.3 channels, ameliorates autoimmune diseases in rodent models, and human phase 1 trials of this agent in healthy volunteers have been completed. In this study, we identified and characterized a large family of Stichodactyla helianthus toxin (ShK)–related peptides in parasitic worms. Based on phylogenetic analysis, 2 worm peptides were selected for study: AcK1, a 51-residue peptide expressed in the anterior secretory glands of the dog-infecting hookworm Ancylostoma caninum and the human-infecting hookworm Ancylostoma ceylanicum, and BmK1, the C-terminal domain of a metalloprotease from the filarial worm Brugia malayi. These peptides in solution adopt helical structures closely resembling that of ShK. At doses in the nanomolar–micromolar range, they block native Kv1.3 in human T cells and cloned Kv1.3 stably expressed in L929 mouse fibroblasts. They preferentially suppress the proliferation of rat CCR7− effector memory T cells without affecting naive and central memory subsets and inhibit the delayed-type hypersensitivity (DTH) response caused by skin-homing effector memory T cells in rats. Further, they suppress IFNγ production by human T lymphocytes. ShK-related peptides in parasitic worms may contribute to the potential beneficial effects of probiotic parasitic worm therapy in human autoimmune diseases.—Chhabra, S., Chang, S. C., Nguyen, H. M., Huq, R., Tanner, M. R., Londono, L. M., Estrada, R., Dhawan, V., Chauhan, S., Upadhyay, S. K., Gindin, M., Hotez, P. J., Valenzuela, J. G., Mohanty, B., Swarbrick, J. D., Wulff, H., Iadonato, S. P., Gutman, G. A., Beeton, C., Pennington, M. W., Norton, R. S., Chandy, K. G. Kv1.3 channel-blocking immunomodulatory peptides from parasitic worms: implications for autoimmune diseases. PMID:24891519

  9. Grand Coulee - Bell 500-kV Transmission Line Project, Draft Environmental Impact Statement

    SciTech Connect

    N /A

    2002-08-09

    BPA is proposing to construct a 500-kilovolt (kV) transmission line that would extend approximately 84 miles between the Grand Coulee 500-kV Switchyard, near Grand Coulee Dam, and the Bell Substation, in Mead just north of Spokane. The new line would cross portions of Douglas, Grant, Lincoln, and Spokane counties. In addition to the transmission line, new equipment would be installed at the substations at each end of the new line and at other facilities. The proposed action would remove an existing 115-kV transmission line and replace it with the new 500-kV line on existing right-of-way for most of its length. Additional right-of-way would be needed in the first 3.5 miles out of the Grand Coulee Switchyard to connect to the existing 115-kV right-of-way. Since the mid-1990s, the transmission path west of Spokane, called the West of Hatwai transmission pathway, has grown increasingly constrained. To date, BPA has been able to manage operation of the path through available operating practices, and customer needed have been met while maintaining the reliability of the path. however, in early 2001, operations showed that the amount of electricity that needs to flow from east to west along this path creates severe transmission congestion. Under these conditions, the system is at risk of overloads and violation of industry safety and reliability standards. The problem is particularly acute in the spring and summer months because of the large amount of power generated by dams east of the path. Large amounts of water cannot be spilled during that time in order for BPA to fulfill its obligation to protect threatened and endangered fish. The amount of power that needs to move through this area during these months at times could exceed the carrying capacity of the existing transmission lines. In additional capacity is not added, BPA will run a significant risk that it will not be able to continue to meet its contractual obligations to deliver power and maintain reliability

  10. Commissioning a 50-100 kV X-ray unit for skin cancer treatment.

    PubMed

    Sheu, Ren-Dih; Powers, Allison; Lo, Yeh-Chi

    2015-01-01

    This study provides the authors' experience along with dosimetric data from the commissioning of two Sensus SRT-100 50-100 kV X-ray units. Data collected during the commissioning process included: a) HVL, b) output (dose rate), c) applicator cone factors, and d) percentage depth dose. A Farmer-type chamber (PTW-N23333), and a thin-window parallel plate ion chamber (PTW-N23342) were used for dose rate measurements and dose profiles were measured with EBT3 GafChromic film. The average HVL values for 50, 70, and 100 kV of the two treatment units were found to be 0.52, 1.15, and 2.20 mm Al, respectively. The HVL's were 5%-9% lower when measured with the Farmer chamber, as compared to measurements with the parallel plate chamber, for energies of 70 and 100 kV. Dose rates were also measured to be 3%-4% lower with the Farmer chamber. The dose rate variation between the two units was found to be 2%-9% for 50, 70, and 100 kV. The dose uniformity over a circle of 2 cm diameter was within 4% in four cardinal directions; however, the dose profiles for the 5 cm applicator were nonuniform, especially in the cathode-anode direction. Measurements indicated as much as 15% lower dose for the 50 kV beam at field edge on the anode side, when normalized to the center. The crossline profile was relatively more symmetric, with a maximum deviation of 10% at the field edge. All ion chamber readings agreed with film measurements within 3%. The nonuniform profile produced by these units may introduce uncertainty in dose rate measurements, especially for larger applicators. Since there is no intrinsic tool (crosshair or field light) for alignment with the beam axis, the user should take care when positioning the chamber for output measurements. The data obtained with a Farmer-type chamber should be used cautiously and as a reference only for the SRT-100 X-ray treatment unit. PMID:26103186

  11. Altered Kv2.1 functioning promotes increased excitability in hippocampal neurons of an Alzheimer's disease mouse model.

    PubMed

    Frazzini, V; Guarnieri, S; Bomba, M; Navarra, R; Morabito, C; Mariggiò, M A; Sensi, S L

    2016-01-01

    Altered neuronal excitability is emerging as an important feature in Alzheimer's disease (AD). Kv2.1 potassium channels are important modulators of neuronal excitability and synaptic activity. We investigated Kv2.1 currents and its relation to the intrinsic synaptic activity of hippocampal neurons from 3xTg-AD (triple transgenic mouse model of Alzheimer's disease) mice, a widely employed preclinical AD model. Synaptic activity was also investigated by analyzing spontaneous [Ca(2+)]i spikes. Compared with wild-type (Non-Tg (non-transgenic mouse model)) cultures, 3xTg-AD neurons showed enhanced spike frequency and decreased intensity. Compared with Non-Tg cultures, 3xTg-AD hippocampal neurons revealed reduced Kv2.1-dependent Ik current densities as well as normalized conductances. 3xTg-AD cultures also exhibited an overall decrease in the number of functional Kv2.1 channels. Immunofluorescence assay revealed an increase in Kv2.1 channel oligomerization, a condition associated with blockade of channel function. In Non-Tg neurons, pharmacological blockade of Kv2.1 channels reproduced the altered pattern found in the 3xTg-AD cultures. Moreover, compared with untreated sister cultures, pharmacological inhibition of Kv2.1 in 3xTg-AD neurons did not produce any significant modification in Ik current densities. Reactive oxygen species (ROS) promote Kv2.1 oligomerization, thereby acting as negative modulator of the channel activity. Glutamate receptor activation produced higher ROS levels in hippocampal 3xTg-AD cultures compared with Non-Tg neurons. Antioxidant treatment with N-Acetyl-Cysteine was found to rescue Kv2.1-dependent currents and decreased spontaneous hyperexcitability in 3xTg-AD neurons. Analogous results regarding spontaneous synaptic activity were observed in neuronal cultures treated with the antioxidant 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox). Our study indicates that AD-related mutations may promote enhanced ROS generation, oxidative

  12. Serum starvation-induced voltage-gated potassium channel Kv7.5 expression and its regulation by Sp1 in canine osteosarcoma cells.

    PubMed

    Lee, Bo Hyung; Ryu, Pan Dong; Lee, So Yeong

    2014-01-01

    The KCNQ gene family, whose members encode Kv7 channels, belongs to the voltage-gated potassium (Kv) channel group. The roles of this gene family have been widely investigated in nerve and muscle cells. In the present study, we investigated several characteristics of Kv7.5, which is strongly expressed in the canine osteosarcoma cell line, CCL-183. Serum starvation upregulated Kv7.5 expression, and the Kv7 channel opener, flupirtine, attenuated cell proliferation by arresting cells in the G0/G1 phase. We also showed that Kv7.5 knockdown helps CCL-183 cells to proliferate. In an effort to find an endogenous regulator of Kv7.5, we used mithramycin A to reduce the level of the transcription factor Sp1, and it strongly inhibited the induction of Kv7.5 in CCL-183 cells. These results suggest that the activation of Kv7.5 by flupirtine may exert an anti-proliferative effect in canine osteosarcoma. Therefore, Kv7.5 is a possible molecular target for canine osteosarcoma therapy. PMID:24434641

  13. Serum Starvation-Induced Voltage-Gated Potassium Channel Kv7.5 Expression and Its Regulation by Sp1 in Canine Osteosarcoma Cells

    PubMed Central

    Lee, Bo Hyung; Ryu, Pan Dong; Lee, So Yeong

    2014-01-01

    The KCNQ gene family, whose members encode Kv7 channels, belongs to the voltage-gated potassium (Kv) channel group. The roles of this gene family have been widely investigated in nerve and muscle cells. In the present study, we investigated several characteristics of Kv7.5, which is strongly expressed in the canine osteosarcoma cell line, CCL-183. Serum starvation upregulated Kv7.5 expression, and the Kv7 channel opener, flupirtine, attenuated cell proliferation by arresting cells in the G0/G1 phase. We also showed that Kv7.5 knockdown helps CCL-183 cells to proliferate. In an effort to find an endogenous regulator of Kv7.5, we used mithramycin A to reduce the level of the transcription factor Sp1, and it strongly inhibited the induction of Kv7.5 in CCL-183 cells. These results suggest that the activation of Kv7.5 by flupirtine may exert an anti-proliferative effect in canine osteosarcoma. Therefore, Kv7.5 is a possible molecular target for canine osteosarcoma therapy. PMID:24434641

  14. Subtype-selective activation of K(v)7 channels by AaTXKβ₂₋₆₄, a novel toxin variant from the Androctonus australis scorpion venom.

    PubMed

    Landoulsi, Zied; Miceli, Francesco; Palmese, Angelo; Amoresano, Angela; Marino, Gennaro; El Ayeb, Mohamed; Taglialatela, Maurizio; Benkhalifa, Rym

    2013-11-01

    K(v)7.4 channel subunits are expressed in central auditory pathways and in inner ear sensory hair cells and skeletal and smooth muscle cells. Openers of K(v)7.4 channels have been suggested to improve hearing loss, systemic or pulmonary arterial hypertension, urinary incontinence, gastrointestinal and neuropsychiatric diseases, and skeletal muscle disorders. Scorpion venoms are a large source of peptides active on K⁺ channels. Therefore, we have optimized a combined purification/screening procedure to identify specific modulator(s) of K(v)7.4 channels from the venom of the North African scorpion Androctonus australis (Aa). We report the isolation and functional characterization of AaTXKβ₂₋₆₄, a novel variant of AaTXKβ₁₋₆₄, in a high-performance liquid chromatography fraction from Aa venom (named P8), which acts as the first peptide activator of K(v)7.4 channels. In particular, in both Xenopus oocytes and mammalian Chinese hamster ovary cells, AaTXKβ₂₋₆₄, but not AaTXKβ₁₋₆₄, hyperpolarized the threshold voltage of current activation and increased the maximal currents of heterologously expressed K(v)7.4 channels. AaTXKβ₂₋₆₄ also activated K(v)7.3, K(v)7.2/3, and K(v)7.5/3 channels, whereas homomeric K(v)1.1, K(v)7.1, and K(v)7.2 channels were unaffected. We anticipate that these results may prove useful in unraveling the novel biologic roles of AaTXKβ₂₋₆₄-sensitive K(v)7 channels and developing novel pharmacologic tools that allow subtype-selective targeting of K(v)7 channels. PMID:24019223

  15. The episodic ataxia type 1 mutation I262T alters voltage-dependent gating and disrupts protein biosynthesis of human Kv1.1 potassium channels

    PubMed Central

    Chen, Szu-Han; Fu, Ssu-Ju; Huang, Jing-Jia; Tang, Chih-Yung

    2016-01-01

    Voltage-gated potassium (Kv) channels are essential for setting neuronal membrane excitability. Mutations in human Kv1.1 channels are linked to episodic ataxia type 1 (EA1). The EA1-associated mutation I262T was identified from a patient with atypical phenotypes. Although a previous report has characterized its suppression effect, several key questions regarding the impact of the I262T mutation on Kv1.1 as well as other members of the Kv1 subfamily remain unanswered. Herein we show that the dominant-negative effect of I262T on Kv1.1 current expression is not reversed by co-expression with Kvβ1.1 or Kvβ2 subunits. Biochemical examinations indicate that I262T displays enhanced protein degradation and impedes membrane trafficking of Kv1.1 wild-type subunits. I262T appears to be the first EA1 mutation directly associated with impaired protein stability. Further functional analyses demonstrate that I262T changes the voltage-dependent activation and Kvβ1.1-mediated inactivation, uncouples inactivation from activation gating, and decelerates the kinetics of cumulative inactivation of Kv1.1 channels. I262T also exerts similar dominant effects on the gating of Kv1.2 and Kv1.4 channels. Together our data suggest that I262T confers altered channel gating and reduced functional expression of Kv1 channels, which may account for some of the phenotypes of the EA1 patient. PMID:26778656

  16. Deletion of Kvβ1.1 subunit leads to electrical and haemodynamic changes causing cardiac hypertrophy in female murine hearts

    PubMed Central

    Tur, Jared; Chapalamadugu, Kalyan C.; Padawer, Timothy; Badole, Sachin L.; Kilfoil, Peter J.; Bhatnagar, Aruni; Tipparaju, Srinivas M.

    2016-01-01

    Cardiovascular disease is the leading cause of death and debility in women in the USA, and cardiac arrhythmias are a major concern. Voltage-gated potassium (Kv) channels along with the binding partners; Kvβ subunits are major regulators of the action potential (AP) shape and duration (APD). The regulation of Kv channels by the Kvβ1 subunit is unknown in female hearts. In the present study, we hypothesized that the Kvβ1 subunit is an important regulator of female cardiac physiology. To test this hypothesis, we ablated (knocked out; KO) the KCNAB1 isoform 1 (Kvβ1.1) subunit in mice and evaluated cardiac function and electrical activity by using ECG, monophasic action potential recordings and echocardiography. Our results showed that the female Kvβ1.1 KO mice developed cardiac hypertrophy, and the hearts were structurally different, with enlargement and increased area. The electrical derangements caused by Kvβ1.1 KO in female mice included long QTc and QRS intervals along with increased APD (APD20–90% repolarization). The male Kvβ1.1 KO mice did not develop cardiac hypertrophy, but they showed long QTc and prolonged APD. Molecular analysis showed that several genes that support cardiac hypertrophy were significantly altered in Kvβ1.1 KO female hearts. In particular, myosin heavy chain αexpression was significantly elevated in Kvβ1.1 KO mouse heart. Using a small interfering RNA strategy, we identified that knockdown of Kvβ1 increases myosin heavy chain αexpression in H9C2 cells. Collectively, changes in molecular and cell signalling pathways clearly point towards a distinct electrical and structural remodelling consistent with cardiac hypertrophy in the Kvβ1.1 KO female mice. PMID:27038296

  17. Cellular Mechanism of Inner Ear Genetic Disease, roles of Kv7.1 (KCNQ1) Channel

    NASA Astrophysics Data System (ADS)

    Mousavi Nik, Atefeh

    Potassium channels are the most diverse and widely distributed membrane protein in all living organisms. They have various roles in the body such as controlling membrane potential, cell volume, and cell migration. Many studies have shown that mutation in these channels is associated with different diseases for example: Hearing Defect, Cardiac Arrhythmia, Episodic Ataxia, Seizure and Neuromyotonia. One of the most important diseases associated with K+ channel mutations is called Jervell and Lange-Nielsen syndrome (JLNS). This disease causes bilateral congenital deafness and the patients also suffer from Long QT and they usually experience syncopal episodes in their life and eventually die as a result of cardiac arrest. The gene KCNQ1 encodes the Kv7.1 voltage gated potassium channel. This channel expresses in apical membrane of marginal cell in stria vasularis of cochlea and secret K+ ion to endolymp to keep the endocochlear potential stable, which is necessary for the inner ear to function properly. Kv7.1 channel also expresses in cardiac myocytes and mutation in this gene is associated with another syndrome called Romano-Ward syndrome (RWS). Although Romano-Ward patients have mutation in KCNQ1, similar to Jervell and Lange-Nielsen patients, they only suffer from cardiac defect, and their hearing is completely normal. Several studies identified that mutations in Kv7.1 gene is associated with JLNS and RWS, but the biophysical and cellular mechanisms of these mutations are still unknown. To determine the cellular mechanisms of JLNS and RWS, and to provide mechanistic insight on the functional outputs of JLNS versus RWS mutations, we generated several mutant forms of the human Kv7.1 ( KCNQ1) clone, using site-directed mutagenesis to define their sub-cellular localization and examined their electrophysiological properties. We identified JLNS and RWS mutations at the S4-S5-linker, the pore loop (P-loop) and the C-terminus of hKv7.1 which have been found to control

  18. Solution structure and phospholipid interactions of the isolated voltage-sensor domain from KvAP

    PubMed Central

    Butterwick, Joel A.; MacKinnon, Roderick

    2010-01-01

    Voltage-sensor domains (VSDs) are specialized transmembrane segments that confer voltage sensitivity to many proteins such as ion channels and enzymes. The activities of these domains are highly dependent on both the chemical and physical properties of the surrounding membrane environment. To learn about VSD-lipid interactions, we used nuclear magnetic resonance (NMR) spectroscopy to determine the structure and phospholipid interface of the VSD from the voltage-dependent K+ channel KvAP. The solution structure of the KvAP VSD solubilized within phospholipid micelles is similar to a previously determined crystal structure solubilized by a non-ionic detergent and complexed with an antibody fragment. Two differences observed include a previously unidentified short amphipathic α-helix that precedes the first transmembrane helix and a subtle rigid body repositioning of the S3-S4 voltage-sensor paddle. Using 15N relaxation experiments, we show that most of the VSD, including the pronounced kink in S3 and the S3-S4 paddle, is relatively rigid on the ps–ns time scale. In contrast, the kink in S3 is mobile on the μs–ms time scale and may act as a hinge in the movement of the paddle during channel gating. We characterized the VSD-phospholipid micelle interactions using nuclear Overhauser effect spectroscopy and show that the micelle uniformly coats the KvAP VSD and approximates the chemical environment of a phospholipid bilayer. Using paramagnetically labeled phospholipids, we show that bilayer-forming lipids interact with the S3 and S4 helices more strongly than with S1 and S2. PMID:20851706

  19. RBE of kV CBCT radiation determined by Monte Carlo DNA damage simulations.

    PubMed

    Kirkby, C; Ghasroddashti, E; Poirier, Y; Tambasco, M; Stewart, R D

    2013-08-21

    Due to the higher LET of kilovoltage (kV) radiation, there is potential for an increase in relative biological effectiveness (RBE) of absorbed doses of radiation from kV cone beam computed tomography (CBCT) sources in reference to megavoltage or Co-60 doses. This work develops a method for accurately coupling a Monte Carlo (MC) radiation transport code (PENELOPE) with the damage simulation (MCDS) to predict relative numbers of DNA double strand breaks (DSBs). The MCDS accounts for slowing down of electrons and delta ray production within the cell nucleus; however, determining the spectrum of electrons incident on the cell nucleus from photons interacting in a larger region of tissue is not trivial. PENELOPE simulations were conducted with a novel tally algorithm invoked where electrons incident on a detection material were tracked and both the incident energy and the final deposited dose were recorded. The DSB yield predicted by a set of MCDS runs of monoenergetic electrons was then looked up in a table and weighted by the specific energy of the incident electron. Our results indicate that the RBE for DSB induction is 1.1 for diagnostic x-rays with energies from 80 to 125 kVp. We found no significant change in RBE with depth or filtration. The predicted absolute DSB yields are about three times lower for cells irradiated under anoxic conditions than the yield in cells irradiated under normoxic (5%) or fully aerobic (100%) conditions. However, oxygen concentration has a negligible (± 0.02) effect on the RBE of kV CBCT x-rays. PMID:23899567

  20. RBE of kV CBCT radiation determined by Monte Carlo DNA damage simulations

    NASA Astrophysics Data System (ADS)

    Kirkby, C.; Ghasroddashti, E.; Poirier, Y.; Tambasco, M.; Stewart, R. D.

    2013-08-01

    Due to the higher LET of kilovoltage (kV) radiation, there is potential for an increase in relative biological effectiveness (RBE) of absorbed doses of radiation from kV cone beam computed tomography (CBCT) sources in reference to megavoltage or Co-60 doses. This work develops a method for accurately coupling a Monte Carlo (MC) radiation transport code (PENELOPE) with the damage simulation (MCDS) to predict relative numbers of DNA double strand breaks (DSBs). The MCDS accounts for slowing down of electrons and delta ray production within the cell nucleus; however, determining the spectrum of electrons incident on the cell nucleus from photons interacting in a larger region of tissue is not trivial. PENELOPE simulations were conducted with a novel tally algorithm invoked where electrons incident on a detection material were tracked and both the incident energy and the final deposited dose were recorded. The DSB yield predicted by a set of MCDS runs of monoenergetic electrons was then looked up in a table and weighted by the specific energy of the incident electron. Our results indicate that the RBE for DSB induction is 1.1 for diagnostic x-rays with energies from 80 to 125 kVp. We found no significant change in RBE with depth or filtration. The predicted absolute DSB yields are about three times lower for cells irradiated under anoxic conditions than the yield in cells irradiated under normoxic (5%) or fully aerobic (100%) conditions. However, oxygen concentration has a negligible (±0.02) effect on the RBE of kV CBCT x-rays.

  1. Transcriptional repression of the M channel subunit Kv7.2 in chronic nerve injury.

    PubMed

    Rose, Kirstin; Ooi, Lezanne; Dalle, Carine; Robertson, Brian; Wood, Ian C; Gamper, Nikita

    2011-04-01

    Neuropathic pain is a severe health problem for which there is a lack of effective therapy. A frequent underlying condition of neuropathic pain is a sustained overexcitability of pain-sensing (nociceptive) sensory fibres. Therefore, the identification of mechanisms for such abnormal neuronal excitability is of utmost importance for understanding neuropathic pain. Despite much effort, an inclusive model explaining peripheral overexcitability is missing. We investigated transcriptional regulation of the Kcnq2 gene, which encodes the Kv7.2 subunit of membrane potential-stabilizing M channel, in peripheral sensory neurons in a model of neuropathic pain-partial sciatic nerve ligation (PSNL). We show that Kcnq2 is the major Kcnq gene transcript in dorsal root ganglion (DRG); immunostaining and patch-clamp recordings from acute ganglionic slices verified functional expression of Kv7.2 in small-diameter nociceptive DRG neurons. Neuropathic injury induced substantial downregulation of Kv7.2 expression. Levels of repressor element 1-silencing transcription factor (REST), which is known to suppress Kcnq2 expression, were upregulated in response to neuropathic injury identifying the likely mechanism of Kcnq2 regulation. Behavioural experiments demonstrated that neuropathic hyperalgesia following PSNL developed faster than the downregulation of Kcnq2 expression could be detected, suggesting that this transcriptional mechanism may contribute to the maintenance rather than the initiation of neuropathic pain. Importantly, the decrease in the peripheral M channel abundance could be functionally compensated by peripherally applied M channel opener flupirtine, which alleviated neuropathic hyperalgesia. Our work suggests a novel mechanism for neuropathic overexcitability and brings focus on M channels and REST as peripheral targets for the treatment of neuropathic pain. PMID:21345591

  2. Collimated electron beam accelerated at 12 kV from a Penning discharge

    SciTech Connect

    Toader, D.; Oane, M.; Ticoş, C. M.

    2015-01-15

    A pulsed electron beam accelerated at 12 kV with a duration of 40 μs per pulse is obtained from a Penning discharge with a hollow anode and two cathodes. The electrons are extracted through a hole in one of the cathodes and focused by a pair of coils. The electron beam has a diameter of a few mm in the cross section, while the beam current reaches peak values of 400 mA, depending on the magnetic field inside the focussing coils. This relatively inexpensive and compact device is suitable for the irradiation of small material samples placed in high vacuum.

  3. Accelerated aging and flashover tests on 138 kV nonceramic line post insulators

    SciTech Connect

    Schneider, H.M.; Guidi, W.W. ); Burnham, J.T. ); Gorur, R.S. ); Hall, J.F. )

    1993-01-01

    The behavior of 138 kV nonceramic line post insulators is investigated by means of clean fog tests conducted before and after aging in a specially designed accelerated aging chamber. The laboratory aging cycles are justified on the basis of actual weather in the coastal regions of Florida. Analytical measurements quantifying the degree of artificial aging are discussed and comparisons of artificial aging with service experience are presented. Observations of audible noise and radio influence voltage during the clean fog tests are reported.

  4. Analysis of a 1200 kV circuit breaker for a gas insulated substation. Final report

    SciTech Connect

    Not Available

    1985-01-01

    This report describes the work carried out to analyze and design a circuit for use in 1200 kV gas insulated substations. The first part of the project was devoted to a thorough analysis of the requirements for the circuit breaker from the standpoint of the electrical system in which it would operate. A conceptual design was selected and all of the components of the circuit breaker were designed, modeled and verified. Finally a plan was prepared for the construction of a complete circuit breaker.

  5. Construction of 8000A class 275kV gas insulated transmission line

    SciTech Connect

    Kuroyanagi, Y.; Toya, A. ); Hayashi, T. ); Araki, T. )

    1990-01-01

    The 8000A class 275kV gas insulated transmission lines (GIL), 138m and 158m long, completed as getaway lines in the Shin Noda substation in 1988 is described. The GIL was designed to match large current capacity. New techniques were applied to construct these lines. Thermal expansion and contraction of a straight section of GIL is absorbed by an angle variation of a deflection unit. Supports were designed to absorb a ground depression. Units of GIL were installed with gantry cranes. Joints of enclosure were welded by an automatic welding machine.

  6. Performance of 115 kV fiberglass transmission crossarms in wet and salt fog environments

    SciTech Connect

    Shah, K.R.; Sarkinen, C.F.; Verma, V.

    1983-04-01

    This paper discusses 60 Hz-dielectric tests performed on 115 kV solid fiberglass transmission crossarm-insulator assemblies to evaluate their performance in wet and salt fog environments. Testing included the following variables: (1) wet and dry conditions, (2) new and used crossarms and insulators, (3) type and number of insulators, and (4) salt fog salinity. Analysis of test results shows that the surface coating of fiberglass insulation is susceptible to degradation in a salt fog environment. Once this coating is eroded and the glass fibers are exposed, carbonized tracking develops. The fiberglass crossarm thus loses its electrical strength.

  7. Design and development of a 5 kV isolated solid state switch

    NASA Technical Reports Server (NTRS)

    Holbrook, R. J.; Scapple, R. Y.; Keister, F. Z.; Gooder, S. T.

    1975-01-01

    This paper describes the design and fabrication of a 5000 volt isolated hybrid switch developed by Hughes Aircraft Company under contract to NASA/Lewis. Hughes did the packaging design and NASA did the circuit design. This unique microcircuit is intended for use as a shorting switch for large extraterrestrial solar cell arrays. The packaging design for the 5 kV isolated hybrid switch is different from most hybrid microcircuits in that it utilizes a compartmentalized plastic case (a portion of which is encapsulated), is not hermetic, and is designed for high voltage operation.

