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Sample records for 2-electrode voltage clamp

  1. Inner Voltage Clamping

    PubMed Central

    Feldberg, Stephen W.; Delgado, Alicia B.

    1978-01-01

    Ketterer, et al. (1971) have suggested that a combination of electrostatic and chemical interactions may cause hydrophobic ions absorbed within a bilayer lipid membrane to reside in two potential wells, each close to a membrane surface. The resulting two planes of charges would define three regions of membrane dielectric: two identical outer regions each between a plane of absorbed charges and the plane of closest approach of ions in the aqueous phase; and the inner region between the two planes of adsorbed charges. The theory describing charge translocation across the inner region is based on a simple three-capacitor model. A significant theoretical conclusion is that the difference between the voltage across the inner region, Vi, and the voltage across the entire membrane, Vm, is directly proportional to the amount of charge that has flowed in a voltage clamp experiment. We demonstrate that we can construct an “inner voltage clamp” that can maintain, with positive feedback, a constant inner voltage, Vi. The manifestation of proper feedback is that the clamp current (after a voltage step) will exhibit pure (i.e., single time-constant) exponential decay, because the voltage dependent rate constants governing translocation will be independent of time. The “pureness” of the exponential is maximized when the standard deviation of the least-square fit of the appropriate exponential equation to the experimental data is minimized. The concomitant feedback is directly related to the capacitances of the inner and outer membrane regions, Ci and Co. Experimental results with tetraphenylborate ion adsorbed in bacterial phosphatidylethanolamine/n-decane bilayers indicate Ci ∼ 5 · 10-7F/cm2 and Co ≈ 5 · 10-5F/cm2. PMID:620078

  2. Axon voltage-clamp simulations. I. Methods and tests.

    PubMed Central

    Moore, J W; Ramón, F; Joyner, R W

    1975-01-01

    This is the first in a series of four papers in which we present the numerical simulation of the application of the voltage clamp technique to excitable cells. In this paper we describe the application of the Crank-Nicolson (1947) method for the solution of the parabolic partial differential equations that describe a cylindrical cell in which the ionic conductances are functions of voltage and time (Hodgkin and Huxley, 1952). This method is compared with other methods in terms of accuracy and speed of solution for a propagated action potential. In addition, differential equations representing a simple voltage-clamp electronic circuit are presented. Using the voltage clamp circuit equations, we simulate the voltage clamp of a single isopotential membrane patch and show how the parameters of the circuit affect the transient response of the patch to a step change in the control potential.The stimulation methods presented in this series of papers allow the evaluation of voltage clamp control of an excitable cell or a syncytium of excitable cells. To the extent that membrane parameters and geometrical factors can be determined, the methods presented here provide solutions for the voltage profile as a function of time. PMID:1174640

  3. An improved vaseline gap voltage clamp for skeletal muscle fibers

    PubMed Central

    1976-01-01

    A Vaseline gap potentiometric recording and voltage clamp method is developed for frog skeletal muscle fibers. The method is based on the Frankenhaeuser-Dodge voltage clamp for myelinated nerve with modifications to improve the frequency response, to compensate for external series resistance, and to compensate for the complex impedance of the current-passing pathway. Fragments of single muscle fibers are plucked from the semitendinosus muscle and mounted while depolarized by a solution like CsF. After Vaseline seals are formed between fluid pools, the fiber ends are cut once again, the central region is rinsed with Ringer solution, and the feedback amplifiers are turned on. Errors in the potential and current records are assessed by direct measurements with microelectrodes. The passive properties of the preparation are simulated by the "disk" equivalent circuit for the transverse tubular system and the derived parameters are similar to previous measurements with microelectrodes. Action potentials at 5 degrees C are long because of the absence of delayed rectification. Their shape is approximately simulated by solving the disk model with sodium permeability in the surface and tubular membranes. Voltage clamp currents consist primarily of capacity currents and sodium currents. The peak inward sodium current density at 5 degrees C is 3.7 mA/cm2. At 5 degrees C the sodium currents are smoothly graded with increasing depolarization and free of notches suggesting good control of the surface membrane. At higher temperatures a small, late extra inward current appears for small depolarizations that has the properties expected for excitation in the transverse tubular system. Comparison of recorded currents with simulations shows that while the transverse tubular system has regenerative sodium currents, they are too small to make important errors in the total current recorded at the surface under voltage clamp at low temperature. The tubules are definitely not under voltage

  4. Extracellular potassium accumulation in voltage-clamped frog ventricular muscle.

    PubMed Central

    Cleemann, L; Morad, M

    1979-01-01

    1. Application of voltage clamp pulses (1--10 sec) to frog ventricular strips causes temporary changes in the extracellular K concentration. 2. The changes in the extracellular K concentration can be estimated from (a) slowly decaying post-clamp after-potentials, (b) changes in the action potential duration, and (c) measurements with a K-selective micro-electrode. 3. The depolarization of the resting potential and the shortening of the action potential are present in approximately the same proportions during voltage-clamp induced extracellular K accumulation and during perfusion with a K-ricn Ringer solution but small consistent differences are noticed. 4. The measurements of the after-potential, the action potential shortening, and the K-electrode response were analysed as indicators of extracellular K+ activity and it was concluded that the after-potential provides the most convenient and reliable estimate of the absolute magnitude of the voltage-clamp induced extracellular K accumulation. 5. The depolarizing after-potentials decay more slowly than the hyperpolarizing after-potentials but it is found that this reflects the selectivity of the membrane to K+ concentrations as predicted by the Nernst or the Goldman equations. 6. Analysis of the redistribution of accumulated K+ from the decay of the after-potential suggests that the major part of the redistribution process can be described by a single time constant (2--4 sec). A much longer time constant is required for a smaller component of the 'tail' in order to bring [K]o to the normal resting state. 7. N-shaped relations similar to the 'steady state' current-voltage relation are obtained when the post-clamp after-potential, the action potential shortening, and the K-electrode response are plotted versus the clamped membrane potential. The maxima of these curves are located around -40 mV and the minima around -20 mV. 8. In spite of a significant outward membrane current (1--1.5 microamperemeter) in the minimum

  5. Potassium Chloride Versus Voltage Clamp Contractures in Ventricular Muscle

    NASA Astrophysics Data System (ADS)

    Morad, M.; Reeck, S.; Rao, M.

    1981-01-01

    In frog ventricle, developed tension was markedly larger in response to depolarization caused by a voltage clamp step than to depolarization induced by high concentrations of potassium chloride. Measurement of extracellular potassium activity at the surface and at the depth of muscle during the development of contractures showed that the diffusion of potassium is much slower than the spread of depolarization through the cross section of muscle. These two observations suggest that competition between the depolarizing and the negative inotropic effects of an increase in the extracellular potassium ion concentration may determine the time course and magnitude of contractile tension in heart muscle.

  6. Application of active electrode compensation to perform continuous voltage-clamp recordings with sharp microelectrodes

    NASA Astrophysics Data System (ADS)

    Gómez-González, J. F.; Destexhe, A.; Bal, T.

    2014-10-01

    Objective. Electrophysiological recordings of single neurons in brain tissues are very common in neuroscience. Glass microelectrodes filled with an electrolyte are used to impale the cell membrane in order to record the membrane potential or to inject current. Their high resistance induces a high voltage drop when passing current and it is essential to correct the voltage measurements. In particular, for voltage clamping, the traditional alternatives are two-electrode voltage-clamp technique or discontinuous single electrode voltage-clamp (dSEVC). Nevertheless, it is generally difficult to impale two electrodes in a same neuron and the switching frequency is limited to low frequencies in the case of dSEVC. We present a novel fully computer-implemented alternative to perform continuous voltage-clamp recordings with a single sharp-electrode. Approach. To reach such voltage-clamp recordings, we combine an active electrode compensation algorithm (AEC) with a digital controller (AECVC). Main results. We applied two types of control-systems: a linear controller (proportional plus integrative controller) and a model-based controller (optimal control). We compared the performance of the two methods to dSEVC using a dynamic model cell and experiments in brain slices. Significance. The AECVC method provides an entirely digital method to perform continuous recording and smooth switching between voltage-clamp, current clamp or dynamic-clamp configurations without introducing artifacts.

  7. Modeling CICR in rat ventricular myocytes: voltage clamp studies

    PubMed Central

    2010-01-01

    Background The past thirty-five years have seen an intense search for the molecular mechanisms underlying calcium-induced calcium-release (CICR) in cardiac myocytes, with voltage clamp (VC) studies being the leading tool employed. Several VC protocols including lowering of extracellular calcium to affect Ca2+ loading of the sarcoplasmic reticulum (SR), and administration of blockers caffeine and thapsigargin have been utilized to probe the phenomena surrounding SR Ca2+ release. Here, we develop a deterministic mathematical model of a rat ventricular myocyte under VC conditions, to better understand mechanisms underlying the response of an isolated cell to calcium perturbation. Motivation for the study was to pinpoint key control variables influencing CICR and examine the role of CICR in the context of a physiological control system regulating cytosolic Ca2+ concentration ([Ca2+]myo). Methods The cell model consists of an electrical-equivalent model for the cell membrane and a fluid-compartment model describing the flux of ionic species between the extracellular and several intracellular compartments (cell cytosol, SR and the dyadic coupling unit (DCU), in which resides the mechanistic basis of CICR). The DCU is described as a controller-actuator mechanism, internally stabilized by negative feedback control of the unit's two diametrically-opposed Ca2+ channels (trigger-channel and release-channel). It releases Ca2+ flux into the cyto-plasm and is in turn enclosed within a negative feedback loop involving the SERCA pump, regulating[Ca2+]myo. Results Our model reproduces measured VC data published by several laboratories, and generates graded Ca2+ release at high Ca2+ gain in a homeostatically-controlled environment where [Ca2+]myo is precisely regulated. We elucidate the importance of the DCU elements in this process, particularly the role of the ryanodine receptor in controlling SR Ca2+ release, its activation by trigger Ca2+, and its refractory characteristics

  8. Solutions for transients in arbitrarily branching cables: III. Voltage clamp problems.

    PubMed Central

    Major, G

    1993-01-01

    Branched cable voltage recording and voltage clamp analytical solutions derived in two previous papers are used to explore practical issues concerning voltage clamp. Single exponentials can be fitted reasonably well to the decay phase of clamped synaptic currents, although they contain many underlying components. The effective time constant depends on the fit interval. The smoothing effects on synaptic clamp currents of dendritic cables and series resistance are explored with a single cylinder + soma model, for inputs with different time courses. "Soma" and "cable" charging currents cannot be separated easily when the soma is much smaller than the dendrites. Subtractive soma capacitance compensation and series resistance compensation are discussed. In a hippocampal CA1 pyramidal neurone model, voltage control at most dendritic sites is extremely poor. Parameter dependencies are illustrated. The effects of series resistance compound those of dendritic cables and depend on the "effective capacitance" of the cell. Plausible combinations of parameters can cause order-of-magnitude distortions to clamp current waveform measures of simulated Schaeffer collateral inputs. These voltage clamp problems are unlikely to be solved by the use of switch clamp methods. PMID:8369450

  9. Sources of errors in different single-electrode voltage-clamp techniques: a computer simulation study.

    PubMed

    Sala, F; Sala, S

    1994-08-01

    The use of voltage clamp with a single electrode has been useful in estimating kinetic parameters for a number of ionic whole-cell currents. There are two main types of such a technique: discontinuous voltage clamp (dSEVC) (Brennecke and Lindemann, 1974), and continuous voltage clamp (cSEVC) (Hamill et al., 1981). We have studied, by means of computer simulations, the performance of both types of clamp on estimating activation kinetics parameters of a typical neuronal Ca2+ current. Deviations from the theoretical values are shown to be sensitive on both set-up and cell properties. Both types of clamp are shown to lose voltage control when either access resistance or absolute membrane conductance are increased. In contrast, changes in membrane capacitance affect differently to the estimates obtained by the two types of clamp. Cell size is also shown to affect cSEVC performance but not that of dSEVC. The nature and magnitude of errors obtained by using both types of clamp in different situations are discussed.

  10. Cell-attached voltage-clamp and current-clamp recording and stimulation techniques in brain slices.

    PubMed

    Perkins, Katherine L

    2006-06-30

    Cell-attached recording provides a way to record the activity of - and to stimulate - neurons in brain slices without rupturing the cell membrane. This review uses theory and experimental data to address the proper application of this technique and the correct interpretation of the data. Voltage-clamp mode is best-suited for recording cell firing activity, and current-clamp mode is best-suited for recording resting membrane potential and synaptic potentials. The magnitude of the seal resistance determines what types of experiments can be accomplished with a cell-attached recording: a loose seal is adequate for recording action potential currents, and a tight seal is required for evoking action potentials in the attached cell and for recording resting and synaptic potentials. When recording action potential currents, if the researcher does not want to change the firing activity of the cell, then it is important that no current passes from the amplifier through the patch resistance. In order to accomplish this condition, the recording pipette should be held at the potential that gives a holding current of 0. An advantage of cell-attached current-clamp over whole-cell recording is that it accurately depicts whether a synaptic potential is hyperpolarizing or depolarizing without the risk of changing its polarity.

  11. Stoichiometry and voltage dependence of the sodium pump in voltage- clamped, internally dialyzed squid giant axon

    PubMed Central

    1989-01-01

    The stoichiometry and voltage dependence of the Na/K pump were studied in internally dialyzed, voltage-clamped squid giant axons by simultaneously measuring, at various membrane potentials, the changes in Na efflux (delta phi Na) and holding current (delta I) induced by dihydrodigitoxigenin (H2DTG). H2DTG stops the Na/K pump without directly affecting other current pathways: (a) it causes no delta I when the pump lacks Na, K, Mg, or ATP, and (b) ouabain causes no delta I or delta phi Na in the presence of saturating H2DTG. External K (Ko) activates Na efflux with Michaelis-Menten kinetics (Km = 0.45 +/- 0.06 mM [SEM]) in Na-free seawater (SW), but with sigmoid kinetics in approximately 400 mM Na SW (Hill coefficient = 1.53 +/- 0.08, K1/2 = 3.92 +/- 0.29 mM). H2DTG inhibits less strongly (Ki = 6.1 +/- 0.3 microM) in 1 or 10 mM K Na-free SW than in 10 mM K, 390 mM Na SW (1.8 +/- 0.2 microM). Dialysis with 5 mM each ATP, phosphoenolpyruvate, and phosphoarginine reduced Na/Na exchange to at most 2% of the H2DTG- sensitive Na efflux. H2DTG sensitive but nonpump current caused by periaxonal K accumulation upon stopping the pump, was minimized by the K channel blockers 3,4-diaminopyridine (1 mM), tetraethylammonium (approximately 200 mM), and phenylpropyltriethylammonium (20-25 mM) whose adequacy was tested by varying [K]o (0-10 mM) with H2DTG present. Two ancillary clamp circuits suppressed stray current from the axon ends. Current and flux measured from the center pool derive from the same membrane area since, over the voltage range -60 to +20 mV, tetrodotoxin-sensitive current and Na efflux into Na-free SW, under K- free conditions, were equal. The stoichiometry and voltage dependence of pump Na/K exchange were examined at near-saturating [ATP], [K]o and [Na]i in both Na-free and 390 mM Na SW. The H2DTG-sensitive F delta phi Na/delta I ratio (F is Faraday's constant) of paired measurements corrected for membrane area match, was 2.86 +/- 0.09 (n = 8) at 0 mV and 3

  12. VOLTAGE CLAMP BEHAVIOR OF IRON-NITRIC ACID SYSTEM AS COMPARED WITH THAT OF NERVE MEMBRANE

    PubMed Central

    Tasaki, I.; Bak, A. F.

    1959-01-01

    The current-voltage relation for the surface layer of an iron wire immersed in nitric acid was investigated by the voltage clamp technique. Comparing the phase of nitric acid to the axoplasm and the metallic phase to the external fluid medium for the nerve fiber, a striking analogy was found between the voltage clamp behavior of the iron-nitric acid system and that of the nerve membrane. The current voltage curve was found to consist of three parts: (a) a straight line representing the behavior of the resting (passive) membrane, (b) a straight line representing the fully excited (active) state, and (c) an intermediate zone connecting (a) and (b). It was shown that in the intermediate zone, the surface of iron consisted of a fully active patch (or patches) surrounded by a remaining resting area. The phenomenon corresponding to "repetitive firing of responses under voltage clamp" in the nerve membrane was demonstrated in the intermediate zone. The behavior of the cobalt electrode system was also investigated by the same technique. An attempt was made to interpret the phenomenon of initiation and abolition of an active potential on the basis of the thermodynamics of irreversible processes. PMID:13654740

  13. The Anion Paradox in Sodium Taste Reception: Resolution by Voltage-Clamp Studies

    NASA Astrophysics Data System (ADS)

    Ye, Qing; Heck, Gerard L.; Desimone, John A.

    1991-11-01

    Sodium salts are potent taste stimuli, but their effectiveness is markedly dependent on the anion, with chloride yielding the greatest response. The cellular mechanisms that mediate this phenomenon are not known. This "anion paradox" has been resolved by considering the field potential that is generated by restricted electrodiffusion of the anion through paracellular shunts between taste-bud cells. Neural responses to sodium chloride, sodium acetate, and sodium gluconate were studied while the field potential was voltage-clamped. Clamping at electronegative values eliminated the anion effect, whereas clamping at electropositive potentials exaggerated it. Thus, field potentials across the lingual epithelium modulate taste reception, indicating that the functional unit of taste reception includes the taste cell and its paracellular microenvironment.