  8. Characterization of a novel high-potency positive modulator of K(v)7 channels.

    PubMed

    Dalby-Brown, William; Jessen, Carsten; Hougaard, Charlotte; Jensen, Marianne L; Jacobsen, Thomas A; Nielsen, Karin S; Erichsen, Helle K; Grunnet, Morten; Ahring, Philip K; Christophersen, Palle; Strøbæk, Dorte; Jørgensen, Susanne

    2013-06-01

    K(v)7 channel activators decrease neuronal excitability and might potentially treat neuronal hyperexcitability disorders like epilepsy and mania. Here we introduce NS15370 ((2-(3,5-difluorophenyl)-N-[6-[(4-fluorophenyl)methylamino]-2-morpholino-3-pyridyl]acetamide)hydrochloride, an in vitro high-potency chemical analogue of retigabine, without effects on GABA(A) receptors. NS15370 activates recombinant homo- and heteromeric K(v)7.2-K(v)7.5 channels in HEK293 cells at sub-micromolar concentrations (EC₅₀~100 nM, as quantified by a fluorescence based Tl⁺-influx assay). In voltage clamp experiments NS15370 exhibits a complex, concentration-dependent mode-of-action: At low concentrations it accelerates voltage-dependent activation rates, slows deactivations, and increases steady-state current amplitudes. Quantified by the peak-tail current method, the V½ value of the steady-state activation curve is shifted towards hyperpolarized potentials at concentrations ~100 times lower than retigabine. However, in contrast to retigabine, NS15370 also introduces a distinct time-dependent current decrease, which eventually, at higher concentrations, causes suppression of the current at depolarized potentials, and an apparent "cross-over" of the voltage-activation curve. In brain slices, NS15370 hyperpolarizes and increases spike frequency adaptation of hippocampal CA1 neurons and the compound reduces the autonomous firing of dopaminergic neurons in the substantia-nigra pars compacta. NS15370 is effective in rodent models of hyperexcitability: (i) it yields full protection against mouse 6 Hz seizures and rat amygdala kindling discharges, two models of partial epilepsia; (ii) it reduces (+)-MK-801 hydrogen maleate (MK-801)-induced hyperactivity as well as chlordiazepoxide (CDP)+d-amphetamine (AMP)-induced hyperactivity, models sensitive to classic anti-psychotic and anti-manic treatments, respectively. Our findings with NS15370 consolidate neuronal K(v)7 channels as targets for

  9. Sidney-North Yuma 230-kV Transmission Line Project, Colorado and Nebraska

    SciTech Connect

    Not Available

    1991-06-01

    This report describes the need for a 230-kV overhead transmission line to supply power from Sidney, Nebraska to eastern Colorado. The alternative scenario compared to construction of the line is No Action. Rejected alternatives include underground lines and different routing paths, with a possible extension to the Sterling area. Both scenarios are evaluated for environmental effects, cost, and consequences for the eastern Colorado region. The proposed route is determined to be the environmentally preferred choice. 120 refs., 6 figs., 13 tabs. (MHB)

  10. Efficient computation of matched solutions of the KV envelopeequation for periodic focusing lattices

    SciTech Connect

    Lund, Steven M.; Chilton, Sven H.; Lee, Edward P.

    2006-01-03

    A new iterative method is developed to numerically calculate the periodic, matched beam envelope solution of the coupled Kapchinskij-Vladimirskij (KV) equations describing the transverse evolution of a beam in a periodic, linear focusing lattice of arbitrary complexity. Implementation of the method is straightforward. It is highly convergent and can be applied to all usual parameterizations of the matched envelope solutions. The method is applicable to all classes of linear focusing lattices without skew couplings, and also applies to parameters where the matched beam envelope is strongly unstable. Example applications are presented for periodic solenoidal and quadrupole focusing lattices. Convergence properties are summarized over a wide range of system parameters.

  11. Metrological traceability for AC High-Voltage in Inmetro up to 40 kV

    NASA Astrophysics Data System (ADS)

    Vitorio, P. C. O.; de Lima, V. R.; Borges Filho, O.; de Souza, L. A. A.; Asencios, O. W. G.

    2016-07-01

    This paper refers to a project carried out in Inmetro aiming to provide internal metrological traceability for 60 Hz AC High-Voltage up to 40 kV. It presents details about the method used, its equations and obtained results. A capacitance and tanb bridge, with a built-in current comparator, was used in combination with two standard capacitors to calibrate a standard potential transformer (PT), both in ratio and phase angle. The results obtained by Inmetro showed good agreement with PTB ones, for the same PT. The maximum estimated uncertainty was 0,0049% for ratio error and 104 μrad for phase angle error.

  12. Development of 10 kV 4H-SiC JBS diode with FGR termination

    NASA Astrophysics Data System (ADS)

    Runhua, Huang; Yonghong, Tao; Pengfei, Cao; Ling, Wang; Gang, Chen; Song, Bai; Rui, Li; Yun, Li; Zhifei, Zhao

    2014-07-01

    The design, fabrication, and electrical characteristics of the 4H-SiC JBS diode with a breakdown voltage higher than 10 kV are presented. 60 floating guard rings have been used in the fabrication. Numerical simulations have been performed to select the doping level and thickness of the drift layer and the effectiveness of the edge termination technique. The n-type epilayer is 100 μm in thickness with a doping of 6 × 1014 cm-3. The on-state voltage was 2.7 V at JF = 13 A/cm2.

  13. Development of flexible joint for 500kV Al-sheathed O. F. cable

    SciTech Connect

    Komaba, T.; Kanai, K. ); Yoshida, S.; Shigetoshi, I.; Amano, K. )

    1992-10-01

    This paper reports on a flexible dimensionaly flush joint for 500kV aluminum-sheathed oil-filled cables capable of being assembled, pulled an installed in a similar manner to cables at site which has been developed. This joint is intended for use at intermediate points on long bridges or tunnels where local assembly and installation of conventional joints would be difficult. In developing the joint various novel and original techniques have been employed, including the flexible flush jointing of segmental conductors, the flexible jointing of paper insulation by a combination of wrapping-back and stepping methods, plus on-site aluminum sheath corrugating and sealing methods.

  14. Long term pre-qualification testing program on a 230kV XLPE cable system

    SciTech Connect

    Champion, T.C.; Agostinelli, F.M.; Rosevear, R.D.

    1994-12-31

    this paper describes the installation, testing, and results of a long term, full scale laboratory evaluation of a 230kV XLPE insulated cable system. System components included two innovative, pre-molded splices a 128 meter (420 foot) cable run, and two silicone oil filled, porcelain cable terminations. Load cycle aging was performed on the cable system over a four year period. After successfully completing the outdoor aging program, the system was exposed to a final impulses breakdown test. Results demonstrated the importance of considering thermomechanical bending in aluminum conductor, XLPE insulated designs. The influence on cable ampacity of solar heating at riser transitions was also demonstrated.

  15. Long term pre-qualification testing program on a 230kV XLPE cable system

    SciTech Connect

    Champion, T.C.; Agostinelli, F.M.; Rosevear, R.D.

    1995-01-01

    This paper describes the installation, testing, and results of a long term, full scale laboratory evaluation of a 230kV XLPE insulated cable system. System components included two innovative, pre-molded splices, a 128 meter (420 foot) cable run, and two silicone oil filled, porcelain cable terminations. Load cycle aging was performed on the cable system over a four year period. After successfully completing the outdoor aging program, the system was exposed to a final impulse breakdown test. Results demonstrated the importance or considering thermomechanical bending in aluminum conductor, XLPE insulated designs. The influence on cable ampacity of solar beating at riser transitions was also demonstrated.

  16. WE-G-18A-02: Calibration-Free Combined KV/MV Short Scan CBCT

    SciTech Connect

    Wu, M; Loo, B; Bazalova, M; Fahrig, R; Star-Lack, J

    2014-06-15

    Purpose: To combine orthogonal kilo-voltage (kV) and Mega-voltage (MV) projection data for short scan cone-beam CT to reduce imaging time on current radiation treatment systems, using a calibration-free gain correction method. Methods: Combining two orthogonal projection data sets for kV and MV imaging hardware can reduce the scan angle to as small as 110° (90°+fan) such that the total scan time is ∼18 seconds, or within a breath hold. To obtain an accurate reconstruction, the MV projection data is first linearly corrected using linear regression using the redundant data from the start and end of the sinogram, and then the combined data is reconstructed using the FDK method. To correct for the different changes of attenuation coefficients in kV/MV between soft tissue and bone, the forward projection of the segmented bone and soft tissue from the first reconstruction in the redundant region are added to the linear regression model. The MV data is corrected again using the additional information from the segmented image, and combined with kV for a second FDK reconstruction. We simulated polychromatic 120 kVp (conventional a-Si EPID with CsI) and 2.5 MVp (prototype high-DQE MV detector) projection data with Poisson noise using the XCAT phantom. The gain correction and combined kV/MV short scan reconstructions were tested with head and thorax cases, and simple contrast-to-noise ratio measurements were made in a low-contrast pattern in the head. Results: The FDK reconstruction using the proposed gain correction method can effectively reduce artifacts caused by the differences of attenuation coefficients in the kV/MV data. The CNRs of the short scans for kV, MV, and kV/MV are 5.0, 2.6 and 3.4 respectively. The proposed gain correction method also works with truncated projections. Conclusion: A novel gain correction and reconstruction method was developed to generate short scan CBCT from orthogonal kV/MV projections. This work is supported by NIH Grant 5R01CA138426-05.

  17. Efficient computation of matched solutions of the KV envelope equations for periodic focusing lattices.

    SciTech Connect

    Lund, S M; Chilton, S H; Lee, E P

    2006-01-17

    A new iterative method is developed to numerically calculate the periodic, matched beam envelope solution of the coupled Kapchinskij-Vladimirskij (KV) equations describing the transverse evolution of a beam in a periodic, linear focusing lattice of arbitrary complexity. Implementation of the method is straightforward. It is highly convergent and can be applied to all usual parameterizations of the matched envelope solutions. The method is applicable to all classes of linear focusing lattices without skew couplings, and also applies to parameters where the matched beam envelope is strongly unstable. Example applications are presented for periodic solenoidal and quadrupole focusing lattices. Convergence properties are summarized over a wide range of system parameters.

  18. Distinct Kv Channel Subtypes Contribute to Differences in Spike Signaling Properties in the Axon Initial Segment and Presynaptic Boutons of Cerebellar Interneurons

    PubMed Central

    Rowan, Matthew J. M.; Tranquil, Elizabeth

    2014-01-01

    The discrete arrangement of voltage-gated K+ (Kv) channels in axons may impart functional advantages in action potential (AP) signaling yet, in compact cell types, the organization of Kv channels is poorly understood. We find that in cerebellar stellate cell interneurons of mice, the composition and influence of Kv channels populating the axon is diverse and depends on location allowing axonal compartments to differentially control APs in a local manner. Kv1 channels determine AP repolarization at the spike initiation site but not at more distal sites, limiting the expression of use-dependent spike broadening to the most proximal axon region, likely a key attribute informing spiking phenotype. Local control of AP repolarization at presynaptic boutons depends on Kv3 channels keeping APs brief, thus limiting Ca2+ influx and synaptic strength. These observations suggest that AP repolarization is tuned by the local influence of distinct Kv channel types, and this organization enhances the functional segregation of axonal compartments. PMID:24806686

  19. Metal artifact correction for x-ray computed tomography using kV and selective MV imaging

    SciTech Connect

    Wu, Meng; Keil, Andreas; Constantin, Dragos; Star-Lack, Josh; Zhu, Lei; Fahrig, Rebecca

    2014-12-15

    Purpose: The overall goal of this work is to improve the computed tomography (CT) image quality for patients with metal implants or fillings by completing the missing kilovoltage (kV) projection data with selectively acquired megavoltage (MV) data that do not suffer from photon starvation. When both of these imaging systems, which are available on current radiotherapy devices, are used, metal streak artifacts are avoided, and the soft-tissue contrast is restored, even for regions in which the kV data cannot contribute any information. Methods: Three image-reconstruction methods, including two filtered back-projection (FBP)-based analytic methods and one iterative method, for combining kV and MV projection data from the two on-board imaging systems of a radiotherapy device are presented in this work. The analytic reconstruction methods modify the MV data based on the information in the projection or image domains and then patch the data onto the kV projections for a FBP reconstruction. In the iterative reconstruction, the authors used dual-energy (DE) penalized weighted least-squares (PWLS) methods to simultaneously combine the kV/MV data and perform the reconstruction. Results: The authors compared kV/MV reconstructions to kV-only reconstructions using a dental phantom with fillings and a hip-implant numerical phantom. Simulation results indicated that dual-energy sinogram patch FBP and the modified dual-energy PWLS method can successfully suppress metal streak artifacts and restore information lost due to photon starvation in the kV projections. The root-mean-square errors of soft-tissue patterns obtained using combined kV/MV data are 10–15 Hounsfield units smaller than those of the kV-only images, and the structural similarity index measure also indicates a 5%–10% improvement in the image quality. The added dose from the MV scan is much less than the dose from the kV scan if a high efficiency MV detector is assumed. Conclusions: The authors have shown that it

  20. Roles of specific Kv channel types in repolarization of the action potential in genetically identified subclasses of pyramidal neurons in mouse neocortex.

    PubMed

    Pathak, Dhruba; Guan, Dongxu; Foehring, Robert C

    2016-05-01

    The action potential (AP) is a fundamental feature of excitable cells that serves as the basis for long-distance signaling in the nervous system. There is considerable diversity in the appearance of APs and the underlying repolarization mechanisms in different neuronal types (reviewed in Bean BP. Nat Rev Neurosci 8: 451-465, 2007), including among pyramidal cell subtypes. In the present work, we used specific pharmacological blockers to test for contributions of Kv1, Kv2, or Kv4 channels to repolarization of single APs in two genetically defined subpopulations of pyramidal cells in layer 5 of mouse somatosensory cortex (etv1 and glt) as well as pyramidal cells from layer 2/3. These three subtypes differ in AP properties (Groh A, Meyer HS, Schmidt EF, Heintz N, Sakmann B, Krieger P. Cereb Cortex 20: 826-836, 2010; Guan D, Armstrong WE, Foehring RC. J Neurophysiol 113: 2014-2032, 2015) as well as laminar position, morphology, and projection targets. We asked what the roles of Kv1, Kv2, and Kv4 channels are in AP repolarization and whether the underlying mechanisms are pyramidal cell subtype dependent. We found that Kv4 channels are critically involved in repolarizing neocortical pyramidal cells. There are also pyramidal cell subtype-specific differences in the role for Kv1 channels. Only Kv4 channels were involved in repolarizing the narrow APs of glt cells. In contrast, in etv1 cells and layer 2/3 cells, the broader APs are partially repolarized by Kv1 channels in addition to Kv4 channels. Consistent with their activation in the subthreshold range, Kv1 channels also regulate AP voltage threshold in all pyramidal cell subtypes. PMID:26864770

  1. The Scorpion Toxin Analogue BmKTX-D33H as a Potential Kv1.3 Channel-Selective Immunomodulator for Autoimmune Diseases.

    PubMed

    Ye, Fang; Hu, Youtian; Yu, Weiwei; Xie, Zili; Hu, Jun; Cao, Zhijian; Li, Wenxin; Wu, Yingliang

    2016-04-01

    The Kv1.3 channel-acting scorpion toxins usually adopt the conserved anti-parallel β-sheet domain as the binding interface, but it remains challenging to discover some highly selective Kv1.3 channel-acting toxins. In this work, we investigated the pharmacological profile of the Kv1.3 channel-acting BmKTX-D33H, a structural analogue of the BmKTX scorpion toxin. Interestingly, BmKTX-D33H, with its conserved anti-parallel β-sheet domain as a Kv1.3 channel-interacting interface, exhibited more than 1000-fold selectivity towards the Kv1.3 channel as compared to other K⁺ channels (including Kv1.1, Kv1.2, Kv1.7, Kv11.1, KCa2.2, KCa2.3, and KCa3.1). As expected, BmKTX-D33H was found to inhibit the cytokine production and proliferation of both Jurkat cells and human T cells in vitro. It also significantly improved the delayed-type hypersensitivity (DTH) responses, an autoreactive T cell-mediated inflammation in rats. Amino acid sequence alignment and structural analysis strongly suggest that the "evolutionary" Gly11 residue of BmKTX-D33H interacts with the turret domain of Kv1 channels; it appears to be a pivotal amino acid residue with regard to the selectivity of BmKTX-D33H towards the Kv1.3 channel (in comparison with the highly homologous scorpion toxins). Together, our data indicate that BmKTX-D33H is a Kv1.3 channel-specific blocker. Finally, the remarkable selectivity of BmKTX-D33H highlights the great potential of evolutionary-guided peptide drug design in future studies. PMID:27104568

  2. A polyether biotoxin binding site on the lipid-exposed face of the pore domain of Kv channels revealed by the marine toxin gambierol

    PubMed Central

    Kopljar, Ivan; Labro, Alain J.; Cuypers, Eva; Johnson, Henry W. B.; Rainier, Jon D.; Tytgat, Jan; Snyders, Dirk J.

    2009-01-01

    Gambierol is a marine polycyclic ether toxin belonging to the group of ciguatera toxins. It does not activate voltage-gated sodium channels (VGSCs) but inhibits Kv1 potassium channels by an unknown mechanism. While testing whether Kv2, Kv3, and Kv4 channels also serve as targets, we found that Kv3.1 was inhibited with an IC50 of 1.2 ± 0.2 nM, whereas Kv2 and Kv4 channels were insensitive to 1 μM gambierol. Onset of block was similar from either side of the membrane, and gambierol did not compete with internal cavity blockers. The inhibition did not require channel opening and could not be reversed by strong depolarization. Using chimeric Kv3.1–Kv2.1 constructs, the toxin sensitivity was traced to S6, in which T427 was identified as a key determinant. In Kv3.1 homology models, T427 and other molecular determinants (L348, F351) reside in a space between S5 and S6 outside the permeation pathway. In conclusion, we propose that gambierol acts as a gating modifier that binds to the lipid-exposed surface of the pore domain, thereby stabilizing the closed state. This site may be the topological equivalent of the neurotoxin site 5 of VGSCs. Further elucidation of this previously undescribed binding site may explain why most ciguatoxins activate VGSCs, whereas others inhibit voltage-dependent potassium (Kv) channels. This previously undescribed Kv neurotoxin site may have wide implications not only for our understanding of channel function at the molecular level but for future development of drugs to alleviate ciguatera poisoning or to modulate electrical excitability in general. PMID:19482941

  3. SU-E-J-31: Monitor Interfractional Variation of Tumor Respiratory Motion Using 4D KV Conebeam Computed Tomography for Stereotactic Body Radiation Therapy of Lung Cancer

    SciTech Connect

    Tai, A; Prior, P; Gore, E; Johnstone, C; Li, X

    2015-06-15

    Purpose: 4DCT has been widely used to generate internal tumor volume (ITV) for a lung tumor for treatment planning. However, lung tumors may show different respiratory motion on the treatment day. The purpose of this study is to evaluate 4D KV conebeam computed tomography (CBCT) for monitoring tumor interfractional motion variation between simulation and each fraction of stereotactic body radiation therapy (SBRT) for lung cancer. Methods: 4D KV CBCT was acquired with the Elekta XVI system. The accuracy of 4D KV CBCT for image-guided radiation therapy (IGRT) was tested with a dynamic thorax motion phantom (CIRS, Virginia) with a linear amplitude of 2 cm. In addition, an adult anthropomorphic phantom (Alderson, Rando) with optically stimulated luminescence (OSL) dosimeters embedded at the center and periphery of a slab of solid water was used to measure the dose of 4D KV CBCT and to compare it with the dose with 3D KV CBCT. The image registration was performed by aligning\\ each phase images of 4D KV CBCT to the planning images and the final couch shifts were calculated as a mean of all these individual shifts along each direction.A workflow was established based on these quality assurance tests for lung cancer patients. Results: 4D KV CBCT does not increase imaging dose in comparison to 3D KV CBCT. Acquisition of 4D KV CBCT is 4 minutes as compared to 2 minutes for 3D KV CBCT. Most of patients showed a small daily variation of tumor respiratory motion about 2 mm. However, some patients may have more than 5 mm variations of tumor respiratory motion. Conclusion: The radiation dose does not increase with 4D KV CBCT. 4D KV CBCT is a useful tool for monitoring interfractional variations of tumor respiratory motion before SBRT of lung cancer patients.

  4. The Scorpion Toxin Analogue BmKTX-D33H as a Potential Kv1.3 Channel-Selective Immunomodulator for Autoimmune Diseases

    PubMed Central

    Ye, Fang; Hu, Youtian; Yu, Weiwei; Xie, Zili; Hu, Jun; Cao, Zhijian; Li, Wenxin; Wu, Yingliang

    2016-01-01

    The Kv1.3 channel-acting scorpion toxins usually adopt the conserved anti-parallel β-sheet domain as the binding interface, but it remains challenging to discover some highly selective Kv1.3 channel-acting toxins. In this work, we investigated the pharmacological profile of the Kv1.3 channel-acting BmKTX-D33H, a structural analogue of the BmKTX scorpion toxin. Interestingly, BmKTX-D33H, with its conserved anti-parallel β-sheet domain as a Kv1.3 channel-interacting interface, exhibited more than 1000-fold selectivity towards the Kv1.3 channel as compared to other K+ channels (including Kv1.1, Kv1.2, Kv1.7, Kv11.1, KCa2.2, KCa2.3, and KCa3.1). As expected, BmKTX-D33H was found to inhibit the cytokine production and proliferation of both Jurkat cells and human T cells in vitro. It also significantly improved the delayed-type hypersensitivity (DTH) responses, an autoreactive T cell-mediated inflammation in rats. Amino acid sequence alignment and structural analysis strongly suggest that the “evolutionary” Gly11 residue of BmKTX-D33H interacts with the turret domain of Kv1 channels; it appears to be a pivotal amino acid residue with regard to the selectivity of BmKTX-D33H towards the Kv1.3 channel (in comparison with the highly homologous scorpion toxins). Together, our data indicate that BmKTX-D33H is a Kv1.3 channel–specific blocker. Finally, the remarkable selectivity of BmKTX-D33H highlights the great potential of evolutionary-guided peptide drug design in future studies. PMID:27104568

  5. Localization and function of the Kv3.1b subunit in the rat medulla oblongata: focus on the nucleus tractus solitarii

    PubMed Central

    Dallas, Mark L; Atkinson, Lucy; Milligan, Carol J; Morris, Neil P; Lewis, David I; Deuchars, Susan A; Deuchars, Jim

    2005-01-01

    The voltage-gated potassium channel subunit Kv3.1 confers fast firing characteristics to neurones. Kv3.1b subunit immunoreactivity (Kv3.1b-IR) was widespread throughout the medulla oblongata, with labelled neurones in the gracile, cuneate and spinal trigeminal nuclei. In the nucleus of the solitary tract (NTS), Kv3.1b-IR neurones were predominantly located close to the tractus solitarius (TS) and could be GABAergic or glutamatergic. Ultrastructurally, Kv3.1b-IR was detected in NTS terminals, some of which were vagal afferents. Whole-cell current-clamp recordings from neurones near the TS revealed electrophysiological characteristics consistent with the presence of Kv3.1b subunits: short duration action potentials (4.2 ± 1.4 ms) and high firing frequencies (68.9 ± 5.3 Hz), both sensitive to application of TEA (0.5 mm) and 4-aminopyridine (4-AP; 30 μm). Intracellular dialysis of an anti-Kv3.1b antibody mimicked and occluded the effects of TEA and 4-AP in NTS and dorsal column nuclei neurones, but not in dorsal vagal nucleus or cerebellar Purkinje cells (which express other Kv3 subunits, but not Kv3.1b). Voltage-clamp recordings from outside-out patches from NTS neurones revealed an outward K+ current with the basic characteristics of that carried by Kv3 channels. In NTS neurones, electrical stimulation of the TS evoked EPSPs and IPSPs, and TEA and 4-AP increased the average amplitude and decreased the paired pulse ratio, consistent with a presynaptic site of action. Synaptic inputs evoked by stimulation of a region lacking Kv3.1b-IR neurones were not affected, correlating the presence of Kv3.1b in the TS with the pharmacological effects. PMID:15528247

  6. The heterogeneity of hyperinsulinaemic hypoglycaemia in 19 patients with Beckwith-Wiedemann syndrome due to KvDMR1 hypomethylation.

    PubMed

    Senniappan, Senthil; Ismail, Dunia; Shipster, Caroleen; Beesley, Clare; Hussain, Khalid

    2015-01-01

    Beckwith-Wiedemann syndrome (BWS) is an overgrowth syndrome caused by multiple epigenetic and genetic changes affecting imprinted genes on chromosome 11p15.5. Hypomethylation of KvDMR1 on the maternal allele is the most common genetic cause, and hyperinsulinaemic hypoglycaemia (HH) is the most common biochemical abnormality. We evaluated the correlation between severity of HH and degree of hypomethylation in BWS. Out of the 19 patients with BWS due to KvDMR1 hypomethylation, 10 patients had no HH, 5 had mild transient HH that resolved spontaneously, and 4 required diazoxide therapy for up to 6 months. There was no correlation between the degree of KvDMR1 hypomethylation and severity of HH in the 6 patients studied. All patients also showed marked clinical heterogeneity with respect to the features of BWS. In patients with BWS due to hypomethylation of KvDMR1, the clinical presentation of HH is quite heterogeneous with no correlation with the degree of KvDMR1 hypomethylation. PMID:24468603

  7. Ca2+/calmodulin regulates Kvβ1.1-mediated inactivation of voltage-gated K+ channels

    PubMed Central

    Swain, Sandip M.; Sahoo, Nirakar; Dennhardt, Sophie; Schönherr, Roland; Heinemann, Stefan H.

    2015-01-01

    A-type K+ channels open on membrane depolarization and undergo subsequent rapid inactivation such that they are ideally suited for fine-tuning the electrical signaling in neurons and muscle cells. Channel inactivation mostly follows the so-called ball-and-chain mechanism, in which the N-terminal structures of either the K+ channel’s α or β subunits occlude the channel pore entry facing the cytosol. Inactivation of Kv1.1 and Kv1.4 channels induced by Kvβ1.1 subunits is profoundly decelerated in response to a rise in the intracellular Ca2+ concentration, thus making the affected channel complexes negative feedback regulators to limit neuronal overexcitation. With electrophysiological and biochemical experiments we show that the Ca2+ dependence is gained by binding of calmodulin to the “chain” segment of Kvβ1.1 thereby compromising the mobility of the inactivation particle. Furthermore, inactivation regulation via Ca2+/calmodulin does not interfere with the β subunit’s enzymatic activity as an NADPH-dependent oxidoreductase, thus rendering the Kvβ1.1 subunit a multifunctional receptor that integrates cytosolic signals to be transduced to altered electrical cellular activity. PMID:26487174

  8. Inhibitors of mitochondrial Kv1.3 channels induce Bax/Bak-independent death of cancer cells

    PubMed Central

    Leanza, Luigi; Henry, Brian; Sassi, Nicola; Zoratti, Mario; Chandy, K George; Gulbins, Erich; Szabò, Ildikò

    2012-01-01

    Overcoming the resistance of tumours to chemotherapy, often due to downregulation of Bax and Bak, represents a significant clinical challenge. It is therefore important to identify novel apoptosis inducers that bypass Bax and Bak. Potassium channels are emerging as oncological targets and a crucial role of mitochondrial Kv1.3 in apoptosis has been demonstrated. Here we report for the first time that Psora-4, PAP-1 and clofazimine, three distinct membrane-permeant inhibitors of Kv1.3, induce death by directly targeting the mitochondrial channel in multiple human and mouse cancer cell lines. Importantly, these drugs activated the intrinsic apoptotic pathway also in the absence of Bax and Bak, a result in agreement with the current mechanistic model for mitochondrial Kv1.3 action. Genetic deficiency or short interfering RNA (siRNA)-mediated downregulation of Kv1.3 abrogated the effects of the drugs. Intraperitoneal injection of clofazimine reduced tumour size by 90% in an orthotopic melanoma B16F10 mouse model in vivo, while no adverse effects were observed in several healthy tissues. The study indicates that inhibition of mitochondrial Kv1.3 might be a novel therapeutic option for the induction of cancer cell death independent of Bax and Bak. PMID:22496117

  9. Cux1 Enables Interhemispheric Connections of Layer II/III Neurons by Regulating Kv1-Dependent Firing.

    PubMed

    Rodríguez-Tornos, Fernanda M; Briz, Carlos G; Weiss, Linnea A; Sebastián-Serrano, Alvaro; Ares, Saúl; Navarrete, Marta; Frangeul, Laura; Galazo, Maria; Jabaudon, Denis; Esteban, José A; Nieto, Marta

    2016-02-01

    Neuronal subtype-specific transcription factors (TFs) instruct key features of neuronal function and connectivity. Activity-dependent mechanisms also contribute to wiring and circuit assembly, but whether and how they relate to TF-directed neuronal differentiation is poorly investigated. Here we demonstrate that the TF Cux1 controls the formation of the layer II/III corpus callosum (CC) projections through the developmental transcriptional regulation of Kv1 voltage-dependent potassium channels and the resulting postnatal switch to a Kv1-dependent firing mode. Loss of Cux1 function led to a decrease in the expression of Kv1 transcripts, aberrant firing responses, and selective loss of CC contralateral innervation. Firing and innervation were rescued by re-expression of Kv1 or postnatal reactivation of Cux1. Knocking down Kv1 mimicked Cux1-mediated CC axonal loss. These findings reveal that activity-dependent processes are central bona fide components of neuronal TF-differentiation programs and establish the importance of intrinsic firing modes in circuit assembly within the neocortex. PMID:26804994

  10. Voltage-sensor conformation shapes the intra-membrane drug binding site that determines gambierol affinity in Kv channels.

    PubMed

    Kopljar, Ivan; Grottesi, Alessandro; de Block, Tessa; Rainier, Jon D; Tytgat, Jan; Labro, Alain J; Snyders, Dirk J

    2016-08-01

    Marine ladder-shaped polyether toxins are implicated in neurological symptoms of fish-borne food poisonings. The toxin gambierol, produced by the marine dinoflagellate Gambierdiscus toxicus, belongs to the group of ladder-shaped polyether toxins and inhibits Kv3.1 channels with nanomolar affinity through a mechanism of gating modification. Binding determinants for gambierol localize at the lipid-exposed interface of the pore forming S5 and S6 segments, suggesting that gambierol binds outside of the permeation pathway. To explore a possible involvement of the voltage-sensing domain (VSD), we made different chimeric channels between Kv3.1 and Kv2.1, exchanging distinct parts of the gating machinery. Our results showed that neither the electro-mechanical coupling nor the S1-S3a region of the VSD affect gambierol sensitivity. In contrast, the S3b-S4 part of the VSD (paddle motif) decreased gambierol sensitivity in Kv3.1 more than 100-fold. Structure determination by homology modeling indicated that the position of the S3b-S4 paddle and its primary structure defines the shape and∖or the accessibility of the binding site for gambierol, explaining the observed differences in gambierol affinity between the channel chimeras. Furthermore, these findings explain the observed difference in gambierol affinity for the closed and open channel configurations of Kv3.1, opening new possibilities for exploring the VSDs as selectivity determinants in drug design. PMID:26956727

  11. The inhibitory effect of propofol on Kv2.1 potassium channel in rat parietal cortical neurons.

    PubMed

    Zhang, Yan-Zhuo; Zhang, Rui; Zeng, Xian-Zhang; Song, Chun-Yu

    2016-03-11

    Excessive K(+) efflux via activated voltage-gated K(+) channels can deplete intracellular K(+) and lead to long-lasting membrane depolarization which will promote neuronal apoptosis during ischemia/hypoxia injury. The Kv2.1 potassium channel was the major component of delayed rectifier potassium current (Ik) in pyramidal neurons in cortex and hippocampus. The neuronal protective effect of propofol has been proved. Delayed rectifier potassium current (Ik) has been shown to have close relationship with neuronal damage. The study was designed to test the inhibitory effect of propofol on Kv2.1 potassium channel in rat parietal cortical neurons. Whole-cell patch clamp recordings and Western blot analysis were used to investigate the electrophysiological function and protein expression of Kv2.1 in rat parietal cortical neurons after propofol treatment. We found that propofol concentration-dependently inhibited Ik in pyramidal neurons. Propofol also caused a downward shift of the I-V curve of Ik at 30μM concentration. Propofol significantly inhibited the expression of Kv2.1 protein level at 30μM, 50μM, 100μM concentration. In conclusion, our data showed that propofol could inhibit Ik, probably via depressing the expression of Kv2.1 protein in rat cerebral parietal cortical neurons. PMID:26828304

  12. Auxiliary KChIP4a Suppresses A-type K+ Current through Endoplasmic Reticulum (ER) Retention and Promoting Closed-state Inactivation of Kv4 Channels*

    PubMed Central

    Tang, Yi-Quan; Liang, Ping; Zhou, Jingheng; Lu, Yanxin; Lei, Lei; Bian, Xiling; Wang, KeWei

    2013-01-01

    In the brain and heart, auxiliary Kv channel-interacting proteins (KChIPs) co-assemble with pore-forming Kv4 α-subunits to form a native K+ channel complex and regulate the expression and gating properties of Kv4 currents. Among the KChIP1–4 members, KChIP4a exhibits a unique N terminus that is known to suppress Kv4 function, but the underlying mechanism of Kv4 inhibition remains unknown. Using a combination of confocal imaging, surface biotinylation, and electrophysiological recordings, we identified a novel endoplasmic reticulum (ER) retention motif, consisting of six hydrophobic and aliphatic residues, 12–17 (LIVIVL), within the KChIP4a N-terminal KID, that functions to reduce surface expression of Kv4-KChIP complexes. This ER retention capacity is transferable and depends on its flanking location. In addition, adjacent to the ER retention motif, the residues 19–21 (VKL motif) directly promote closed-state inactivation of Kv4.3, thus leading to an inhibition of channel current. Taken together, our findings demonstrate that KChIP4a suppresses A-type Kv4 current via ER retention and enhancement of Kv4 closed-state inactivation. PMID:23576435

  13. Association of the Kv1 family of K+ channels and their functional blueprint in the properties of auditory neurons as revealed by genetic and functional analyses.

    PubMed

    Wang, Wenying; Kim, Hyo Jeong; Lv, Ping; Tempel, Bruce; Yamoah, Ebenezer N

    2013-10-01

    Developmental plasticity in spiral ganglion neurons (SGNs) ensues from profound alterations in the functional properties of the developing hair cell (HC). For example, prehearing HCs are spontaneously active. However, at the posthearing stage, HC membrane properties transition to graded receptor potentials. The dendrotoxin (DTX)-sensitive Kv1 channel subunits (Kv1.1, 1.2, and 1.6) shape the firing properties and membrane potential of SGNs, and the expression of the channel undergoes developmental changes. Because of the stochastic nature of Kv subunit heteromultimerization, it has been difficult to determine physiologically relevant subunit-specific interactions and their functions in the underlying mechanisms of Kv1 channel plasticity in SGNs. Using Kcna2 null mutant mice, we demonstrate a surprising paradox in changes in the membrane properties of SGNs. The resting membrane potential of Kcna2(-/-) SGNs was significantly hyperpolarized compared with that of age-matched wild-type (WT) SGNs. Analyses of outward currents in the mutant SGNs suggest an apparent approximately twofold increase in outward K(+) currents. We show that in vivo and in vitro heteromultimerization of Kv1.2 and Kv1.4 α-subunits underlies the striking and unexpected alterations in the properties of SGNs. The results suggest that heteromeric interactions of Kv1.2 and Kv1.4 dominate the defining features of Kv1 channels in SGNs. PMID:23864368

  14. Novel Roles for Kv7 Channels in Shaping Histamine-Induced Contractions and Bradykinin-Dependent Relaxations in Pig Coronary Arteries

    PubMed Central

    Chen, Xingjuan; Li, Wennan; Hiett, S. Christopher; Obukhov, Alexander G.

    2016-01-01

    Voltage-gated Kv7 channels are inhibited by agonists of Gq-protein-coupled receptors, such as histamine. Recent works have provided evidence that inhibition of vascular Kv7 channels may trigger vessel contractions. In this study, we investigated how Kv7 activity modulates the histamine-induced contractions in “healthy” and metabolic syndrome (MetS) pig right coronary arteries (CAs). We performed isometric tension and immunohistochemical studies with domestic, lean Ossabaw, and MetS Ossabaw pig CAs. We found that neither the Kv7.2/Kv7.4/Kv7.5 activator ML213 nor the general Kv7 inhibitor XE991 altered the tension of CA rings under preload, indicating that vascular Kv7 channels are likely inactive in the preloaded rings. Conversely, ML213 potently dilated histamine-pre-contracted CAs, suggesting that Kv7 channels are activated during histamine applications and yet partially inhibited by histamine. Immunohistochemistry analysis revealed strong Kv7.4 immunostaining in the medial and intimal layers of the CA wall, whereas Kv7.5 immunostaining intensity was strong in the intimal but weak in the medial layers. The medial Kv7 immunostaining was significantly weaker in MetS Ossabaw CAs as compared to lean Ossabaw or domestic CAs. Consistently, histamine-pre-contracted MetS Ossabaw CAs exhibited attenuated ML213-dependent dilations. In domestic pig CAs, where medial Kv7 immunostaining intensity was stronger, histamine-induced contractions spontaneously decayed to ~31% of the peak amplitude within 4 minutes. Oppositely, in Ossabaw CAs, where Kv7 immunostaining intensity was weaker, the histamine-induced contractions were more sustained. XE991 pretreatment significantly slowed the decay rate of histamine-induced contractions in domestic CAs, supporting the hypothesis that increased Kv7 activity correlates with a faster rate of histamine-induced contraction decay. Alternatively, XE991 significantly decreased the amplitude of bradykinin-dependent dilations in pre

  15. A Gating Model for the Archeal Voltage-Dependent K+ Channel KvAP in DPhPC and POPE:POPG decane lipid bilayers

    PubMed Central

    Schmidt, Daniel; Cross, Sam R.; MacKinnon, Roderick

    2009-01-01

    Voltage-dependent K+ (Kv) channels form the basis of the excitability of nerves and muscles. KvAP is a well-characterized archeal Kv channel that has been widely used to investigate many aspects of Kv channel biochemistry, biophysics and structure. In this study a minimal kinetic gating model for KvAP function in two different phospholipid decane bilayers is developed. In most aspects KvAP gating is similar to the well-studied eukaryotic Shaker Kv channel: conformational changes occur within four voltage sensors followed by pore opening. Unlike Shaker, KvAP possesses an inactivated state that is accessible from the pre-open state of the channel. Changing the lipid composition of the membrane influences multiple gating transitions in the model, but most dramatically the rate of recovery from inactivation. Inhibition by the voltage sensor toxin VSTx1 is most easily explained if VSTx1 binds only to the depolarized conformation of the voltage sensor. By delaying the voltage sensor’s return to the hyperpolarized conformation VSTx1 favors the inactivated state of KvAP. PMID:19481093

  16. Alternative splicing modulates Kv channel clustering through a molecular ball and chain mechanism

    NASA Astrophysics Data System (ADS)

    Zandany, Nitzan; Marciano, Shir; Magidovich, Elhanan; Frimerman, Teddy; Yehezkel, Rinat; Shem-Ad, Tzilhav; Lewin, Limor; Abdu, Uri; Orr, Irit; Yifrach, Ofer

    2015-03-01

    Ion channel clustering at the post-synaptic density serves a fundamental role in action potential generation and transmission. Here, we show that interaction between the Shaker Kv channel and the PSD-95 scaffold protein underlying channel clustering is modulated by the length of the intrinsically disordered C terminal channel tail. We further show that this tail functions as an entropic clock that times PSD-95 binding. We thus propose a ‘ball and chain’ mechanism to explain Kv channel binding to scaffold proteins, analogous to the mechanism describing channel fast inactivation. The physiological relevance of this mechanism is demonstrated in that alternative splicing of the Shaker channel gene to produce variants of distinct tail lengths resulted in differential channel cell surface expression levels and clustering metrics that correlate with differences in affinity of the variants for PSD-95. We suggest that modulating channel clustering by specific spatial-temporal spliced variant targeting serves a fundamental role in nervous system development and tuning.

  17. Design and Evaluation of 275 kV-3 kA HTS Power Cable

    NASA Astrophysics Data System (ADS)

    Yagi, M.; Mukoyama, S.; Mitsuhashi, T.; Jun, T.; Liu, J.; Nakayama, R.; Hayakawa, N.; Wang, X.; Ishiyama, A.; Amemiya, N.; Hasegawa, T.; Saitoh, T.; Ohkuma, T.; Maruyama, O.

    A 275 kV 3 kA high temperature superconducting (HTS) cable has been developed in the Materials & Power Applications of Coated Conductors (M-PACC) project. The cable is expected to be put to practical use as the backbone power line in the future because the capacity of 1.5 GW is about the same as overhead transmission lines. The 30 m cable has been designed on the basis of design values that had been obtained by various voltage tests, AC loss measurement tests, short circuit tests, and other elementary tests. Cable insulation was determined by the design stresses and test conditions based on IEC, JEC (Japan electrical standards), and other HTS demonstrations. This cable was also designed to withstand the short circuit test of 63 kA for 0.6 seconds and to have low losses, including AC loss and dielectric loss of 0.8 W/m at 3kA, 275 kV. Based on the design, a 30 m cable was manufactured, and short samples during this manufacturing process were confirmed to have the designed characteristics. Furukawa Electric prepared a demonstration of the 30 m cable with two terminations and a cable joint. The long-term test under a current of 3 kA, and test voltage determined from 30 years of insulation degradation has been conducted since November 2012 at Shenyang in China.

  18. Chromatic Aberration Correction for Atomic Resolution TEM Imaging from 20 to 80 kV

    NASA Astrophysics Data System (ADS)

    Linck, Martin; Hartel, Peter; Uhlemann, Stephan; Kahl, Frank; Müller, Heiko; Zach, Joachim; Haider, Max.; Niestadt, Marcel; Bischoff, Maarten; Biskupek, Johannes; Lee, Zhongbo; Lehnert, Tibor; Börrnert, Felix; Rose, Harald; Kaiser, Ute

    2016-08-01

    Atomic resolution in transmission electron microscopy of thin and light-atom materials requires a rigorous reduction of the beam energy to reduce knockon damage. However, at the same time, the chromatic aberration deteriorates the resolution of the TEM image dramatically. Within the framework of the SALVE project, we introduce a newly developed Cc/Cs corrector that is capable of correcting both the chromatic and the spherical aberration in the range of accelerating voltages from 20 to 80 kV. The corrector allows correcting axial aberrations up to fifth order as well as the dominating off-axial aberrations. Over the entire voltage range, optimum phase-contrast imaging conditions for weak signals from light atoms can be adjusted for an optical aperture of at least 55 mrad. The information transfer within this aperture is no longer limited by chromatic aberrations. We demonstrate the performance of the microscope using the examples of 30 kV phase-contrast TEM images of graphene and molybdenum disulfide, showing unprecedented contrast and resolution that matches image calculations.

  19. Evolutionary conservation of Kv3.1 in the barn owl Tyto alba.

    PubMed

    Kullmann, Lars; Schlüter, Tina; Wagner, Hermann; Nothwang, Hans Gerd

    2013-01-01

    For prey capture in the dark, the barn owl Tyto alba has evolved into an auditory specialist with an exquisite capability of sound localization. Adaptations include asymmetrical ears, enlarged auditory processing centers, the utilization of minute interaural time differences, and phase locking along the entire hearing range up to 10 kHz. Adaptations on the molecular level have not yet been investigated. Here, we tested the hypothesis that divergence in the amino acid sequence of the voltage-gated K(+) channel Kv3.1 contributes to the accuracy and high firing rates of auditory neurons in the barn owl. We therefore cloned both splice variants of Kcnc1, the gene encoding Kv3.1. Both splice variants, Kcnc1a and Kcnc1b, encode amino acids identical to those of the chicken, an auditory generalist. Expression analyses confirmed neural-restricted expression of the channel. In summary, our data reveal strong evolutionary conservation of Kcnc1 in the barn owl and point to other genes involved in auditory specializations of this animal. The data also demonstrate the feasibility to address neuroethological questions in organisms with no reference genome by molecular approaches. This will open new avenues for neuroethologists working in these organisms. PMID:23615168

  20. Engineering of beam direct conversion for a 120-kV, 1-MW ion beam

    NASA Technical Reports Server (NTRS)

    Barr, W. L.; Doggett, J. N.; Hamilton, G. W.; Kinney, J. D.; Moir, R. W.

    1977-01-01

    Practical systems for beam direct conversion are required to recover the energy from ion beams at high efficiency and at very high beam power densities in the environment of a high-power neutral-injection system. Such an experiment is now in progress using a 120-kV beam with a maximum total current of 20 A. After neutralization, the H(+) component to be recovered will have a power of approximately 1 MW. A system testing these concepts has been designed and tested at 15 kV, 2 kW in preparation for the full-power tests. The engineering problems involved in the full-power tests affect electron suppression, gas pumping, voltage holding, diagnostics, and measurement conditions. Planning for future experiments at higher power includes the use of cryopumping and electron suppression by a magnetic field rather than by an electrostatic field. Beam direct conversion for large fusion experiments and reactors will save millions of dollars in the cost of power supplies and electricity and will dispose of the charged beam under conditions that may not be possible by other techniques.

  1. Next generation KATRIN high precision voltage divider for voltages up to 65kV

    NASA Astrophysics Data System (ADS)

    Bauer, S.; Berendes, R.; Hochschulz, F.; Ortjohann, H.-W.; Rosendahl, S.; Thümmler, T.; Schmidt, M.; Weinheimer, C.

    2013-10-01

    The KATRIN (KArlsruhe TRItium Neutrino) experiment aims to determine the mass of the electron antineutrino with a sensitivity of 200 meV by precisely measuring the electron spectrum of the tritium beta decay. This will be done by the use of a retarding spectrometer of the MAC-E-Filter type. To achieve the desired sensitivity the stability of the retarding potential of -18.6 kV has to be monitored with a precision of 3 ppm over at least two months. Since this is not feasible with commercial devices, two ppm-class high voltage dividers were developed, following the concept of the standard divider for DC voltages of up to 100 kV of the Physikalisch-Technische Bundesanstalt (PTB). In order to reach such high accuracies different effects have to be considered. The two most important ones are the temperature dependence of resistance and leakage currents, caused by insulators or corona discharges. For the second divider improvements were made concerning the high-precision resistors and the thermal design of the divider. The improved resistors are the result of a cooperation with the manufacturer. The design improvements, the investigation and the selection of the resistors, the built-in ripple probe and the calibrations at PTB will be reported here. The latter demonstrated a stability of about 0.1 ppm/month over a period of two years.

  2. A compact 45 kV curve tracer with picoampere current measurement capability

    NASA Astrophysics Data System (ADS)

    Sullivan, W. W.; Mauch, D.; Bullick, A.; Hettler, C.; Neuber, A.; Dickens, J.

    2013-03-01

    This paper discusses a compact high voltage curve tracer for high voltage semiconductor device characterization. The system sources up to 3 mA at up to 45 kV in dc conditions. It measures from 328 V to 60 kV with 15 V resolution and from 9.4 pA to 4 mA with 100 fA minimum resolution. Control software for the system is written in Microsoft Visual C# and features real-time measurement control and IV plotting, arc-protection and detection, an electrically isolated universal serial bus interface, and easy data exporting capabilities. The system has survived numerous catastrophic high voltage device-under-test arcing failures with no loss of measurement capability or system damage. Overall sweep times are typically under 2 min, and the curve tracer system was used to characterize the blocking performance of high voltage ceramic capacitors, high voltage silicon carbide photoconductive semiconductor switches, and high voltage coaxial cable.

  3. Optimizing 50kV hydrogen diagnostic neutral beam performance for active spectroscopy in MST

    NASA Astrophysics Data System (ADS)

    Feng, X.; Boguski, J.; Craig, D.; den Hartog, D. J.; Munaretto, S.; Nornberg, M. D.; Olivia, S.

    2015-11-01

    The 50 kV hydrogen diagnostic neutral beam on MST provides local measurements of impurity ion emission through charge exchange recombination spectroscopy (CHERS) and of core-localized magnetic field through the motional Stark effect (MSE). The beam, which was designed to provide 5A of neutral current at 50 kV to meet these needs, is currently on a test stand to accommodate diagnosis, in order to increase the reliability of beam formation, sustain a steady current of 5 amps for 20ms, and optimize the primary energy fraction. The reliability of arc formation was increased from 40% to 80% success rate with increase of cathode gas pressure from 150kPa to 200kPa, and the stability of the arc current is improved with a decrease of the insulation magnetic field. A calorimeter with 5 thermocouples is installed to measure the horizontal and vertical beam profiles as well as beam divergence. Beam energy components are quantified through Doppler-shift spectroscopy. Preliminary simulation results of the beam using the ALCBEAM code as well as a description of how changes to the beam performance can affect CHERS and MSE measurements are presented. This work is supported by the U.S. DOE.

  4. A novel optimization design for 3.3 kV injection-enhanced gate transistor

    NASA Astrophysics Data System (ADS)

    Xiaoli, Tian; Weili, Chu; Jiang, Lu; Shuojin, Lu; Qiaoqun, Yu; Yangjun, Zhu

    2014-01-01

    This paper introduces a homemade injection-enhanced gate transistor (IEGT) with blocking voltage up to 3.7 kV. An advanced cell structure with dummy trench and a large cell pitch is adopted in the IEGT. The carrier concentration at the N-emitter side is increased by the larger cell pitch of the IEGT and it enhances the P—i—N effect within the device. The result shows that the IEGT has a remarkablely low on-state forward voltage drop (VCE(sat)) compared to traditional trench IGBT structures. However, too large cell pitch decreases the channel density of the trench IEGT and increases the voltage drop across the channel, finally it will increase the VCE(sat) of the IEGT. Therefore, the cell pitch selection is the key parameter consideration in the design of the IEGT. In this paper, a cell pitch selection method and the optimal value of 3.3 kV IEGT are presented by simulations and experimental results.

  5. Optically isolated, 2 kHz repetition rate, 4 kV solid-state pulse trigger generator

    NASA Astrophysics Data System (ADS)

    Barnett, D. H.; Parson, J. M.; Lynn, C. F.; Kelly, P. M.; Taylor, M.; Calico, S.; Scott, M. C.; Dickens, J. C.; Neuber, A. A.; Mankowski, J. J.

    2015-03-01

    This paper presents the design and operation characteristics of a solid-state high voltage pulse generator. Its primary utilization is aimed at triggering a gaseous spark gap with high repeatability. Specifically, the trigger generator is designed to achieve a risetime on the order of 0.1 kV/ns to trigger the first stage, trigatron spark gap of a 10-stage, 500 kV Marx generator. The major design components are comprised of a 60 W constant current DC-DC converter for high voltage charging, a single 4 kV thyristor, a step-up pulse transformer, and magnetic switch for pulse steepening. A risetime of <30 ns and pulse magnitude of 4 kV is achieved matching the simulated performance of the design.

  6. Optically isolated, 2 kHz repetition rate, 4 kV solid-state pulse trigger generator.

    PubMed

    Barnett, D H; Parson, J M; Lynn, C F; Kelly, P M; Taylor, M; Calico, S; Scott, M C; Dickens, J C; Neuber, A A; Mankowski, J J

    2015-03-01

    This paper presents the design and operation characteristics of a solid-state high voltage pulse generator. Its primary utilization is aimed at triggering a gaseous spark gap with high repeatability. Specifically, the trigger generator is designed to achieve a risetime on the order of 0.1 kV/ns to trigger the first stage, trigatron spark gap of a 10-stage, 500 kV Marx generator. The major design components are comprised of a 60 W constant current DC-DC converter for high voltage charging, a single 4 kV thyristor, a step-up pulse transformer, and magnetic switch for pulse steepening. A risetime of <30 ns and pulse magnitude of 4 kV is achieved matching the simulated performance of the design. PMID:25832253

  7. Development of 66kV XLPE submarine cable using optical fiber as a mechanical-damage-detection-sensor

    SciTech Connect

    Nishimoto, Toshio; Miyahara, Tsutomu; Takehana, Hajime; Tateno, Fuminori

    1995-10-01

    Submarine cables are exposed to great risk of serious mechanical damage by ship anchors or equipment used for fishing. Detection of such damage in a submarine cable is a very useful technology for improving the reliability of a submarine cable transmission line. A mechanical-damage-detection-sensor using optical fiber was developed. A prototype 66kV XLPE submarine cable incorporating the sensor was manufactured for trial, and the ability of a sensor was confirmed by compression test. Actual 66kV XLPE submarine cable incorporating the sensor was manufactured for trial, and the ability of a sensor was confirmed by compression test. Actual 66kV XLPE submarine cable with the sensor was manufactured and installed as an operating transmission line in Japan.

  8. Convergent phosphomodulation of the major neuronal dendritic potassium channel Kv4.2 by pituitary adenylate cyclase-activating polypeptide.

    PubMed

    Gupte, Raeesa P; Kadunganattil, Suraj; Shepherd, Andrew J; Merrill, Ronald; Planer, William; Bruchas, Michael R; Strack, Stefan; Mohapatra, Durga P

    2016-02-01

    The endogenous neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) is secreted by both neuronal and non-neuronal cells in the brain and spinal cord, in response to pathological conditions such as stroke, seizures, chronic inflammatory and neuropathic pain. PACAP has been shown to exert various neuromodulatory and neuroprotective effects. However, direct influence of PACAP on the function of intrinsically excitable ion channels that are critical to both hyperexcitation as well as cell death, remain largely unexplored. The major dendritic K(+) channel Kv4.2 is a critical regulator of neuronal excitability, back-propagating action potentials in the dendrites, and modulation of synaptic inputs. We identified, cloned and characterized the downstream signaling originating from the activation of three PACAP receptor (PAC1) isoforms that are expressed in rodent hippocampal neurons that also exhibit abundant expression of Kv4.2 protein. Activation of PAC1 by PACAP leads to phosphorylation of Kv4.2 and downregulation of channel currents, which can be attenuated by inhibition of either PKA or ERK1/2 activity. Mechanistically, this dynamic downregulation of Kv4.2 function is a consequence of reduction in the density of surface channels, without any influence on the voltage-dependence of channel activation. Interestingly, PKA-induced effects on Kv4.2 were mediated by ERK1/2 phosphorylation of the channel at two critical residues, but not by direct channel phosphorylation by PKA, suggesting a convergent phosphomodulatory signaling cascade. Altogether, our findings suggest a novel GPCR-channel signaling crosstalk between PACAP/PAC1 and Kv4.2 channel in a manner that could lead to neuronal hyperexcitability. PMID:26456351

  9. Granzyme B-Induced Neurotoxicity Is Mediated via Activation of PAR-1 Receptor and Kv1.3 Channel

    PubMed Central

    Wang, Tongguang; Lee, Myoung-Hwa; Choi, Elliot; Pardo-Villamizar, Carlos A.; Lee, Sung Bin; Yang, In Hong; Calabresi, Peter A.; Nath, Avindra

    2012-01-01

    Increasing evidence supports a critical role of T cells in neurodegeneration associated with acute and subacute brain inflammatory disorders. Granzyme B (GrB), released by activated T cells, is a cytotoxic proteinase which may induce perforin-independent neurotoxicity. Here, we studied the mechanism of perforin-independent GrB toxicity by treating primary cultured human neuronal cells with recombinant GrB. GrBactivated the protease-activated receptor (PAR)-1 receptor on the neuronal cell surface leading to decreased intracellular cyclic AMP levels. This was followed by increased expression and translocation of the voltage gated potassium channel, Kv1.3 to the neuronal cell membrane. Similar expression of Kv1.3 was also seen in neurons of the cerebral cortex adjacent to active inflammatory lesions in patients with multiple sclerosis. Kv1.3 expression was followed by activation of Notch-1 resulting in neurotoxicity. Blocking PAR-1, Kv1.3 or Notch-1 activation using specific pharmacological inhibitors or siRNAs prevented GrB-induced neurotoxicity. Furthermore, clofazimine protected against GrB-induced neurotoxicity in rat hippocampus, in vivo. These observations indicate that GrB released from T cells induced neurotoxicity by interacting with the membrane bound Gi-coupled PAR-1 receptor and subsequently activated Kv1.3 and Notch-1. These pathways provide novel targets to treat T cell-mediated neuroinflammatory disorders. Kv1.3 is of particular interest since it is expressed on the cell surface, only under pathological circumstances, and early in the cascade of events making it an attractive therapeutic target. PMID:22952817

  10. Curcumin serves as a human kv1.3 blocker to inhibit effector memory T lymphocyte activities.

    PubMed

    Lian, Yi-Tian; Yang, Xiao-Fang; Wang, Zhao-Hui; Yang, Yong; Yang, Ying; Shu, Yan-Wen; Cheng, Long-Xian; Liu, Kun

    2013-09-01

    Curcumin, the principal active component of turmeric, has long been used to treat various diseases in India and China. Recent studies show that curcumin can serve as a therapeutic agent for autoimmune diseases via a variety of mechanisms. Effector memory T cells (T(EM), CCR7⁻ CD45RO⁺ T lymphocyte) have been demonstrated to play a crucial role in the pathogenesis of T cell-mediated autoimmune diseases, such as multiple sclerosis (MS) or rheumatoid arthritis (RA). Kv1.3 channels are predominantly expressed in T(EM) cells and control T(EM) activities. In the present study, we examined the effect of curcumin on human Kv1.3 (hKv1.3) channels stably expressed in HEK-293 cells and its ability to inhibit proliferation and cytokine secretion of T(EM) cells isolated from patients with MS or RA. Curcumin exhibited a direct blockage of hKv1.3 channels in a time-dependent and concentration-dependent manner. Moreover, the activation curve was shifted to a more positive potential, which was consistent with an open-channel blockade. Paralleling hKv1.3 inhibition, curcumin significantly inhibited proliferation and interferon-γ secretion of T(EM) cells. Our findings demonstrate that curcumin is able to inhibit proliferation and proinflammatory cytokine secretion of T(EM) cells probably through inhibition of hKv1.3 channels, which contributes to the potency of curcumin for the treatment of autoimmune diseases. This is probably one of pharmacological mechanisms of curcumin used to treat autoimmune diseases. PMID:23132777