  14. Voltage- and space-clamp errors associated with the measurement of electrotonically remote synaptic events.

    PubMed

    Spruston, N; Jaffe, D B; Williams, S H; Johnston, D

    1993-08-01

    1. The voltage- and space-clamp errors associated with the use of a somatic electrode to measure current from dendritic synapses are evaluated using both equivalent-cylinder and morphologically realistic models of neuronal dendritic trees. 2. As a first step toward understanding the properties of synaptic current distortion under voltage-clamp conditions, the attenuation of step and sinusoidal voltage changes are evaluated in equivalent cylinder models. Demonstration of the frequency-dependent attenuation of voltage in the cable is then used as a framework for understanding the distortion of synaptic currents generated at sites remote from the somatic recording electrode and measured in the voltage-clamp recording configuration. 3. Increases in specific membrane resistivity (Rm) are shown to reduce steady-state voltage attenuation, while producing only minimal reduction in attenuation of transient voltage changes. Experimental manipulations that increase Rm therefore improve the accuracy of estimates of reversal potential for electrotonically remote synapses, but do not significantly reduce the attenuation of peak current. In addition, increases in Rm have the effect of slowing the kinetics of poorly clamped synaptic currents. 4. The effects of the magnitude of the synaptic conductance and its kinetics on the measured synaptic currents are also examined and discussed. The error in estimating parameters from measured synaptic currents is greatest for synapses with fast kinetics and large conductances. 5. A morphologically realistic model of a CA3 pyramidal neuron is used to demonstrate the generality of the conclusions derived from equivalent cylinder models. The realistic model is also used to fit synaptic currents generated by stimulation of mossy fiber (MF) and commissural/associational (C/A) inputs to CA3 neurons and to estimate the amount of distortion of these measured currents. 6. Anatomic data from the CA3 pyramidal neuron model are used to construct a

  15. Voltage-clamp frequency domain analysis of NMDA-activated neurons.

    PubMed

    Moore, L E; Hill, R H; Grillner, S

    1993-02-01

    1. Voltage and current-clamp steps were added to a sum of sine waves to measure the tetrodotoxin-insensitive membrane properties of neurons in the intact lamprey spinal cord. A systems analysis in the frequency domain was carried out on two types of cells that have very different morphologies in order to investigate the structural dependence of their electrophysiological properties. The method explicitly takes into account the geometrical shapes of (i) nearly spherical dorsal cells with one or two processes and (ii) motoneurons and interneurons that have branched dendritic structures. Impedance functions were analysed to obtain the cable properties of these in situ neurons. These measurements show that branched neurons are not isopotential and, therefore, a conventional voltage-clamp analysis is not valid. 2. The electrophysiological data from branched neurons were curve-fitted with a lumped soma-equivalent cylinder model consisting of eight equal compartments coupled to an isopotential cell body to obtain membrane parameters for both passive and active properties. The analysis provides a quantitative description of both the passive electrical properties imposed by the geometrical structure of neurons and the voltage-dependent ionic conductances determined by ion channel kinetics. The model fitting of dorsal cells was dominated by a one-compartment resistance and capacitance in parallel (RC) corresponding to the spherical, non-branched shape of these cells. Branched neurons required a model that contained both an RC compartment and a cable that reflected the structure of the cells. At rest, the electrotonic length of the cable was about two. Uniformly distributed voltage-dependent ionic conductance sites were adequate to describe the data at different membrane potentials. 3. The frequency domain admittance method in conjunction with a step voltage clamp was used to control and measure the oscillatory behavior induced by N-methyl-D-aspartate (NMDA) on lamprey spinal

  16. Action of certain tropine esters on voltage-clamped lobster axon.

    PubMed

    Blaustein, M P

    1968-03-01

    Tropine p-tolylacetate (TPTA) and its quaternary analogue, tropine p-tolylacetate methiodide (TPTA MeI) decrease the early transient (Na) and late (K) currents in the voltage-clamped lobster giant axon. These agents, which block the nerve action potential, reduce the maximum Na and K conductance increases associated with membrane depolarization. They also slow the rate at which the sodium conductance is increased and shift the (normalized) membrane conductance vs. voltage curves in the direction of depolarization along the voltage axis. All these effects are qualitatively similar to those resulting from the action of procaine on the voltage-clamped axon. One unusual effect of the tropine esters, noticeable particularly at large depolarization steps, is that they cause the late, K current to reach a peak and then fall off with increasing pulse duration. This effect has not been reported to occur as a result of procaine action. Tropine p-chlorophenyl acetate (TPClphiA), which differs from TPTA only by the substitution of a p-Cl for a p-CH(3) group on the benzene ring, had a negligible effect on axonal excitability.

  17. High spatial resolution single multiwalled carbon nanotube electrode for stimulation, recording, and whole cell voltage clamping of electrically active cells

    NASA Astrophysics Data System (ADS)

    de Asis, Edward D.; Leung, Joseph; Wood, Sally; Nguyen, Cattien V.

    2009-10-01

    We report the stimulation, recording, and voltage clamp of muscle fibers using a 30 nm diameter single multiwalled carbon nanotube electrode (sMWNT electrode) tip. Because of the lower access resistance, the sMWNT electrode conducts extracellular and intracellular stimulation more efficiently compared to glass micropipettes. The sMWNT electrode records field potentials and action potentials and performs whole cell voltage clamping of single fibers.

  18. Measurement of Outer Hair Cell Electromotility Using a Fast Voltage Clamp

    NASA Astrophysics Data System (ADS)

    Evans, Michael G.; Fettiplace, Robert

    2009-02-01

    Isolated outer hair cells from rat pups (P9-P16) were voltage clamped using the whole-cell recording technique, and changes in cell length in response to step voltage changes were measured using a photodiode pair. Cell length changes were rapid and could be reasonably well fitted by single exponential functions with time constants around 0.1-0.2 ms, although particularly with larger steps double exponentials gave a better fit. Replacement of intracellular chloride by sulphate shifted the voltage-dependence of the motility to more depolarised potentials but did not alter the time course of the length changes. Exposure to low calcium (0.1 mM) solution, with or without 0.2 mM dihydrostreptomycin, also had no obvious effect on the time course of motility.

  19. Numerical analysis of the voltage-clamp technique applied to frog neuromuscular junctions.

    PubMed Central

    Torres, M E; Sevcik, C; Parthe, V

    1982-01-01

    The nonlinear cable equation was solved numerically by means of an implicit procedure. The correlation between end-plate length and fiber diameter was determined in frog (Rana pipiens) sartorius muscles stained with gold chloride (Löwit, 1875). The diameter of the fibers stained by the Löwit method was 80 (74-85) micron (median and its 95% confidence interval for 52 fibers), the length of the end plates in the same fibers was 382 (353-417) micron. The fibers simulated were 80 micron in diameter. To solve the equation the muscle fibers were represented by 500 segments 20 micron long, and the equation was solved in steps of 10 microseconds; a double exponential function was incorporated to the first seven segments to represent the neuromuscular junction. The potential of the first segment of the cable was set to the clamping level and the membrane potential of the remaining segments calculated. The current needed to hold the first segment was estimated by adding the current flowing through the first segment to the current flowing from it to the second segment. Our results indicate that the lack of space clamp in the point voltage-clamp studies of the frog neuromuscular junction introduces serious errors in the estimates of the end-plate conductance value, the kinetics of the conductance changes, and the reversal potential of the end-plate currents. The possibility of an efficient voltage-clamp technique is also explored. Our calculations suggest that the study of end-plate current and conductance is possible with little error if the end-plate potential is controlled at both ends of the synaptic area simultaneously. Images FIGURE 1 PMID:6981435

  20. Simultaneous measurement of changes in current and tracer flux in voltage-clamped squid giant axon.

    PubMed

    Rakowski, R F

    1989-04-01

    A method is described for the simultaneous measurement of changes in membrane current and unidirectional radiotracer flux in internally dialyzed voltage-clamped squid giant axons. The small currents that are produced by electrogenic transport processes or steady-state ionic currents can be resolved using this method. Because the use of grounded guard electrodes in the end pools is not, by itself, an adequate means of eliminating end-effects, two ancillary end pool clamp circuits are described to eliminate extraneous current flow from the ends of the axon. The end pool voltage-clamp circuits serve to minimize net current flow between the end pools and center pool, and employ stable, low-impedance calomel electrodes to monitor the potentials of the end and center pools. The adequacy of the method is demonstrated by experiments in which unidirectional 22Na efflux and current, flowing through tetrodotoxin (TTX)-sensitive Na channels into Na-free seawater, under K-free conditions, are shown to be equal. The equality of unidirectional TTX-sensitive flux and current is maintained over the entire range of membrane potentials examined (-60 to +20 mV). The method has been applied to a series of experiments in which the voltage dependence and stoichiometry of the Na/K pump have been measured (Rakowski et al., 1989), and can be applied in general to the simultaneous measurement of changes in current and flux of other electrogenic transport processes, and of currents through ionic channels that open under steady-state conditions.

  1. An analysis of the dose-response relationship at voltage-clamped frog neuromuscular junctions.

    PubMed Central

    Dionne, V E; Steinbach, J H; Stevens, C F

    1978-01-01

    1. Frog neuromuscular junctions were viewed with Nomarski optics and voltage clamped. Agonist was applied ionophoretically and agonist concentrations were measured using a micro-electrode sensitive to quaternary amines. 2. The dose-response relationship was studied using the agonists carbamylcholine, suberyldicholine and hydroxyphenyl-propyltrimethylammonium. 3. With all of these agonists, it appeared that the ACh receptor could be active when either one or two agonist binding sites were occupied. The receptor was much more likely to be active when both sites were occupied. Agonist dissociation constants and receptor activation probabilities were estimated by non-linear regression techniques for several possible receptor activation schemes. PMID:309004

  2. A fast transient outward current in the rat sympathetic neurone studied under voltage-clamp conditions.

    PubMed Central

    Belluzzi, O; Sacchi, O; Wanke, E

    1985-01-01

    Post-ganglionic neurones of the isolated rat superior cervical ganglion were voltage clamped at 37 degrees C using separate intracellular voltage and current micro-electrodes. Control experiments in current clamp suggested that the neurone is electrotonically compact, the soma and the proximal dendritic membranes being under good spatial voltage uniformity. Depolarizing voltage steps from membrane potentials near -50 mV evoked: (i) a voltage-dependent inward Na+ current, (ii) an inward Ca2+ current, (iii) a voltage-dependent outward K+ current, (iv) a Ca2+-activated K+ outward current. Depolarizations from holding potentials more negative than -60 mV elicited, besides the currents mentioned above, a fast transient outward current IA which peaked in 1-2.5 ms and then decayed to zero following an exponential time course. The IA current was shown to be primarily, if not exclusively, carried by K+. It was unaffected by removal of external Ca2+ or addition of Cd2+ and was weakly blocked by tetraethylammonium ions and partially by 4-aminopyridine. The IA current showed a linear instantaneous current-voltage relationship. Its activation ranged from -60 to 0 mV with a mid-point at -30 mV. The A conductance could be described in terms of a simple Boltzmann distribution for a single gating particle with a valency of +3. Both the development and removal of inactivation followed a single exponential time course with a voltage-dependent time constant which was large near the resting potential (42 ms at -70 mV) and small (11 ms) near -100 and -40 mV. Steady-state inactivation h infinity ranged from -100 to -50 mV, with a mid-point at -78 mV, suggesting that approximately 50% of the IA channels are available at the physiological resting potential. Action potentials elicited from various holding potentials showed maximal repolarization rates dependent on the holding potential itself. This voltage dependence was found to be in reasonably good agreement with that of h infinity curve

  3. Nile blue fluorescence signals from cut single muscle fibers under voltage or current clamp conditions

    PubMed Central

    1978-01-01

    A method is presented for recording extrinsic optical signals from segments of single skeletal muscle fibers under current or voltage clamp conditions. Such segments, which are cut from intact fibers, are maintained in a relaxed state, while exhbiting otherwise normal physiological properties, including healthy delayed rectifier currents. Extrinsic fluorescence changes are demonstrated, using the permeant potentiometric probe, Nile Blue A. These changes vary nonlinearly with the controlled surface membrane potential, in a manner which suggests that they arise from potential changes in the sarcoplasmic reticulum. According to this interpretation, a simple model based on the gating charge movement implicated in excitation-contraction coupling, provides a self-consistent description of the voltage dependence of the signal that requires no additional parameters. PMID:310445

  4. Barbiturates block sodium and potassium conductance increases in voltage-clamped lobster axons.

    PubMed

    Blaustein, M P

    1968-03-01

    Sodium pentobarbital and sodium thiopental decrease both the peak initial (Na) and late steady-state (K) currents and reduce the maximum sodium and potassium conductance increases in voltage-clamped lobster giant axons. These barbiturates also slow the rate at which the sodium conductance turns on, and shift the normalized sodium conductance vs. voltage curves in the direction of depolarization along the voltage axis. Since pentobarbital (pK(a) = 8.0) blocks the action potential more effectively at pH 8.5 than at pH 6.7, the anionic form of the drug appears to be active. The data suggest that these drugs affect the axon membrane directly, rather than secondarily through effects on intermediary metabolism. It is suggested that penetration of the lipid layer of the membrane by the nonpolar portion of the barbiturate molecules may cause the decrease in membrane conductances, while electrostatic interactions involving the anionic group on the barbiturate, divalent cations, and "fixed charges" in the membrane could account for the slowing of the rate of sodium conductance turn-on and the shift of the normalized conductance curves along the voltage axis.

  5. Voltage clamp methods for the study of membrane currents and SR Ca2+ release in adult skeletal muscle fibres

    PubMed Central

    Hernández-Ochoa, Erick O.; Schneider, Martin F.

    2012-01-01

    Skeletal muscle excitation-contraction (E-C)1 coupling is a process composed of multiple sequential stages, by which an action potential triggers sarcoplasmic reticulum (SR)2 Ca2+ release and subsequent contractile activation. The various steps in the E-C coupling process in skeletal muscle can be studied using different techniques. The simultaneous recordings of sarcolemmal electrical signals and the accompanying elevation in myoplasmic Ca2+, due to depolarization-initiated SR Ca2+ release in skeletal muscle fibres, have been useful to obtain a better understanding of muscle function. In studying the origin and mechanism of voltage dependency of E-C coupling a variety of different techniques have been used to control the voltage in adult skeletal fibres. Pioneering work in muscles isolated from amphibians or crustaceans used microelectrodes or ‘high resistance gap’ techniques to manipulate the voltage in the muscle fibres. The development of the patch clamp technique and its variant, the whole-cell clamp configuration that facilitates the manipulation of the intracellular environment, allowed the use of the voltage clamp techniques in different cell types, including skeletal muscle fibres. The aim of this article is to present an historical perspective of the voltage clamp methods used to study skeletal muscle E-C coupling as well as to describe the current status of using the whole-cell patch clamp technique in studies in which the electrical and Ca2+ signalling properties of mouse skeletal muscle membranes are being investigated. PMID:22306655

  6. Voltage-dependent facilitation of Ca2+ entry in voltage-clamped, aequorin-injected molluscan neurons.

    PubMed Central

    Eckert, R; Tillotson, D; Ridgway, E B

    1977-01-01

    Voltage-clamp experiments were performed on giant neurons of the nudibranch Anisodoris nobilis injected with the Ca-sensitive photoprotein, aequorin. Depolarization beyond -10 to +5 m V produced an aequorin signal, the amplitude of which depended on the extracellular Ca2+ concentration, the amplitude of the depolarization, and its duration. In paired pulse experiments, the amplitude of the aequorin signal produced in response to the second of two identical depolarizing pulses was larger than that produced during the first, resulting from an increased entry of Ca2+ during the second pulse. The increment in Ca conductance inferred from the augmented signal during the second pulse was independent of Ca2+ influx during the first pulse but, instead, was related to the amplitude and duration of the first pulse. PMID:266215

  7. Supercharging accelerates T-tubule membrane potential changes in voltage clamped frog skeletal muscle fibers.

    PubMed

    Kim, A M; Vergara, J L

    1998-10-01

    In voltage-clamp studies of single frog skeletal muscle fibers stained with the potentiometric indicator 1-(3-sulfonatopropyl)-4-[beta[2-(di-n-octylamino)-6-naphthyl] vinyl]pyridinium betaine (di-8 ANEPPS), fluorescence transients were recorded in response to both supercharging and step command pulses. Several illumination paradigms were utilized to study global and localized regions of the transverse tubule system (T-system). The rising phases of transients obtained from global illumination regions showed distinct accelerations when supercharging pulses were applied (95% of steady-state fluorescence achieved in 1.5 ms with supercharging pulses versus 14.6 ms with step pulses). When local transients were recorded at the edge of the muscle fiber, their kinetics resembled those of the applied waveform, but a similar relationship was not observed in transients from regions near the edge chosen to minimize the surface membrane contribution. We developed a model of the T-system capable of simulating membrane potential changes as a function of time and distance along the T-system cable and the associated fluorescence changes in regions corresponding to the experimental illumination strategies. A critical parameter was the access resistance term, for which values of 110-150 Omega.cm2 were adequate to fit the data. The results suggest that the primary mechanism through which supercharging pulses boost the kinetics of T-system voltage changes most likely involves their compensating the voltage attenuation across the access resistance at the mouth of the T-tubule.

  8. How the early voltage clamp studies of José del Castillo inform "modern" neuroscience.

    PubMed

    Zottoli, Steven J

    2012-10-01

    The description of ionic currents that flow across the membrane of the squid giant axon during an action potential sparked an interest in determining whether there were similar currents in vertebrates. The preparation of choice was the node of Ranvier in single myelinated fibers in frog. José del Castillo spent 3 years on the United States mainland from 1956 to 1959. During that time, he collaborated with Jerome Y. Lettvin and John W. Moore. I discuss how these individuals met one another and some of their scientific discoveries using the voltage clamp to study squid giant axons and frog nodes. Much of this work was conducted at the Marine Biological Laboratory in Woods Hole, MA, and I attempt to convey a sense of the unique scientific "melting pot" that existed at the Marine Biological Laboratory and the broader effect that del Castillo had on "modern" neuroscience.