  11. Long-term stability and mechanical characteristics of kV digital imaging system for proton radiotherapy

    PubMed Central

    Zhu, Mingyao; Botticello, Thomas; Lu, Hsiao-Ming; Winey, Brian

    2014-01-01

    Purpose: To quantitatively evaluate the long-term image panel positioning stability and gantry angle dependence for gantry-mounted kV imaging systems. Methods: For patient setup digital imaging systems in isocentric rotating proton beam delivery facilities, physical crosshairs are commonly inserted into the snout to define the kV x-ray beam isocenter. Utilizing an automatic detection algorithm, the authors analyzed the crosshair center positions in 2744 patient setup kV images acquired with the four imagers in two treatment rooms from January 2012 to January 2013. The crosshair position was used as a surrogate for imaging panel position, and its long-term stability at the four cardinal angles and the panel flex dependency on gantry angle was investigated. Results: The standard deviation of the panel position distributions was within 0.32 mm (with the range of variation less than ± 1.4 mm) in both the X-Z plane and Y direction. The mean panel inplane rotations were not more than 0.51° for the four panels at the cardinal angles, with standard deviations ≤0.26°. The panel position variations with gantry rotation due to gravity (flex) were within ±4 mm, and were panel-specific. Conclusions: The authors demonstrated that the kV image panel positions in our proton treatment system were highly reproducible at the cardinal angles over 13 months and also that the panel positions can be correlated to gantry angles. This result indicates that the kV image panel positions are stable over time; the amount of panel sag is predictable during gantry rotation and the physical crosshair for kV imaging may eventually be removed, with the imaging beam isocenter position routinely verified by adequate quality assurance procedures and measurements. PMID:24694126

  12. Functionally Active T1-T1 Interfaces Revealed by the Accessibility of Intracellular Thiolate Groups in Kv4 Channels

    PubMed Central

    Wang, Guangyu; Shahidullah, Mohammad; Rocha, Carmen A.; Strang, Candace; Pfaffinger, Paul J.; Covarrubias, Manuel

    2005-01-01

    Gating of voltage-dependent K+ channels involves movements of membrane-spanning regions that control the opening of the pore. Much less is known, however, about the contributions of large intracellular channel domains to the conformational changes that underlie gating. Here, we investigated the functional role of intracellular regions in Kv4 channels by probing relevant cysteines with thiol-specific reagents. We find that reagent application to the intracellular side of inside-out patches results in time-dependent irreversible inhibition of Kv4.1 and Kv4.3 currents. In the absence or presence of Kv4-specific auxiliary subunits, mutational and electrophysiological analyses showed that none of the 14 intracellular cysteines is essential for channel gating. C110, C131, and C132 in the intersubunit interface of the tetramerization domain (T1) are targets responsible for the irreversible inhibition by a methanethiosulfonate derivative (MTSET). This result is surprising because structural studies of Kv4-T1 crystals predicted protection of the targeted thiolate groups by constitutive high-affinity Zn2+ coordination. Also, added Zn2+ or a potent Zn2+ chelator (TPEN) does not significantly modulate the accessibility of MTSET to C110, C131, or C132; and furthermore, when the three critical cysteines remained as possible targets, the MTSET modification rate of the activated state is ∼200-fold faster than that of the resting state. Biochemical experiments confirmed the chemical modification of the intact α-subunit and the purified tetrameric T1 domain by MTS reagents. These results conclusively demonstrate that the T1–T1 interface of Kv4 channels is functionally active and dynamic, and that critical reactive thiolate groups in this interface may not be protected by Zn2+ binding. PMID:15955876

  13. Inhalational anaesthetics and n-alcohols share a site of action in the neuronal Shaw2 Kv channel

    PubMed Central

    Bhattacharji, Aditya; Klett, Nathan; Go, Ramon Christopher V; Covarrubias, Manuel

    2010-01-01

    Background and purpose: Neuronal ion channels are key targets of general anaesthetics and alcohol, and binding of these drugs to pre-existing and relatively specific sites is thought to alter channel gating. However, the underlying molecular mechanisms of this action are still poorly understood. Here, we investigated the neuronal Shaw2 voltage-gated K+ (Kv) channel to ask whether the inhalational anaesthetic halothane and n-alcohols share a binding site near the activation gate of the channel. Experimental approach: Focusing on activation gate mutations that affect channel modulation by n-alcohols, we investigated n-alcohol-sensitive and n-alcohol-resistant Kv channels heterologously expressed in Xenopus oocytes to probe the functional modulation by externally applied halothane using two-electrode voltage clamping and a gas-tight perfusion system. Key results: Shaw2 Kv channels are reversibly inhibited by halothane in a dose-dependent and saturable manner (K0.5= 400 µM; nH= 1.2). Also, discrete mutations in the channel's S4S5 linker are sufficient to reduce or confer inhibition by halothane (Shaw2-T330L and Kv3.4-G371I/T378A respectively). Furthermore, a point mutation in the S6 segment of Shaw2 (P410A) converted the halothane-induced inhibition into halothane-induced potentiation. Lastly, the inhibition resulting from the co-application of n-butanol and halothane is consistent with the presence of overlapping binding sites for these drugs and weak binding cooperativity. Conclusions and implications: These observations strongly support a molecular model of a general anaesthetic binding site in the Shaw2 Kv channel. This site may involve the amphiphilic interface between the S4S5 linker and the S6 segment, which plays a pivotal role in Kv channel activation. PMID:20136839

  14. Hyperexcitability and reduced low threshold potassium currents in auditory neurons of mice lacking the channel subunit Kv1.1

    PubMed Central

    Brew, Helen M; Hallows, Janice L; Tempel, Bruce L

    2003-01-01

    A low voltage-activated potassium current, IKL, is found in auditory neuron types that have low excitability and precisely preserve the temporal pattern of activity present in their presynaptic inputs. The gene Kcnal codes for Kv1.1 potassium channel subunits, which combine in expression systems to produce channel tetramers with properties similar to those of IKL, including sensitivity to dendrotoxin (DTX). Kv1.1 is strongly expressed in neurons with IKL, including auditory neurons of the medial nucleus of the trapezoid body (MNTB). We therefore decided to investigate how the absence of Kv1.1 affected channel properties and function in MNTB neurons from mice lacking Kcnal. We used the whole cell version of the patch clamp technique to record from MNTB neurons in brainstem slices from Kcnal-null (−/−) mice and their wild-type (+/+) and heterozygous (+/−) littermates. There was an IKL in voltage-clamped −/− MNTB neurons, but it was about half the amplitude of the IKL in +/+ neurons, with otherwise similar properties. Consistent with this, −/− MNTB neurons were more excitable than their +/+ counterparts; they fired more than twice as many action potentials (APs) during current steps, and the threshold current amplitude required to generate an AP was roughly halved. +/− MNTB neurons had excitability and IKL amplitudes identical to the +/+ neurons. The IKL remaining in −/− neurons was blocked by DTX, suggesting the underlying channels contained subunits Kv1.2 and/or Kv1.6 (also DTX-sensitive). DTX increased excitability further in the already hyperexcitable −/− MNTB neurons, suggesting that −/−IKL limited excitability despite its reduced amplitude in the absence of Kv1.1 subunits. PMID:12611922

  15. Long-term stability and mechanical characteristics of kV digital imaging system for proton radiotherapy

    SciTech Connect

    Zhu, Mingyao Botticello, Thomas; Lu, Hsiao-Ming; Winey, Brian

    2014-04-15

    Purpose: To quantitatively evaluate the long-term image panel positioning stability and gantry angle dependence for gantry-mounted kV imaging systems. Methods: For patient setup digital imaging systems in isocentric rotating proton beam delivery facilities, physical crosshairs are commonly inserted into the snout to define the kV x-ray beam isocenter. Utilizing an automatic detection algorithm, the authors analyzed the crosshair center positions in 2744 patient setup kV images acquired with the four imagers in two treatment rooms from January 2012 to January 2013. The crosshair position was used as a surrogate for imaging panel position, and its long-term stability at the four cardinal angles and the panel flex dependency on gantry angle was investigated. Results: The standard deviation of the panel position distributions was within 0.32 mm (with the range of variation less than ± 1.4 mm) in both the X-Z plane and Y direction. The mean panel inplane rotations were not more than 0.51° for the four panels at the cardinal angles, with standard deviations ≤0.26°. The panel position variations with gantry rotation due to gravity (flex) were within ±4 mm, and were panel-specific. Conclusions: The authors demonstrated that the kV image panel positions in our proton treatment system were highly reproducible at the cardinal angles over 13 months and also that the panel positions can be correlated to gantry angles. This result indicates that the kV image panel positions are stable over time; the amount of panel sag is predictable during gantry rotation and the physical crosshair for kV imaging may eventually be removed, with the imaging beam isocenter position routinely verified by adequate quality assurance procedures and measurements.

  16. Innovation on Energy Power Technology (6)Development of 550kV Gas Insulated Switchgear and its Practical Application

    NASA Astrophysics Data System (ADS)

    Takatuka, Keizo; Tsuchie, Ei

    Since the first prototype GIS was constructed in Japan in 1968, rapid progress and wide extension were made during a few years. In 1973, the first 550kV hybrid GIS was constructed and after three years, in 1976, the world first 550kV full GIS was completed in Ohi nuclear power plant. During this remarkable short period of GIS development, effort of utilities and Mitsubishi Electric Corporation, as a manufacturer, are mentioned. Especially, some of technical subjects to the large sized equipments were picked up and that solutions were reported to be correct judging from up-to-date knowledges.

  17. Interaction of DPP10a with Kv4.3 channel complex results in a sustained current component of human transient outward current Ito.

    PubMed

    Turnow, K; Metzner, K; Cotella, D; Morales, M J; Schaefer, M; Christ, T; Ravens, U; Wettwer, E; Kämmerer, S

    2015-03-01

    The sustained component of the K(+) outward current in human atrial myocytes is believed to be due to the slowly inactivating ultra-rapid potassium current I Kur and not to the fast inactivating transient outward current Ito. Here we provide evidence for contribution of Ito to this late current due to the effects of dipeptidyl peptidase-like protein (DPP) 10 (DPP10a) interacting with Kv4.3 channels. We studied the late current component of Ito in human atrial myocytes and CHO cells co-expressing Kv4.3 or Kv4.3/KChIP2 (control) and DPP proteins using voltage-clamp technique and a pharmacological approach. A voltage dependent and slowly inactivating late current (43% of peak amplitude) could be observed in atrial myocytes. We found a similar current in CHO cells expressing Kv4.3/KChIP2 + DPP10a, but not in cells co-expressing Kv4.3 + DPP or Kv4.3/KChIP2 + DPP6-S. Assuming that DPP10a influences atrial Ito, we detected DPP10 expression of three alternatively spliced mRNAs, DPP10 protein and colocalization of Kv4.3 and DPP10 proteins in human atrial myocytes. DPP10a did not affect properties of expressed Kv1.5 excluding a contribution to the sustained IKur in atrial cells. To test for the contribution of Kv4-based Ito on sustained K(+) outward currents in human atrial myocytes, we used 4-AP to block IKur, in combination with Heteropoda toxin 2 to block Kv4 channels. We could clearly separate an Ito fraction of about 19% contributing to the late current in atrial myocytes. Thus, the interaction of DPP10a, expressed in human atrium, with Kv4.3 channels generates a sustained current component of Ito, which may affect late repolarization phase of atrial action potentials. PMID:25600224

  18. Potent Suppression of Kv1.3 Potassium Channel and IL-2 Secretion by Diphenyl Phosphine Oxide-1 in Human T Cells

    PubMed Central

    Zhao, Ning; Dong, Qian; Du, Li-Li; Fu, Xiao-Xing; Du, Yi-Mei; Liao, Yu-Hua

    2013-01-01

    Diphenyl phosphine oxide-1 (DPO-1) is a potent Kv1.5 channel inhibitor that has therapeutic potential for the treatment of atrial fibrillation. Many other Kv1.5 channel blockers also potently inhibit the Kv1.3 channel, but whether DPO-1 blocks Kv1.3 channels has not been investigated. The Kv1.3 channel is highly expressed in activated T cells, which is considered a favorable target for immunomodulation. Accordingly, we hypothesized that DPO-1 may exert immunosuppressive and anti-inflammatory effects by inhibiting Kv1.3 channel activity. In this study, DPO-1 blocked Kv1.3 current in a voltage-dependent and concentration-dependent manner, with IC50 values of 2.58 µM in Jurkat cells and 3.11 µM in human peripheral blood T cells. DPO-1 also accelerated the inactivation rate and negatively shifted steady-state inactivation. Moreover, DPO-1 at 3 µM had no apparent effect on the Ca2+ activated potassium channel (KCa) current in both Jurkat cells and human peripheral blood T cells. In Jurkat cells, pre-treatment with DPO-1 for 24 h decreased Kv1.3 current density, and protein expression by 48±6% and 60±9%, at 3 and 10 µM, respectively (both p<0.05). In addition, Ca2+ influx to Ca2+-depleted cells was blunted and IL-2 production was also reduced in activated Jurkat cells. IL-2 secretion was also inhibited by the Kv1.3 inhibitors margatoxin and charybdotoxin. Our results demonstrate for the first time that that DPO-1, at clinically relevant concentrations, blocks Kv1.3 channels, decreases Kv1.3 channel expression and suppresses IL-2 secretion. Therefore, DPO-1 may be a useful treatment strategy for immunologic disorders. PMID:23717641

  19. High power cable with internal water cooling 400 kV

    NASA Astrophysics Data System (ADS)

    Rasquin, W.; Harjes, B.

    1982-08-01

    Due to the concentration of electricity production in large power plants, the need of higher power transmissions, and the protection of environment, developement of a 400 kV water cooled cable in the power range of 1 to 5 GVA was undertaken. The fabrication and testing of equipment, engineering of cable components, fabrication of a test cable, development of cable terminal laboratory, testing of test cable, field testing of test cable, fabrication of industrial cable laboratory, testing of industrial cable, field testing of industrial cable, and system analysis for optimization were prepared. The field testing was impossible to realize. However, it is proved that a cable consisting of an internal stainless steel water cooled tube, covered by stranded copper profiles, insulated with heavy high quality paper, and protected by an aluminum cover can be produced, withstand tests accordingly to IEC/VDE recommendations, and is able to fulfill all exploitation conditions.

  20. A 6.4 kV pulse generator with transformers

    NASA Astrophysics Data System (ADS)

    Bastein, W. L.

    The possibility of producing a 6.4 kV voltage pulse using transformers was investigated. The pulse has to have a rise and fall time of 100 ns, a pulse duration of 2 to 50 microns, a repetition frequency of 2500 Hz, and a ripple of less than 10 V. It is shown that it is in principle possible, using transformers for the low frequency and high frequency parts of the pulse, to produce a pulse which fulfills the slope requirements. However, it is difficult to make and tune the circuits to flatten the pulse sufficiently. The losses are so high that it is recommended to investigate the possibility of producing the pulse with a number of MOSFET's in series.

  1. Lightning overvoltage protection of the paddock 362-145 kV Gas-Insulated Substation

    SciTech Connect

    Elahi, H.; Sublich, M. ); Anderson, M.E.; Nelson, B.D. )

    1990-01-01

    Backflashovers close to the Paddiock 362-145 kV Gas-Insulated Substation (GIS) have been analyzed with the Electro-Magnetic Transient Program (EMTP) using a frequency dependent multi-conductor system. The severity of the lightning stroke currents were derived based on recent recordings in the eastern United States. Impacts of corona attenuation and distortion were accounted for using a shunt linear model approach. Turn-up effects of both line insulator flashover voltages and surge arrester protective characteristics were represented based on manufacturer's volt-time curves. Wave shaping effects of substation capacitances (ie., PT's transformers, CCPD's) were also modeled. Results show the importance of various modeling details in determining the overvoltages inside the GIS due to close backflashovers, which are caused by lightning strokes with varying intensity. These results are aimed at better evaluation of lightning protection requirements for GIS protected by metal-oxide surge arresters.

  2. Comparative evaluation by laboratory aging of 15 and 35 kV extruded dielectric cables

    SciTech Connect

    Katz, C.; Dyndul, J. ); Walker, M. )

    1990-04-01

    Utility engineers encounter a significant problem in discerning the accuracy of claims made of superior cable quality and in identifying cables which will provide reliable performance over their anticipated life. The authors in two independent investigations of 15 and 35 kV cables have compared the performance of a number of cables made with different compounds, by different manufacturers. They show that judging cables by their unaged voltage breakdown characteristics alone can be very misleading; that in the long run, dry cured cables, aged in moist environments, rapidly lose their original dielectric strength advantage; that certain ethylene propylene rubber formulations degrade as fast as crosslinked polyethylene; that the best overall results during aging are obtained with tree resistant polyethylene insulated cables and that from a voltage breakdown point of view water tree length is more influential than number of water trees.

  3. Using a 400 kV Van de Graaff accelerator to teach physics at West Point

    NASA Astrophysics Data System (ADS)

    Marble, D. K.; Bruch, S. E.; Lainis, T.

    1997-02-01

    A small accelerator visitation laboratory is being built at the United States Military Academy using two 400 kV Van de Graaff accelerators. This laboratory will provide quality teaching experiments and increased research opportunities for both faculty and cadets as well as enhancing the department's ability to teach across the curriculum by using nuclear techniques to solve problems in environmental engineering, material science, archeology, art, etc. This training enhances a students ability to enter non-traditional fields that are becoming a large part of the physics job market. Furthermore, a small accelerator visitation laboratory for high school students can stimulate student interest in science and provide an effective means of communicating the scientific method to a general audience. A discussion of the USMA facility, class experiments and student research projects will be presented.

  4. Capabilities and some applications of the LLNL 100-kV electric gun

    SciTech Connect

    Osher, J.E.; Gathers, G.R.; Chau, H.H.; Weingart, R.C.

    1989-06-05

    The LLNL 100-kV electric gun is an experimental device for launching thin flyer plates at velocities as high as 20 km/s to study impact damage and shock-wave physics. The hypervelocity impact studies reported here include a damage study of spalling damage in aluminum, a study of one-dimensional shock-wave attenuation in materials, use of the well-characterized one-dimensional shock waves for equation-of-state measurements in various materials, and use of thin flyer plates at velocities up to 18 km/s to produce reverse ballistic impact on target objects such as rods to measure damage and fragmentation. Our studies include both experimental results and correlation with numerical calculations using a code such as DYNA2D. 2 refs., 9 figs.

  5. Simulation of ion beam transport through the 400 Kv ion implanter at Michigan Ion Beam Laboratory

    SciTech Connect

    Naab, F. U.; Toader, O. F.; Was, G. S.

    2013-04-19

    The Michigan Ion Beam Laboratory houses a 400 kV ion implanter. An application that simulates the ion beam trajectories through the implanter from the ion source to the target was developed using the SIMION Registered-Sign code. The goals were to have a tool to develop an intuitive understanding of abstract physics phenomena and diagnose ion trajectories. Using this application, new implanter users of different fields in science quickly understand how the machine works and quickly learn to operate it. In this article we describe the implanter simulation application and compare the parameters of the implanter components obtained from the simulations with the measured ones. The overall agreement between the simulated and measured values of magnetic fields and electric potentials is {approx}10%.

  6. Weld-Windsor 115-kV Transmission Line Project, Weld County, Colorado

    SciTech Connect

    1996-05-01

    The Western Area Power Administration is proposing to rebuild a 3.0 mile segment of the existing Flatiron-Weld 115-kV transmission line in Weld County. The line would be reconductored with new conductor on new wood pole double circuit structures. The new structures would support a double circuit transmission line configuration. The first circuit would be owned by Western and the second by Public Service Company of Colorado (PSCO). Alternatives considered included no action, constructing PSCO`s circuit on new right-of-way, and reconductoring Western`s existing line on the same structures. The proposed action was selected because it provided an opportunity to share structures with PSCO and, overall, would minimize costs and environmental impacts. The environmental assessment identifies minor effects on existing natural or human resources and minor benefits for agricultural operations.

  7. Study on 12kV outdoor vacuum switch with replaceable HRC element drop out fuse

    SciTech Connect

    Wang Jiimei

    1996-12-31

    A new type of vacuum interrupter for 12kV outdoor vacuum switch was experimentally studied, the envelope of which was made of porcelain with petticoat flange for outdoor insulation. In order to produce an axial magnetic field and improve the capacity of transfer current in the vacuum interrupter, an iron plate of horse-shoe construction ingeniously designed was chosen as an electrode. The drop-out fuse with replaceable sand-filled HRC element in series with the vacuum switch is a new conception of design to increase breaking capacity. However, it is a vacuum switch of newly designed to form {open_quotes}a vacuum switch and drop-out type fuse combination{close_quotes}.

  8. AC and lightning performance of fiberglass crossarms aged in 115 kV transmission line

    SciTech Connect

    Grzybowski, S. . Dept. of Electrical and Computer Engineering); Jenkins, E.B. . Generation and Transmission Group)

    1993-10-01

    This paper presents the results of an investigation of the electrical performance of 115 kV transmission line fiberglass cross-arm used by Mississippi Power and Light Company. A transmission line fiberglass crossarm removed from service and companion cross-arms outdoors but not in service were examined. The evaluation of electrical performance was based on flashover voltage value at AC voltage and standard lightning impulses as well as under dry and wet conditions. The tests were performed in the Mississippi State University High Voltage Laboratory. The obtained flashover voltages show no large differences in electrical strength of fiberglass crossarms removed from service and those stored outdoors. The Added CFO voltage by fiberglass crossarm to the porcelain suspension insulators is presented versus the length of the fiberglass crossarm for dry and wet conditions.

  9. A non-canonical di-acidic signal at the C-terminus of Kv1.3 determines anterograde trafficking and surface expression.

    PubMed

    Martínez-Mármol, Ramón; Pérez-Verdaguer, Mireia; Roig, Sara R; Vallejo-Gracia, Albert; Gotsi, Pelagia; Serrano-Albarrás, Antonio; Bahamonde, María Isabel; Ferrer-Montiel, Antonio; Fernández-Ballester, Gregorio; Comes, Núria; Felipe, Antonio

    2013-12-15

    Impairment of Kv1.3 expression at the cell membrane in leukocytes and sensory neuron contributes to the pathophysiology of autoimmune diseases and sensory syndromes. Molecular mechanisms underlying Kv1.3 channel trafficking to the plasma membrane remain elusive. We report a novel non-canonical di-acidic signal (E483/484) at the C-terminus of Kv1.3 essential for anterograde transport and surface expression. Notably, homologous motifs are conserved in neuronal Kv1 and Shaker channels. Biochemical analysis revealed interactions with the Sec24 subunit of the coat protein complex II. Disruption of this complex retains the channel at the endoplasmic reticulum. A molecular model of the Kv1.3-Sec24a complex suggests salt-bridges between the di-acidic E483/484 motif in Kv1.3 and the di-basic R750/752 sequence in Sec24. These findings identify a previously unrecognized motif of Kv channels essential for their expression on the cell surface. Our results contribute to our understanding of how Kv1 channels target to the cell membrane, and provide new therapeutic strategies for the treatment of pathological conditions. PMID:24144698

  10. Closed-state inactivation involving an internal gate in Kv4.1 channels modulates pore blockade by intracellular quaternary ammonium ions.

    PubMed

    Fineberg, Jeffrey D; Szanto, Tibor G; Panyi, Gyorgy; Covarrubias, Manuel

    2016-01-01

    Voltage-gated K(+) (Kv) channel activation depends on interactions between voltage sensors and an intracellular activation gate that controls access to a central pore cavity. Here, we hypothesize that this gate is additionally responsible for closed-state inactivation (CSI) in Kv4.x channels. These Kv channels undergo CSI by a mechanism that is still poorly understood. To test the hypothesis, we deduced the state of the Kv4.1 channel intracellular gate by exploiting the trap-door paradigm of pore blockade by internally applied quaternary ammonium (QA) ions exhibiting slow blocking kinetics and high-affinity for a blocking site. We found that inactivation gating seemingly traps benzyl-tributylammonium (bTBuA) when it enters the central pore cavity in the open state. However, bTBuA fails to block inactivated Kv4.1 channels, suggesting gated access involving an internal gate. In contrast, bTBuA blockade of a Shaker Kv channel that undergoes open-state P/C-type inactivation exhibits fast onset and recovery inconsistent with bTBuA trapping. Furthermore, the inactivated Shaker Kv channel is readily blocked by bTBuA. We conclude that Kv4.1 closed-state inactivation modulates pore blockade by QA ions in a manner that depends on the state of the internal activation gate. PMID:27502553

  11. Dendritic Kv3.3 potassium channels in cerebellar purkinje cells regulate generation and spatial dynamics of dendritic Ca2+ spikes.

    PubMed

    Zagha, Edward; Manita, Satoshi; Ross, William N; Rudy, Bernardo

    2010-06-01

    Purkinje cell dendrites are excitable structures with intrinsic and synaptic conductances contributing to the generation and propagation of electrical activity. Voltage-gated potassium channel subunit Kv3.3 is expressed in the distal dendrites of Purkinje cells. However, the functional relevance of this dendritic distribution is not understood. Moreover, mutations in Kv3.3 cause movement disorders in mice and cerebellar atrophy and ataxia in humans, emphasizing the importance of understanding the role of these channels. In this study, we explore functional implications of this dendritic channel expression and compare Purkinje cell dendritic excitability in wild-type and Kv3.3 knockout mice. We demonstrate enhanced excitability of Purkinje cell dendrites in Kv3.3 knockout mice, despite normal resting membrane properties. Combined data from local application pharmacology, voltage clamp analysis of ionic currents, and assessment of dendritic Ca(2+) spike threshold in Purkinje cells suggest a role for Kv3.3 channels in opposing Ca(2+) spike initiation. To study the physiological relevance of altered dendritic excitability, we measured [Ca(2+)](i) changes throughout the dendritic tree in response to climbing fiber activation. Ca(2+) signals were specifically enhanced in distal dendrites of Kv3.3 knockout Purkinje cells, suggesting a role for dendritic Kv3.3 channels in regulating propagation of electrical activity and Ca(2+) influx in distal dendrites. These findings characterize unique roles of Kv3.3 channels in dendrites, with implications for synaptic integration, plasticity, and human disease. PMID:20357073

  12. A Potent and Selective Peptide Blocker of the Kv1.3 Channel: Prediction from Free-Energy Simulations and Experimental Confirmation

    PubMed Central

    Rashid, M. Harunur; Heinzelmann, Germano; Huq, Redwan; Tajhya, Rajeev B.; Chang, Shih Chieh; Chhabra, Sandeep; Pennington, Michael W.; Beeton, Christine; Norton, Raymond S.; Kuyucak, Serdar

    2013-01-01

    The voltage-gated potassium channel Kv1.3 is a well-established target for treatment of autoimmune diseases. ShK peptide from a sea anemone is one of the most potent blockers of Kv1.3 but its application as a therapeutic agent for autoimmune diseases is limited by its lack of selectivity against other Kv channels, in particular Kv1.1. Accurate models of Kv1.x-ShK complexes suggest that specific charge mutations on ShK could considerably enhance its specificity for Kv1.3. Here we evaluate the K18A mutation on ShK, and calculate the change in binding free energy associated with this mutation using the path-independent free energy perturbation and thermodynamic integration methods, with a novel implementation that avoids convergence problems. To check the accuracy of the results, the binding free energy differences were also determined from path-dependent potential of mean force calculations. The two methods yield consistent results for the K18A mutation in ShK and predict a 2 kcal/mol gain in Kv1.3/Kv1.1 selectivity free energy relative to wild-type peptide. Functional assays confirm the predicted selectivity gain for ShK[K18A] and suggest that it will be a valuable lead in the development of therapeutics for autoimmune diseases. PMID:24244345

  13. A potent and Kv1.3-selective analogue of the scorpion toxin HsTX1 as a potential therapeutic for autoimmune diseases

    NASA Astrophysics Data System (ADS)

    Rashid, M. Harunur; Huq, Redwan; Tanner, Mark R.; Chhabra, Sandeep; Khoo, Keith K.; Estrada, Rosendo; Dhawan, Vikas; Chauhan, Satendra; Pennington, Michael W.; Beeton, Christine; Kuyucak, Serdar; Norton, Raymond S.

    2014-03-01

    HsTX1 toxin, from the scorpion Heterometrus spinnifer, is a 34-residue, C-terminally amidated peptide cross-linked by four disulfide bridges. Here we describe new HsTX1 analogues with an Ala, Phe, Val or Abu substitution at position 14. Complexes of HsTX1 with the voltage-gated potassium channels Kv1.3 and Kv1.1 were created using docking and molecular dynamics simulations, then umbrella sampling simulations were performed to construct the potential of mean force (PMF) of the ligand and calculate the corresponding binding free energy for the most stable configuration. The PMF method predicted that the R14A mutation in HsTX1 would yield a > 2 kcal/mol gain for the Kv1.3/Kv1.1 selectivity free energy relative to the wild-type peptide. Functional assays confirmed the predicted selectivity gain for HsTX1[R14A] and HsTX1[R14Abu], with an affinity for Kv1.3 in the low picomolar range and a selectivity of more than 2,000-fold for Kv1.3 over Kv1.1. This remarkable potency and selectivity for Kv1.3, which is significantly up-regulated in activated effector memory cells in humans, suggest that these analogues represent valuable leads in the development of therapeutics for autoimmune diseases.

  14. Closed-state inactivation involving an internal gate in Kv4.1 channels modulates pore blockade by intracellular quaternary ammonium ions

    PubMed Central

    Fineberg, Jeffrey D.; Szanto, Tibor G.; Panyi, Gyorgy; Covarrubias, Manuel

    2016-01-01

    Voltage-gated K+ (Kv) channel activation depends on interactions between voltage sensors and an intracellular activation gate that controls access to a central pore cavity. Here, we hypothesize that this gate is additionally responsible for closed-state inactivation (CSI) in Kv4.x channels. These Kv channels undergo CSI by a mechanism that is still poorly understood. To test the hypothesis, we deduced the state of the Kv4.1 channel intracellular gate by exploiting the trap-door paradigm of pore blockade by internally applied quaternary ammonium (QA) ions exhibiting slow blocking kinetics and high-affinity for a blocking site. We found that inactivation gating seemingly traps benzyl-tributylammonium (bTBuA) when it enters the central pore cavity in the open state. However, bTBuA fails to block inactivated Kv4.1 channels, suggesting gated access involving an internal gate. In contrast, bTBuA blockade of a Shaker Kv channel that undergoes open-state P/C-type inactivation exhibits fast onset and recovery inconsistent with bTBuA trapping. Furthermore, the inactivated Shaker Kv channel is readily blocked by bTBuA. We conclude that Kv4.1 closed-state inactivation modulates pore blockade by QA ions in a manner that depends on the state of the internal activation gate. PMID:27502553

  15. A potent and Kv1.3-selective analogue of the scorpion toxin HsTX1 as a potential therapeutic for autoimmune diseases

    PubMed Central

    Rashid, M. Harunur; Huq, Redwan; Tanner, Mark R.; Chhabra, Sandeep; Khoo, Keith K.; Estrada, Rosendo; Dhawan, Vikas; Chauhan, Satendra; Pennington, Michael W.; Beeton, Christine; Kuyucak, Serdar; Norton, Raymond S.

    2014-01-01

    HsTX1 toxin, from the scorpion Heterometrus spinnifer, is a 34-residue, C-terminally amidated peptide cross-linked by four disulfide bridges. Here we describe new HsTX1 analogues with an Ala, Phe, Val or Abu substitution at position 14. Complexes of HsTX1 with the voltage-gated potassium channels Kv1.3 and Kv1.1 were created using docking and molecular dynamics simulations, then umbrella sampling simulations were performed to construct the potential of mean force (PMF) of the ligand and calculate the corresponding binding free energy for the most stable configuration. The PMF method predicted that the R14A mutation in HsTX1 would yield a > 2 kcal/mol gain for the Kv1.3/Kv1.1 selectivity free energy relative to the wild-type peptide. Functional assays confirmed the predicted selectivity gain for HsTX1[R14A] and HsTX1[R14Abu], with an affinity for Kv1.3 in the low picomolar range and a selectivity of more than 2,000-fold for Kv1.3 over Kv1.1. This remarkable potency and selectivity for Kv1.3, which is significantly up-regulated in activated effector memory cells in humans, suggest that these analogues represent valuable leads in the development of therapeutics for autoimmune diseases. PMID:24676092

  16. Characterization of scattered radiation in kV CBCT images using Monte Carlo simulations

    SciTech Connect

    Jarry, Genevieve; Graham, Sean A.; Moseley, Douglas J.; Jaffray, David J.; Siewerdsen, Jeffrey H.; Verhaegen, Frank

    2006-11-15

    Kilovoltage (kV) cone beam computed tomography (CBCT) images suffer from a substantial scatter contribution. In this study, Monte Carlo (MC) simulations are used to evaluate the scattered radiation present in projection images. These predicted scatter distributions are also used as a scatter correction technique. Images were acquired using a kV CBCT bench top system. The EGSnrc MC code was used to model the flat panel imager, the phantoms, and the x-ray source. The x-ray source model was validated using first and second half-value layers (HVL) and profile measurements. The HVLs and the profile were found to agree within 3% and 6%, respectively. MC simulated and measured projection images for a cylindrical water phantom and for an anthropomorphic head phantom agreed within 8% and 10%. A modified version of the DOSXYZnrc MC code was used to score phase space files with identified scattered and primary particles behind the phantoms. The cone angle, the source-to-detector distance, the phantom geometry, and the energy were varied to determine their effect on the scattered radiation distribution. A scatter correction technique was developed in which the MC predicted scatter distribution is subtracted from the projections prior to reconstruction. Preliminary testing of the procedure was done with an anthropomorphic head phantom and a contrast phantom. Contrast and profile measurements were obtained for the scatter corrected and noncorrected images. An improvement of 3% for contrast between solid water and a liver insert and 11% between solid water and a Teflon insert were obtained and a significant reduction in cupping and streaking artifacts was observed.

  17. SU-E-J-15: Automatically Detect Patient Treatment Position and Orientation in KV Portal Images

    SciTech Connect

    Qiu, J; Yang, D

    2015-06-15

    Purpose: In the course of radiation therapy, the complex information processing workflow will Result in potential errors, such as incorrect or inaccurate patient setups. With automatic image check and patient identification, such errors could be effectively reduced. For this purpose, we developed a simple and rapid image processing method, to automatically detect the patient position and orientation in 2D portal images, so to allow automatic check of positions and orientations for patient daily RT treatments. Methods: Based on the principle of portal image formation, a set of whole body DRR images were reconstructed from multiple whole body CT volume datasets, and fused together to be used as the matching template. To identify the patient setup position and orientation shown in a 2D portal image, the 2D portal image was preprocessed (contrast enhancement, down-sampling and couch table detection), then matched to the template image so to identify the laterality (left or right), position, orientation and treatment site. Results: Five day’s clinical qualified portal images were gathered randomly, then were processed by the automatic detection and matching method without any additional information. The detection results were visually checked by physicists. 182 images were correct detection in a total of 200kV portal images. The correct rate was 91%. Conclusion: The proposed method can detect patient setup and orientation quickly and automatically. It only requires the image intensity information in KV portal images. This method can be useful in the framework of Electronic Chart Check (ECCK) to reduce the potential errors in workflow of radiation therapy and so to improve patient safety. In addition, the auto-detection results, as the patient treatment site position and patient orientation, could be useful to guide the sequential image processing procedures, e.g. verification of patient daily setup accuracy. This work was partially supported by research grant from

  18. Final environmental assessment: Los Reales 115 kV transmission line alternative routes

    SciTech Connect

    Not Available

    1989-10-01

    The Central Arizona Project (CAP) was authorized as a part of the Colorado River Basin Project Act (Public Law 90-537) on September 30, 1968. The primary purpose of the CAP is to furnish water for agricultural, municipal, and industrial uses in central and southern Arizona, and western New Mexico. Due to its magnitude, the CAP is divided into several major features serving separate but interrelated functions. The Tucson Aqueduct Phase B pumping plants were designed and constructed to operate on a looped power system. The entire looped power system, including two switching stations and connecting 115-kv transmission lines, was identified in the FEIS and approved for construction in the Secretary of Interior's Record of Decision dated September 24, 1985. The loop begins in the vicinity of the Twin Peaks Pumping Plant -- the northernmost Phase B pumping station, at the Rattlesnake Switching Station. All of the looped power system has been constructed with the exception of the switching station and portion of transmission line proposed to be constructed in this project. Without construction of this final portion of the looped power system, the Phase B pumping plants will not be able to operate normally without negatively affecting nearby power sources. The CAP will also not be able to provide the reliability necessary for municipal water systems dependent upon CAP water. The purpose of this EA is to describe impacts that would result from relocating the Los Reales 115-kV transmission line, and possibly the switching station, originally identified in the FEIS. It should be mentioned the Department of Energy will complete a separate NEPA review.

  19. Novel phenotype associated with a mutation in the KCNA1(Kv1.1) gene

    PubMed Central

    D'Adamo, Maria C.; Gallenmüller, Constanze; Servettini, Ilenio; Hartl, Elisabeth; Tucker, Stephen J.; Arning, Larissa; Biskup, Saskia; Grottesi, Alessandro; Guglielmi, Luca; Imbrici, Paola; Bernasconi, Pia; Di Giovanni, Giuseppe; Franciolini, Fabio; Catacuzzeno, Luigi; Pessia, Mauro; Klopstock, Thomas

    2015-01-01

    Episodic ataxia type 1 (EA1) is an autosomal dominant K+ channelopathy which manifests with short attacks of cerebellar ataxia and dysarthria, and may also show interictal myokymia. Episodes can be triggered by emotional or physical stress, startle response, sudden postural change or fever. Here we describe a 31-year-old man displaying markedly atypical symptoms, including long-lasting attacks of jerking muscle contractions associated with hyperthermia, severe migraine, and a relatively short-sleep phenotype. A single nucleotide change in KCNA1 (c.555C>G) was identified that changes a highly conserved residue (p.C185W) in the first transmembrane segment of the voltage-gated K+ channel Kv1.1. The patient is heterozygous and the mutation was inherited from his asymptomatic mother. Next generation sequencing revealed no variations in the CACNA1A, CACNB4, KCNC3, KCNJ10, PRRT2 or SCN8A genes of either the patient or mother, except for a benign variant in SLC1A3. Functional analysis of the p.C185W mutation in KCNA1 demonstrated a deleterious dominant-negative phenotype where the remaining current displayed slower activation kinetics, subtle changes in voltage-dependence and faster recovery from slow inactivation. Structural modeling also predicts the C185W mutation to be functionally deleterious. This description of novel clinical features, associated with a Kv1.1 mutation highlights a possibly unrecognized relationship between K+ channel dysfunction, hyperthermia and migraine in EA1, and suggests that thorough assessments for these symptoms should be carefully considered for all patients affected by EA1. PMID:25642194

  20. Jingzhaotoxin-35, a novel gating-modifier toxin targeting both Nav1.5 and Kv2.1 channels.

    PubMed

    Wei, Peng; Xu, Changxi; Wu, Qiaoqi; Huang, Lang; Liang, Songping; Yuan, Chunhua

    2014-12-15

    Jingzhaotoxin-35 (JZTX-35), a 36-residue polypeptide, was purified from the venom of the Chinese tarantula Chilobrachys jingzhao. JZTX-35 inhibited Nav1.5 and Kv2.1 currents with the IC50 value of 1.07 μM and 3.62 μM, respectively, but showed no significant effect on either Na(+) currents or Ca(2+) currents evoked in hippocampal neurons. It shifted the activation of the Nav1.5 and Kv2.1 channels to more depolarized voltages, and markedly shifted the steady-state inactivation of Nav1.5 currents toward more hyperpolarized potentials. Moreover, JZTX-35 can bind to a close state of Nav1.5 and Kv2.1 channels. These results indicate that JZTX-35 is a new gating modifier toxin. JZTX-35 shares high sequence similarity with Jingzhaotoxins (JZTXs) targeting Nav1.5 or Kv2.1 channels, but they showed different ion channel selectivity. Structure-function analysis in this study would provide important clues for the exploration of ion channel selectivity of JZTXs. PMID:25449098

  1. 75 FR 32357 - Gallatin National Forest; Montana; Jack Rabbit to Big Sky Meadow Village 161 kV Transmission Line...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-08

    ... Forest Service Gallatin National Forest; Montana; Jack Rabbit to Big Sky Meadow Village 161 kV... Four Corners, west of Bozeman, Montana, to a new substation near Big Sky Meadow Village in Big Sky... facility between the Jack Rabbit substation and the Meadow Village substation, along with building a...

  2. 76 FR 56791 - Notice of Availability of Record of Decision for the Tropic To Hatch (Garkane) 138 kV...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-09-14

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF THE INTERIOR Bureau of Land Management Notice of Availability of Record of Decision for the Tropic To Hatch (Garkane... (ROD) for the Tropic to Hatch (Garkane) 138 kilovolt (kV) Transmission Line Environmental...

  3. Size-based protocol optimization using automatic tube current modulation and automatic kV selection in computed tomography.

    PubMed

    MacDougall, Robert D; Kleinman, Patricia L; Callahan, Michael J

    2016-01-01

    Size-based diagnostic reference ranges (DRRs) for contrast-enhanced pediatric abdominal computed tomography (CT) have been published in order to establish practical upper and lower limits of CTDI, DLP, and SSDE. Based on these DRRs, guidelines for establishing size-based SSDE target levels from the SSDE of a standard adult by applying a linear correction factor have been published and provide a great reference for dose optimization initiatives. The necessary step of designing manufacturer-specific CT protocols to achieve established SSDE targets is the responsibility of the Qualified Medical Physicist. The task is straightforward if fixed-mA protocols are used, however, more difficult when automatic exposure control (AEC) and automatic kV selection are considered. In such cases, the physicist must deduce the operation of AEC algorithms from technical documentation or through testing, using a wide range of phantom sizes. Our study presents the results of such testing using anthropomorphic phantoms ranging in size from the newborn to the obese adult. The effect of each user-controlled parameter was modeled for a single-manufacturer AEC algorithm (Siemens CARE Dose4D) and automatic kV selection algorithm (Siemens CARE kV). Based on the results presented in this study, a process for designing mA-modulated, pediatric abdominal CT protocols that achieve user-defined SSDE and kV targets is described. PMID:26894344

  4. Kansas Volunteer Tutor Utilization: Training of Resources. KV-TUTOR. 310-ABE Funded 1984-1985 Volunteer Coordinator Manual.

    ERIC Educational Resources Information Center

    Wichita Public Schools, KS.

    This manual is intended to assist volunteer coordinators involved in the Kansas Volunteer Tutor Utilization (KV-TUTOR) project. The first section, which deals with planning, covers management by objectives and a clarification system to test objectives that is denoted by the acronym SMAC (specific, measurable, achievable, compatible). A section on…

  5. The endocannabinoid 2-AG controls skeletal muscle cell differentiation via CB1 receptor-dependent inhibition of Kv7 channels

    PubMed Central

    Iannotti, Fabio A.; Silvestri, Cristoforo; Mazzarella, Enrico; Martella, Andrea; Calvigioni, Daniela; Piscitelli, Fabiana; Ambrosino, Paolo; Petrosino, Stefania; Czifra, Gabriella; Bíró, Tamás; Harkany, Tibor; Taglialatela, Maurizio; Di Marzo, Vincenzo

    2014-01-01

    Little is known of the involvement of endocannabinoids and cannabinoid receptors in skeletal muscle cell differentiation. We report that, due to changes in the expression of genes involved in its metabolism, the levels of the endocannabinoid 2-arachidonoylglycerol (2-AG) are decreased both during myotube formation in vitro from murine C2C12 myoblasts and during mouse muscle growth in vivo. The endocannabinoid, as well as the CB1 agonist arachidonoyl-2-chloroethylamide, prevent myotube formation in a manner antagonized by CB1 knockdown and by CB1 antagonists, which, per se, instead stimulate differentiation. Importantly, 2-AG also inhibits differentiation of primary human satellite cells. Muscle fascicles from CB1 knockout embryos contain more muscle fibers, and postnatal mice show muscle fibers of an increased diameter relative to wild-type littermates. Inhibition of Kv7.4 channel activity, which plays a permissive role in myogenesis and depends on phosphatidylinositol 4,5-bisphosphate (PIP2), underlies the effects of 2-AG. We find that CB1 stimulation reduces both total and Kv7.4-bound PIP2 levels in C2C12 cells and inhibits Kv7.4 currents in transfected CHO cells. We suggest that 2-AG is an endogenous repressor of myoblast differentiation via CB1-mediated inhibition of Kv7.4 channels. PMID:24927567

  6. A designer ligand specific for Kv1.3 channels from a scorpion neurotoxin-based library

    PubMed Central

    Takacs, Zoltan; Toups, Megan; Kollewe, Astrid; Johnson, Erik; Cuello, Luis G.; Driessens, Gregory; Biancalana, Matthew; Koide, Akiko; Ponte, Cristiano G.; Perozo, Eduardo; Gajewski, Thomas F.; Suarez-Kurtz, Guilherme; Koide, Shohei; Goldstein, Steve A. N.

    2009-01-01

    Venomous animals immobilize prey using protein toxins that act on ion channels and other targets of biological importance. Broad use of toxins for biomedical research, diagnosis, and therapy has been limited by inadequate target discrimination, for example, among ion channel subtypes. Here, a synthetic toxin is produced by a new strategy to be specific for human Kv1.3 channels, critical regulators of immune T cells. A phage display library of 11,200 de novo proteins is designed using the α-KTx scaffold of 31 scorpion toxin sequences known or predicted to bind to potassium channels. Mokatoxin-1 (moka1) is isolated by affinity selection on purified target. Moka1 blocks Kv1.3 at nanomolar levels that do not inhibit Kv1.1, Kv1.2, or KCa1.1. As a result, moka1 suppresses CD3/28-induced cytokine secretion by T cells without cross-reactive gastrointestinal hyperactivity. The 3D structure of moka1 rationalizes its specificity and validates the engineering approach, revealing a unique interaction surface supported on an α-KTx scaffold. This scaffold-based/target-biased strategy overcomes many obstacles to production of selective toxins. PMID:20007782

  7. A novel epileptic encephalopathy mutation in KCNB1 disrupts Kv2.1 ion selectivity, expression, and localization

    PubMed Central

    Thiffault, Isabelle; Speca, David J.; Austin, Daniel C.; Cobb, Melanie M.; Eum, Kenneth S.; Safina, Nicole P.; Grote, Lauren; Farrow, Emily G.; Miller, Neil; Soden, Sarah; Kingsmore, Stephen F.

    2015-01-01

    The epileptic encephalopathies are a group of highly heterogeneous genetic disorders. The majority of disease-causing mutations alter genes encoding voltage-gated ion channels, neurotransmitter receptors, or synaptic proteins. We have identified a novel de novo pathogenic K+ channel variant in an idiopathic epileptic encephalopathy family. Here, we report the effects of this mutation on channel function and heterologous expression in cell lines. We present a case report of infantile epileptic encephalopathy in a young girl, and trio-exome sequencing to determine the genetic etiology of her disorder. The patient was heterozygous for a de novo missense variant in the coding region of the KCNB1 gene, c.1133T>C. The variant encodes a V378A mutation in the α subunit of the Kv2.1 voltage-gated K+ channel, which is expressed at high levels in central neurons and is an important regulator of neuronal excitability. We found that expression of the V378A variant results in voltage-activated currents that are sensitive to the selective Kv2 channel blocker guangxitoxin-1E. These voltage-activated Kv2.1 V378A currents were nonselective among monovalent cations. Striking cell background–dependent differences in expression and subcellular localization of the V378A mutation were observed in heterologous cells. Further, coexpression of V378A subunits and wild-type Kv2.1 subunits reciprocally affects their respective trafficking characteristics. A recent study reported epileptic encephalopathy-linked missense variants that render Kv2.1 a tonically activated, nonselective cation channel that is not voltage activated. Our findings strengthen the correlation between mutations that result in loss of Kv2.1 ion selectivity and development of epileptic encephalopathy. However, the strong voltage sensitivity of currents from the V378A mutant indicates that the loss of voltage-sensitive gating seen in all other reported disease mutants is not required for an epileptic encephalopathy

  8. A fiducial detection algorithm for real-time image guided IMRT based on simultaneous MV and kV imaging

    PubMed Central

    Mao, Weihua; Riaz, Nadeem; Lee, Louis; Wiersma, Rodney; Xing, Lei

    2008-01-01

    The advantage of highly conformal dose techniques such as 3DCRT and IMRT is limited by intrafraction organ motion. A new approach to gain near real-time 3D positions of internally implanted fiducial markers is to analyze simultaneous onboard kV beam and treatment MV beam images (from fluoroscopic or electronic portal image devices). Before we can use this real-time image guidance for clinical 3DCRT and IMRT treatments, four outstanding issues need to be addressed. (1) How will fiducial motion blur the image and hinder tracking fiducials? kV and MV images are acquired while the tumor is moving at various speeds. We find that a fiducial can be successfully detected at a maximum linear speed of 1.6 cm∕s. (2) How does MV beam scattering affect kV imaging? We investigate this by varying MV field size and kV source to imager distance, and find that common treatment MV beams do not hinder fiducial detection in simultaneous kV images. (3) How can one detect fiducials on images from 3DCRT and IMRT treatment beams when the MV fields are modified by a multileaf collimator (MLC)? The presented analysis is capable of segmenting a MV field from the blocking MLC and detecting visible fiducials. This enables the calculation of nearly real-time 3D positions of markers during a real treatment. (4) Is the analysis fast enough to track fiducials in nearly real time? Multiple methods are adopted to predict marker positions and reduce search regions. The average detection time per frame for three markers in a 1024×768 image was reduced to 0.1 s or less. Solving these four issues paves the way to tracking moving fiducial markers throughout a 3DCRT or IMRT treatment. Altogether, these four studies demonstrate that our algorithm can track fiducials in real time, on degraded kV images (MV scatter), in rapidly moving tumors (fiducial blurring), and even provide useful information in the case when some fiducials are blocked from view by the MLC. This technique can provide a gating signal

  9. A fiducial detection algorithm for real-time image guided IMRT based on simultaneous MV and kV imaging

    SciTech Connect

    Mao Weihua; Riaz, Nadeem; Lee, Louis; Wiersma, Rodney; Xing Lei

    2008-08-15

    The advantage of highly conformal dose techniques such as 3DCRT and IMRT is limited by intrafraction organ motion. A new approach to gain near real-time 3D positions of internally implanted fiducial markers is to analyze simultaneous onboard kV beam and treatment MV beam images (from fluoroscopic or electronic portal image devices). Before we can use this real-time image guidance for clinical 3DCRT and IMRT treatments, four outstanding issues need to be addressed. (1) How will fiducial motion blur the image and hinder tracking fiducials? kV and MV images are acquired while the tumor is moving at various speeds. We find that a fiducial can be successfully detected at a maximum linear speed of 1.6 cm/s. (2) How does MV beam scattering affect kV imaging? We investigate this by varying MV field size and kV source to imager distance, and find that common treatment MV beams do not hinder fiducial detection in simultaneous kV images. (3) How can one detect fiducials on images from 3DCRT and IMRT treatment beams when the MV fields are modified by a multileaf collimator (MLC)? The presented analysis is capable of segmenting a MV field from the blocking MLC and detecting visible fiducials. This enables the calculation of nearly real-time 3D positions of markers during a real treatment. (4) Is the analysis fast enough to track fiducials in nearly real time? Multiple methods are adopted to predict marker positions and reduce search regions. The average detection time per frame for three markers in a 1024x768 image was reduced to 0.1 s or less. Solving these four issues paves the way to tracking moving fiducial markers throughout a 3DCRT or IMRT treatment. Altogether, these four studies demonstrate that our algorithm can track fiducials in real time, on degraded kV images (MV scatter), in rapidly moving tumors (fiducial blurring), and even provide useful information in the case when some fiducials are blocked from view by the MLC. This technique can provide a gating signal or

  10. A fiducial detection algorithm for real-time image guided IMRT based on simultaneous MV and kV imaging.

    PubMed

    Mao, Weihua; Riaz, Nadeem; Lee, Louis; Wiersma, Rodney; Xing, Lei

    2008-08-01

    The advantage of highly conformal dose techniques such as 3DCRT and IMRT is limited by intrafraction organ motion. A new approach to gain near real-time 3D positions of internally implanted fiducial markers is to analyze simultaneous onboard kV beam and treatment MV beam images (from fluoroscopic or electronic portal image devices). Before we can use this real-time image guidance for clinical 3DCRT and IMRT treatments, four outstanding issues need to be addressed. (1) How will fiducial motion blur the image and hinder tracking fiducials? kV and MV images are acquired while the tumor is moving at various speeds. We find that a fiducial can be successfully detected at a maximum linear speed of 1.6 cm/s. (2) How does MV beam scattering affect kV imaging? We investigate this by varying MV field size and kV source to imager distance, and find that common treatment MV beams do not hinder fiducial detection in simultaneous kV images. (3) How can one detect fiducials on images from 3DCRT and IMRT treatment beams when the MV fields are modified by a multileaf collimator (MLC)? The presented analysis is capable of segmenting a MV field from the blocking MLC and detecting visible fiducials. This enables the calculation of nearly real-time 3D positions of markers during a real treatment. (4) Is the analysis fast enough to track fiducials in nearly real time? Multiple methods are adopted to predict marker positions and reduce search regions. The average detection time per frame for three markers in a 1024 x 768 image was reduced to 0.1 s or less. Solving these four issues paves the way to tracking moving fiducial markers throughout a 3DCRT or IMRT treatment. Altogether, these four studies demonstrate that our algorithm can track fiducials in real time, on degraded kV images (MV scatter), in rapidly moving tumors (fiducial blurring), and even provide useful information in the case when some fiducials are blocked from view by the MLC. This technique can provide a gating signal or

  11. Theoretical investigation of the design and performance of a dual energy (kV and MV) radiotherapy imager

    SciTech Connect

    Liu, Langechuan; Antonuk, Larry E. El-Mohri, Youcef; Zhao, Qihua; Jiang, Hao

    2015-04-15

    Purpose: In modern radiotherapy treatment rooms, megavoltage (MV) portal imaging and kilovoltage (kV) cone-beam CT (CBCT) imaging are performed using various active matrix flat-panel imager (AMFPI) designs. To expand the clinical utility of MV and kV imaging, MV AMFPIs incorporating thick, segmented scintillators and, separately, kV imaging using a beam’s eye view geometry have been investigated by a number of groups. Motivated by these previous studies, it is of interest to explore to what extent it is possible to preserve the benefits of kV and MV imaging using a single AMFPI design, given the considerably different x ray energy spectra used for kV and MV imaging. In this paper, considerations for the design of such a dual energy imager are explored through examination of the performance of a variety of hypothetical AMFPIs based on x ray converters employing segmented scintillators. Methods: Contrast, noise, and contrast-to-noise ratio performances were characterized through simulation modeling of CBCT imaging, while modulation transfer function, Swank factor, and signal performance were characterized through simulation modeling of planar imaging. The simulations were based on a previously reported hybrid modeling technique (accounting for both radiation and optical effects), augmented through modeling of electronic additive noise. All designs employed BGO scintillator material with thicknesses ranging from 0.25 to 4 cm and element-to-element pitches ranging from 0.508 to 1.016 mm. A series of studies were performed under both kV and MV imaging conditions to determine the most advantageous imager configuration (involving front or rear x ray illumination and use of a mirror or black reflector), converter design (pitch and thickness), and operating mode (pitch-binning combination). Results: Under the assumptions of the present study, the most advantageous imager design was found to employ rear illumination of the converter in combination with a black reflector

  12. Increase of Kv3.1b expression in avian auditory brainstem neurons correlates with synaptogenesis in vivo and in vitro.