  9. Ultraviolet photoalteration of ion channels in voltage-clamped lobster giant axons.

    PubMed

    Oxford, G S; Pooler, J P

    1975-01-01

    An analysis of the ultraviolet light-induced changes in ionic conductances of lobster giant axon membranes has been carried out using the double sucrose gap voltage-clamp technique. The predominant effect of monochromatic light from a xenon arc source in the 255 to 305 nm region is an irreversible reduction in the magnitude of sodium conductance, without change in sodium channel activation or inactivation kinetics. A considerably smaller reduction in the magnitude of potassium conductance occurs, with some slowing of potassium channel activation kinetics. Leakage conductance is essentially not altered. The fall in sodium conductance follows an exponential time course toward a zero asymptote. The rate constant for conductance decrease was used as an assay for the wavelength dependence. The sodium conductance was maximally sensitive at 290 nm. It is suggested that individual sodium channels are closed upon absorption of single photons by aromatic amino acid residues in membrane proteins.

  10. Supercharging accelerates T-tubule membrane potential changes in voltage clamped frog skeletal muscle fibers.

    PubMed Central

    Kim, A M; Vergara, J L

    1998-01-01

    In voltage-clamp studies of single frog skeletal muscle fibers stained with the potentiometric indicator 1-(3-sulfonatopropyl)-4-[beta[2-(di-n-octylamino)-6-naphthyl] vinyl]pyridinium betaine (di-8 ANEPPS), fluorescence transients were recorded in response to both supercharging and step command pulses. Several illumination paradigms were utilized to study global and localized regions of the transverse tubule system (T-system). The rising phases of transients obtained from global illumination regions showed distinct accelerations when supercharging pulses were applied (95% of steady-state fluorescence achieved in 1.5 ms with supercharging pulses versus 14.6 ms with step pulses). When local transients were recorded at the edge of the muscle fiber, their kinetics resembled those of the applied waveform, but a similar relationship was not observed in transients from regions near the edge chosen to minimize the surface membrane contribution. We developed a model of the T-system capable of simulating membrane potential changes as a function of time and distance along the T-system cable and the associated fluorescence changes in regions corresponding to the experimental illumination strategies. A critical parameter was the access resistance term, for which values of 110-150 Omega.cm2 were adequate to fit the data. The results suggest that the primary mechanism through which supercharging pulses boost the kinetics of T-system voltage changes most likely involves their compensating the voltage attenuation across the access resistance at the mouth of the T-tubule. PMID:9746552

  11. An accurate online calibration system based on combined clamp-shape coil for high voltage electronic current transformers

    SciTech Connect

    Li, Zhen-hua; Li, Hong-bin; Zhang, Zhi

    2013-07-15

    Electronic transformers are widely used in power systems because of their wide bandwidth and good transient performance. However, as an emerging technology, the failure rate of electronic transformers is higher than that of traditional transformers. As a result, the calibration period needs to be shortened. Traditional calibration methods require the power of transmission line be cut off, which results in complicated operation and power off loss. This paper proposes an online calibration system which can calibrate electronic current transformers without power off. In this work, the high accuracy standard current transformer and online operation method are the key techniques. Based on the clamp-shape iron-core coil and clamp-shape air-core coil, a combined clamp-shape coil is designed as the standard current transformer. By analyzing the output characteristics of the two coils, the combined clamp-shape coil can achieve verification of the accuracy. So the accuracy of the online calibration system can be guaranteed. Moreover, by employing the earth potential working method and using two insulating rods to connect the combined clamp-shape coil to the high voltage bus, the operation becomes simple and safe. Tests in China National Center for High Voltage Measurement and field experiments show that the proposed system has a high accuracy of up to 0.05 class.

  12. An accurate online calibration system based on combined clamp-shape coil for high voltage electronic current transformers.

    PubMed

    Li, Zhen-hua; Li, Hong-bin; Zhang, Zhi

    2013-07-01

    Electronic transformers are widely used in power systems because of their wide bandwidth and good transient performance. However, as an emerging technology, the failure rate of electronic transformers is higher than that of traditional transformers. As a result, the calibration period needs to be shortened. Traditional calibration methods require the power of transmission line be cut off, which results in complicated operation and power off loss. This paper proposes an online calibration system which can calibrate electronic current transformers without power off. In this work, the high accuracy standard current transformer and online operation method are the key techniques. Based on the clamp-shape iron-core coil and clamp-shape air-core coil, a combined clamp-shape coil is designed as the standard current transformer. By analyzing the output characteristics of the two coils, the combined clamp-shape coil can achieve verification of the accuracy. So the accuracy of the online calibration system can be guaranteed. Moreover, by employing the earth potential working method and using two insulating rods to connect the combined clamp-shape coil to the high voltage bus, the operation becomes simple and safe. Tests in China National Center for High Voltage Measurement and field experiments show that the proposed system has a high accuracy of up to 0.05 class.

  13. An accurate online calibration system based on combined clamp-shape coil for high voltage electronic current transformers

    NASA Astrophysics Data System (ADS)

    Li, Zhen-hua; Li, Hong-bin; Zhang, Zhi

    2013-07-01

    Electronic transformers are widely used in power systems because of their wide bandwidth and good transient performance. However, as an emerging technology, the failure rate of electronic transformers is higher than that of traditional transformers. As a result, the calibration period needs to be shortened. Traditional calibration methods require the power of transmission line be cut off, which results in complicated operation and power off loss. This paper proposes an online calibration system which can calibrate electronic current transformers without power off. In this work, the high accuracy standard current transformer and online operation method are the key techniques. Based on the clamp-shape iron-core coil and clamp-shape air-core coil, a combined clamp-shape coil is designed as the standard current transformer. By analyzing the output characteristics of the two coils, the combined clamp-shape coil can achieve verification of the accuracy. So the accuracy of the online calibration system can be guaranteed. Moreover, by employing the earth potential working method and using two insulating rods to connect the combined clamp-shape coil to the high voltage bus, the operation becomes simple and safe. Tests in China National Center for High Voltage Measurement and field experiments show that the proposed system has a high accuracy of up to 0.05 class.

  14. Voltage-clamp studies of transient inward current and mechanical oscillations induced by ouabain in ferret papillary muscle.

    PubMed

    Karagueuzian, H S; Katzung, B G

    1982-06-01

    1. We studied the effects of a toxic concentration of ouabain on transmembrane electrical activity and on mechanical behaviour of right ventricular papillary muscles from ferrets in a single sucrose-gap using current clamp and voltage clamp.2. Ouabain (1.4-1.8 muM) induced oscillatory after-potentials and after-concentrations in current-clamp experiments. Voltage clamp showed that the oscillatory after-potential was caused by a transient inward current, similar to that in Purkinje fibres.3. The transient current had a sigmoidal dependence on the preceding (activating) voltage step V1, with a treshold around -13 mV and a plateau between +10 and 20 mV. There was a decline in current amplitude for more positive clamps. When activated by a fixed V1 voltage step, and measured at different repolarization levels V2, the transient current manifested an inverse dependence on V2 between -50 and -10 mV. No outward transient current could be detected. Total replacement of Na in the bathing medium by Tris or by sucrose abolished the transient current.4. Ouabain caused an increase of phasic (twitch) tension responses to voltage steps at all potentials without shifting the curve relating these variables on the voltage axis. The drug evoked an even greater increase in the tonic tension responses.5. After prolonged exposure, oscillatory mechanical responses were frequently recorded during positive voltage steps. Unlike the after-contraction, these mechanical fluctuations were not consistently damped and were not accompanied by detectable synchronous current fluctuations. Catecholamines and dibutyryl cyclic AMP markedly reduced the amplitude of the tonic contraction and the mechanical oscillations but increased their frequency. Caffeine had no effect on the tonic contraction amplitude but abolished the fluctuations.6. These results support the proposal that Ca is transiently released from the overloaded sarcoplasmic reticulum in ouabain-intoxicated muscle and may evoke oscillatory

  15. Characterization of heterologously expressed transporter genes by patch- and voltage-clamp methods: application to cyclic nucleotide-dependent responses.

    PubMed

    Lemtiri-Chlieh, Fouad; Ali, Rashid

    2013-01-01

    The application of patch- and voltage-clamp methods to study ion transport can be limited by many -hurdles: the size of the cells to be patched and/or stabbed, the subcellular localization of the molecule of interest, and its density of expression that could be too low even in their own native environment. Functional expression of genes using recombinant DNA technology not only overcomes those hurdles but also affords additional and elegant investigations such as single-point mutation studies and subunit -associations/regulations. In this chapter, we give a step-by-step description of two electrophysiological methods, patch clamp and two-electrode voltage clamp (TEVC), that are routinely used in combination with heterologous gene expression to assist researchers interested in the identification and characterization of ion transporters. We describe how to (1) obtain and maintain the cells suitable for the use with each of the above-mentioned methods (i.e., HEK-293 cells and yeast spheroplasts to use with the patch-clamp methodology and Xenopus laevis oocytes with TEVC), (2) transfect/inject them with the gene of interest, and (3) record ion transport activities.

  16. Determination of ionic permeability coefficients of the plasma membrane of Xenopus laevis oocytes under voltage clamp.

    PubMed Central

    Costa, P F; Emilio, M G; Fernandes, P L; Ferreira, H G; Ferreira, K G

    1989-01-01

    1. A method of estimating absolute ionic permeability coefficients which does not depend on the use of impermeant substitutes is reported. 2. The method is based on a pump leak model of the Xenopus laevis oocyte membrane. The procedure consists of measuring, in the same experiment, the pump current and the currents generated under voltage clamp by the partial substitution of one or two ions at a time. For each experimental condition, the measured currents are substituted in a Goldman-Hodgkin-Katz type equation with two unknowns (the permeability coefficients). The set of equations thus generated enables the computation of all the ionic permeability coefficients. 3. The Xenopus oocyte membrane (stages IV and V, Dumont, 1972) has been found to be permeable to conventional ion substitutes such as N-methyl-D-glucamine (NMG), sulphate, isethionate and gluconate. 4. The values for sodium, potassium and chloride permeability coefficients obtained from sixty-eight pooled experiments were, respectively, 5.44, 17.41 and 1.49 x 10(-8) cm s-1. 5. The diffusional currents for sodium, potassium and chloride computed from the experiments referred to above were, respectively, -1.16, 0.69 and -0.038 microA cm-2. 6. A stoichiometry of the Na+-K+ pump exchange of 3/1.8 was computed. 7. The intracellular concentrations of sodium, potassium and chloride ions, as determined by ion-selective microelectrodes, were, respectively, 10.1 +/- 0.66 mM (n = 12), 109.5 +/- 3.3 mM (n = 13) and 37.7 +/- 1.18 mM (n = 19), corresponding to equilibrium potentials of 61, -95 and -28 mV. 8. Since chloride is not at equilibrium across the membrane, we propose that there is an inward uphill Cl- transport. PMID:2600847

  17. Verification of the windings axial clamping forces for high voltage power transformers by using passively mode-locked fiber lasers

    NASA Astrophysics Data System (ADS)

    Şchiopu, IonuÅ£ Romeo; ǎgulinescu, Andrei, Dr; Iordǎnescu, Raluca; Marinescu, Andrei

    2015-02-01

    The current paper describes an optoelectronic method for direct monitoring of the axial clamping forces both in static and in dynamic duty. As advantages of this method we can state that it can be applied both to new and refurbished transformers without performing constructive changes or affecting in any way the transformer safety in operation. For monitoring the axial clamping forces for high-voltage (HV) power transformers, we use an optical fiber that we integrate into the laser cavity of a passively mode-locked fiber laser (PMFL). To each axial clamp corresponds a solitonic optical spectrum that is changed at the periodical passing of the fundamental soliton pulse through the sensitive fiber inside the transformer. Moreover, as a specific characteristic, the laser stability is unique for each set of axial clamping forces. Other important advantages of using an optical fiber as compared to the classical approach in which electronic sensors are used consist in the good reliability and insulator properties of the optical fiber, avoiding any risk of fire or damage of the transformer.

  18. Local, stochastic release of Ca2+ in voltage-clamped rat heart cells: visualization with confocal microscopy.

    PubMed Central

    López-López, J R; Shacklock, P S; Balke, C W; Wier, W G

    1994-01-01

    1. Confocal microscopy and the fluorescent Ca2+ indicator fluo-3 (K+ salt) were used to measure cytosolic free calcium ion concentration ([Ca2+]) during excitation-contraction (E-C) coupling in single, voltage-clamped, rat cardiac ventricular cells. 2. Local [Ca2+]i transients were measured nearly simultaneously in different, separate, subcellular volumes of approximately 2.0 microns 3. During depolarization, local [Ca2+]i transients were distinctly different from each other and from whole-cell [Ca2+]i transients. These differences were particularly apparent during small depolarizations, and were substantially reduced by ryanodine. 3. Components of the local [Ca2+]i transients, particularly those evoked by small depolarizations, were closely similar, in time course and amplitude, to spontaneous local [Ca2+]i transients, or 'sparks' (which have been shown previously to be Ca2+ released from sarcoplasmic reticulum). 4. Analysis of local [Ca2+]i transients in the spatial frequency domain (power spectrum) revealed that high power at spatial frequencies of 0.05-0.2 microns-1 was always associated with spontaneous calcium 'sparks' and with local [Ca2+]i transients evoked by small depolarizing pulses (e.g. to -31 mV). Evoked local [Ca2+]o transients in the presence of ryanodine, and those evoked by depolarization to very positive clamp-pulse potentials (+45 mV), were associated with considerably lower power at this frequency. 5. The results suggest that whole-cell [Ca2+]i transients evoked by voltage-clamp depolarization, and thus by L-type Ca2+ current, are comprised of local [Ca2+]i transients that are similar to the spontaneous calcium 'sparks'. At very positive clamp-pulse potentials, however, the electrically evoked local [Ca2+]i transients may be smaller, perhaps as a result of smaller unitary L-type Ca2+ current. Images Figure 1 Figure 2 Figure 3 PMID:7853223

  19. Evidence by a voltage clamp study of an electrically mediated block to polyspermy in the egg of the ascidian Phallusia mammillata.

    PubMed

    Goudeau, H; Depresle, Y; Rosa, A; Goudeau, M

    1994-12-01

    Eggs of the ascidian Phallusia mammillata were voltage clamped (from -100 to +60 mV) and inseminated with a low or heavy sperm concentration. From inseminations with low sperm concentration (1 x 10(6) sp/ml), we found that fertilization currents occurred between -100 and +40 mV: they were always inward and displayed an analogous pattern whatever the clamped voltage. We established that the percentages of inseminated eggs that produced a fertilization current varied as a function of the clamped voltage. These percentages were not statistically different from 100% at clamped voltages between -100 and -30 mV, they decreased to 68 and 56% at clamped Vm of -10 and 0 mV, respectively, but were not statistically different from 0% at clamped Vm between +10 and +40 mV. We never obtained any egg electrical response at a clamped voltage of +50 mV. Almost all eggs (96%) which responded electrically were penetrated by one or several spermatozoa. These eggs were resuming meiosis (81 to 50%) at values of clamped Vm between -100 and 0 mV, respectively. At clamped Vm between +10 and +50 mV, the percentages of eggs resuming meiosis were not statistically different from 0. These results indicate that in P. mammillata eggs, the occurrence of an electrical response is voltage dependent and consequently that the initial depolarizing shift of the fertilization potential constitutes a fast block to polyspermy. However, in this species, the sperm penetration is not voltage dependent, since it occurred at clamped Vm from -100 to +40 mV. On the other hand, when eggs were clamped from -100 to +60 mV and inseminated with a heavy sperm concentration (2 x 10(7) sp/ml), the curves expressing, respectively, the percentages of eggs which responded electrically, the percentages of eggs which were penetrated by one or several spermatozoa, and the percentages of eggs resuming meiosis, as functions of the clamped Vm, were shifted by approximately 35 mV toward more positive voltages, compared to the

  20. Voltage-clamp predictions by gompertz kinetics model relating squid-axon Na+-gating and ionic currents.

    PubMed

    Easton, Dexter M

    2005-10-01

    Gompertz kinetics is a simple, realistic, accurate, and computationally parsimonious alternative for prediction of macroscopic changes in Na+ conductance during voltage clamp. Conductance delay and time course depend on initial amplitudes and decay rates of surrogates for the macroscopic gating currents. The model is tested by the fit to published data of other authors. The proposed physical basis for the model is that membrane potential perturbation triggers motion of charged "gating" components of the axon membrane at rapid (activating) and at slow (inactivating) rates. The resulting distortion increases and more slowly diminishes the probability that conduction channels will be open.

  1. Electrophysiological Characterization of Na,K-ATPases Expressed in Xenopus laevis Oocytes Using Two-Electrode Voltage Clamping.

    PubMed

    Hilbers, Florian; Poulsen, Hanne

    2016-01-01

    The transport of three Na(+) per two K(+) means that the Na,K-ATPase is electrogenic, and though the currents generated by the ion pump are small compared to ion channel currents, they can be measured using electrophysiology, both steady-state pumping and individual steps in the transport cycle. Various electrophysiological techniques have been used to study the endogenous pumps of the squid giant axon and of cardiac myocytes from for example rabbits. Here, we describe the characterization of heterologously expressed Na,K-ATPases using two-electrode voltage clamping (TEVC) and oocytes from the Xenopus laevis frog as the model cell. With this system, the effects of particular mutations can be studied, including the numerous mutations that in later years have been found to cause human diseases.

  2. Correlation of 125I-LSD autoradiographic labeling with serotonin voltage clamp responses in Aplysia neurons

    SciTech Connect

    Evans, M.L.; Kadan, M.J.; Hartig, P.R.; Carpenter, D.O. )

    1991-05-01

    Autoradiographic receptor binding studies using 125I-LSD (2-(125I)lysergic acid diethyamide) revealed intense labelling on the soma of a symmetrically located pair of cells in the abdominal ganglion of Aplysia californica. This binding was blocked by micromolar concentrations of serotonin and lower concentrations of the serotonergic antagonists, cyproheptadine and mianserin. Electrophysiological investigation of responses to serotonin of neurons in the left upper quadrant, where one of the labeled neurons is located, revealed a range of serotonin responses. Cells L3 and L6 have a K+ conductance increase in response to serotonin that is not blocked by cyproheptadine or mianserin. Cells L2 and L4 have a biphasic response to serotonin: a Na+ conductance increase, which can be blocked by cyproheptadine and mianserin, followed by a voltage dependent Ca2+ conductance which is blocked by Co2+ but not the serotonergic antagonists. Cell L1, and its symmetrical pair, R1, have in addition to the Na+ and Ca2+ responses observed in L2 and L4, a Cl- conductance increase blocked by LSD, cyproheptadine and mianserin. LSD had little effect on the other responses. The authors conclude that the symmetrically located cells L1 and R1 have a Cl- channel linked to a cyproheptadine- and mianserin-sensitive serotonin receptor that is selectively labelled by 125I-LSD. This receptor has many properties in common with the mammalian serotonin 1C receptor.

  3. Modeling Ca(2+) currents and buffered diffusion of Ca(2+) in human β-cells during voltage clamp experiments.

    PubMed

    Félix-Martínez, Gerardo J; Godínez-Fernández, J Rafael

    2015-12-01

    Macroscopic Ca(2+) currents of the human β-cells were characterized using the Hodgkin-Huxley formalism. Expressions describing the Ca(2+)-dependent inactivation process of the L-type Ca(2+) channels in terms of the concentration of Ca(2+) were obtained. By coupling the modeled Ca(2+) currents to a three-dimensional model of buffered diffusion of Ca(2+), we simulated the Ca(2+) transients formed in the immediate vicinity of the cell membrane during voltage clamp experiments performed in high buffering conditions. Our modeling approach allowed us to consider the distribution of the Ca(2+) sources over the cell membrane. The effect of exogenous (EGTA) and endogenous Ca(2+) buffers on the temporal course of the Ca(2+) transients was evaluated. We show that despite the high Ca(2+) buffering capacity, nanodomains are formed in the submembrane space, where a peak Ca(2+) concentration between ∼76 and 143 µM was estimated from our simulations. In addition, the contribution of each Ca(2+) current to the formation of the Ca(2+) nanodomains was also addressed. Here we provide a general framework to incorporate the spatial aspects to the models of the pancreatic β-cell, such as a more detailed and realistic description of Ca(2+) dynamics in response to electrical activity in physiological conditions can be provided by future models.

  4. Two-suction-electrode voltage-clamp analysis of the sustained calcium current in cat sensory neurones.

    PubMed Central

    Taylor, W R

    1988-01-01

    1. The kinetics of the sustained calcium current were examined in cat dorsal root ganglion (DRG) neurones, using a two-suction-electrode voltage clamp. It was shown that this current could be examined with minimal contamination from other ionic currents. Experiments were performed at 20 degrees C, with a concentration of 10 mM-calcium externally. 2. The transient calcium current was eliminated by using a holding potential of -50 mV. The sustained calcium current showed no evidence of steady-state inactivation at potentials between -90 and -30 mV. 3. The activation and deactivation time course of the calcium current was described by a double-exponential function. The activation process was examined without interference from significant inactivation under the conditions used. 4. The steady-state activation of the calcium channels was approximated by a two-step activation process. Both reactions were voltage sensitive, the first having an equivalent valency of 3.4 +/- 0.1 electronic charges (e-), and the second with an equivalent valency of 1.0 +/- 0.2 e-. On average, half-maximal channel activation occurred at +10 mV. 5. The fast and the slow time constants of the exponential relaxations differed by a factor of 4-10 and both showed significant voltage dependence. Both the fast and slow time constants were greatest at potentials where approximately half the available channels were activated in the steady state. The slow time constant measured from activation and deactivation appeared to be independent of the starting potential. 6. The fractional amplitude of the slow exponential component of the tail currents was 0.09 +/- 0.01 at -70 mV and increased steadily at more positive potentials passing through a clear maximum of 0.59 +/- 0.03 at -10 mV. 7. Reducing the temperature decreased the magnitude of the peak inward current, with an apparent activation energy (Ea) of 67 kJ/mol. The slow time constants measured from activation and deactivation were also reduced at lower

  5. Kinetics of calcium-dependent inactivation of calcium current in voltage-clamped neurones of Aplysia californica.

    PubMed Central

    Chad, J; Eckert, R; Ewald, D

    1984-01-01

    Ca currents flowing during voltage-clamp depolarizations were examined in axotomized Aplysia neurones under conditions that virtually eliminated other currents. Moderate to large currents exhibited a two-component time course of relaxation that can be approximated reasonably well by the sum of two exponentials. The rapid phase (tau 1 approximately equal to 70 ms at 0 mV) plus the slower phase (tau 2 approximately equal to 300 ms at 0 mV) ride upon a steady, non-inactivating current, I infinity. Conditions that diminish the peak current amplitude, such as reduced stimulus depolarization, inactivation remaining from a prior depolarization, or partial blockade of the Ca conductance by Cd, slowed both phases of inactivation, and all selectively eliminated the tau 1 phase, such that weak currents exhibited only the slower phase of decline. Injection of EGTA slowed both phases of inactivation, decreased the extent of the tau 1 phase, and increased the intensity of I infinity and of the current during the tau 2 phase. For a given voltage, the rate of inactivation increased as the peak current strength was increased, and decreased as the peak current strength was decreased. For a given peak current the rate of inactivation decreased as depolarization was increased. The relation of inactivation to prior Ca2+ entry was essentially linear for small currents, but decreased in slope with time during strong currents. The relation also became shallower with increasing depolarization, suggesting an apparent decrease in the efficacy of Ca in causing inactivation at more positive potentials. The basic kinetics of Ca current inactivation along with experimentally induced changes in those kinetics were simulated with a binding-site model in which inactivation develops during current flow as a function of the entry and accumulation of free Ca2+. This demonstrated that a single Ca-mediated process can account for the two-component time course of inactivation, and that the nearly bi

  6. High-resolution measurement and calibration of Ca(2+)-transients using Indo-1 in guinea-pig atrial myocytes under voltage clamp.

    PubMed

    Callewaert, G; Lipp, P; Pott, L; Carmeliet, E

    1991-04-01

    Spherical atrial myocytes obtained by enzymatic dispersion of hearts from adult guinea-pigs were loaded with the fluorescent Ca(2+)-indicator Indo-1 via patch-clamp pipettes. The dialysing solution additionally contained citrate (60 mM) as low-affinity ('linear') Ca(2+)-chelating compound in order to slow intracellular Ca(2+)-transients. Changes in Indo-1 fluorescence under voltage-clamp due to Ca(2+)-entry and/or release from the SR were calibrated using an in vivo procedure to determine the limiting fluorescence ratios. Sample recordings will be presented to demonstrate that components of a [Ca2+]i-transient due to entry via L-type Ca(2+)-channels and due to Ca(2+)-release from the SR can be directly visualized.

  7. Analysis of the effects of calcium or magnesium on voltage-clamp currents in perfused squid axons bathed in solutions of high potassium.

    PubMed

    Rojas, E; Taylor, R E; Atwater, I; Bezanilla, F

    1969-10-01

    Isolated axons from the squid, Dosidicus gigas, were internally perfused with potassium fluoride solutions. Membrane currents were measured following step changes of membrane potential in a voltage-clamp arrangement with external isosmotic solution changes in the order: potassium-free artificial seawater; potassium chloride; potassium chloride containing 10, 25, 40 or 50, mM calcium or magnesium; and potassium-free artificial seawater. The following results suggest that the currents measured under voltage clamp with potassium outside and inside can be separated into two components and that one of them, the predominant one, is carried through the potassium system. (a) Outward currents in isosmotic potassium were strongly and reversibly reduced by tetraethylammonium chloride. (b) Without calcium or magnesium a progressive increase in the nontime-dependent component of the currents (leakage) occurred. (c) The restoration of calcium or magnesium within 15-30 min decreases this leakage. (d) With 50 mM divalent ions the steady-state current-voltage curve was nonlinear with negative resistance as observed in intact axons in isosmotic potassium. (e) The time-dependent components of the membrane currents were not clearly affected by calcium or magnesium. These results show a strong dependence of the leakage currents on external calcium or magnesium concentration but provide no support for the involvement of calcium or magnesium in the kinetics of the potassium system.

  8. A combined patch-clamp and electrorotation study of the voltage- and frequency-dependent membrane capacitance caused by structurally dissimilar lipophilic anions.

    PubMed

    Zimmermann, D; Kiesel, M; Terpitz, U; Zhou, A; Reuss, R; Kraus, J; Schenk, W A; Bamberg, E; Sukhorukov, V L

    2008-01-01

    Interactions of structurally dissimilar anionic compounds with the plasma membrane of HEK293 cells were analyzed by patch clamp and electrorotation. The combined approach provides complementary information on the lipophilicity, preferential affinity of the anions to the inner/outer membrane leaflet, adsorption depth and transmembrane mobility. The anionic species studied here included the well-known lipophilic anions dipicrylamine (DPA(-)), tetraphenylborate (TPB(-)) and [W(2)(CO)(10)(S(2)CH)](-), the putative lipophilic anion B(CF(3))(4)(-) and three new heterocyclic W(CO)(5) derivatives. All tested anions partitioned strongly into the cell membrane, as indicated by the capacitance increase in patch-clamped cells. The capacitance increment exhibited a bell-shaped dependence on membrane voltage. The midpoint potentials of the maximum capacitance increment were negative, indicating the exclusion of lipophilic anions from the outer membrane leaflet. The adsorption depth of the large organic anions DPA(-), TPB(-) and B(CF(3))(4)(-) increased and that of W(CO)(5) derivatives decreased with increasing concentration of mobile charges. In agreement with the patch-clamp data, electrorotation of cells treated with DPA(-) and W(CO)(5) derivatives revealed a large dispersion of membrane capacitance in the kilohertz to megahertz range due to the translocation of mobile charges. In contrast, in the presence of TPB(-) and B(CF(3))(4)(-) no mobile charges could be detected by electrorotation, despite their strong membrane adsorption. Our data suggest that the presence of oxygen atoms in the outer molecular shell is an important factor for the fast translocation ability of lipophilic anions.

  9. Fast activating voltage- and calcium-dependent potassium (BK) conductance promotes bursting in pituitary cells: a dynamic clamp study

    PubMed Central

    Tabak, Joël; Tomaiuolo, Maurizio; Gonzalez-Iglesias, Arturo E.; Milescu, Lorin S.; Bertram, Richard

    2011-01-01

    The electrical activity pattern of endocrine pituitary cells regulates their basal secretion level. Rat somatotrophs and lactotrophs exhibit spontaneous bursting and have high basal levels of hormone secretion, while gonadotrophs exhibit spontaneous spiking and have low basal hormone secretion. It has been proposed that the difference in electrical activity between bursting somatotrophs and spiking gonadotrophs is due to the presence of large conductance potassium (BK) channels on somatotrophs but not on gonadotrophs. This is one example where the role of an ion channel type may be clearly established. We demonstrate here that BK channels indeed promote bursting activity in pituitary cells. Blocking BK channels in bursting lacto-somatotroph GH4C1 cells changes their firing activity to spiking, while further adding an artificial BK conductance via dynamic clamp restores bursting. Importantly, this burst-promoting effect requires a relatively fast BK activation/deactivation, as predicted by computational models. We also show that adding a fast activating BK conductance to spiking gonadotrophs converts the activity of these cells to bursting. Together, our results suggest that differences in BK channel expression may underlie the differences in electrical activity and basal hormone secretion levels among pituitary cell types and that the rapid rate of BK channel activation is key to its role in burst promotion. PMID:22090511

  10. Rigid clamp

    DOEpatents

    Benavides, Gilbert L.; Burt, Jack D.

    1994-01-01

    The invention relates to a clamp mechanism that can be used to attach or temporarily support objects inside of tubular goods. The clamp mechanism can also be modified so that it grips objects. The clamp has a self-centering feature to accommodate out-of-roundness or other internal defections in tubular objects such as pipe. A plurality of clamping shoes are expanded by a linkage which is preferably powered by a motor to contact the inside of a pipe. The motion can be reversed and jaw elements can be connected to the linkage so as to bring the jaws together to grab an object.

  11. Sperm Patch-Clamp

    PubMed Central

    Lishko, Polina; Clapham, David E.; Navarro, Betsy; Kirichok, Yuriy

    2014-01-01

    Sperm intracellular pH and calcium concentration ([Ca2+]i) are two central factors that control sperm activity within the female reproductive tract. As such, the ion channels of the sperm plasma membrane that alter intracellular sperm [Ca2+] and pH play important roles in sperm physiology and the process of fertilization. Indeed, sperm ion channels regulate sperm motility, control sperm chemotaxis toward the egg in some species, and may trigger the acrosome reaction. Until recently, our understanding of these important molecules was rudimentary due to the inability to patch-clamp spermatozoa and directly record the activity of these ion channels under voltage clamp. Recently, we overcame this technical barrier and developed a method for reproducible application of the patch-clamp technique to mouse and human spermatozoa. This chapter covers important aspects of application of the patch-clamp technique to spermatozoa, such as selection of the electrophysiological equipment, isolation of spermatozoa for patch-clamp experiments, formation of the gigaohm seal with spermatozoa, and transition into the whole-cell mode of recording. We also discuss potential pitfalls in application of the patch-clamp technique to flagellar ion channels. PMID:23522465

  12. Reconstitution of a Transmembrane Protein, the Voltage-gated Ion Channel, KvAP, into Giant Unilamellar Vesicles for Microscopy and Patch Clamp Studies

    PubMed Central

    Garten, Matthias; Aimon, Sophie; Bassereau, Patricia; Toombes, Gilman E. S.

    2015-01-01

    Giant Unilamellar Vesicles (GUVs) are a popular biomimetic system for studying membrane associated phenomena. However, commonly used protocols to grow GUVs must be modified in order to form GUVs containing functional transmembrane proteins. This article describes two dehydration-rehydration methods — electroformation and gel-assisted swelling — to form GUVs containing the voltage-gated potassium channel, KvAP. In both methods, a solution of protein-containing small unilamellar vesicles is partially dehydrated to form a stack of membranes, which is then allowed to swell in a rehydration buffer. For the electroformation method, the film is deposited on platinum electrodes so that an AC field can be applied during film rehydration. In contrast, the gel-assisted swelling method uses an agarose gel substrate to enhance film rehydration. Both methods can produce GUVs in low (e.g., 5 mM) and physiological (e.g., 100 mM) salt concentrations. The resulting GUVs are characterized via fluorescence microscopy, and the function of reconstituted channels measured using the inside-out patch-clamp configuration. While swelling in the presence of an alternating electric field (electroformation) gives a high yield of defect-free GUVs, the gel-assisted swelling method produces a more homogeneous protein distribution and requires no special equipment. PMID:25650630

  13. Effects of deuterium oxide on the rate and dissociation constants for saxitoxin and tetrodotoxin action. Voltage-clamp studies on frog myelinated nerve

    PubMed Central

    1981-01-01

    The actions of tetrodotoxin (TTX) and saxitoxin (STX) in normal water and in deuterium oxide (D2O) have been studied in frog myelinated nerve. Substitution of D2O for H2O in normal Ringer's solution has no effect on the potency of TTX in blocking action potentials but increases the potency of STX by approximately 50%. Under voltage clamp, the steady-state inhibition of sodium currents by 1 nM STX is doubled in D2O as a result of a halving of the rate of dissociation of STX from the sodium channel; the rate of block by STX is not measurably changed by D2O. Neither steady-state inhibition nor the on- or off-rate constants of TTX are changed by D2O substitution. The isotopic effects on STX binding are observed less than 10 min after the toxin has been added to D2O, thus eliminating the possibility that slow-exchange (t 1/2 greater than 10 h) hydrogen-binding sites on STX are involved. The results are consistent with a hypothesis that attributes receptor-toxin stabilization to isotopic changes of hydrogen bonding; this interpretation suggests that hydrogen bonds contribute more to the binding of STX than to that of TTX at the sodium channel. PMID:6268735

  14. Unloaded speed of shortening in voltage-clamped intact skeletal muscle fibers from wt, mdx, and transgenic minidystrophin mice using a novel high-speed acquisition system.

    PubMed

    Friedrich, O; Weber, C; von Wegner, F; Chamberlain, J S; Fink, R H A

    2008-06-01

    Skeletal muscle unloaded shortening has been indirectly determined in the past. Here, we present a novel high-speed optical tracking technique that allows recording of unloaded shortening in single intact, voltage-clamped mammalian skeletal muscle fibers with 2-ms time resolution. L-type Ca(2+) currents were simultaneously recorded. The time course of shortening was biexponential: a fast initial phase, tau(1), and a slower successive phase, tau(2,) with activation energies of 59 kJ/mol and 47 kJ/mol. Maximum unloaded shortening speed, v(u,max), was faster than that derived using other techniques, e.g., approximately 14.0 L(0) s(-1) at 30 degrees C. Our technique also allowed direct determination of shortening acceleration. We applied our technique to single fibers from C57 wild-type, dystrophic mdx, and minidystrophin-expressing mice to test whether unloaded shortening was affected in the pathophysiological mechanism of Duchenne muscular dystrophy. v(u,max) and a(u,max) values were not significantly different in the three strains, whereas tau(1) and tau(2) were increased in mdx fibers. The results were complemented by myosin heavy and light chain (MLC) determinations that showed the same myosin heavy chain IIA profiles in the interossei muscles from the different strains. In mdx muscle, MLC-1f was significantly increased and MLC-2f and MLC-3f somewhat reduced. Fast initial active shortening seems almost unaffected in mdx muscle.