    PubMed

    Kuenzel, Thomas; Wirth, Marcus J; Luksch, Harald; Wagner, Hermann; Mey, Jörg

    2009-12-11

    In the auditory system voltage-activated currents mediated by potassium channels Kv1.1 and Kv3.1b and their interaction with sodium inward currents play a crucial role for computational function. However, it is unresolved how these potassium channels are developmentally regulated. We have therefore combined a biochemical investigation of Kv1.1 and Kv3.1b protein expression with electrophysiological recordings of membrane currents to characterize neuronal differentiation in the auditory brain stem of the chick. Differentiation in vitro was compared with cells prepared from corresponding embryonic stages in vivo. Using a computer model based on the empirical data we were then able to predict physiological properties of developing auditory brain stem neurons. In vivo Kv3.1b expression increased strongly between E10 and E14, a time of functional synaptogenesis in the auditory brainstem. We also found this increase of expression in vitro, again coinciding with synaptogenesis in the cultures. Whole-cell patch recordings revealed a corresponding increase of the (Kv3.1-like) high threshold potassium current. In contrast, Kv1.1 protein expression failed to increase in vitro, and changes in (Kv1.1-like) low threshold potassium current with time in culture were not significant. Electrophysiological recordings revealed that sodium inward currents increased with cultivation time. Thus, our data suggest that Kv3.1b expression occurs with the onset of functional synaptogenesis, while a different signal, absent from cultures of dissociated auditory brain stem, is needed for Kv1.1 expression. A biophysical model constructed with parameters from our recordings was used to investigate the functional impact of the currents mediated by these channels. We found that during development both high and low threshold potassium currents need to be increased in a concerted manner with the sodium conductance for the neurons to exhibit fast and phasic action potential firing and a narrow time

  13. Role of N-Terminal Domain and Accessory Subunits in Controlling Deactivation-Inactivation Coupling of Kv4.2 Channels

    PubMed Central

    Barghaan, Jan; Tozakidou, Magdalini; Ehmke, Heimo; Bähring, Robert

    2008-01-01

    We examined the relationship between deactivation and inactivation in Kv4.2 channels. In particular, we were interested in the role of a Kv4.2 N-terminal domain and accessory subunits in controlling macroscopic gating kinetics and asked if the effects of N-terminal deletion and accessory subunit coexpression conform to a kinetic coupling of deactivation and inactivation. We expressed Kv4.2 wild-type channels and N-terminal deletion mutants in the absence and presence of Kv channel interacting proteins (KChIPs) and dipeptidyl aminopeptidase-like proteins (DPPs) in human embryonic kidney 293 cells. Kv4.2-mediated A-type currents at positive and deactivation tail currents at negative membrane potentials were recorded under whole-cell voltage-clamp and analyzed by multi-exponential fitting. The observed changes in Kv4.2 macroscopic inactivation kinetics caused by N-terminal deletion, accessory subunit coexpression, or a combination of the two maneuvers were compared with respective changes in deactivation kinetics. Extensive correlation analyses indicated that modulatory effects on deactivation closely parallel respective effects on inactivation, including both onset and recovery kinetics. Searching for the structural determinants, which control deactivation and inactivation, we found that in a Kv4.2Δ2–10 N-terminal deletion mutant both the initial rapid phase of macroscopic inactivation and tail current deactivation were slowed. On the other hand, the intermediate and slow phase of A-type current decay, recovery from inactivation, and tail current decay kinetics were accelerated in Kv4.2Δ2–10 by KChIP2 and DPPX. Thus, a Kv4.2 N-terminal domain, which may control both inactivation and deactivation, is not necessary for active modulation of current kinetics by accessory subunits. Our results further suggest distinct mechanisms for Kv4.2 gating modulation by KChIPs and DPPs. PMID:17981906

  14. KV1 channels identified in rodent myelinated axons, linked to Cx29 in innermost myelin: support for electrically active myelin in mammalian saltatory conduction.

    PubMed

    Rash, John E; Vanderpool, Kimberly G; Yasumura, Thomas; Hickman, Jordan; Beatty, Jonathan T; Nagy, James I

    2016-04-01

    Saltatory conduction in mammalian myelinated axons was thought to be well understood before recent discoveries revealed unexpected subcellular distributions and molecular identities of the K(+)-conductance pathways that provide for rapid axonal repolarization. In this study, we visualize, identify, localize, quantify, and ultrastructurally characterize axonal KV1.1/KV1.2 channels in sciatic nerves of rodents. With the use of light microscopic immunocytochemistry and freeze-fracture replica immunogold labeling electron microscopy, KV1.1/KV1.2 channels are localized to three anatomically and compositionally distinct domains in the internodal axolemmas of large myelinated axons, where they form densely packed "rosettes" of 9-nm intramembrane particles. These axolemmal KV1.1/KV1.2 rosettes are precisely aligned with and ultrastructurally coupled to connexin29 (Cx29) channels, also in matching rosettes, in the surrounding juxtaparanodal myelin collars and along the inner mesaxon. As >98% of transmembrane proteins large enough to represent ion channels in these specialized domains, ∼500,000 KV1.1/KV1.2 channels define the paired juxtaparanodal regions as exclusive membrane domains for the voltage-gated K(+)conductance that underlies rapid axonal repolarization in mammals. The 1:1 molecular linkage of KV1 channels to Cx29 channels in the apposed juxtaparanodal collars, plus their linkage to an additional 250,000-400,000 Cx29 channels along each inner mesaxon in every large-diameter myelinated axon examined, supports previously proposed K(+)conductance directly from juxtaparanodal axoplasm into juxtaparanodal myeloplasm in mammalian axons. With neither Cx29 protein nor myelin rosettes detectable in frog myelinated axons, these data showing axon-to-myelin linkage by abundant KV1/Cx29 channels in rodent axons support renewed consideration of an electrically active role for myelin in increasing both saltatory conduction velocity and maximum propagation frequency in

  15. Variability of Potassium Channel Blockers in Mesobuthus eupeus Scorpion Venom with Focus on Kv1.1

    PubMed Central

    Kuzmenkov, Alexey I.; Vassilevski, Alexander A.; Kudryashova, Kseniya S.; Nekrasova, Oksana V.; Peigneur, Steve; Tytgat, Jan; Feofanov, Alexey V.; Kirpichnikov, Mikhail P.; Grishin, Eugene V.

    2015-01-01

    The lesser Asian scorpion Mesobuthus eupeus (Buthidae) is one of the most widely spread and dispersed species of the Mesobuthus genus, and its venom is actively studied. Nevertheless, a considerable amount of active compounds is still under-investigated due to the high complexity of this venom. Here, we report a comprehensive analysis of putative potassium channel toxins (KTxs) from the cDNA library of M. eupeus venom glands, and we compare the deduced KTx structures with peptides purified from the venom. For the transcriptome analysis, we used conventional tools as well as a search for structural motifs characteristic of scorpion venom components in the form of regular expressions. We found 59 candidate KTxs distributed in 30 subfamilies and presenting the cysteine-stabilized α/β and inhibitor cystine knot types of fold. M. eupeus venom was then separated to individual components by multistage chromatography. A facile fluorescent system based on the expression of the KcsA-Kv1.1 hybrid channels in Escherichia coli and utilization of a labeled scorpion toxin was elaborated and applied to follow Kv1.1 pore binding activity during venom separation. As a result, eight high affinity Kv1.1 channel blockers were identified, including five novel peptides, which extend the panel of potential pharmacologically important Kv1 ligands. Activity of the new peptides against rat Kv1.1 channel was confirmed (IC50 in the range of 1–780 nm) by the two-electrode voltage clamp technique using a standard Xenopus oocyte system. Our integrated approach is of general utility and efficiency to mine natural venoms for KTxs. PMID:25792741

  16. Targeting the voltage sensor of Kv7.2 voltage-gated K+ channels with a new gating-modifier.

    PubMed

    Peretz, Asher; Pell, Liat; Gofman, Yana; Haitin, Yoni; Shamgar, Liora; Patrich, Eti; Kornilov, Polina; Gourgy-Hacohen, Orit; Ben-Tal, Nir; Attali, Bernard

    2010-08-31

    The pore and gate regions of voltage-gated cation channels have been often targeted with drugs acting as channel modulators. In contrast, the voltage-sensing domain (VSD) was practically not exploited for therapeutic purposes, although it is the target of various toxins. We recently designed unique diphenylamine carboxylates that are powerful Kv7.2 voltage-gated K(+) channel openers or blockers. Here we show that a unique Kv7.2 channel opener, NH29, acts as a nontoxin gating modifier. NH29 increases Kv7.2 currents, thereby producing a hyperpolarizing shift of the activation curve and slowing both activation and deactivation kinetics. In neurons, the opener depresses evoked spike discharges. NH29 dampens hippocampal glutamate and GABA release, thereby inhibiting excitatory and inhibitory postsynaptic currents. Mutagenesis and modeling data suggest that in Kv7.2, NH29 docks to the external groove formed by the interface of helices S1, S2, and S4 in a way that stabilizes the interaction between two conserved charged residues in S2 and S4, known to interact electrostatically, in the open state of Kv channels. Results indicate that NH29 may operate via a voltage-sensor trapping mechanism similar to that suggested for scorpion and sea-anemone toxins. Reflecting the promiscuous nature of the VSD, NH29 is also a potent blocker of TRPV1 channels, a feature similar to that of tarantula toxins. Our data provide a structural framework for designing unique gating-modifiers targeted to the VSD of voltage-gated cation channels and used for the treatment of hyperexcitability disorders. PMID:20713704

  17. Lovastatin blocks Kv1.3 channel in human T cells: a new mechanism to explain its immunomodulatory properties

    PubMed Central

    Zhao, Ning; Dong, Qian; Qian, Cheng; Li, Sen; Wu, Qiong-Feng; Ding, Dan; Li, Jing; Wang, Bin-Bin; Guo, Ke-fang; Xie, Jiang-jiao; Cheng, Xiang; Liao, Yu-Hua; Du, Yi-Mei

    2015-01-01

    Lovastatin is a member of Statins, which are beneficial in a lot of immunologic cardiovascular diseases and T cell-mediated autoimmune diseases. Kv1.3 channel plays important roles in the activation and proliferation of T cells, and have become attractive target for immune-related disorders. The present study was designed to examine the block effect of Lovastatin on Kv1.3 channel in human T cells, and to clarify its new immunomodulatory mechanism. We found that Lovastatin inhibited Kv1.3 currents in a concentration- and voltage-dependent manner, and the IC50 for peak, end of the pulse was 39.81 ± 5.11, 6.92 ± 0.95 μM, respectively. Lovastatin also accelerated the decay rate of current inactivation and negatively shifted the steady-state inactivation curves concentration-dependently, without affecting the activation curve. However, 30 μM Lovastatin had no apparent effect on KCa current in human T cells. Furthermore, Lovastatin inhibited Ca2+ influx, T cell proliferation as well as IL-2 production. The activities of NFAT1 and NF-κB p65/50 were down-regulated by Lovastatin, too. At last, Mevalonate application only partially reversed the inhibition of Lovastatin on IL-2 secretion, and the siRNA against Kv1.3 also partially reduced this inhibitory effect of Lovastatin. In conclusion, Lovastatin can exert immunodulatory properties through the new mechanism of blocking Kv1.3 channel. PMID:26616555

  18. Evaluation of gambierol and its analogs for their inhibition of human Kv1.2 and cytotoxicity.

    PubMed

    Konoki, Keiichi; Suga, Yuto; Fuwa, Haruhiko; Yotsu-Yamashita, Mari; Sasaki, Makoto

    2015-02-01

    Gambierol and its heptacyclic and tetracyclic analogs were tested for inhibitory activity against the human voltage-gated potassium channel Kv1.2 (hKv1.2), which was stably expressed in Chinese hamster ovary (CHO) cells. Gambierol, the heptacyclic analog, and the tetracyclic analog inhibited the potassium current evoked by a step pulse from -80mV to 40mV. The IC50 values for the three compounds were 0.75±0.15nM, 7.6±1.2nM, and 28±4.0nM (the mean±SEM, n=3), respectively. The cytotoxic activity was examined in order to assess a relationship between cytotoxicity and inhibition of the hKv1.2. The IC50 values for gambierol, the heptacyclic analog, and the tetracyclic analog in the wild-type CHO cells were 95±7.1μM, 6.5±0.8μM (the mean±SEM, n=3), and >100μM (n=3), respectively, whereas those in the CHO cells stably expressing hKv1.2 were 78±5.8μM, 6.0±1.0μM (the mean±SEM, n=3), and >100μM (n=3). These results suggested that cytotoxicity is not triggered by inhibition of the human Kv1.2. The electrophysiological recording at the resting potential in the presence of gambierol, the heptacyclic analog, and the tetracyclic analog revealed the dose-dependent leak current, which was largest when the heptacyclic analog was administered to the cells. We thus propose that the leak current induced by these compounds might cause a fatal effect on the cultured cells. PMID:25556093

  19. Voltage-Gated Potassium Channels Kv1.3--Potentially New Molecular Target in Cancer Diagnostics and Therapy.

    PubMed

    Teisseyre, Andrzej; Gąsiorowska, Justyna; Michalak, Krystyna

    2015-01-01

    Voltage-gated potassium channels, Kv1.3, which were discovered in 1984, are integral membrane proteins which are activated ("open") upon change of the cell membrane potential, enabling a passive flux of potassium ions across the cell membrane. The channels are expressed in many different tissues, both normal and cancer. Since 2005 it has been known that the channels are expressed not only in the plasma membrane, but also in the inner mitochondrial membrane. The activity of Kv1.3 channels plays an important role, among others, in setting the cell resting membrane potential, cell proliferation, apoptosis and volume regulation. For some years, these channels have been considered a potentially new molecular target in both the diagnostics and therapy of some cancer diseases. This review article focuses on: 1) changes of expression of the channels in cancer disorders with special regard to correlations between the channels' expression and stage of the disease, 2) influence of inhibitors of Kv1.3 channels on proliferation and apoptosis of cancer cells, 3) possible future applications of Kv1.3 channels' inhibitors in therapy of some cancer diseases. In the last section, the results of studies performed in our Laboratory of Bioelectricity on the influence of selected biologically active plant-derived compounds from the groups of flavonoids and stilbenes and their natural and synthetic derivatives on the activity of Kv1.3 channels in normal and cancer cells are reviewed. A possible application of some compounds from these groups to support therapy of cancer diseases, such as breast, colon and lymph node cancer, and melanoma or chronic lymphocytic leukemia (B-CLL), is announced. PMID:26467143

  20. Implementing chemically amplified resist to 10kV raster e-beam process in photomask manufacturing

    NASA Astrophysics Data System (ADS)

    Kim, Sook-Kyeong; Kim, Byung-Gook; Moon, Seong-Yong; Choi, Sung-Woon; Han, Woo-Sung

    2005-06-01

    CAR(Chemically amplified resist) is widely used in 50keV VSB (Variable Shaped Beam) e-beam process in photomask manufacturing due to its advantage of high sensitivity which gives to reduced writing time compared to non-CAR. The 10kV raster e-beam system, however, is spread out already worldwide and plays a important role till now in middle grade mask-making. Conventionally the non-CAR like ZEP7000 has been applied to the 10kV raster e-beam system and it gives good performance for raster scan e-beam system. In mass production, sometimes, maintaining two kinds of resist simultaneously of CAR and non-CAR are inefficient strategy to the mask house which has limited resources. This situation makes the authors to apply CAR to the 10kV raster e-beam process. Generally, the grid of 10kV raster e-beam(MEBES) is large and limited compared to the current VSB grid. Historically, many layout data is designed already based on the large limited grid and this gives to limited sizing value. Moreover, it is difficult to control exposure dose in raster e-beam system and control bias with develop time in CAR process. These situations make more difficult CAR application to raster e-beam system under the simple mask data preparation strategy. In this paper, some critical problems will be discussed in isofocal process making for raster scan e-beam system. Advantage and disadvantage will be also discussed through the comparison of basic parameters such as dose margin, develop margin, and the fogging effect between the CAR and non-CAR process in 10kV raster e-beam process.

  1. In vitro and intrathecal siRNA mediated KV1.1 knock-down in primary sensory neurons

    PubMed Central

    Baker, Mark D.; Chen, Ya-Chun; Shah, Syed U.; Okuse, Kenji

    2011-01-01

    KV1.1 is a Shaker homologue K+ channel that contributes to the juxta-paranodal membrane conductance in myelinated axons, and is blocked by fampridine (4-aminopyridine), used to treat the symptoms of multiple sclerosis. The present experiments investigate KV1.1 function in primary sensory neurons and A-fibres, and help define its characteristics as a drug-target using sequence specific small-interfering RNAs (siRNAs). siRNA (71 nM) was used to knock-down functional expression of KV1.1 in sensory neurons (> 25 μm in apparent diameter) in culture, and was also delivered intrathecally in vivo (9.3 μg). K+ channel knock-down in sensory neurons was found to make the voltage-threshold for action potential generation significantly more negative than in control (p = 0.02), led to the breakdown of accommodation and promoted spontaneous action potential firing. Exposure to dendrotoxin-K (DTX-K, 10–100 nM) also selectively abolished K+ currents at negative potentials and made voltage-threshold more negative, consistent with KV1.1 controlling excitability close to the nominal resting potential of the neuron cell body, near − 60 mV. Introduction of one working siRNA sequence into the intrathecal space in vivo was associated with a small increase in the amplitude of the depolarising after-potential in sacral spinal roots (p < 0.02), suggesting a reduction in the number of working K+ channels in internodal axon membrane. Our study provides evidence that KV1.1 contributes to the control of peripheral sensory nerve excitability, and suggests that its characteristics as a putative drug target can be assessed by siRNA transfection in primary sensory neurons in vitro and in vivo. PMID:21903165

  2. Contribution of the Kv3.1 potassium channel to high-frequency firing in mouse auditory neurones

    PubMed Central

    Wang, Lu-Yang; Gan, Li; Forsythe, Ian D; Kaczmarek, Leonard K

    1998-01-01

    Using a combination of patch-clamp, in situ hybridization and computer simulation techniques, we have analysed the contribution of potassium channels to the ability of a subset of mouse auditory neurones to fire at high frequencies.Voltage-clamp recordings from the principal neurones of the medial nucleus of the trapezoid body (MNTB) revealed a low-threshold dendrotoxin (DTX)-sensitive current (ILT) and a high-threshold DTX-insensitive current (IHT).IHT displayed rapid activation and deactivation kinetics, and was selectively blocked by a low concentration of tetraethylammonium (TEA; 1 mm).The physiological and pharmacological properties of IHT very closely matched those of the Shaw family potassium channel Kv3.1 stably expressed in a CHO cell line.An mRNA probe corresponding to the C-terminus of the Kv3.1 channel strongly labelled MNTB neurones, suggesting that this channel is expressed in these neurones.TEA did not alter the ability of MNTB neurones to follow stimulation up to 200 Hz, but specifically reduced their ability to follow higher frequency impulses.A computer simulation, using a model cell in which an outward current with the kinetics and voltage dependence of the Kv3.1 channel was incorporated, also confirmed that the Kv3.1- like current is essential for cells to respond to a sustained train of high-frequency stimuli.We conclude that in mouse MNTB neurones the Kv3.1 channel contributes to the ability of these cells to lock their firing to high-frequency inputs. PMID:9547392

  3. The membrane protein MiRP3 regulates Kv4.2 channels in a KChIP-dependent manner

    PubMed Central

    Levy, Daniel I; Cepaitis, Egle; Wanderling, Sherry; Toth, Peter T; Archer, Stephen L; Goldstein, Steve A N

    2010-01-01

    MiRP3, the single-span membrane protein encoded by KCNE4, is localized by immunofluorescence microscopy to the transverse tubules of murine cardiac myocytes. MiRP3 is found to co-localize with Kv4.2 subunits that contribute to cardiac transient outward potassium currents (Ito). Whole-cell, voltage-clamp recordings of human MiRP3 and Kv4.2 expressed in a clonal cell line (tsA201) reveal MiRP3 to modulate Kv4.2 current activation, inactivation and recovery from inactivation. MiRP3 shifts the half-maximal voltage for activation (V1/2) ∼20 mV and slows time to peak ∼100%. In addition, MiRP3 slows inactivation ∼100%, speeds recovery from inactivation ∼30%, and enhances restored currents so they ‘overshoot’ baseline levels. The cytoplasmic accessory subunit KChIP2 also assembles with Kv4.2 in tsA201 cells to increase peak current, shift V1/2 ∼5 mV, slow time to peak ∼10%, slow inactivation ∼100%, and speed recovery from inactivation ∼250% without overshoot. Simultaneous expression of all three subunits yields a biophysical profile unlike either accessory subunit alone, abolishes MiRP3-induced overshoot, and allows biochemical isolation of the ternary complex. Thus, regional heterogeneity in cardiac expression of MiRP3, Kv4.2 and KChIP2 in health and disease may establish the local attributes and magnitude of cardiac Ito. PMID:20498229

  4. Ternary Kv4.2 channels recapitulate voltage-dependent inactivation kinetics of A-type K+ channels in cerebellar granule neurons

    PubMed Central

    Amarillo, Yimy; De Santiago-Castillo, Jose A; Dougherty, Kevin; Maffie, Jonathon; Kwon, Elaine; Covarrubias, Manuel; Rudy, Bernardo

    2008-01-01

    Kv4 channels mediate most of the somatodendritic subthreshold operating A-type current (ISA) in neurons. This current plays essential roles in the regulation of spike timing, repetitive firing, dendritic integration and plasticity. Neuronal Kv4 channels are thought to be ternary complexes of Kv4 pore-forming subunits and two types of accessory proteins, Kv channel interacting proteins (KChIPs) and the dipeptidyl-peptidase-like proteins (DPPLs) DPPX (DPP6) and DPP10. In heterologous cells, ternary Kv4 channels exhibit inactivation that slows down with increasing depolarization. Here, we compared the voltage dependence of the inactivation rate of channels expressed in heterologous mammalian cells by Kv4.2 proteins with that of channels containing Kv4.2 and KChIP1, Kv4.2 and DPPX-S, or Kv4.2, KChIP1 and DPPX-S, and found that the relation between inactivation rate and membrane potential is distinct for these four conditions. Moreover, recordings from native neurons showed that the inactivation kinetics of the ISA in cerebellar granule neurons has voltage dependence that is remarkably similar to that of ternary Kv4 channels containing KChIP1 and DPPX-S proteins in heterologous cells. The fact that this complex and unique behaviour (among A-type K+ currents) is observed in both the native current and the current expressed in heterologous cells by the ternary complex containing Kv4, DPPX and KChIP proteins supports the hypothesis that somatically recorded native Kv4 channels in neurons include both types of accessory protein. Furthermore, quantitative global kinetic modelling showed that preferential closed-state inactivation and a weakly voltage-dependent opening step can explain the slowing of the inactivation rate with increasing depolarization. Therefore, it is likely that preferential closed-state inactivation is the physiological mechanism that regulates the activity of both ternary Kv4 channel complexes and native ISA-mediating channels. PMID:18276729

  5. Metal artifact correction for x-ray computed tomography using kV and selective MV imaging

    PubMed Central

    Wu, Meng; Keil, Andreas; Constantin, Dragos; Star-Lack, Josh; Zhu, Lei; Fahrig, Rebecca

    2014-01-01

    Purpose: The overall goal of this work is to improve the computed tomography (CT) image quality for patients with metal implants or fillings by completing the missing kilovoltage (kV) projection data with selectively acquired megavoltage (MV) data that do not suffer from photon starvation. When both of these imaging systems, which are available on current radiotherapy devices, are used, metal streak artifacts are avoided, and the soft-tissue contrast is restored, even for regions in which the kV data cannot contribute any information. Methods: Three image-reconstruction methods, including two filtered back-projection (FBP)-based analytic methods and one iterative method, for combining kV and MV projection data from the two on-board imaging systems of a radiotherapy device are presented in this work. The analytic reconstruction methods modify the MV data based on the information in the projection or image domains and then patch the data onto the kV projections for a FBP reconstruction. In the iterative reconstruction, the authors used dual-energy (DE) penalized weighted least-squares (PWLS) methods to simultaneously combine the kV/MV data and perform the reconstruction. Results: The authors compared kV/MV reconstructions to kV-only reconstructions using a dental phantom with fillings and a hip-implant numerical phantom. Simulation results indicated that dual-energy sinogram patch FBP and the modified dual-energy PWLS method can successfully suppress metal streak artifacts and restore information lost due to photon starvation in the kV projections. The root-mean-square errors of soft-tissue patterns obtained using combined kV/MV data are 10–15 Hounsfield units smaller than those of the kV-only images, and the structural similarity index measure also indicates a 5%–10% improvement in the image quality. The added dose from the MV scan is much less than the dose from the kV scan if a high efficiency MV detector is assumed. Conclusions: The authors have shown that it

  6. Anti-proliferative effect of Kv1.3 blockers in A549 human lung adenocarcinoma in vitro and in vivo.

    PubMed

    Jang, Soo Hwa; Choi, Seon Young; Ryu, Pan Dong; Lee, So Yeong

    2011-01-25

    Voltage-gated potassium (Kv) channels are widely expressed in the plasma membranes of numerous cells and contribute to a variety of cellular functions in both excitable neuronal cells and non-excitable epithelial cells. Recently, it has been demonstrated that Kv channels are associated with the proliferation of several types of cancer cells. In the present study, we investigated the effects of suppression of Kv1.3 expression on cell proliferation and cell cycle progression in human lung adenocarcinoma, A549 cells. Treatment with margatoxin (MgTX), a selective blocker of Kv1.3 or short hairpin RNA (shRNA) against Kv1.3, significantly blocked A549 cells' proliferation. In addition, selective inhibition of Kv1.3 significantly increased expression level of p21(Waf1/Cip1) and significantly decreased the expression level of Cdk4 and cyclin D3. We also applied the MgTX into a xenograft model using nude mice, and MgTX caused a reduction of tumor volume when it was injected into the tumor tissues. These results suggest that Kv1.3 may serve as a novel therapeutic target for lung adenocarcinoma therapy. PMID:21087602

  7. SU-E-T-140: Dynamic Wave Arc Trajectory Verification Using KV X-Ray Fluoroscopy

    SciTech Connect

    Burghelea, M; Poels, K; Depuydt, T; Tournel, K; Verellen, D; De Ridder, M

    2014-06-01

    Purpose: Purpose: This study investigates the geometric accuracy of simultaneous Gantry/Ring rotation during Dynamic Wave Arc (DWA) delivery. Methods: The Vero SBRT system consists of a 6MV LINAC mounted on an O-ring gantry that can rotate around the vertical axis (±60°), similar to couch rotation on C-arm gantries. To provide CBCT and fluoroscopy imaging functionalities, two orthogonal kV imaging units are attached to the O-ring at −45°/+45° from the beam axis.Dynamic Wave Arc maximizes Vero's motion capabilities by employing synchronized gantry and ring motion on a complex non-coplanar trajectory in combination with aperture based optimized MLC segments.Four wave arc trajectories (T1-4) were delivered using a cubic phantom with a configuration of five lead beads. O-ring gantry position information was retrieved through continuous dual-source kV X-ray image acquisition during DWA. An in-house algorithm read in the image set, extracted the projected marker positions and determined the angulation through reconstruction of the beam source position. The geometric error was quantified as the distance between the independently detected positions from kV-images and reference trajectory derived from the treatment plan in the Ring-Gantry space. Results: The average displacement between the 3D gantry/ring positions reconstructed from the fluoroscopy images and the reference trajectory was 0.346 mm (SD 0,171) for T1. A mean offset of 0.348 mm (SD 0,182) and 0.357 mm (SD 0.194) was observed for trajectory T2(2segmens) and T3(4segments), respectively. The saw shape T4 presented a mean geometric error of 0.363 (SD 0.156). The overall systematic error of 0.350 was caused by the difference between planned reference trajectory created by linear interpolation between CP, and the machine delivery following a spline curve. Conclusion: An independent geometric QA approach has been developed for DWA delivery verification, successfully applied on diverse trajectories and disclosed

  8. A New Regulatory Mechanism for Kv7.2 Protein During Neuropathy: Enhanced Transport from the Soma to Axonal Terminals of Injured Sensory Neurons.

    PubMed

    Cisneros, Elsa; Roza, Carolina; Jackson, Nieka; López-García, José Antonio

    2015-01-01

    Kv7.2 channel expression has been reported to decrease in dorsal root ganglia (DRG) following the induction of a peripheral neuropathy while other experiments show that Kv7.2 accumulates in peripheral neuromas. The mechanisms underlying these novel expression patterns are poorly understood. Here we use immunofluorescence methods to analyze Kv7.2 protein expression changes in sensory neurons following peripheral axotomy and the potential role of axonal transport. Results indicate that DRG neurons express Kv7.2 in ~16% of neurons and that this number decreases by about 65% after axotomy. Damaged neurons were identified in DRG by application of the tracer Fluoro-ruby at the site of injury during surgery. Reduction of Kv7.2 expression was particularly strong in damaged neurons although some loss was also found in putative uninjured neurons. In parallel to the decrease in the soma of axotomized sensory neurons, Kv7.2 accumulated at neuromatose fiber endings. Blockade of axonal transport with either vinblastine (VLB) or colchicine (COL) abolished Kv7.2 redistribution in neuropathic animals. Channel distribution rearrangements did not occur following induction of inflammation in the hind paw. Behavioral tests indicate that protein rearrangements within sensory afferents are essential to the development of allodynia under neuropathic conditions. These results suggest that axotomy enhances axonal transport in injured sensory neurons, leading to a decrease of somatic expression of Kv7.2 protein and a concomitant accumulation in damaged fiber endings. Localized changes in channel expression patterns under pathological conditions may create novel opportunities for Kv7.2 channel openers to act as analgesics. PMID:26696829

  9. Structure of a Ca2+/CaM:Kv7.4 (KCNQ4) B helix complex provides insight into M-current modulation

    PubMed Central

    Xu, Qiang; Chang, Aram; Tolia, Alexandra; Minor, Daniel L.