  15. Clamp usable as jig and lifting clamp

    DOEpatents

    Tsuyama, Yoshizo

    1976-01-01

    There is provided a clamp which is well suited for use as a lifting clamp for lifting and moving materials of assembly in a shipyard, etc. and as a pulling jig in welding and other operations. The clamp comprises a clamp body including a shackle for engagement with a pulling device and a slot for receiving an article, and a pair of jaws provided on the leg portions of the clamp body on the opposite sides of the slot to grip the article in the slot, one of said jaws consisting of a screw rod and the other jaw consisting of a swivel jaw with a spherical surface, whereby when the article clamped in the slot by the pair of jaws tends to slide in any direction with respect to the clamp body, the article is more positively gripped by the pair of jaws.

  16. Coating Solution for High-Voltage Cathode: AlF3 Atomic Layer Deposition for Freestanding LiCoO2 Electrodes with High Energy Density and Excellent Flexibility.

    PubMed

    Zhou, Yun; Lee, Younghee; Sun, Huaxing; Wallas, Jasmine M; George, Steven M; Xie, Ming

    2017-03-22

    Freestanding LiCoO2/multiwall carbon nanotube/nanocellulose fibril (LCO-MWCNT-NCF) electrodes are fabricated by a vacuum filtration technique. The electrode has a high LCO loading of 20 mg/cm(2) with excellent flexibility, uniform material distribution, and low surface resistivity. When coated with 2 ALD cycles of AlF3, LCO-MWCNT-NCF has a high specific capacity of 216 mAh/g at 4.7 V. The freestanding AlF3-coated electrode preserves 75.7% of its initial capacity after 100 cycles and 70% after 160 cycles of charge discharge. In contrast, electrodes coated with 2 ALD cycles of Al2O3 cannot be cycled above 4.5 V. By elimination of the unnecessary weight of current collector, and increasing in the working voltage simultaneously, this freestanding LCO-MWCNT-NCF electrode can significantly improve the gravimetric and volumetric energy density of lithium ion batteries.

  17. Effects of Conformational Peptide Probe DP4 on Bidirectional Signaling between DHPR and RyR1 Calcium Channels in Voltage-Clamped Skeletal Muscle Fibers

    PubMed Central

    Olojo, Rotimi O.; Hernández-Ochoa, Erick O.; Ikemoto, Noriaki; Schneider, Martin F.

    2011-01-01

    In skeletal muscle, excitation-contraction coupling involves the activation of dihydropyridine receptors (DHPR) and type-1 ryanodine receptors (RyR1) to produce depolarization-dependent sarcoplasmic reticulum Ca2+ release via orthograde signaling. Another form of DHPR-RyR1 communication is retrograde signaling, in which RyRs modulate the gating of DHPR. DP4 (domain peptide 4), is a peptide corresponding to residues Leu2442-Pro2477 of the central domain of the RyR1 that produces RyR1 channel destabilization. Here we explore the effects of DP4 on orthograde excitation-contraction coupling and retrograde RyR1-DHPR signaling in isolated murine muscle fibers. Intracellular dialysis of DP4 increased the peak amplitude of Ca2+ release during step depolarizations by 64% without affecting its voltage-dependence or kinetics, and also caused a similar increase in Ca2+ release during an action potential waveform. DP4 did not modify either the amplitude or the voltage-dependence of the intramembrane charge movement. However, DP4 augmented DHPR Ca2+ current density without affecting its voltage-dependence. Our results demonstrate that the conformational changes induced by DP4 regulate both orthograde E-C coupling and retrograde RyR1-DHPR signaling. PMID:21575570

  18. Force-Measuring Clamps

    NASA Technical Reports Server (NTRS)

    Nunnelee, Mark

    2003-01-01

    Force-measuring clamps have been invented to facilitate and simplify the task of measuring the forces or pressures applied to clamped parts. There is a critical need to measure clamping forces or pressures in some applications for example, while bonding sensors to substrates or while clamping any sensitive or delicate parts. Many manufacturers of adhesives and sensors recommend clamping at specific pressures while bonding sensors or during adhesive bonding between parts in general. In the absence of a force-measuring clamp, measurement of clamping force can be cumbersome at best because of the need for additional load sensors and load-indicating equipment. One prior method of measuring clamping force involved the use of load washers or miniature load cells in combination with external power sources and load-indicating equipment. Calibrated spring clamps have also been used. Load washers and miniature load cells constitute additional clamped parts in load paths and can add to the destabilizing effects of loading mechanisms. Spring clamps can lose calibration quickly through weakening of the springs and are limited to the maximum forces that the springs can apply. The basic principle of a force-measuring clamp can be implemented on a clamp of almost any size and can enable measurement of a force of almost any magnitude. No external equipment is needed because the component(s) for transducing the clamping force and the circuitry for supplying power, conditioning the output of the transducers, and displaying the measurement value are all housed on the clamp. In other words, a force-measuring clamp is a complete force-application and force-measurement system all in one package. The advantage of unitary packaging of such a system is that it becomes possible to apply the desired clamping force or pressure with precision and ease.

  19. Estimating the number of channels in patch-clamp recordings: application to kinetic analysis of multichannel data from voltage-operated channels.

    PubMed Central

    Baumgartner, W; Hohenthanner, K; Höfer, G F; Groschner, K; Romanin, C

    1997-01-01

    Important kinetic information of voltage-operated ion channels can be obtained by estimating the open probability, the availability, and the first latency, and by applying run analysis. In the case of multichannel patches, estimation of the number of available channels is a prerequisite for the above analysis. Here we describe a method for calculation of the a posteriori probability of the number of available channels in each sweep by using the Bayes formula. This probability serves as a measure for the number of channels and allows for first latency determination and run analysis. The methods described were applied to simulated and experimental data obtained from L-type Ca2+ channel recordings. PMID:9138562

  20. Ionic currents influencing spontaneous firing and pacemaker frequency in dopamine neurons of the ventrolateral periaqueductal gray and dorsal raphe nucleus (vlPAG/DRN): A voltage-clamp and computational modelling study.

    PubMed

    Dougalis, Antonios G; Matthews, Gillian A C; Liss, Birgit; Ungless, Mark A

    2017-04-03

    Dopamine (DA) neurons of the ventrolateral periaqueductal gray (vlPAG) and dorsal raphe nucleus (DRN) fire spontaneous action potentials (APs) at slow, regular patterns in vitro but a detailed account of their intrinsic membrane properties responsible for spontaneous firing is currently lacking. To resolve this, we performed a voltage-clamp electrophysiological study in brain slices to describe their major ionic currents and then constructed a computer model and used simulations to understand the mechanisms behind autorhythmicity in silico. We found that vlPAG/DRN DA neurons exhibit a number of voltage-dependent currents activating in the subthreshold range including, a hyperpolarization-activated cation current (IH), a transient, A-type, potassium current (IA), a background, 'persistent' (INaP) sodium current and a transient, low voltage activated (LVA) calcium current (ICaLVA). Brain slice pharmacology, in good agreement with computer simulations, showed that spontaneous firing occurred independently of IH, IA or calcium currents. In contrast, when blocking sodium currents, spontaneous firing ceased and a stable, non-oscillating membrane potential below AP threshold was attained. Using the DA neuron model we further show that calcium currents exhibit little activation (compared to sodium) during the interspike interval (ISI) repolarization while, any individual potassium current alone, whose blockade positively modulated AP firing frequency, is not required for spontaneous firing. Instead, blockade of a number of potassium currents simultaneously is necessary to eliminate autorhythmicity. Repolarization during ISI is mediated initially via the deactivation of the delayed rectifier potassium current, while a sodium background 'persistent' current is essentially indispensable for autorhythmicity by driving repolarization towards AP threshold.

  1. Force-Measuring Clamp

    NASA Technical Reports Server (NTRS)

    Nunnelee, Mark (Inventor)

    2004-01-01

    A precision clamp that accurately measures force over a wide range of conditions is described. Using a full bridge or other strain gage configuration. the elastic deformation of the clamp is measured or detected by the strain gages. Thc strain gages transmit a signal that corresponds to the degree of stress upon the clamp. Thc strain gage signal is converted to a numeric display. Calibration is achieved by ero and span potentiometers which enable accurate measurements by the force-measuring clamp.

  2. Voltage clamp measurements of the hyperpolarization-activated inward current I(f) in single cells from rabbit sino-atrial node.

    PubMed Central

    van Ginneken, A C; Giles, W

    1991-01-01

    1. The kinetics and ion transfer characteristics of the hyperpolarization-activated inward current, I(f), have been studied in single cells obtained by enzymatic dispersion from the rabbit sino-atrial (S-A) node. These experiments were done to assess the role of I(f) in the generation of the pacemaker depolarization in the S-A node. 2. The activation and the deactivation of I(f) in these single cells are accompanied by significant conductance increases and decreases respectively, confirming earlier findings from multicellular man-made strips of rabbit S-A node, and from mammalian Purkinje fibres. 3. The steady-state activation of I(f) lies between -40 and -120 mV, and its voltage dependence can be described by a Boltzmann relation with the half-activation point at approximately -70 mV. 4. The delay or sigmoidicity in both the onset of I(f) and the deactivation of the tail currents can be accounted for semi-quantitatively by using a second-order Hodgkin-Huxley kinetic scheme. 5. The reversal potential for I(f) is -24 +/- 2 mV (mean +/- S.E.M., n = 6). It does not change significantly as a function of the amount of I(f) which is activated, indicating that ion accumulation or depletion phenomena are not important variables controlling the time course of I(f), or its selectivity. 6. The fully-activated current-voltage relationship for I(f) is approximately linear with a slope conductance of 12.0 +/- 0.88 nS per cell (mean +/- S.E.M., n = 6). 7. A simple mathematical model based on the measured values of maximum conductance, reversal potential, and kinetics of I(f) has been developed to simulate the size and time course of I(f) during typical spontaneous pacemaker activity in rabbit sino-atrial node cells. The calculations show that I(f) can change significantly during pacing and suggest that this current change is, at least in part, responsible for the pacemaker depolarization. Images Fig. 1 PMID:1708824

  3. Photovoltaic panel clamp

    SciTech Connect

    Mittan, Margaret Birmingham; Miros, Robert H. J.; Brown, Malcolm P; Stancel, Robert

    2012-06-05

    A photovoltaic panel clamp includes an upper and lower section. The interface between the assembled clamp halves and the module edge is filled by a flexible gasket material, such as EPDM rubber. The gasket preferably has small, finger like protrusions that allow for easy insertion onto the module edge while being reversed makes it more difficult to remove them from the module once installed. The clamp includes mounting posts or an integral axle to engage a bracket. The clamp also may include a locking tongue to secure the clamp to a bracket.

  4. Photovoltaic panel clamp

    SciTech Connect

    Brown, Malcolm P.; Mittan, Margaret Birmingham; Miros, Robert H. J.; Stancel, Robert

    2013-03-19

    A photovoltaic panel clamp includes an upper and lower section. The interface between the assembled clamp halves and the module edge is filled by a flexible gasket material, such as EPDM rubber. The gasket preferably has small, finger like protrusions that allow for easy insertion onto the module edge while being reversed makes it more difficult to remove them from the module once installed. The clamp includes mounting posts or an integral axle to engage a bracket. The clamp also may include a locking tongue to secure the clamp to a bracket.

  5. Clamping in Boltzmann machines.

    PubMed

    Livesey, M

    1991-01-01

    A certain assumption that appears in the proof of correctness of the standard Boltzmann machine learning procedure is investigated. The assumption, called the clamping assumption, concerns the behavior of a Boltzmann machine when some of its units are clamped to a fixed state. It is argued that the clamping assumption is essentially an assertion of the time reversibility of a certain Markov chain underlying the behavior of the Boltzmann machine. As such, the clamping assumption is generally false, though it is certainly true of the Boltzmann machines themselves. The author also considers how the concept of the Boltzmann machine may be generalized while retaining the validity of the clamping assumption.

  6. Construction of a low-frequency high-power piezoelectric transformer with a specified step-up voltage transformation ratio using two identical bolt-clamped Langevin-type transducers

    NASA Astrophysics Data System (ADS)

    Adachi, Kazunari; Konno, Takuma; Kosugi, Satoshi

    2015-06-01

    We propose a low-frequency piezoelectric transformer comprising two identical bolt-clamped Langevin-type transducers (BLTs) and a stepped horn with a half-wavelength straight extension. The transformer can realize a specified step-up voltage transformation ratio as determined by the cross-sectional area ratio of the horn whose both ends the two BLTs are connected to, and the driving frequency at which the specified transformation ratio is realized can be set near its mechanical resonance. Thus, it can be mechanically held firmly at its vibratory node without affecting the mechanical vibration mode or resulting in a loss of energy. After relevant finite-element simulations, experiments were conducted for a trial-fabricated transformer of the above type. As a result, the experimental results predicted by the simulations were obtained in step-up operation. The influence of the load resistance on the deviation of the driving frequency from its total mechanical resonance of 53.1 kHz was found to be less than 130 Hz (0.24% of the resonance frequency) only. High-power performance of the piezoelectric transformer was also demonstrated.

  7. Radial wedge flange clamp

    DOEpatents

    Smith, Karl H.

    2002-01-01

    A radial wedge flange clamp comprising a pair of flanges each comprising a plurality of peripheral flat wedge facets having flat wedge surfaces and opposed and mating flat surfaces attached to or otherwise engaged with two elements to be joined and including a series of generally U-shaped wedge clamps each having flat wedge interior surfaces and engaging one pair of said peripheral flat wedge facets. Each of said generally U-shaped wedge clamps has in its opposing extremities apertures for the tangential insertion of bolts to apply uniform radial force to said wedge clamps when assembled about said wedge segments.

  8. Quick action clamp

    NASA Technical Reports Server (NTRS)

    Calco, Frank S. (Inventor)

    1991-01-01

    A quick release toggle clamp that utilizes a spring that requires a deliberate positive action for disengagement is presented. The clamp has a sliding bolt that provides a latching mechanism. The bolt is moved by a handle that tends to remain in an engaged position while under tension.

  9. Laser beam guard clamps

    SciTech Connect

    Dickson, Richard K.

    2010-09-07

    A quick insert and release laser beam guard panel clamping apparatus having a base plate mountable on an optical table, a first jaw affixed to the base plate, and a spring-loaded second jaw slidably carried by the base plate to exert a clamping force. The first and second jaws each having a face acutely angled relative to the other face to form a V-shaped, open channel mouth, which enables wedge-action jaw separation by and subsequent clamping of a laser beam guard panel inserted through the open channel mouth. Preferably, the clamping apparatus also includes a support structure having an open slot aperture which is positioned over and parallel with the open channel mouth.

  10. A monogenean without clamps

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ectoparasites face a daily challenge: to remain attached to their host. Polyopisthocotylean monogeneans attach to the surface of fish gills by highly specialized structures, the sclerotized clamps. In the original description of the protomicrocotylid species Lethacotyle fijiensis, described 50 years...

  11. Piezoresistive cantilever force-clamp system

    SciTech Connect

    Park, Sung-Jin; Petzold, Bryan C.; Pruitt, Beth L.; Goodman, Miriam B.

    2011-04-15

    We present a microelectromechanical device-based tool, namely, a force-clamp system that sets or ''clamps'' the scaled force and can apply designed loading profiles (e.g., constant, sinusoidal) of a desired magnitude. The system implements a piezoresistive cantilever as a force sensor and the built-in capacitive sensor of a piezoelectric actuator as a displacement sensor, such that sample indentation depth can be directly calculated from the force and displacement signals. A programmable real-time controller operating at 100 kHz feedback calculates the driving voltage of the actuator. The system has two distinct modes: a force-clamp mode that controls the force applied to a sample and a displacement-clamp mode that controls the moving distance of the actuator. We demonstrate that the system has a large dynamic range (sub-nN up to tens of {mu}N force and nm up to tens of {mu}m displacement) in both air and water, and excellent dynamic response (fast response time, <2 ms and large bandwidth, 1 Hz up to 1 kHz). In addition, the system has been specifically designed to be integrated with other instruments such as a microscope with patch-clamp electronics. We demonstrate the capabilities of the system by using it to calibrate the stiffness and sensitivity of an electrostatic actuator and to measure the mechanics of a living, freely moving Caenorhabditis elegans nematode.

  12. Clamping characteristics study on different types of clamping unit

    SciTech Connect

    Jiao, Zhiwei; Liu, Haichao; Xie, Pengcheng; Yang, Weimin

    2015-05-22

    Plastic products are becoming more and more widely used in aerospace, IT, digital electronics and many other fields. With the development of technology, the requirement of product precision is getting higher and higher. However, type and working performance of clamping unit play a decisive role in product precision. Clamping characteristics of different types of clamping unit are discussed in this article, which use finite element numerical analysis method through the software ABAQUS to study the clamping uniformity, and detect the clamping force repeatability precision. The result shows that compared with toggled three-platen clamping unit, clamping characteristics of internal circulation two-platen clamping unit are better, for instance, its mold cavity deformation and force that bars and mold parting surface suffered are more uniform, and its clamping uniformity and repeatability precision is also better.

  13. Clamp for detonating fuze

    NASA Technical Reports Server (NTRS)

    Holderman, E. J.

    1968-01-01

    Quick acting clamp provides physical support for a closely confined detonating fuse in an application requiring removal and replacement at frequent intervals during test. It can be designed with a base of any required strength and configuration to permit the insertion of an object.

  14. Microchip amplifier for in vitro, in vivo, and automated whole cell patch-clamp recording

    PubMed Central

    Kolb, Ilya; Kodandaramaiah, Suhasa B.; Chubykin, Alexander A.; Yang, Aimei; Bear, Mark F.; Boyden, Edward S.; Forest, Craig R.

    2014-01-01

    Patch clamping is a gold-standard electrophysiology technique that has the temporal resolution and signal-to-noise ratio capable of reporting single ion channel currents, as well as electrical activity of excitable single cells. Despite its usefulness and decades of development, the amplifiers required for patch clamping are expensive and bulky. This has limited the scalability and throughput of patch clamping for single-ion channel and single-cell analyses. In this work, we have developed a custom patch-clamp amplifier microchip that can be fabricated using standard commercial silicon processes capable of performing both voltage- and current-clamp measurements. A key innovation is the use of nonlinear feedback elements in the voltage-clamp amplifier circuit to convert measured currents into logarithmically encoded voltages, thereby eliminating the need for large high-valued resistors, a factor that has limited previous attempts at integration. Benchtop characterization of the chip shows low levels of current noise [1.1 pA root mean square (rms) over 5 kHz] during voltage-clamp measurements and low levels of voltage noise (8.2 μV rms over 10 kHz) during current-clamp measurements. We demonstrate the ability of the chip to perform both current- and voltage-clamp measurement in vitro in HEK293FT cells and cultured neurons. We also demonstrate its ability to perform in vivo recordings as part of a robotic patch-clamping system. The performance of the patch-clamp amplifier microchip compares favorably with much larger commercial instrumentation, enabling benchtop commoditization, miniaturization, and scalable patch-clamp instrumentation. PMID:25429119

  15. Cantilever clamp fitting

    NASA Technical Reports Server (NTRS)

    Melton, Patrick B. (Inventor)

    1989-01-01

    A device is disclosed for sealing and clamping a cylindrical element which is to be attached to an object such as a wall, a pressurized vessel or another cylindrical element. The device includes a gland having an inner cylindrical wall, which is threaded at one end and is attached at a bendable end to a deformable portion, which in turn is attached to one end of a conical cantilever structure. The other end of the cantilever structure connects at a bendable area to one end of an outer cylindrical wall. The opposite end of cylindrical wall terminates in a thickened portion, the radially outer surface of which is adapted to accommodate a tool for rotating the gland. The terminal end of cylindrical wall also includes an abutment surface, which is adapted to engage a seal, which in turn engages a surface of a receiver. The receiver further includes a threaded portion for engagement with the threaded portion of gland whereby a tightening rotation of gland relative to receiver will cause relative movement between cylindrical walls and of gland. This movement causes a rotation of the conical structure and thus a bending action at bending area and at the bending end of the upper end of inner cylindrical wall. These rotational and bending actions result in a forcing of the deformable portion radially inwardly so as to contact and deform a pipe. This forcible contact creates a seal between gland and pipe, and simultaneously clamps the pipe in position.

  16. Whole-GUV patch-clamping.

    PubMed

    Garten, Matthias; Mosgaard, Lars D; Bornschlögl, Thomas; Dieudonné, Stéphane; Bassereau, Patricia; Toombes, Gilman E S

    2017-01-10

    Studying how the membrane modulates ion channel and transporter activity is challenging because cells actively regulate membrane properties, whereas existing in vitro systems have limitations, such as residual solvent and unphysiologically high membrane tension. Cell-sized giant unilamellar vesicles (GUVs) would be ideal for in vitro electrophysiology, but efforts to measure the membrane current of intact GUVs have been unsuccessful. In this work, two challenges for obtaining the "whole-GUV" patch-clamp configuration were identified and resolved. First, unless the patch pipette and GUV pressures are precisely matched in the GUV-attached configuration, breaking the patch membrane also ruptures the GUV. Second, GUVs shrink irreversibly because the membrane/glass adhesion creating the high-resistance seal (>1 GΩ) continuously pulls membrane into the pipette. In contrast, for cell-derived giant plasma membrane vesicles (GPMVs), breaking the patch membrane allows the GPMV contents to passivate the pipette surface, thereby dynamically blocking membrane spreading in the whole-GMPV mode. To mimic this dynamic passivation mechanism, beta-casein was encapsulated into GUVs, yielding a stable, high-resistance, whole-GUV configuration for a range of membrane compositions. Specific membrane capacitance measurements confirmed that the membranes were truly solvent-free and that membrane tension could be controlled over a physiological range. Finally, the potential for ion transport studies was tested using the model ion channel, gramicidin, and voltage-clamp fluorometry measurements were performed with a voltage-dependent fluorophore/quencher pair. Whole-GUV patch-clamping allows ion transport and other voltage-dependent processes to be studied while controlling membrane composition, tension, and shape.

  17. Single electrode dynamic clamp with StdpC

    PubMed Central

    Samu, David; Marra, Vincenzo; Kemenes, Ildiko; Crossley, Michael; Kemenes, György; Staras, Kevin; Nowotny, Thomas

    2012-01-01

    Dynamic clamp is a powerful approach for electrophysiological investigations allowing researchers to introduce artificial electrical components into target neurons to simulate ionic conductances, chemical or electrotonic inputs or connections to other cells. Due to the rapidly changing and potentially large current injections during dynamic clamp, problematic voltage artifacts appear on the electrode used to inject dynamic clamp currents into a target neuron. Dynamic clamp experiments, therefore, typically use two separate electrodes in the same cell, one for recording membrane potential and one for injecting currents. The requirement for two independent electrodes has been a limiting factor for the use of dynamic clamp in applications where dual recordings of this kind are difficult or impossible to achieve. The recent development of an active electrode compensation (AEC) method has overcome some of these prior limitations, permitting artifact-free dynamic clamp experimentation with a single electrode. Here we describe an AEC method for the free dynamic clamp software StdpC. The AEC component of StdpC is the first such system implemented for the use of non-expert users and comes with a set of semi-automated configuration and calibration procedures that facilitate its use. We briefly introduce the AEC method and its implementation in StdpC and then validate it with an electronic model cell and in two different biological preparations. PMID:22898473

  18. Single electrode dynamic clamp with StdpC.

    PubMed

    Samu, David; Marra, Vincenzo; Kemenes, Ildiko; Crossley, Michael; Kemenes, György; Staras, Kevin; Nowotny, Thomas

    2012-10-15

    Dynamic clamp is a powerful approach for electrophysiological investigations allowing researchers to introduce artificial electrical components into target neurons to simulate ionic conductances, chemical or electrotonic inputs or connections to other cells. Due to the rapidly changing and potentially large current injections during dynamic clamp, problematic voltage artifacts appear on the electrode used to inject dynamic clamp currents into a target neuron. Dynamic clamp experiments, therefore, typically use two separate electrodes in the same cell, one for recording membrane potential and one for injecting currents. The requirement for two independent electrodes has been a limiting factor for the use of dynamic clamp in applications where dual recordings of this kind are difficult or impossible to achieve. The recent development of an active electrode compensation (AEC) method has overcome some of these prior limitations, permitting artifact-free dynamic clamp experimentation with a single electrode. Here we describe an AEC method for the free dynamic clamp software StdpC. The AEC component of StdpC is the first such system implemented for the use of non-expert users and comes with a set of semi-automated configuration and calibration procedures that facilitate its use. We briefly introduce the AEC method and its implementation in StdpC and then validate it with an electronic model cell and in two different biological preparations.

  19. Saddle Clamp With Captive Components

    NASA Technical Reports Server (NTRS)

    Belrose, Charles R.

    1993-01-01

    Saddle clamp modified to prevent parts from falling off when installed or removed. Allows easy access for tightening or loosening bolts, and retains alignment with tube mounted in it when opened. All parts are held captive - bolts by retaining washers, floating nuts by pressing and swaging, and upper clamp band by tether. Upper and lower bolt flanges offset from each other to ensure access.

  20. Plastic Clamp Retains Clevis Pin

    NASA Technical Reports Server (NTRS)

    Cortes, R. G.

    1983-01-01

    Plastic clamp requires no special installation or removal tools. Clamp slips easily over end of pin. Once engaged in groove, holds pin securely. Installed and removed easily without special tools - screwdriver or putty knife adequate for prying out of groove. Used to retain bearings, rollers pulleys, other parts that rotate. Applications include slowly and intermittently rotating parts in appliances.

  1. Internal V-Band Clamp

    DOEpatents

    Vaughn, Mark R.; Hafenrichter, Everett S.; Chapa, Agapito C.; Harris, Steven M.; Martinez, Marcus J.; Baty, Roy S.

    2006-02-28

    A system for clamping two tubular members together in an end-to-end relationship uses a split ring with a V-shaped outer rim that can engage a clamping surface on each member. The split ring has a relaxed closed state where the ends of the ring are adjacent and the outside diameter of the split ring is less than the minimum inside diameter of the members at their ends. The members are clamped when the split ring is spread into an elastically stretched position where the ring rim is pressed tightly against the interior surfaces of the members. Mechanisms are provided for removing the spreader so the split ring will return to the relaxed state, releasing the clamped members.

  2. Reconstruction of Cell Surface Densities of Ion Pumps, Exchangers, and Channels from mRNA Expression, Conductance Kinetics, Whole-Cell Calcium, and Current-Clamp Voltage Recordings, with an Application to Human Uterine Smooth Muscle Cells.

    PubMed

    Atia, Jolene; McCloskey, Conor; Shmygol, Anatoly S; Rand, David A; van den Berg, Hugo A; Blanks, Andrew M

    2016-04-01

    Uterine smooth muscle cells remain quiescent throughout most of gestation, only generating spontaneous action potentials immediately prior to, and during, labor. This study presents a method that combines transcriptomics with biophysical recordings to characterise the conductance repertoire of these cells, the 'conductance repertoire' being the total complement of ion channels and transporters expressed by an electrically active cell. Transcriptomic analysis provides a set of potential electrogenic entities, of which the conductance repertoire is a subset. Each entity within the conductance repertoire was modeled independently and its gating parameter values were fixed using the available biophysical data. The only remaining free parameters were the surface densities for each entity. We characterise the space of combinations of surface densities (density vectors) consistent with experimentally observed membrane potential and calcium waveforms. This yields insights on the functional redundancy of the system as well as its behavioral versatility. Our approach couples high-throughput transcriptomic data with physiological behaviors in health and disease, and provides a formal method to link genotype to phenotype in excitable systems. We accurately predict current densities and chart functional redundancy. For example, we find that to evoke the observed voltage waveform, the BK channel is functionally redundant whereas hERG is essential. Furthermore, our analysis suggests that activation of calcium-activated chloride conductances by intracellular calcium release is the key factor underlying spontaneous depolarisations.

  3. Reconstruction of Cell Surface Densities of Ion Pumps, Exchangers, and Channels from mRNA Expression, Conductance Kinetics, Whole-Cell Calcium, and Current-Clamp Voltage Recordings, with an Application to Human Uterine Smooth Muscle Cells

    PubMed Central

    Atia, Jolene; McCloskey, Conor; Shmygol, Anatoly S.; Rand, David A.; van den Berg, Hugo A.; Blanks, Andrew M.

    2016-01-01

    Uterine smooth muscle cells remain quiescent throughout most of gestation, only generating spontaneous action potentials immediately prior to, and during, labor. This study presents a method that combines transcriptomics with biophysical recordings to characterise the conductance repertoire of these cells, the ‘conductance repertoire’ being the total complement of ion channels and transporters expressed by an electrically active cell. Transcriptomic analysis provides a set of potential electrogenic entities, of which the conductance repertoire is a subset. Each entity within the conductance repertoire was modeled independently and its gating parameter values were fixed using the available biophysical data. The only remaining free parameters were the surface densities for each entity. We characterise the space of combinations of surface densities (density vectors) consistent with experimentally observed membrane potential and calcium waveforms. This yields insights on the functional redundancy of the system as well as its behavioral versatility. Our approach couples high-throughput transcriptomic data with physiological behaviors in health and disease, and provides a formal method to link genotype to phenotype in excitable systems. We accurately predict current densities and chart functional redundancy. For example, we find that to evoke the observed voltage waveform, the BK channel is functionally redundant whereas hERG is essential. Furthermore, our analysis suggests that activation of calcium-activated chloride conductances by intracellular calcium release is the key factor underlying spontaneous depolarisations. PMID:27105427

  4. Near-uv photon efficiency in a TiO2 electrode - Application to hydrogen production from solar energy

    NASA Technical Reports Server (NTRS)

    Desplat, J.-L.

    1976-01-01

    An n-type (001) TiO2 electrode irradiated at 365 nm was tested under anodic polarization. A saturation current independent of pH and proportional to light intensity has been observed. Accurate measurements of the incident power lead to a 60 per cent photon efficiency. A photoelectrochemical cell built with such an electrode, operated under solar irradiation without concentration, produced an electrolysis current of 0.7 mA/sq cm without applied voltage.

  5. Split-tapered joint clamping device

    DOEpatents

    Olsen, Max J.; Schwartz, Jr., John F.

    1988-01-01

    This invention relates to a clamping device for removably attaching a tool element to a bracket element wherein a bracket element is disposed in a groove in the tool and a clamping member is disposed in said groove and in engagement with a clamping face of the bracket and a wall of the groove and with the clamping member having pivot means engaging the bracket and about which the clamping member rotates.

  6. Note: High-efficiency energy harvester using double-clamped piezoelectric beams

    SciTech Connect

    Zheng, Yingmei; Wu, Xuan; Parmar, Mitesh; Lee, Dong-weon

    2014-02-15

    In this study, an improvement in energy conversion efficiency has been reported, which is realized by using a double-clamped piezoelectric beam, based on uniaxial stretching strain. The buckling mechanism is applied to maximize axial stress in the double-clamped beam. The voltage generated by using the double-clamped piezoelectric beam is higher than that generated by using other conventional structures, such as bending cantilevers coated/sandwiched with piezoelectric film, which is proven both theoretically and experimentally. The power generation efficiency is enhanced by further optimizing the double-clamped structure. The optimized high-efficiency energy harvester utilizing double-clamped piezoelectric beams generates a peak output power of 80 μW, under an acceleration of 0.1g.

  7. Note: high-efficiency energy harvester using double-clamped piezoelectric beams.

    PubMed

    Zheng, Yingmei; Wu, Xuan; Parmar, Mitesh; Lee, Dong-weon

    2014-02-01

    In this study, an improvement in energy conversion efficiency has been reported, which is realized by using a double-clamped piezoelectric beam, based on uniaxial stretching strain. The buckling mechanism is applied to maximize axial stress in the double-clamped beam. The voltage generated by using the double-clamped piezoelectric beam is higher than that generated by using other conventional structures, such as bending cantilevers coated/sandwiched with piezoelectric film, which is proven both theoretically and experimentally. The power generation efficiency is enhanced by further optimizing the double-clamped structure. The optimized high-efficiency energy harvester utilizing double-clamped piezoelectric beams generates a peak output power of 80 μW, under an acceleration of 0.1g.

  8. Note: High-efficiency energy harvester using double-clamped piezoelectric beams

    NASA Astrophysics Data System (ADS)

    Zheng, Yingmei; Wu, Xuan; Parmar, Mitesh; Lee, Dong-weon

    2014-02-01

    In this study, an improvement in energy conversion efficiency has been reported, which is realized by using a double-clamped piezoelectric beam, based on uniaxial stretching strain. The buckling mechanism is applied to maximize axial stress in the double-clamped beam. The voltage generated by using the double-clamped piezoelectric beam is higher than that generated by using other conventional structures, such as bending cantilevers coated/sandwiched with piezoelectric film, which is proven both theoretically and experimentally. The power generation efficiency is enhanced by further optimizing the double-clamped structure. The optimized high-efficiency energy harvester utilizing double-clamped piezoelectric beams generates a peak output power of 80 μW, under an acceleration of 0.1g.

  9. Excitation-contraction coupling in voltage clamped uterine smooth muscle

    PubMed Central

    Mironneau, Jean

    1973-01-01

    1. The relationship between ionic currents and contraction has been investigated in uterine strips of pregnant rat by means of a double sucrose gap apparatus combined with an optical method which permits the measurement of the contraction of the small muscular bundle where potential and current are recorded. 2. Effects of duration, size and frequency of imposed potentials upon contraction have been studied. The uterine muscle shows summation and tetanus phenomena. Tension elicited by depolarizing pulses of different durations and amplitudes can be considered as made of two components. 3. The first component of the contraction evoked by short depolarizing steps (about 50 ms) depends on the slow inward current. This contraction is abolished by manganese and lanthanum ions and by compound D 600. The amplitude of the tension can be related to the external calcium concentration and consequently to the calcium influx. The slow inward current is supposed to release a part of the bound calcium without excluding, however, a direct activation of myofibrils. 4. The second component of the contraction is observed in manganese containing solution with depolarizations longer than 200 ms and without inward current. Such a component of tension suggests the possibility of release of calcium from intracellular stores which could be located in the sarcoplasmic membrane of the uterine smooth muscle. PMID:4796671

  10. The RFC clamp loader: structure and function.

    PubMed

    Yao, Nina Y; O'Donnell, Mike

    2012-01-01

    The eukaryotic RFC clamp loader couples the energy of ATP hydrolysis to open and close the circular PCNA sliding clamp onto primed sites for use by DNA polymerases and repair factors. Structural studies reveal clamp loaders to be heteropentamers. Each subunit contains a region of homology to AAA+ proteins that defines two domains. The AAA+ domains form a right-handed spiral upon binding ATP. This spiral arrangement generates a DNA binding site within the center of RFC. DNA enters the central chamber through a gap between the AAA+ domains of two subunits. Specificity for a primed template junction is achieved by a third domain that blocks DNA, forcing it to bend sharply. Thus only DNA with a flexible joint can bind the central chamber. DNA entry also requires a slot in the PCNA clamp, which is opened upon binding the AAA+ domains of the clamp loader. ATP hydrolysis enables clamp closing and ejection of RFC, completing the clamp loading reaction.

  11. High-speed pressure clamp.

    PubMed

    Besch, Stephen R; Suchyna, Thomas; Sachs, Frederick

    2002-10-01

    We built a high-speed, pneumatic pressure clamp to stimulate patch-clamped membranes mechanically. The key control element is a newly designed differential valve that uses a single, nickel-plated piezoelectric bending element to control both pressure and vacuum. To minimize response time, the valve body was designed with minimum dead volume. The result is improved response time and stability with a threefold decrease in actuation latency. Tight valve clearances minimize the steady-state air flow, permitting us to use small resonant-piston pumps to supply pressure and vacuum. To protect the valve from water contamination in the event of a broken pipette, an optical sensor detects water entering the valve and increases pressure rapidly to clear the system. The open-loop time constant for pressure is 2.5 ms for a 100-mmHg step, and the closed-loop settling time is 500-600 micros. Valve actuation latency is 120 micros. The system performance is illustrated for mechanically induced changes in patch capacitance.