    2012-01-01

    Calmodulin (CaM) is an important regulator of Kv7.x (KCNQx) voltage-gated potassium channels. Channels from this family produce neuronal M-currents and cardiac and auditory IKS currents, and harbor mutations that cause arrhythmias, epilepsy, and deafness. Despite extensive functional characterization, biochemical and structural details of the interaction between CaM and the channel have remained elusive. Here, we show that both apo-CaM and Ca2+/CaM bind to the C-terminal tail of the neuronal channel Kv7.4 (KCNQ4), which is involved both hearing and mechanosensation. Interactions between apo-CaM and the Kv7.4 tail involve two C-terminal tail segments, known as the A and B segments, whereas the interaction between Ca2+/CaM and the Kv7.4 C-terminal tail requires only the B segment. Biochemical studies show that the calcium dependence of the CaM:B segment interaction is conserved in all Kv7 subtypes. X-ray crystallographic determination of the structure of the Ca2+/CaM:Kv7.4 B segment complex shows that Ca2+/CaM wraps around the Kv7.4 B segment, which forms an α-helix, in an antiparallel orientation that embodies a variation of the classic 1-14 Ca2+/CaM interaction motif. Taken together with the context of prior studies, our data suggest a model for modulation of neuronal Kv7 channels involving a calcium-dependent conformational switch from an apo-CaM form that bridges the A and B segments to a Ca2+/CaM form bound to the B-helix. The structure presented here also provides a context for a number of disease causing mutations and for further dissection of the mechanisms by which CaM controls Kv7 function. PMID:23178170

  10. Effects of neferine on Kv4.3 channels expressed in HEK293 cells and ex vivo electrophysiology of rabbit hearts

    PubMed Central

    Wang, Chen; Chen, Yu-fang; Quan, Xiao-qing; Wang, Huan; Zhang, Rui; Xiao, Jun-hua; Wang, Jia-ling; Zhang, Cun-tai; Xiang, Ji-zhou; Tang, Qiang

    2015-01-01

    Aim: Neferine is an isoquinoline alkaloid isolated from seed embryos of Nelumbo nucifera (Gaertn), which has a variety of biological activities. In this study we examined the effects of neferine on Kv4.3 channels, a major contributor to the transient outward current (Ito) in rabbit heart, and on ex vivo electrophysiology of rabbit hearts. Methods: Whole-cell Kv4.3 currents were recorded in HEK293 cells expressing human cardiac Kv4.3 channels using patch-clamp technique. Arterially perfused wedges of rabbit left ventricles (LV) were prepared, and transmembrane action potentials were simultaneously recorded from epicardial (Epi) and endocardial (Endo) sites with floating microelectrodes together with transmural electrocardiography (ECG). Results: Neferine (0.1–100 μmol/L) dose-dependently and reversibly inhibited Kv4.3 currents (the IC50 value was 8.437 μmol/L, and the maximal inhibition at 100 μmol/L was 44.12%). Neferine (10 μmol/L) caused a positive shift of the steady-state activation curve of Kv4.3 currents, and a negative shift of the steady-state inactivation curve. Furthermore, neferine (10 μmol/L) accelerated the inactivation but not the activation of Kv4.3 currents, and markedly slowed the recovery of Kv4.3 currents from inactivation. Neferine-induced blocking of Kv4.3 currents was frequency-dependent. In arterially perfused wedges of rabbit LV, neferine (1, 3, and 10 μmol/L) dose-dependently prolonged the QT intervals and action potential durations (APD) at both Epi and Endo sites, and caused dramatic increase of APD10 at Epi sites. Conclusion: Neferine inhibits Kv4.3 channels likely by blocking the open state and inactivating state channels, which contributes to neferine-induced dramatic increase of APD10 at Epi sites of rabbit heart. PMID:26592512

  11. A New Regulatory Mechanism for Kv7.2 Protein During Neuropathy: Enhanced Transport from the Soma to Axonal Terminals of Injured Sensory Neurons

    PubMed Central

    Cisneros, Elsa; Roza, Carolina; Jackson, Nieka; López-García, José Antonio

    2015-01-01

    Kv7.2 channel expression has been reported to decrease in dorsal root ganglia (DRG) following the induction of a peripheral neuropathy while other experiments show that Kv7.2 accumulates in peripheral neuromas. The mechanisms underlying these novel expression patterns are poorly understood. Here we use immunofluorescence methods to analyze Kv7.2 protein expression changes in sensory neurons following peripheral axotomy and the potential role of axonal transport. Results indicate that DRG neurons express Kv7.2 in ~16% of neurons and that this number decreases by about 65% after axotomy. Damaged neurons were identified in DRG by application of the tracer Fluoro-ruby at the site of injury during surgery. Reduction of Kv7.2 expression was particularly strong in damaged neurons although some loss was also found in putative uninjured neurons. In parallel to the decrease in the soma of axotomized sensory neurons, Kv7.2 accumulated at neuromatose fiber endings. Blockade of axonal transport with either vinblastine (VLB) or colchicine (COL) abolished Kv7.2 redistribution in neuropathic animals. Channel distribution rearrangements did not occur following induction of inflammation in the hind paw. Behavioral tests indicate that protein rearrangements within sensory afferents are essential to the development of allodynia under neuropathic conditions. These results suggest that axotomy enhances axonal transport in injured sensory neurons, leading to a decrease of somatic expression of Kv7.2 protein and a concomitant accumulation in damaged fiber endings. Localized changes in channel expression patterns under pathological conditions may create novel opportunities for Kv7.2 channel openers to act as analgesics. PMID:26696829

  12. Specification of skeletal muscle differentiation by repressor element-1 silencing transcription factor (REST)-regulated Kv7.4 potassium channels.

    PubMed

    Iannotti, Fabio Arturo; Barrese, Vincenzo; Formisano, Luigi; Miceli, Francesco; Taglialatela, Maurizio

    2013-02-01

    Changes in the expression of potassium (K(+)) channels is a pivotal event during skeletal muscle differentiation. In mouse C(2)C(12) cells, similarly to human skeletal muscle cells, myotube formation increased the expression of K(v)7.1, K(v)7.3, and K(v)7.4, the last showing the highest degree of regulation. In C(2)C(12) cells, K(v)7.4 silencing by RNA interference reduced the expression levels of differentiation markers (myogenin, myosin heavy chain, troponinT-1, and Pax3) and impaired myotube formation and multinucleation. In K(v)7.4-silenced cells, the differentiation-promoting effect of the K(v)7 activator N-(2-amino-4-(4-fluorobenzylamino)-phenyl)-carbamic acid ethyl ester (retigabine) was abrogated. Expression levels for the repressor element-1 silencing transcription factor (REST) declined during myotube formation. Transcript levels for K(v)7.4, as well as for myogenin, troponinT-1, and Pax3, were reduced by REST overexpression and enhanced upon REST suppression by RNA interference. Four regions containing potential REST-binding sites in the 5' untranslated region and in the first intron of the K(v)7.4 gene were identified by bioinformatic analysis. Chromatin immunoprecipitation assays showed that REST binds to these regions, exhibiting a higher efficiency in myoblasts than in myotubes. These data suggest that K(v)7.4 plays a permissive role in skeletal muscle differentiation and highlight REST as a crucial transcriptional regulator for this K(+) channel subunit. PMID:23242999

  13. SU-E-J-109: Testing the KV Imaging Center Congruence with Radiation Isocenter of Small MLC and SRS Cone Field On Two Machines

    SciTech Connect

    Fu,; Chen, Y; Yu, Y; Liu, H

    2014-06-01

    Purpose: Orthogonal kV image pairs are used for target localization when fiducial markers are implanted. CBCT is used to verify cone SRS setup. Therefore it is necessary to evaluate the isocenter congruence between radiation fields and kV imaging center. This study used a simple method to evaluate the isocenter congruence, and compared the results for MLC and cone fields on two different Linacs. Methods: Varian OBI block was attached on the couch. It has a central 1mm BB with markers on three surfaces to align with laser. KV and MV images were taken at four cardinal angles. A 3x3cm2 MLC field and a 20mm cone field were irradiated respectively. On each kV image, the distance from BB center to the kV graticule center were measured. On the MV image of MLC field, the center of radiation field was determined manually, while for cone field, the Varian AM maintenance software was used to analyze the distance between BB and radiation field. The subtraction of the two distances gives the discrepancy between kV and radiation centers. Each procedure was repeated on five days at Trilogy and TrueBeam respectively. Results: The maximum discrepancy was found in the longitudinal direction at 180° gantry angel. It was 1.5±0.1mm for Trilogy and 0.6±0.1mm for TrueBeam. For Trilogy, although radiation center wobbled only 0.7mm and image center wobbled 0.8mm, they wobbled to the opposite direction. KV Pair using gantry 180° should be avoided in this case. Cone vs. kV isocenter has less discrepancy than MLC for Trilogy. Conclusion: Radiation isocenter of MLC and cone field is different, so is between Trilogy and TrueBeam. The method is simple and reproducible to check kV and radiation isocenter congruence.

  14. Proposal to amend existing operating permit for the Ault-Craig 345-kV and Hayden-Archer 230-kV transmission lines, Routt, Jackson and Larimer Counties, Colorado

    SciTech Connect

    1997-08-01

    The Western Area Power Administration, Rocky Mountain Region, is proposing to amend an existing US Forest Service operating permit for the Ault-Craig 345-kV and Hayden-Archer 230-kV transmission lines, which are located in Routt, jackson, and Larimer counties, Colorado. These transmission lines cross portions of the Roosevelt and Routt National Forests. The long-term use authorization Western is requesting from the Forest Service would be for the life of the Ault-Craig and Hayden-Archer transmission lines. This environmental assessment addresses those access road and right-of-way maintenance activities identified by Western that would be performed on Forest Service managed lands during the next approximately five years.

  15. Coexpression of auxiliary subunits KChIP and DPPL in potassium channel Kv4-positive nociceptors and pain-modulating spinal interneurons.

    PubMed

    Cheng, Chau-Fu; Wang, Wan-Chen; Huang, Chia-Yi; Du, Po-Hau; Yang, Jung-Hui; Tsaur, Meei-Ling

    2016-03-01

    Subthreshold A-type K(+) currents (ISA s) have been recorded from the somata of nociceptors and spinal lamina II excitatory interneurons, which sense and modulate pain, respectively. Kv4 channels are responsible for the somatodendritic ISA s. Accumulative evidence suggests that neuronal Kv4 channels are ternary complexes including pore-forming Kv4 subunits and two types of auxiliary subunits: K(+) channel-interacting proteins (KChIPs) and dipeptidyl peptidase-like proteins (DPPLs). Previous reports have shown Kv4.3 in a subset of nonpeptidergic nociceptors and Kv4.2/Kv4.3 in certain spinal lamina II excitatory interneurons. However, whether and which KChIP and DPPL are coexpressed with Kv4 in these ISA -expressing pain-related neurons is unknown. In this study we mapped the protein distribution of KChIP1, KChIP2, KChIP3, DPP6, and DPP10 in adult rat dorsal root ganglion (DRG) and spinal cord by immunohistochemistry. In the DRG, we found colocalization of KChIP1, KChIP2, and DPP10 in the somatic surface and cytoplasm of Kv4.3(+) nociceptors. KChIP3 appears in most Aβ and Aδ sensory neurons as well as a small population of peptidergic nociceptors, whereas DPP6 is absent in sensory neurons. In the spinal cord, KChIP1 is coexpressed with Kv4.3 in the cell bodies of a subset of lamina II excitatory interneurons, while KChIP1, KChIP2, and DPP6 are colocalized with Kv4.2 and Kv4.3 in their dendrites. Within the dorsal horn, besides KChIP3 in the inner lamina II and lamina III, we detected DPP10 in most projection neurons, which transmit pain signal to brain. The results suggest the existence of Kv4/KChIP/DPPL ternary complexes in ISA -expressing nociceptors and pain-modulating spinal interneurons. PMID:26239200

  16. Characterization of SEMA3A-Encoded Semaphorin as a Naturally Occurring Kv4.3 Protein Inhibitor and its Contribution to Brugada Syndrome

    PubMed Central

    Boczek, Nicole J.; Ye, Dan; Johnson, Eric K.; Wang, Wei; Crotti, Lia; Tester, David J.; Dagradi, Federica; Mizusawa, Yuka; Torchio, Margherita; Alders, Marielle; Giudicessi, John R.; Wilde, Arthur A.; Schwartz, Peter J.; Nerbonne, Jeanne M.; Ackerman, Michael J.

    2016-01-01

    Rational SEMA3A-encoded semaphorin is a chemorepellent that disrupts neural patterning in the nervous and cardiac systems. In addition, SEMA3A has an amino acid motif that is analogous to hanatoxin, an inhibitor of voltage-gated K+ channels. SEMA3A knockout mice exhibit an abnormal ECG pattern and are prone to ventricular arrhythmias and sudden cardiac death. Objective To determine whether SEMA3A is a naturally occurring protein inhibitor of Kv4.3 (Ito) channels and its potential contribution to Brugada syndrome (BrS). Methods and Results Kv4.3, Nav1.5, Cav1.2, or Kv4.2 were co-expressed or perfused with SEMA3A in HEK293 cells and electrophysiological properties were examined via whole-cell patch clamp technique. SEMA3A selectively altered Kv4.3 by significantly reducing peak current density without perturbing Kv4.3 cell-surface protein expression. SEMA3A also reduced Ito current density in cardiomyocytes derived from human induced pluripotent stem cells. Disruption of a putative toxin binding domain on Kv4.3 was used to assess physical interactions between SEMA3A and Kv4.3. These findings in combination with co-immunoprecipitations of SEMA3A and Kv4.3 revealed a potential direct binding interaction between these proteins. Comprehensive mutational analysis of SEMA3A was performed on 198 unrelated SCN5A-genotype negative patients with BrS and two rare SEMA3A missense mutations were identified. The SEMA3A mutations disrupted SEMA3A’s ability to inhibit Kv4.3 channels, resulting in a significant gain of Kv4.3 current compared to WT-SEMA3A. Conclusions This study is the first to demonstrate semaphorin3A as a naturally occurring protein that selectively inhibits Kv4.3 and SEMA3A as a possible BrS-susceptibility gene through a Kv4.3 gain-of-function mechanism. PMID:24963029

  17. Differential Regulation of Action Potential Shape and Burst-Frequency Firing by BK and Kv2 Channels in Substantia Nigra Dopaminergic Neurons

    PubMed Central

    Kimm, Tilia; Khaliq, Zayd M.

    2015-01-01

    Little is known about the voltage-dependent potassium currents underlying spike repolarization in midbrain dopaminergic neurons. Studying mouse substantia nigra pars compacta dopaminergic neurons both in brain slice and after acute dissociation, we found that BK calcium-activated potassium channels and Kv2 channels both make major contributions to the depolarization-activated potassium current. Inhibiting Kv2 or BK channels had very different effects on spike shape and evoked firing. Inhibiting Kv2 channels increased spike width and decreased the afterhyperpolarization, as expected for loss of an action potential-activated potassium conductance. BK inhibition also increased spike width but paradoxically increased the afterhyperpolarization. Kv2 channel inhibition steeply increased the slope of the frequency–current (f–I) relationship, whereas BK channel inhibition had little effect on the f–I slope or decreased it, sometimes resulting in slowed firing. Action potential clamp experiments showed that both BK and Kv2 current flow during spike repolarization but with very different kinetics, with Kv2 current activating later and deactivating more slowly. Further experiments revealed that inhibiting either BK or Kv2 alone leads to recruitment of additional current through the other channel type during the action potential as a consequence of changes in spike shape. Enhancement of slowly deactivating Kv2 current can account for the increased afterhyperpolarization produced by BK inhibition and likely underlies the very different effects on the f–I relationship. The cross-regulation of BK and Kv2 activation illustrates that the functional role of a channel cannot be defined in isolation but depends critically on the context of the other conductances in the cell. SIGNIFICANCE STATEMENT This work shows that BK calcium-activated potassium channels and Kv2 voltage-activated potassium channels both regulate action potentials in dopamine neurons of the substantia nigra

  18. Kir1.1 (ROMK) and Kv7.1 (KCNQ1/KvLQT1) are essential for normal gastric acid secretion: importance of functional Kir1.1.

    PubMed

    Vucic, Esad; Alfadda, Tariq; MacGregor, Gordon G; Dong, Ke; Wang, Tong; Geibel, John P

    2015-07-01

    Potassium channels comprise the apical leak pathway supplying extracellular K(+) for exchange with protons by the gastric H(+), K(+)-ATPase and provide potential therapeutic targets for inhibiting gastric acid secretion. The Kir1.1 (ROMK) potassium channel mediates the high capacity K(+) recycling necessary for NaCl reabsorption in the thick ascending limb of the kidney, and this channel exhibits functional and regulatory characteristic well suited for K(+) recycling by gastric parietal cells. We report here that Kir1.1 channels are required for gastric acid secretion and that this channel participates with Kv7.1 (KCNQ1/KvLQT1) in the potassium recycling process. We show that Kir1.1 colocalizes with the β-subunit of H(+), K(+)-ATPase in gastric parietal cells of Kir1.1 wild-type mice. In Kir1.1-deficient mice, gastric mucosal morphology, as well as parietal cell number, proliferation index, and ultrastructure were normal but secretagogue-stimulated gastric acid secretion in whole stomach and perfused gastric glands was absent. Luminal application of potassium-restored acid secretion in perfused gastric glands from Kir1.1-deficient as well as barium-blocked wild-type mice. In wild-type mice, both luminal Tertiapin-Q, an inhibitor of Kir1.1, as well as XE991, an inhibitor of Kv7.1, reduced proton secretion. We propose that Kir1.1 and Kv7.1 channels collaborate in potassium and current recycling across the apical pole of parietal cells. PMID:25127675

  19. N-Terminally extended analogues of the K⁺ channel toxin from Stichodactyla helianthus as potent and selective blockers of the voltage-gated potassium channel Kv1.3.

    PubMed

    Chang, Shih C; Huq, Redwan; Chhabra, Sandeep; Beeton, Christine; Pennington, Michael W; Smith, Brian J; Norton, Raymond S

    2015-06-01

    The voltage-gated potassium channel Kv1.3 is an important target for the treatment of autoimmune diseases and asthma. Blockade of Kv1.3 by the sea anemone peptide K⁺-channel toxin from Stichodactyla helianthus (ShK) inhibits the proliferation of effector memory T lymphocytes and ameliorates autoimmune diseases in animal models. However, the lack of selectivity of ShK for Kv1.3 over the Kv1.1 subtype has driven a search for Kv1.3-selective analogues. In the present study, we describe N-terminally extended analogues of ShK that contain a negatively-charged Glu, designed to mimic the phosphonate adduct in earlier Kv1.3-selective analogues, and consist entirely of common protein amino acids. Molecular dynamics simulations indicated that a Trp residue at position [-3] of the tetrapeptide extension could form stable interactions with Pro377 of Kv1.3 and best discriminates between Kv1.3 and Kv1.1. This led to the development of ShK with an N-terminal Glu-Trp-Ser-Ser extension ([EWSS]ShK), which inhibits Kv1.3 with an IC₅₀ of 34 pm and is 158-fold selective for Kv1.3 over Kv1.1. In addition, [EWSS]ShK is more than 2900-fold more selective for Kv1.3 over Kv1.2 and KCa3.1 channels. As a highly Kv1.3-selective analogue of ShK based entirely on protein amino acids, which can be produced by recombinant expression, this peptide is a valuable addition to the complement of therapeutic candidates for the treatment of autoimmune diseases. PMID:25864722

  20. Performance characteristics of the Atlas 60 kV, 60 kJ plastic capacitors

    SciTech Connect

    Reass, W.; Bennet, G.; Bowman, D.; Lopez, E.; Monroe, M.; Parsons, W.

    1997-12-01

    This paper provides the performance data of Atlas plastic capacitors as supplied by Maxwell Technologies and Aerovox Corporation. The fiberglass cases at 13 inches high by 29 inches wide and 28 inches in depth with a 2 inch by 18 inch bushing on each end. Two styles of the 33.5uF capacitors have been evaluated for Atlas use, a conventional paper-foil and a self-healing metalized-paper and plastic dielectric design. A test program to capacitor failure, is being used to evaluate capacitor lifetime at full voltage (60 kV) and a nominal 15% reversal. With the Atlas parameters, peak currents of {approximately} 340 kA are realized. In anticipation of faults, capacitors are capable, specified, and tested for 700 kA performance. Accurate methods are also utilized to determine capacitor inductance, less than 20 nH. The results of the various capacitor testing programs will be presented in addition to future directives for their R and D efforts.

  1. Analysis of native biological surfaces using a 100kV Massive Gold Cluster Source

    PubMed Central

    Fernandez-Lima, Francisco A.; Post, Jeremy; DeBord, John D.; Eller, Michael J.; Verkhoturov, Stanislav V.; Della-Negra, Serge; Woods, Amina S.; Schweikert, Emile A.

    2011-01-01

    In the present work, the advantages of a new, 100kV platform equipped with a massive gold cluster source for the analysis of native biological surfaces are shown. Inspection of the molecular ion emission as a function of projectile size demonstrate a secondary ion yield increase of ~100x for 520 keV Au400+4 as compared to 130 keV Au3+1 and 43 keV C60. In particular, yields of tens of percent of molecular ions per projectile impact for the most abundant components can be observed with the 520 keV Au400+4 probe, respectively. A comparison between 520 keV Au400+4 ToF-SIMS and MALDI-MS data showed a similar pattern and similar relative intensities of lipids’ components across a rat brain sagittal section. The abundant secondary ion yields of analyte-specific ions makes 520 keV Au4004+ projectiles an attractive probe for sub-μm molecular mapping of native surfaces. PMID:21967684

  2. The 345 kV underground/underwater Long Island Sound cable project

    SciTech Connect

    Grzan, J.; Hahn, E.I. ); Casalaina, R.V.; Kansog, J.O.C. )

    1993-07-01

    A high voltage underground/underwater cable system was installed to increase the transmission capacity from the mainland of New York to Long Island. In terms of weight and diameter, the self-contained, fluid-filled (SCFF) cable used for the underwater portion of the project is the largest underwater cable in the world. The use of high-pressure, fluid-filled (HPFF) pipe-type cable on the land portion represents the largest application of paper-polypropylene-paper (PPP) insulated cable in the United States. State-of-the-art technologies were implemented in the use of fiber optic cables for relay protection and SCADA/RTU, temperature monitoring and leak detection systems, SF[sub 6] gas-insulated substations, and underwater cable laying and embedment techniques. This paper discusses the design and installation of a 750 MVA, 43 km (26.6 mi), 345 kV underground/underwater electric transmission system installed by the New York Power Authority (NYPA).

  3. Experimental Studies on Cryogenic System for 22.9 KV Hts Cable Sytem

    NASA Astrophysics Data System (ADS)

    Sohn, S. H.; Lim, J. H.; Yang, H. S.; Kim, D. L.; Ryoo, H. S.; Kim, C. D.; Kim, D. H.; Lee, S. K.; Hwang, S. D.

    2008-03-01

    In terms of high transmission capacity with lower voltage, a high temperature superconducting (HTS) cable system is a very attractive challenge for utilities. However, the concomitant cryogenic system for the HTS cable system is one of the tantalizing problems in the operation. The reliability and maintainability of cryogenic system are the key issues to apply it to the real electric power grid. Korea Electric Power Corporation (KEPCO) is making an attempt to verify the applicability of the HTS cable system to improve the efficiency of electric power industry. Since May 2006, a 22.9-kV, 50-MVA, 3-phase, 100-m class HTS cable system with an open cooling system has been operated at the KEPCO Gochang test yard. Concurrently, another HTS cable verification test with the same electrical specification and an hybrid cooling system has been carried out by LS Cable (LSC) Ltd in close proximity to the KEPCO's HTS cable system within Gochang test yard. KEPCO conducts the operation of the open cooling system, and is evaluating the hybrid system of LSC with respect to facility performance and usability. This paper compares the cryogenic performance of both HTS cable systems and discusses cooling test results such as step response.

  4. Reclamation and revegetation along a 250-mile 500 kV power line corridor

    SciTech Connect

    Griffith, E.F.

    1990-12-31

    Reclamation and revegetation success along the Montana Power Company Colstrip to Deepdale 500 kV powerline was monitored from 1985 through 1989. Followup revegetation was hampered by inadequate road and tower site earth work as a result of premature release of the reclamation bond. Drought conditions in 1985 and 1988 seriously affected revegetation efforts, especially on difficult sites. Weed control improved markedly, and all ROW areas are weed free except those next to infested range and cropland where a major cooperative effort is needed to control weeds. Access roads in previously unroaded areas resulted in changes of access, management practices, and cattle grazing behavior, all of which adversely affected revegetation success. Erosion, exacerbated by undersized culverts, the presence of very erodible soils, the occasional untrained contractor, is now the major ROW maintenance concern. Observation of the revegetation efforts clearly shows the need for adequate and appropriate reclamation by qualified equipment operators up front, and a patient, measured effort of continued revegetation, weed control and erosion control over the long term. The bond period of 5 years may not be long enough to achieve the 90 percent revegetation success required by the permit. Future projects guidelines should consider look to a flexible timeframe and more realistic vegetation requirements to account for the effects of non-construction related impacts.

  5. Low stored energy 100 kV regulator for ion sources at LANSCE

    SciTech Connect

    Jacobson, E.G.; Haffner, R.L.; Ingalls, W.B.; Meyer, B.J.; Stelzer, J.E.

    1998-12-31

    To minimize accelerating column damage caused by uncontrolled energy release during arc-downs, it is desirable to minimize the available stored electrical energy. For the Los Alamos Neutron Science Center (LANSCE) H{sup {minus}} ion sources, the stored energy includes, in addition to the charge in the power supply output capacitance, the charge on the electronics racks. They are supported and insulated from ground by PVC pipe and have a capacitance to ground of approximately 900 pf. In 1988 (LANSCE) personnel designed a high-voltage current source using a low-stored-energy power supply and planar triode with the goal of eliminating uncontrolled release of charge stored in the power supply. Construction and testing were performed intermittently as resources permitted until 1993. When work on the Short Pulse Spallation Source (SPSS) started on the LANSCE Ion Source Test Stand (ISTS) it was recognized that a higher current power supply would be needed and work resumed on the regulator circuitry. A 120 kV power supply having low output capacitance, and a planar triode have been used to supply 40 mA, 120 Hz, 12% duty-factor current for the ISTS beam. The triode`s cathode current is controlled by circuitry operating both at power-supply voltage level and at ground level via a fiber optic link. Voltage droop is approximately 600 V during the 1 ms beam pulse. The authors present the status of the regulator and its special challenges.

  6. Characterized Discharge Current Waveforms of Metal Oxide Surge Arresters on 77kV Power Systems

    NASA Astrophysics Data System (ADS)

    Miyachi, Iwao; Yoda, Masayuki; Ueda, Toshiaki; Kanehara, Kazuto

    The behaviors of metal oxide surge arresters installed at substation terminal, generally without spark gaps are well-known by the excellent performances of automatic fault clearance and of reliable overvoltage protection of substation apparatus. As the result of four-year research experiences on the 77kV operating power systems including observed substations T and F halfway between the protection terminals, authors have proved the characterized discharge current of those arresters can be divided into three types of waveforms such as instantaneous heavy pulse, low level lingering continuance and medium continued damping oscillation. The negative lightning discharge with subsequent multiple strokes against de-energized power lines may be the distinct and frequent causes of damping current oscillation of surge arresters. Moreover, the negative discharge current on the sound phase has been encountered not a little in the case of related double-line-to-ground fault. In this paper, the combination of those parameters is clearly expressed through several representing figures of arrester current waveforms together with those of corresponding voltage of surge fronts and 60Hz voltage transients if available.