  12. Advanced motor driven clamped borehole seismic receiver

    DOEpatents

    Engler, Bruce P.; Sleefe, Gerard E.; Striker, Richard P.

    1993-01-01

    A borehole seismic tool including a borehole clamp which only moves perpendicular to the borehole. The clamp is driven by an electric motor, via a right angle drive. When used as a seismic receiver, the tool has a three part housing, two of which are hermetically sealed. Accelerometers or geophones are mounted in one hermetically sealed part, the electric meter in the other hermetically sealed part, and the clamp and right angle drive in the third part. Preferably the tool includes cable connectors at both ends. Optionally a shear plate can be added to the clamp to extend the range of the tool.

  13. Advanced motor driven clamped borehole seismic receiver

    DOEpatents

    Engler, B.P.; Sleefe, G.E.; Striker, R.P.

    1993-02-23

    A borehole seismic tool is described including a borehole clamp which only moves perpendicular to the borehole. The clamp is driven by an electric motor, via a right angle drive. When used as a seismic receiver, the tool has a three part housing, two of which are hermetically sealed. Accelerometers or geophones are mounted in one hermetically sealed part, the electric motor in the other hermetically sealed part, and the clamp and right angle drive in the third part. Preferably the tool includes cable connectors at both ends. Optionally a shear plate can be added to the clamp to extend the range of the tool.

  14. MATLAB implementation of a dynamic clamp with bandwidth >125 KHz capable of generating INa at 37°C

    PubMed Central

    Clausen, Chris; Valiunas, Virginijus; Brink, Peter R.; Cohen, Ira S.

    2012-01-01

    We describe the construction of a dynamic clamp with bandwidth >125 KHz that utilizes a high performance, yet low cost, standard home/office PC interfaced with a high-speed (16 bit) data acquisition module. High bandwidth is achieved by exploiting recently available software advances (code-generation technology, optimized real-time kernel). Dynamic-clamp programs are constructed using Simulink, a visual programming language. Blocks for computation of membrane currents are written in the high-level matlab language; no programming in C is required. The instrument can be used in single- or dual-cell configurations, with the capability to modify programs while experiments are in progress. We describe an algorithm for computing the fast transient Na+ current (INa) in real time, and test its accuracy and stability using rate constants appropriate for 37°C. We then construct a program capable of supplying three currents to a cell preparation: INa, the hyperpolarizing-activated inward pacemaker current (If), and an inward-rectifier K+ current (IK1). The program corrects for the IR drop due to electrode current flow, and also records all voltages and currents. We tested this program on dual patch-clamped HEK293 cells where the dynamic clamp controls a current-clamp amplifier and a voltage-clamp amplifier controls membrane potential, and current-clamped HEK293 cells where the dynamic clamp produces spontaneous pacing behavior exhibiting Na+ spikes in otherwise passive cells. PMID:23224681

  15. The Monogenean Which Lost Its Clamps

    PubMed Central

    Justine, Jean-Lou; Rahmouni, Chahrazed; Gey, Delphine; Schoelinck, Charlotte; Hoberg, Eric P.

    2013-01-01

    Ectoparasites face a daily challenge: to remain attached to their hosts. Polyopisthocotylean monogeneans usually attach to the surface of fish gills using highly specialized structures, the sclerotized clamps. In the original description of the protomicrocotylid species Lethacotyle fijiensis, described 60 years ago, the clamps were considered to be absent but few specimens were available and this observation was later questioned. In addition, genera within the family Protomicrocotylidae have either clamps of the “gastrocotylid” or the “microcotylid” types; this puzzled systematists because these clamp types are characteristic of distinct, major groups. Discovery of another, new, species of the genus Lethacotyle, has allowed us to explore the nature of the attachment structures in protomicrocotylids. Lethacotyle vera n. sp. is described from the gills of the carangid Caranx papuensis off New Caledonia. It is distinguished from Lethacotyle fijiensis, the only other species of the genus, by the length of the male copulatory spines. Sequences of 28S rDNA were used to build a tree, in which Lethacotyle vera grouped with other protomicrocotylids. The identity of the host fish was confirmed with COI barcodes. We observed that protomicrocotylids have specialized structures associated with their attachment organ, such as lateral flaps and transverse striations, which are not known in other monogeneans. We thus hypothesized that the clamps in protomicrocotylids were sequentially lost during evolution, coinciding with the development of other attachment structures. To test the hypothesis, we calculated the surfaces of clamps and body in 120 species of gastrocotylinean monogeneans, based on published descriptions. The ratio of clamp surface: body surface was the lowest in protomicrocotylids. We conclude that clamps in protomicrocotylids are vestigial organs, and that occurrence of “gastrocotylid” and simpler “microcotylid” clamps within the same family are

  16. The monogenean which lost its clamps.

    PubMed

    Justine, Jean-Lou; Rahmouni, Chahrazed; Gey, Delphine; Schoelinck, Charlotte; Hoberg, Eric P

    2013-01-01

    Ectoparasites face a daily challenge: to remain attached to their hosts. Polyopisthocotylean monogeneans usually attach to the surface of fish gills using highly specialized structures, the sclerotized clamps. In the original description of the protomicrocotylid species Lethacotyle fijiensis, described 60 years ago, the clamps were considered to be absent but few specimens were available and this observation was later questioned. In addition, genera within the family Protomicrocotylidae have either clamps of the "gastrocotylid" or the "microcotylid" types; this puzzled systematists because these clamp types are characteristic of distinct, major groups. Discovery of another, new, species of the genus Lethacotyle, has allowed us to explore the nature of the attachment structures in protomicrocotylids. Lethacotyle vera n. sp. is described from the gills of the carangid Caranx papuensis off New Caledonia. It is distinguished from Lethacotyle fijiensis, the only other species of the genus, by the length of the male copulatory spines. Sequences of 28S rDNA were used to build a tree, in which Lethacotyle vera grouped with other protomicrocotylids. The identity of the host fish was confirmed with COI barcodes. We observed that protomicrocotylids have specialized structures associated with their attachment organ, such as lateral flaps and transverse striations, which are not known in other monogeneans. We thus hypothesized that the clamps in protomicrocotylids were sequentially lost during evolution, coinciding with the development of other attachment structures. To test the hypothesis, we calculated the surfaces of clamps and body in 120 species of gastrocotylinean monogeneans, based on published descriptions. The ratio of clamp surface: body surface was the lowest in protomicrocotylids. We conclude that clamps in protomicrocotylids are vestigial organs, and that occurrence of "gastrocotylid" and simpler "microcotylid" clamps within the same family are steps in an

  17. Implementation of a fast 16-Bit dynamic clamp using LabVIEW-RT.

    PubMed

    Kullmann, Paul H M; Wheeler, Diek W; Beacom, Joshua; Horn, John P

    2004-01-01

    The dynamic-clamp method provides a powerful electrophysiological tool for creating virtual ionic conductances in living cells and studying their influence on membrane potential. Here we describe G-clamp, a new way to implement a dynamic clamp using the real-time version of the Lab-VIEW programming environment together with a Windows host, an embedded microprocessor that runs a real-time operating system and a multifunction data-acquisition board. The software includes descriptions of a fast voltage-dependent sodium conductance, delayed rectifier, M-type and A-type potassium conductances, and a leak conductance. The system can also read synaptic conductance waveforms from preassembled data files. These virtual conductances can be reliably implemented at speeds < or =43 kHz while simultaneously saving two channels of data with 16-bit precision. G-clamp also includes utilities for measuring current-voltage relations, synaptic strength, and synaptic gain. Taking an approach built on a commercially available software/hardware platform has resulted in a system that is easy to assemble and upgrade. In addition, the graphical programming structure of LabVIEW should make it relatively easy for others to adapt G-clamp for new experimental applications.

  18. Molecular Mechanisms of DNA Polymerase Clamp Loaders

    NASA Astrophysics Data System (ADS)

    Kelch, Brian; Makino, Debora; Simonetta, Kyle; O'Donnell, Mike; Kuriyan, John

    Clamp loaders are ATP-driven multiprotein machines that couple ATP hydrolysis to the opening and closing of a circular protein ring around DNA. This ring-shaped clamp slides along DNA, and interacts with numerous proteins involved in DNA replication, DNA repair and cell cycle control. Recently determined structures of clamp loader complexes from prokaryotic and eukaryotic DNA polymerases have revealed exciting new details of how these complex AAA+ machines perform this essential clamp loading function. This review serves as background to John Kuriyan's lecture at the 2010 Erice School, and is not meant as a comprehensive review of the contributions of the many scientists who have advanced this field. These lecture notes are derived from recent reviews and research papers from our groups.

  19. Dynamic clamp with StdpC software.

    PubMed

    Kemenes, Ildikó; Marra, Vincenzo; Crossley, Michael; Samu, Dávid; Staras, Kevin; Kemenes, György; Nowotny, Thomas

    2011-03-01

    Dynamic clamp is a powerful method that allows the introduction of artificial electrical components into target cells to simulate ionic conductances and synaptic inputs. This method is based on a fast cycle of measuring the membrane potential of a cell, calculating the current of a desired simulated component using an appropriate model and injecting this current into the cell. Here we present a dynamic clamp protocol using free, fully integrated, open-source software (StdpC, for spike timing-dependent plasticity clamp). Use of this protocol does not require specialist hardware, costly commercial software, experience in real-time operating systems or a strong programming background. The software enables the configuration and operation of a wide range of complex and fully automated dynamic clamp experiments through an intuitive and powerful interface with a minimal initial lead time of a few hours. After initial configuration, experimental results can be generated within minutes of establishing cell recording.

  20. Soft-Switched Neutral-Point-Clamped Single-Phase Boost Rectifier

    NASA Astrophysics Data System (ADS)

    Itoh, Ryozo; Ishizaka, Kouichi

    A soft-switched neutral-point-clamped single-phase boost rectifier capable of compensating the imbalance load voltage is studied. This is based on a single-phase rectifier, in which an inductor is placed in series with the AC supply to resonate with a capacitor connected across the DC output of a full-bridge rectifier and the switching transition is mainly governed by a series resonance. The experimental prototype using insulated-gate bipolar transistors is implemented to investigate the operation under the charge control. The experimental results confirm that the rectifier has a neutral-point-clamp feature providing a good quality AC current.

  1. Diverless pipeline repair clamp: Phase 1

    SciTech Connect

    Miller, J.E.; Knott, B. )

    1991-12-01

    Offshore oil and gas developments are underway for water depths beyond which divers can function. The economic lifelines of these projects are the pipelines which will transport the products to shore. In preparation for the day when one of these pipelines will require repair because of a leak, the Pipeline Research Committee of the American Gas Association is funding research directed at developing diverless pipeline repair capabilities. Several types of damage are possible, ranging from latent weld defects on one end of the spectrum to damage resulting in parting of the pipe at the other end. This study is specifically directed toward laying the groundwork for development of a diverless pipeline repair clamp for use in repair of leaks resulting from minor pipe defects. The incentive for a clamp type repair is costs. When compared to replacing a section of pipe, either by welding or by mechanical means, the clamp type repair requires much less disturbance of the pipe, less time, fewer operations and less equipment. This report summarizes (1) capabilities of remotely operated vehicles (ROV's) and associated systems, (2) highlights areas for further research and development, (3) describes the required capabilities of the diverless repairclamp, (4) investigates some alternatives to the diverless clamp, (5) overviews the state of the art in leak repair clamps, and (6) critiques several possible generic clamp concepts.

  2. Sound absorption by clamped poroelastic plates.

    PubMed

    Aygun, H; Attenborough, K

    2008-09-01

    Measurements and predictions have been made of the absorption coefficient and the surface acoustic impedance of poroelastic plates clamped in a large impedance tube and separated from the rigid termination by an air gap. The measured and predicted absorption coefficient and surface impedance spectra exhibit low frequency peaks. The peak frequencies observed in the absorption coefficient are close to those predicted and measured in the deflection spectra of the clamped poroelastic plates. The influences of the rigidity of the clamping conditions and the width of the air gap have been investigated. Both influences are found to be important. Increasing the rigidity of clamping reduces the low frequency absorption peaks compared with those measured for simply supported plates or plates in an intermediate clamping condition. Results for a closed cell foam plate and for two open cell foam plates made from recycled materials are presented. For identical clamping conditions and width of air gap, the results for the different materials differ as a consequence mainly of their different elasticity, thickness, and cell structure.

  3. 21 CFR 882.4460 - Neurosurgical head holder (skull clamp).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Neurosurgical head holder (skull clamp). 882.4460... holder (skull clamp). (a) Identification. A neurosurgical head holder (skull clamp) is a device used to clamp the patient's skull to hold head and neck in a particular position during surgical procedures....

  4. 21 CFR 882.4460 - Neurosurgical head holder (skull clamp).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Neurosurgical head holder (skull clamp). 882.4460... holder (skull clamp). (a) Identification. A neurosurgical head holder (skull clamp) is a device used to clamp the patient's skull to hold head and neck in a particular position during surgical procedures....

  5. 21 CFR 882.4460 - Neurosurgical head holder (skull clamp).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Neurosurgical head holder (skull clamp). 882.4460... holder (skull clamp). (a) Identification. A neurosurgical head holder (skull clamp) is a device used to clamp the patient's skull to hold head and neck in a particular position during surgical procedures....

  6. 21 CFR 882.4460 - Neurosurgical head holder (skull clamp).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Neurosurgical head holder (skull clamp). 882.4460... holder (skull clamp). (a) Identification. A neurosurgical head holder (skull clamp) is a device used to clamp the patient's skull to hold head and neck in a particular position during surgical procedures....

  7. 21 CFR 882.4460 - Neurosurgical head holder (skull clamp).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Neurosurgical head holder (skull clamp). 882.4460... holder (skull clamp). (a) Identification. A neurosurgical head holder (skull clamp) is a device used to clamp the patient's skull to hold head and neck in a particular position during surgical procedures....

  8. Calcium-activated conductance in skate electroreceptors: current clamp experiments.

    PubMed

    Clusin, W T; Bennett, M V

    1977-02-01

    When current clamped, skate electroreceptor epithelium produces large action potentials in response to stimuli that depolarize the lumenal faces of the receptor cells. With increasing stimulus strength these action potentials become prolonged. When the peak voltage exceeds about 140 mV the repolarizing phase is blocked until the end of the stimulus. Perfusion experiments show that the rising phase of the action potential results from an increase in calcium permeability in the lumenal membranes. Perfusion of the lumen with cobalt or with a zero calcium solution containing EGTA blocks the action potential. Perfusion of the lumen with a solution containing 10 mM Ca and 20 mM EGTA initially slows the repolarizing process at all voltages and lowers the potential at which it is blocked. With prolonged perfusion, repolarization is blocked at all voltages. When excitability is abolished by perfusion with cobalt, or with a zero calcium solution containing EGTA, no delayed rectification occurs. We suggest that repolarization during the action potential depends on an influx of calcium into the cytoplasm, and that the rate of repolarization depends on the magnitude of the inward calcium current. Increasingly large stimuli reduce the rate of repolarization by reducing the driving force for calcium, and then block repolarization by causing the lumenal membrane potential to exceed ECa. Changes in extracellular calcium affect repolarization in a manner consistent with the resulting change in ECa.

  9. An Ultrasonic Clamp for Bloodless Partial Nephrectomy

    NASA Astrophysics Data System (ADS)

    Lafon, Cyril; Bouchoux, Guillaume; Murat, François Joseph; Birer, Alain; Theillère, Yves; Chapelon, Jean Yves; Cathignol, Dominique

    2007-05-01

    Maximum conservation of the kidney is preferable through partial nephrectomy for patients at risk of disease recurrence of renal cancers. Haemostatic tools are needed in order to achieve bloodless surgery and reduce post surgery morbidity. Two piezo-ceramic transducers operating at a frequency of 4 MHz were mounted on each arm of a clamp. When used for coagulation purposes, two transducers situated on opposite arms of the clamp were driven simultaneously. Heat delivery was optimized as each transducers mirrored back to targeted tissues the wave generated by the opposite transducer. Real-time treatment monitoring with an echo-based technique was also envisaged with this clamp. Therapy was periodically interrupted so one transducer could generate a pulse. The echo returning from the opposite transducer was treated. Coagulation necroses were obtained in vitro on substantial thicknesses (23-38mm) of pig liver over exposure durations ranging from 30s to 130s, and with acoustic intensities of less than 15W/cm2 per transducer. Both kidneys of two pigs were treated in vivo with the clamp (14.5W/cm2 for 90s), and the partial nephrectomies performed proved to be bloodless. In vitro and in vivo, wide transfixing lesions corresponded to an echo energy decrease superior to -10dB and parabolic form of the time of flight versus treatment time. In conclusion, this ultrasound clamp has proven to be an excellent mean for achieving monitored haemostasis in kidney.

  10. The pathophysiology of aortic cross-clamping.

    PubMed

    Zammert, Martin; Gelman, Simon

    2016-09-01

    During open aortic surgery, interrupting the blood flow through the aorta by applying a cross-clamp is often a key step to allow for surgical repair. As a consequence, ischemia is induced in parts of the body distal to the clamp site. This significant alteration in the blood flow is almost always associated with hemodynamic changes. Upon release of the cross-clamp, the blood flow is restored, triggering an ischemia-reperfusion response, leading to many pathophysiological processes such as inflammation, humoral changes, and metabolite circulation that could lead to injury in many organ systems and may significantly influence the postoperative outcome. It is therefore important to understand these processes and how they can be treated in order to allow for safe surgical aortic repairs while ensuring the best possible outcomes.