  7. Assessment of 69 kV Underground Cable Thermal Ratings using Distributed Temperature Sensing

    NASA Astrophysics Data System (ADS)

    Stowers, Travis

    Underground transmission cables in power systems are less likely to experience electrical faults, however, resulting outage times are much greater in the event that a failure does occur. Unlike overhead lines, underground cables are not self-healing from flashover events. The faulted section must be located and repaired before the line can be put back into service. Since this will often require excavation of the underground duct bank, the procedure to repair the faulted section is both costly and time consuming. These added complications are the prime motivators for developing accurate and reliable ratings for underground cable circuits. This work will review the methods by which power ratings, or ampacity, for underground cables are determined and then evaluate those ratings by making comparison with measured data taken from an underground 69 kV cable, which is part of the Salt River Project (SRP) power subtransmission system. The process of acquiring, installing, and commissioning the temperature monitoring system is covered in detail as well. The collected data are also used to evaluate typical assumptions made when determining underground cable ratings such as cable hot-spot location and ambient temperatures. Analysis results show that the commonly made assumption that the deepest portion of an underground power cable installation will be the hot-spot location does not always hold true. It is shown that distributed cable temperature measurements can be used to locate the proper line segment to be used for cable ampacity calculations.

  8. A 800 kV compact peaking capacitor for nanosecond generator.

    PubMed

    Jia, Wei; Chen, Zhiqiang; Tang, Junping; Chen, Weiqing; Guo, Fan; Sun, Fengrong; Li, Junna; Qiu, Aici

    2014-09-01

    An extremely compact high voltage peaking capacitor is developed. The capacitor has a pancake structure with a diameter of 315 mm, a thickness of 59 mm, and a mass of 6.1 kg. The novel structural design endows the capacitor with a better mechanical stability and reliability under hundreds of kilovolts pulse voltage and an inner gas pressure of more than 1.5 MPa. The theoretical value of the capacitor self-inductance is near to 17 nH. Proved by series of electrical experiments, the capacitor can endure a high-voltage pulse with a rise time of about 20 ns, a half-width duration of around 25 ns, and an amplitude of up to 800 kV in a single shot model. When the capacitor was used in an electromagnetic pulse simulator as a peaking capacitor, the rise time of the voltage pulse can be reduced from 20 ns to less than 3 ns. The practical value of the capacitor's inductance deduced from the experimental date is no more than 25 nH. PMID:25273753

  9. Field evaluation of 69-kV outdoor Polysil reg sign insulators

    SciTech Connect

    Richenbacher, A.G. )

    1990-03-01

    This report, together with previous interim reports, documents and summarizes the field performance of 69 kV Polysil{reg sign} insulators during the field trial period from January, 1983 to December, 1988. These insulators were manufactured for the Electric Power Research Institute by Lindsey Industries in 1979. A description of the insulator development and manufacturing process is contained in the EPRI Final Report EL1281-1. Following their manufacture, the insulators were delivered, in the form of test racks of 17 Polysil{reg sign} insulators and one porcelain insulator, to twelve United States utilities and the Instituto de Investigaciones Electricas (IIE) in Mexico. These racks were subsequently installed and energized at twenty-five test sites during the latter half of 1979 and early 1980 by the project participants for the purpose of analyzing the outdoor field performance of these insulators and comparing the relative effect on performance of variations in composition, coating, electrical grading method, and shape represented by individual insulators within the test racks. This report documents the effects of the various Polysil{reg sign} insulator parameters on electrical performance in the field over a specific period of 6 years. However, the insulators had been energized for approximately 3 years prior to the initiation of this project and, although specific performance data is not available for that time period, the overall effects of field exposure for that additional time period (total of 9 years) are seen in the results of this report. 3 refs., 23 figs., 9 tabs.

  10. Screening and cDNA Cloning of Kv1 Potassium Channel Toxins in Sea Anemones

    PubMed Central

    Yamaguchi, Yoshikazu; Hasegawa, Yuichi; Honma, Tomohiro; Nagashima, Yuji; Shiomi, Kazuo

    2010-01-01

    When 21 species of sea anemones were screened for Kv1 potassium channel toxins by competitive inhibition of the binding of 125I-α-dendrotoxin to rat synaptosomal membranes, 11 species (two species of Actiniidae, one species of Hormathiidae, five species of Stichodactylidae and three species of Thalassianthidae) were found to be positive. Furthermore, full-length cDNAs encoding type 1 potassium channel toxins from three species of Stichodactylidae and three species of Thalassianthidae were cloned by a combination of RT-PCR, 3′RACE and 5′RACE. The precursors of these six toxins are commonly composed of signal peptide, propart and mature peptide portions. As for the mature peptide (35 amino acid residues), the six toxins share more than 90% sequence identities with one another and with κ1.3-SHTX-She1a (Shk) from Stichodactyla helianthus but only 34–63% identities with the other type 1 potassium channel toxins. PMID:21339955

  11. New Waddell-Westwing 230-kV transmission project, Maricopa County, Arizona: Environmental assessment

    SciTech Connect

    Not Available

    1988-10-01

    The Western Area Power Administration (Western) proposes to construct a new 230 kilovolt (kV) transmission line in Maricopa County, Arizona, that would extend from the New Waddell Dam on the Agua Fria River, to the Westwing Substation, about 10.5 miles southwest of the dam. The project area is roughly 45 miles northwest of the Phoenix metropolitan area. Western will complete the National Environmental Policy Act of 1969 (NEPA) process for the line. A final environmental impact statement (EIS) was prepared by BuRec to describe the environmental impacts associated with alternatives for the construction and operation of the Regulatory Storage Division of the Central Arizona Project (CAP). The EIS was supported by 23 technical reports that covered planning, design, public involvement, social and environmental impact assessment, economics, and hydrological analysis. Since the proposed transmission system differs only slightly from that described in the EIS, Western incorporates by reference the EIS and its 23 supporting technical reports into this environmental assessment (EA). In August 1985, BuRec completed an EA on modifications to the New Waddell Dam Project design plans and issued a FONSI on these modifications; the EA did not cover the transmission line. The purpose of this EA is to document the potential environmental effects of the proposed construction of the transmission line and switchyards in order to determine if an EIS is needed or if a finding of no significant impact (FONSI) is indicated. 28 refs., 5 figs., 1 tab.

  12. An Improved Thermal Conductivity Polyurethane Composite for a Space Borne 20KV Power Supply

    NASA Technical Reports Server (NTRS)

    Shapiro, Andrew A.; Haque, Inam

    2005-01-01

    This effort was designed to find a way to reduce the temperature rise of critical components of a 20KV High Voltage Power Supply (HVPS) by improving the overall thermal conductivity of the encapsulated modules. Three strategies were evaluated by developing complete procedures, preparing samples, and performing tests. The three strategies were: 1. Improve the thermal conductivity of the polyurethane encapsulant through the addition of thermally conductive powder while minimizing impact on other characteristics of the encapsulant. 2. Improve the thermal conductivity of the polyurethane encapsulated assembly by the addition of a slab of thermally conductive, electrically insulating material, which is to act as a heat spreader. 3. Employ a more thermally conductive substrate (Al203) with the existing encapsulation scheme. The materials were chosen based on the following criteria: high dielectric breakdown strength; high thermal conductivity, ease of manufacturing, high compliance, and other standard space qualified materials properties (low out-gassing, etc.). An optimized cure was determined by a statistical design of experiments for both filled and unfilled materials. The materials were characterized for the desired properties and a complete process was developed and tested. The thermal performance was substantially improved and the strategies may be used for space flight.

  13. Reconstitution of a Kv Channel into Lipid Membranes for Structural and Functional Studies

    PubMed Central

    Shi, Liang; Jiang, Qiu-Xing

    2013-01-01

    To study the lipid-protein interaction in a reductionistic fashion, it is necessary to incorporate the membrane proteins into membranes of well-defined lipid composition. We are studying the lipid-dependent gating effects in a prototype voltage-gated potassium (Kv) channel, and have worked out detailed procedures to reconstitute the channels into different membrane systems. Our reconstitution procedures take consideration of both detergent-induced fusion of vesicles and the fusion of protein/detergent micelles with the lipid/detergent mixed micelles as well as the importance of reaching an equilibrium distribution of lipids among the protein/detergent/lipid and the detergent/lipid mixed micelles. Our data suggested that the insertion of the channels in the lipid vesicles is relatively random in orientations, and the reconstitution efficiency is so high that no detectable protein aggregates were seen in fractionation experiments. We have utilized the reconstituted channels to determine the conformational states of the channels in different lipids, record electrical activities of a small number of channels incorporated in planar lipid bilayers, screen for conformation-specific ligands from a phage-displayed peptide library, and support the growth of 2D crystals of the channels in membranes. The reconstitution procedures described here may be adapted for studying other membrane proteins in lipid bilayers, especially for the investigation of the lipid effects on the eukaryotic voltage-gated ion channels. PMID:23892292

  14. Synthesis and biological evaluation of negative allosteric modulators of the Kv11.1(hERG) channel.

    PubMed

    Yu, Zhiyi; van Veldhoven, Jacobus P D; 't Hart, Ingrid M E; Kopf, Adrian H; Heitman, Laura H; IJzerman, Adriaan P

    2015-12-01

    We synthesized and evaluated a series of compounds for their allosteric modulation at the Kv11.1 (hERG) channel. Most compounds were negative allosteric modulators of [(3)H]dofetilide binding to the channel, in particular 7f, 7h-j and 7p. Compounds 7f and 7p were the most potent negative allosteric modulators amongst all ligands, significantly increasing the dissociation rate of dofetilide in the radioligand kinetic binding assay, while remarkably reducing the affinities of dofetilide and astemizole in a competitive displacement assay. Additionally, both 7f and 7p displayed peculiar displacement characteristics with Hill coefficients significantly distinct from unity as shown by e.g., dofetilide, further indicative of their allosteric effects on dofetilide binding. Our findings in this investigation yielded several promising negative allosteric modulators for future functional and clinical research with respect to their antiarrhythmic propensities, either alone or in combination with known Kv11.1 blockers. PMID:26519929

  15. Docking ellipticine to the V-VI transmembrane domain of the Kv11.1 potassium channel

    NASA Astrophysics Data System (ADS)

    Lipscomb, Dawn; Brancaleon, Lorenzo; Gentile, S.

    2011-03-01

    Ellipticines such as 9-methoxy-N-2-methylellipticinium acetate (MMEA) and 9-hydroxy-N-2-methylellipticinium acetate (NMEA, Celiptium ) are antineoplastic drugs exerting their selective cytotoxicity against leukemia and endometrial carcinoma. Ellipticine's action is also related to severe physical side effects, but the link between undesired effects and pharmacological application is not well understood. We investigated the binding of Ellipticine derivatives with the Kv11.1 potassium ion channel using Autodock and revealed that hydroxyellipticinium derivatives provide binding configurations with Kv11.1, but the energy, location and estimated dissociation constant varied. The binding energy is as follows: Chloroceliptium (-6.60 kcal/mol) Celiptium (- 6.37 kcal / mol) > Methoxyceliptium (- 6.20 kcal / mol) Datelliptium (-6.08 kcal/mol). The data shows that some configurations enable these molecules to bridge among channel subunits, thus potentially inhibiting the flow of ions.

  16. Altered distribution of juxtaparanodal kv1.2 subunits mediates peripheral nerve hyperexcitability in type 2 diabetes mellitus.

    PubMed

    Zenker, Jennifer; Poirot, Olivier; de Preux Charles, Anne-Sophie; Arnaud, Estelle; Médard, Jean-Jacques; Lacroix, Catherine; Kuntzer, Thierry; Chrast, Roman

    2012-05-30

    Peripheral nerve hyperexcitability (PNH) is one of the distal peripheral neuropathy phenotypes often present in patients affected by type 2 diabetes mellitus (T2DM). Through in vivo and ex vivo electrophysiological recordings in db/db mice, a model of T2DM, we observed that, in addition to reduced nerve conduction velocity, db/db mice also develop PNH. By using pharmacological inhibitors, we demonstrated that the PNH is mediated by the decreased activity of K(v)1-channels. In agreement with these data, we observed that the diabetic condition led to a reduced presence of the K(v)1.2-subunits in juxtaparanodal regions of peripheral nerves in db/db mice and in nerve biopsies from T2DM patients. Together, these observations indicate that the T2DM condition leads to potassium channel-mediated PNH, thus identifying them as a potential drug target to treat some of the DPN related symptoms. PMID:22649228

  17. 1.9 kV AlGaN/GaN Lateral Schottky Barrier Diodes on Silicon

    DOE PAGESBeta

    Zhu, Mingda; Song, Bo; Qi, Meng; Hu, Zongyang; Nomoto, Kazuki; Yan, Xiaodong; Cao, Yu; Johnson, Wayne; Kohn, Erhard; Jena, Debdeep; et al

    2015-02-16

    In this letter, we present AlGaN/GaN lateral Schottky barrier diodes on silicon with recessed anodes and dual field plates. A low specific on-resistance RON,SP (5.12 mΩ · cm2), a low turn-on voltage (<0.7 V) and a high reverse breakdown voltage BV (>1.9 kV), were simultaneously achieved in devices with a 25 μm anode/cathode separation, resulting in a power figure-of-merit (FOM) BV2/RON,SP of 727 MW·cm2. The record high breakdown voltage of 1.9 kV is attributed to the dual field plate structure.

  18. Secondary structure, stability and tetramerisation of recombinant K(V)1.1 potassium channel cytoplasmic N-terminal fragment.

    PubMed

    Abbott, G W; Bloemendal, M; Van Stokkum, I H; Mercer, E A; Miller, R T; Sewing, S; Wolters, M; Pongs, O; Srai, S K

    1997-08-15

    The recombinant N-terminal fragment (amino acids 14-162) of a tetrameric voltage-gated potassium channel (K(V)1.1) has been studied using spectroscopic techniques. Evidence is presented that it forms a tetramer in aqueous solution, whereas when solubilised in 1% Triton X-100 it remains monomeric. The secondary structure content of both monomeric and tetrameric K(V)1.1 N-terminal fragment has been estimated from FTIR and CD spectroscopy to be 20-25% alpha-helix, 20-25% beta-sheet, 20% turns and 30-40% random coil. Solubilisation of the protein in detergent is shown by hydrogen-deuterium exchange analysis to alter tertiary structure rather than secondary structure and this may be the determining factor in tetramerisation ability. Using molecular modelling we propose a supersecondary structure consisting of two structural domains. PMID:9300810

  19. 1.9 kV AlGaN/GaN Lateral Schottky Barrier Diodes on Silicon

    SciTech Connect

    Zhu, Mingda; Song, Bo; Qi, Meng; Hu, Zongyang; Nomoto, Kazuki; Yan, Xiaodong; Cao, Yu; Johnson, Wayne; Kohn, Erhard; Jena, Debdeep; Xing, Grace Huili

    2015-02-16

    In this letter, we present AlGaN/GaN lateral Schottky barrier diodes on silicon with recessed anodes and dual field plates. A low specific on-resistance RON,SP (5.12 mΩ · cm2), a low turn-on voltage (<0.7 V) and a high reverse breakdown voltage BV (>1.9 kV), were simultaneously achieved in devices with a 25 μm anode/cathode separation, resulting in a power figure-of-merit (FOM) BV2/RON,SP of 727 MW·cm2. The record high breakdown voltage of 1.9 kV is attributed to the dual field plate structure.

  20. Development and laboratory testing of a 138-kV PPP-insulated joint for commercial application

    SciTech Connect

    Walldorf, S.P. ); Chu, H.; Elbadaly, H. )

    1990-04-01

    This paper describes the design, development and laboratory testing of a high voltage PPP (paper polypropylene/paper laminate) insulated joint for commercial application on 138-kV PPP-insulated cable. The design approach taken is conservative and addresses the typical variations in field conditions and in skill and workmanship of the splicing. Joint construction details, including choice of connector, taping structure, and joint mechanical reinforcement, are discussed. The test criteria are described and results are presented.

  1. Verifications on the Installed Number of Surge Arresters for 66-500kV Power Systems in Japan

    NASA Astrophysics Data System (ADS)

    Shirakawa, Shingo; Kobayashi, Takayuki; Tanae, Hiroshi; Takamatsu, Tadashi; Kumai, Toshiya; Imura, Hajime; Nishimura, Seisuke

    Surge arresters have contributed to supply of electric power by suppressing lightning surge on transmission lines in case of lightning phenomena. Surge arresters using zinc oxide (ZnO) elements are suitable for insulation coordination, and are enable to reduce LIWV (Lightning Impulse Withstand Voltage) and construction cost of power systems. This paper describes applications and results of surge arresters to verify effectiveness of surge protection for 66-500kV power systems in Japan.

  2. Blocking KV1.3 channels inhibits Th2 lymphocyte function and treats a rat model of asthma.

    PubMed

    Koshy, Shyny; Huq, Redwan; Tanner, Mark R; Atik, Mustafa A; Porter, Paul C; Khan, Fatima S; Pennington, Michael W; Hanania, Nicola A; Corry, David B; Beeton, Christine

    2014-05-01

    Allergic asthma is a chronic inflammatory disease of the airways. Of the different lower airway-infiltrating immune cells that participate in asthma, T lymphocytes that produce Th2 cytokines play important roles in pathogenesis. These T cells are mainly fully differentiated CCR7(-) effector memory T (TEM) cells. Targeting TEM cells without affecting CCR7(+) naïve and central memory (TCM) cells has the potential of treating TEM-mediated diseases, such as asthma, without inducing generalized immunosuppression. The voltage-gated KV1.3 potassium channel is a target for preferential inhibition of TEM cells. Here, we investigated the effects of ShK-186, a selective KV1.3 channel blocker, for the treatment of asthma. A significant proportion of T lymphocytes in the lower airways of subjects with asthma expressed high levels of KV1.3 channels. ShK-186 inhibited the allergen-induced activation of peripheral blood T cells from those subjects. Immunization of F344 rats against ovalbumin followed by intranasal challenges with ovalbumin induced airway hyper-reactivity, which was reduced by the administration of ShK-186. ShK-186 also reduced total immune infiltrates in the bronchoalveolar lavage and number of infiltrating lymphocytes, eosinophils, and neutrophils assessed by differential counts. Rats with the ovalbumin-induced model of asthma had elevated levels of the Th2 cytokines IL-4, IL-5, and IL-13 measured by ELISA in their bronchoalveolar lavage fluids. ShK-186 administration reduced levels of IL-4 and IL-5 and induced an increase in the production of IL-10. Finally, ShK-186 inhibited the proliferation of lung-infiltrating ovalbumin-specific T cells. Our results suggest that KV1.3 channels represent effective targets for the treatment of allergic asthma. PMID:24644290

  3. Blocking KV1.3 Channels Inhibits Th2 Lymphocyte Function and Treats a Rat Model of Asthma*

    PubMed Central

    Koshy, Shyny; Huq, Redwan; Tanner, Mark R.; Atik, Mustafa A.; Porter, Paul C.; Khan, Fatima S.; Pennington, Michael W.; Hanania, Nicola A.; Corry, David B.; Beeton, Christine

    2014-01-01

    Allergic asthma is a chronic inflammatory disease of the airways. Of the different lower airway-infiltrating immune cells that participate in asthma, T lymphocytes that produce Th2 cytokines play important roles in pathogenesis. These T cells are mainly fully differentiated CCR7− effector memory T (TEM) cells. Targeting TEM cells without affecting CCR7+ naïve and central memory (TCM) cells has the potential of treating TEM-mediated diseases, such as asthma, without inducing generalized immunosuppression. The voltage-gated KV1.3 potassium channel is a target for preferential inhibition of TEM cells. Here, we investigated the effects of ShK-186, a selective KV1.3 channel blocker, for the treatment of asthma. A significant proportion of T lymphocytes in the lower airways of subjects with asthma expressed high levels of KV1.3 channels. ShK-186 inhibited the allergen-induced activation of peripheral blood T cells from those subjects. Immunization of F344 rats against ovalbumin followed by intranasal challenges with ovalbumin induced airway hyper-reactivity, which was reduced by the administration of ShK-186. ShK-186 also reduced total immune infiltrates in the bronchoalveolar lavage and number of infiltrating lymphocytes, eosinophils, and neutrophils assessed by differential counts. Rats with the ovalbumin-induced model of asthma had elevated levels of the Th2 cytokines IL-4, IL-5, and IL-13 measured by ELISA in their bronchoalveolar lavage fluids. ShK-186 administration reduced levels of IL-4 and IL-5 and induced an increase in the production of IL-10. Finally, ShK-186 inhibited the proliferation of lung-infiltrating ovalbumin-specific T cells. Our results suggest that KV1.3 channels represent effective targets for the treatment of allergic asthma. PMID:24644290

  4. Draft Genome Sequence of Klebsiella variicola Strain KV321 Isolated from Rhizosphere Soil of Pisolithus tinctorius-Eucalyptus Mycorrhiza.

    PubMed

    Jiang, Shao-Feng; Liu, Yi; Xiao, Mi-Yun; Ruan, Chu-Jin; Lu, Zu-Jun

    2016-01-01

    The draft genome sequences of Klebsiella variicola strain KV321, which was isolated from rhizosphere soil of Pisolithus tinctorius-Eucalyptus mycorrhiza, are reported here. The genome sequences contain genes involved in ABC transporter function in multiple-antibiotic drug resistance and colonization. This genomic analysis will help understand the genomic basis of K. variicola virulence genes and how the genes play a part in its interaction with other living organisms. PMID:27445373

  5. Temperature Effects on Kinetics of KV11.1 Drug Block Have Important Consequences for In Silico Proarrhythmic Risk Prediction.

    PubMed

    Windley, Monique J; Mann, Stefan A; Vandenberg, Jamie I; Hill, Adam P

    2016-07-01

    Drug block of voltage-gated potassium channel subtype 11.1 human ether-a-go-go related gene (Kv11.1) (hERG) channels, encoded by the KCNH2 gene, is associated with reduced repolarization of the cardiac action potential and is the predominant cause of acquired long QT syndrome that can lead to fatal cardiac arrhythmias. Current safety guidelines require that potency of KV11.1 block is assessed in the preclinical phase of drug development. However, not all drugs that block KV11.1 are proarrhythmic, meaning that screening on the basis of equilibrium measures of block can result in high attrition of potentially low-risk drugs. The basis of the next generation of drug-screening approaches is set to be in silico risk prediction, informed by in vitro mechanistic descriptions of drug binding, including measures of the kinetics of block. A critical issue in this regard is characterizing the temperature dependence of drug binding. Specifically, it is important to address whether kinetics relevant to physiologic temperatures can be inferred or extrapolated from in vitro data gathered at room temperature in high-throughout systems. Here we present the first complete study of the temperature-dependent kinetics of block and unblock of a proarrhythmic drug, cisapride, to KV11.1. Our data highlight a complexity to binding that manifests at higher temperatures and can be explained by accumulation of an intermediate, non-blocking encounter-complex. These results suggest that for cisapride, physiologically relevant kinetic parameters cannot be simply extrapolated from those measured at lower temperatures; rather, data gathered at physiologic temperatures should be used to constrain in silico models that may be used for proarrhythmic risk prediction. PMID:27190211

  6. Draft Genome Sequence of Klebsiella variicola Strain KV321 Isolated from Rhizosphere Soil of Pisolithus tinctorius-Eucalyptus Mycorrhiza

    PubMed Central

    Jiang, Shao-feng; Liu, Yi; Xiao, Mi-yun; Ruan, Chu-jin

    2016-01-01

    The draft genome sequences of Klebsiella variicola strain KV321, which was isolated from rhizosphere soil of Pisolithus tinctorius-Eucalyptus mycorrhiza, are reported here. The genome sequences contain genes involved in ABC transporter function in multiple-antibiotic drug resistance and colonization. This genomic analysis will help understand the genomic basis of K. variicola virulence genes and how the genes play a part in its interaction with other living organisms. PMID:27445373

  7. Molecular template for a voltage sensor in a novel K+ channel. I. Identification and functional characterization of KvLm, a voltage-gated K+ channel from Listeria monocytogenes.

    PubMed

    Santos, Jose S; Lundby, Alicia; Zazueta, Cecilia; Montal, Mauricio

    2006-09-01

    The fundamental principles underlying voltage sensing, a hallmark feature of electrically excitable cells, are still enigmatic and the subject of intense scrutiny and controversy. Here we show that a novel prokaryotic voltage-gated K(+) (Kv) channel from Listeria monocytogenes (KvLm) embodies a rudimentary, yet robust, sensor sufficient to endow it with voltage-dependent features comparable to those of eukaryotic Kv channels. The most conspicuous feature of the KvLm sequence is the nature of the sensor components: the motif is recognizable; it appears, however, to contain only three out of eight charged residues known to be conserved in eukaryotic Kv channels and accepted to be deterministic for folding and sensing. Despite the atypical sensor sequence, flux assays of KvLm reconstituted in liposomes disclosed a channel pore that is highly selective for K(+) and is blocked by conventional Kv channel blockers. Single-channel currents recorded in symmetric K(+) solutions from patches of enlarged Escherichia coli (spheroplasts) expressing KvLm showed that channel open probability sharply increases with depolarization, a hallmark feature of Kv channels. The identification of a voltage sensor module in KvLm with a voltage dependence comparable to that of other eukaryotic Kv channels yet encoded by a sequence that departs significantly from the consensus sequence of a eukaryotic voltage sensor establishes a molecular blueprint of a minimal sequence for a voltage sensor. PMID:16908725

  8. Molecular determinants for the tarantula toxin jingzhaotoxin-I interacting with potassium channel Kv2.1.

    PubMed

    Tao, Huai; Wu, Yuanyuan; Deng, Meichun; He, Juan; Wang, Meichi; Xiao, Yucheng; Liang, Songping

    2013-03-01

    With high binding affinity and distinct pharmacological functions, animal toxins are powerful ligands to investigate the structure-function relationships of voltage-gated ion channels. Jingzhaotoxin-I (JZTX-I) is an important neurotoxin from the tarantula Chilobrachys jingzhao venom that inhibits both sodium and potassium channels. In our previous work, JZTX-I, as a gating modifier, is able to inhibit activation of the potassium channel subtype Kv2.1. However, its binding site on Kv2.1 remains unknown. In this study, using Ala-scanning mutagenesis strategy, we demonstrated that four residues (I273, F274, E277, and K280) in S3b-S4 motif contributed to the formation of JZTX-I binding site. The mutations I273A, F274A, E277A, and K280A reduced toxin binding affinity by 6-, 10-, 8-, and 7-fold, respectively. Taken together with our previous data that JZTX-I accelerated channel deactivation, these results suggest that JZTX-I inhibits Kv2.1 activation by docking onto the voltage sensor paddle and trapping the voltage sensor in the closed state. PMID:23246579

  9. Methylation of KvDMR1 involved in regulating the imprinting of CDKN1C gene in cattle.

    PubMed

    Wang, Mengnan; Li, Dongjie; Zhang, Mingyue; Yang, Wenzhi; Cui, Yali; Li, Shijie

    2015-08-01

    The CDKN1C gene encodes a cyclin-dependent kinase inhibitor and is one of the key genes involved in the development of Beckwith-Wiedemann syndrome and cancer. In this study, using a direct sequencing approach based on a single nucleotide polymorphism (SNP) at genomic DNA and cDNA levels, we show that CDKN1C exhibits monoallelic expression in all seven studied organs (heart, liver, spleen, lung, kidney, muscle and subcutaneous fat) in cattle. To investigate how methylation regulates imprinting of CDKN1C in cattle, allele-specific methylation patterns in two putative differential methylation regions (DMRs), the CDKN1C DMR and KvDMR1, were analyzed in three tissues (liver, spleen and lung) using bisulfite sequencing PCR. Our results show that in the CDKN1C DMR both parental alleles were unmethylated in all three analyzed tissues. In contrast, KvDMR1 was differentially methylated between the two parental alleles in the same tissues. Statistical analysis showed that there is a significant difference in the methylation level between the two parental alleles (P < 0.01), confirming that this region is the DMR of KvDMR1 and that it may be correlated with CDKN1C imprinting. PMID:26059028

  10. Development of 6.6 kV/600 A superconducting fault current limiter using coated conductors

    NASA Astrophysics Data System (ADS)

    Yazawa, T.; Koyanagi, K.; Takahashi, M.; Toba, K.; Takigami, H.; Urata, M.; Iijima, Y.; Saitoh, T.; Amemiya, N.; Shiohara, Y.; Ito, T.

    2009-10-01

    As one of the programs in the Ministry of Economy, Trade and Industry (METI) project regarding R&D on superconducting coated conductor, three-phase superconducting fault current limiter (SFCL) for 6.6