  11. Compact, Stiff, Remotely-Actuable Quick-Release Clamp

    NASA Technical Reports Server (NTRS)

    Tsai, Ted W. (Inventor)

    2000-01-01

    The present invention provides a clamp that is compact and lightweight, yet provides high holding strength and stiffness or rigidity. The clamp uses a unique double slant interface design which provides mechanical advantages to resist forces applied to the clamp member as the load increases. The clamp allows for rapid and remote-activated release of the clamp jaws by applying only a small operating force to an over-center lock/release mechanism, such as by pulling a manual tether.

  12. Functional dissection of synaptic circuits: in vivo patch-clamp recording in neuroscience

    PubMed Central

    Tao, Can; Zhang, Guangwei; Xiong, Ying; Zhou, Yi

    2015-01-01

    Neuronal activity is dominated by synaptic inputs from excitatory or inhibitory neural circuits. With the development of in vivo patch-clamp recording, especially in vivo voltage-clamp recording, researchers can not only directly measure neuronal activity, such as spiking responses or membrane potential dynamics, but also quantify synaptic inputs from excitatory and inhibitory circuits in living animals. This approach enables researchers to directly unravel different synaptic components and to understand their underlying roles in particular brain functions. Combining in vivo patch-clamp recording with other techniques, such as two-photon imaging or optogenetics, can provide even clearer functional dissection of the synaptic contributions of different neurons or nuclei. Here, we summarized current applications and recent research progress using the in vivo patch-clamp recording method and focused on its role in the functional dissection of different synaptic inputs. The key factors of a successful in vivo patch-clamp experiment and possible solutions based on references and our experiences were also discussed. PMID:26052270

  13. Clamping instability and van der Waals forces in carbon nanotube mechanical resonators.

    PubMed

    Aykol, Mehmet; Hou, Bingya; Dhall, Rohan; Chang, Shun-Wen; Branham, William; Qiu, Jing; Cronin, Stephen B

    2014-05-14

    We investigate the role of weak clamping forces, typically assumed to be infinite, in carbon nanotube mechanical resonators. Due to these forces, we observe a hysteretic clamping and unclamping of the nanotube device that results in a discrete drop in the mechanical resonance frequency on the order of 5-20 MHz, when the temperature is cycled between 340 and 375 K. This instability in the resonant frequency results from the nanotube unpinning from the electrode/trench sidewall where it is bound weakly by van der Waals forces. Interestingly, this unpinning does not affect the Q-factor of the resonance, since the clamping is still governed by van der Waals forces above and below the unpinning. For a 1 μm device, the drop observed in resonance frequency corresponds to a change in nanotube length of approximately 50-65 nm. On the basis of these findings, we introduce a new model, which includes a finite tension around zero gate voltage due to van der Waals forces and shows better agreement with the experimental data than the perfect clamping model. From the gate dependence of the mechanical resonance frequency, we extract the van der Waals clamping force to be 1.8 pN. The mechanical resonance frequency exhibits a striking temperature dependence below 200 K attributed to a temperature-dependent slack arising from the competition between the van der Waals force and the thermal fluctuations in the suspended nanotube.

  14. MATLAB implementation of a dynamic clamp with bandwidth of >125 kHz capable of generating I Na at 37 °C.

    PubMed

    Clausen, Chris; Valiunas, Virginijus; Brink, Peter R; Cohen, Ira S

    2013-04-01

    We describe the construction of a dynamic clamp with a bandwidth of >125 kHz that utilizes a high-performance, yet low-cost, standard home/office PC interfaced with a high-speed (16 bit) data acquisition module. High bandwidth is achieved by exploiting recently available software advances (code-generation technology and optimized real-time kernel). Dynamic-clamp programs are constructed using Simulink, a visual programming language. Blocks for computation of membrane currents are written in the high-level MATLAB language; no programming in C is required. The instrument can be used in single- or dual-cell configurations, with the capability to modify programs while experiments are in progress. We describe an algorithm for computing the fast transient Na(+) current (I Na) in real time and test its accuracy and stability using rate constants appropriate for 37 °C. We then construct a program capable of supplying three currents to a cell preparation: I Na, the hyperpolarizing-activated inward pacemaker current (I f) and an inward-rectifier K(+) current (I K1). The program corrects for the IR drop due to electrode current flow and also records all voltages and currents. We tested this program on dual patch-clamped HEK293 cells where the dynamic clamp controls a current-clamp amplifier and a voltage-clamp amplifier controls membrane potential, and current-clamped HEK293 cells where the dynamic clamp produces spontaneous pacing behavior exhibiting Na(+) spikes in otherwise passive cells.

  15. Kinetic analysis of PCNA clamp binding and release in the clamp loading reaction catalyzed by Saccharomyces cerevisiae replication factor C

    PubMed Central

    Marzahn, Melissa R.; Hayner, Jaclyn N.; Meyer, Jennifer A.; Bloom, Linda B.

    2014-01-01

    DNA polymerases require a sliding clamp to achieve processive DNA synthesis. The toroidal clamps are loaded onto DNA by clamp loaders, members of the AAA+ family of ATPases. These enzymes utilize the energy of ATP binding and hydrolysis to perform a variety of cellular functions. In this study, a clamp loader-clamp binding assay was developed to measure the rates of ATP-dependent clamp binding and ATP-hydrolysis-dependent clamp release for the S. cerevisiae clamp loader (RFC) and clamp (PCNA). Pre-steady-state kinetics of PCNA binding showed that although ATP binding to RFC increases affinity for PCNA, ATP binding rates and ATP-dependent conformational changes in RFC are fast relative to PCNA binding rates. Interestingly, RFC binds PCNA faster than the Escherichia coli γ complex clamp loader binds the β-clamp. In the process of loading clamps on DNA, RFC maintains contact with PCNA while PCNA closes, as the observed rate of PCNA closing is faster than the rate of PCNA release, precluding the possibility of an open clamp dissociating from DNA. Rates of clamp closing and release are not dependent on the rate of the DNA binding step and are also slower than reported rates of ATP hydrolysis, showing that these rates reflect unique intramolecular reaction steps in the clamp loading pathway. PMID:25450506

  16. Kinetic analysis of PCNA clamp binding and release in the clamp loading reaction catalyzed by Saccharomyces cerevisiae replication factor C.

    PubMed

    Marzahn, Melissa R; Hayner, Jaclyn N; Meyer, Jennifer A; Bloom, Linda B

    2015-01-01

    DNA polymerases require a sliding clamp to achieve processive DNA synthesis. The toroidal clamps are loaded onto DNA by clamp loaders, members of the AAA+family of ATPases. These enzymes utilize the energy of ATP binding and hydrolysis to perform a variety of cellular functions. In this study, a clamp loader-clamp binding assay was developed to measure the rates of ATP-dependent clamp binding and ATP-hydrolysis-dependent clamp release for the Saccharomyces cerevisiae clamp loader (RFC) and clamp (PCNA). Pre-steady-state kinetics of PCNA binding showed that although ATP binding to RFC increases affinity for PCNA, ATP binding rates and ATP-dependent conformational changes in RFC are fast relative to PCNA binding rates. Interestingly, RFC binds PCNA faster than the Escherichia coli γ complex clamp loader binds the β-clamp. In the process of loading clamps on DNA, RFC maintains contact with PCNA while PCNA closes, as the observed rate of PCNA closing is faster than the rate of PCNA release, precluding the possibility of an open clamp dissociating from DNA. Rates of clamp closing and release are not dependent on the rate of the DNA binding step and are also slower than reported rates of ATP hydrolysis, showing that these rates reflect unique intramolecular reaction steps in the clamp loading pathway.

  17. Π-Clamp-mediated cysteine conjugation.

    PubMed

    Zhang, Chi; Welborn, Matthew; Zhu, Tianyu; Yang, Nicole J; Santos, Michael S; Van Voorhis, Troy; Pentelute, Bradley L

    2016-02-01

    Site-selective functionalization of complex molecules is one of the most significant challenges in chemistry. Typically, protecting groups or catalysts must be used to enable the selective modification of one site among many that are similarly reactive, and general strategies that selectively tune the local chemical environment around a target site are rare. Here, we show a four-amino-acid sequence (Phe-Cys-Pro-Phe), which we call the 'π-clamp', that tunes the reactivity of its cysteine thiol for site-selective conjugation with perfluoroaromatic reagents. We use the π-clamp to selectively modify one cysteine site in proteins containing multiple endogenous cysteine residues. These examples include antibodies and cysteine-based enzymes that would be difficult to modify selectively using standard cysteine-based methods. Antibodies modified using the π-clamp retained binding affinity to their targets, enabling the synthesis of site-specific antibody-drug conjugates for selective killing of HER2-positive breast cancer cells. The π-clamp is an unexpected approach to mediate site-selective chemistry and provides new avenues to modify biomolecules for research and therapeutics.

  18. Patch-clamp amplifiers on a chip.

    PubMed

    Weerakoon, Pujitha; Culurciello, Eugenio; Yang, Youshan; Santos-Sacchi, Joseph; Kindlmann, Peter J; Sigworth, Fred J

    2010-10-15

    We present the first, fully integrated, two-channel implementation of a patch-clamp measurement system. With this "PatchChip" two simultaneous whole-cell recordings can be obtained with rms noise of 8pA in a 10kHz bandwidth. The capacitance and series-resistance of the electrode can be compensated up to 10pF and 100MΩ respectively under computer control. Recordings of hERG and Na(v) 1.7 currents demonstrate the system's capabilities, which are on par with large, commercial patch-clamp instrumentation. By reducing patch-clamp amplifiers to a millimeter size micro-chip, this work paves the way to the realization of massively parallel, high-throughput patch-clamp systems for drug screening and ion-channel research. The PatchChip is implemented in a 0.5μm silicon-on-sapphire process; its size is 3×3mm(2) and the power consumption is 5mW per channel with a 3.3V power supply.

  19. Dynamics and Stability of Pinned-Clamped and Clamped-Pinned Cylindrical Shells Conveying Fluid

    NASA Astrophysics Data System (ADS)

    Misra, A. K.; Wong, S. S. T.; Païdoussis, M. P.

    2001-11-01

    The paper examines the dynamics and stability of fluid-conveying cylindrical shells having pinned-clamped or clamped-pinned boundary conditions, where ``pinned'' is an abbreviation for ``simply supported''. Flügge's equations are used to describe the shell motion, while the fluid-dynamic perturbation pressure is obtained utilizing the linearized potential flow theory. The solution is obtained using two methods - the travelling wave method and the Fourier-transform approach. The results obtained by both methods suggest that the negative damping of the clamped-pinned systems and positive damping of the pinned-clamped systems, observed by previous investigators for any arbitrarily small flow velocity, are simply numerical artefacts; this is reinforced by energy considerations, in which the work done by the fluid on the shell is shown to be zero. Hence, it is concluded that both systems are conservative.

  20. Characterization of solid electrode materials using chronoamperometry: A study of the alkaline γ-MnO 2 electrode

    NASA Astrophysics Data System (ADS)

    Malloy, Aaron P.; Donne, Scott W.

    Large voltage step chronoamperometry is shown to be a time-efficient means to examine solid electrode materials compared with conventional electrochemical methods such as linear sweep voltammetry (LSV) and step potential electrochemical spectroscopy (SPECS), all the while providing comparable information concerning the rate capability of a material and its capacity. The applicability of the technique is demonstrated through a study of the alkaline γ-MnO 2 electrode. By sampling the current (and hence the charge) at various times after the chronoamperometric voltage step, the compatibility between chronoamperometry and LSV is disclosed. Furthermore, modelling of the chronoamperometric data using two curves based on a spherical diffusion model representing fast and slow discharge processes are found to be statistically suitable. From this modelling, values of A√ D (where A is the electrochemically active surface area and D is the diffusion coefficient) for the two processes are 3.89 × 10 -4 and 0.70 × 10 -4 cm 3 s -1/2 g -1, respectively, both of which are comparable with A√ D data extracted from a SPECS experiment on an identical electrode.

  1. VOLTAGE REGULATOR

    DOEpatents

    Von Eschen, R.L.; Scheele, P.F.

    1962-04-24

    A transistorized voltage regulator which provides very close voitage regulation up to about 180 deg F is described. A diode in the positive line provides a constant voltage drop from the input to a regulating transistor emitter. An amplifier is coupled to the positive line through a resistor and is connected between a difference circuit and the regulating transistor base which is negative due to the difference in voltage drop across thc diode and the resistor so that a change in the regulator output causes the amplifier to increase or decrease the base voltage and current and incrcase or decrease the transistor impedance to return the regulator output to normal. (AEC)

  2. Can robots patch-clamp as well as humans? Characterization of a novel sodium channel mutation.

    PubMed

    Estacion, M; Choi, J S; Eastman, E M; Lin, Z; Li, Y; Tyrrell, L; Yang, Y; Dib-Hajj, S D; Waxman, S G

    2010-06-01

    Ion channel missense mutations cause disorders of excitability by changing channel biophysical properties. As an increasing number of new naturally occurring mutations have been identified, and the number of other mutations produced by molecular approaches such as in situ mutagenesis has increased, the need for functional analysis by patch-clamp has become rate limiting. Here we compare a patch-clamp robot using planar-chip technology with human patch-clamp in a functional assessment of a previously undescribed Nav1.7 sodium channel mutation, S211P, which causes erythromelalgia. This robotic patch-clamp device can increase throughput (the number of cells analysed per day) by 3- to 10-fold. Both modes of analysis show that the mutation hyperpolarizes activation voltage dependence (8 mV by manual profiling, 11 mV by robotic profiling), alters steady-state fast inactivation so that it requires an additional Boltzmann function for a second fraction of total current (approximately 20% manual, approximately 40% robotic), and enhances slow inactivation (hyperpolarizing shift--15 mV by human,--13 mV robotic). Manual patch-clamping demonstrated slower deactivation and enhanced (approximately 2-fold) ramp response for the mutant channel while robotic recording did not, possibly due to increased temperature and reduced signal-to-noise ratio on the robotic platform. If robotic profiling is used to screen ion channel mutations, we recommend that each measurement or protocol be validated by initial comparison to manual recording. With this caveat, we suggest that, if results are interpreted cautiously, robotic patch-clamp can be used with supervision and subsequent confirmation from human physiologists to facilitate the initial profiling of a variety of electrophysiological parameters of ion channel mutations.

  3. Mechanical properties of a lap joint under uniform clamping pressure

    NASA Technical Reports Server (NTRS)

    Diller, S. V.; Metherell, A. F.

    1969-01-01

    Equations were derived for the load deflection relations, the energy dissipation per cycle, and the instantaneous rate of dissipation for a lap joint idealized as two overlapping plates clamped together under a uniform clamping pressure.

  4. Towards a Dynamic Clamp for Neurochemical Modalities

    PubMed Central

    Rivera, Catalina Maria; Kwon, Hyuck-Jin; Hashmi, Ali; Yu, Gan; Zhao, Jiheng; Gao, Jianlong; Xu, Jie; Xue, Wei; Dimitrov, Alexander G.

    2015-01-01

    The classic dynamic clamp technique uses a real-time electrical interface between living cells and neural simulations in order to investigate hypotheses about neural function and structure. One of the acknowledged drawbacks of that technique is the limited control of the cells' chemical microenvironment. In this manuscript, we use a novel combination of nanosensor and microfluidic technology and microfluidic and neural simulations to add sensing and control of chemical concentrations to the dynamic clamp technique. Specifically, we use a microfluidic lab-on-a-chip to generate distinct chemical concentration gradients (ions or neuromodulators), to register the concentrations with embedded nanosensors and use the processed signals as an input to simulations of a neural cell. The ultimate goal of this project is to close the loop and provide sensor signals to the microfluidic lab-on-a-chip to mimic the interaction of the simulated cell with other cells in its chemical environment. PMID:25946635

  5. Clamp on ultrasonic instruments in subsea applications

    SciTech Connect

    Haugen, S.; Hodgson, S.; Upchurch, J.; McMahan, J.; Hazelrigg, K.; Mundorff, J.

    1995-12-01

    Monitoring of solids flow in pipelines has until recently required pipework intervention in order to insert probes and switches. Both sand particles and cleaning pigs are by nature destructive to these directly exposed detectors due to erosion and inertia. The instruments presented in this paper provide a superior alternative in ultrasonic clamp-on technology requiring only superficial pipework installation. Wells can now be operated efficiently in a way that minimizes the risk of erosion and collapse. Pigging operations can be monitored both by on-line pig detection and debris monitoring thereby allowing a safe and optimized survey to take place. The non-intrusive clamp-on concept raises standards in instrument reliability, durability and measurement performance.

  6. Mechanical and metallurgical properties of carotid artery clamps.

    PubMed

    Dujovny, M; Kossovsky, N; Kossowsky, R; Segal, R; Diaz, F G; Kaufman, H; Perlin, A; Cook, E E

    1985-11-01

    The mechanical and metallurgical properties of carotid artery clamps were evaluated. The pressure plate retreat propensity, metallurgical composition, surface morphology, magnetic properties, and corrosion resistance of the Crutchfield, Selverstone, Salibi, and Kindt clamps were tested. None of the clamps showed evidence of pressure plate retreat. The clamps differed significantly in their composition, surface cleanliness, magnetic properties, and corrosion resistance. The Crutchfield clamp was the only one manufactured from an ASTM-ANSI-approved implantable stainless steel (AISI 316) and the only clamp in which the surfaces were clean and free of debris. The Selverstone clamp was made principally from AISI 304 stainless steel, as was one Salibi clamp. The pressure plate on another Salibi clamp was made from a 1% chromium and 1% manganese steel. Machining and surface debris consisting principally of aluminum, silicon, and sulfur was abundant on the Selverstone and Salibi clamps. The Kindt clamp was manufactured from AISI 301 stainless steel with a silicate-aluminized outer coating. The Crutchfield and Selverstone clamps were essentially nonferromagnetic, whereas the Salibi and Kindt clamps were sensitive to magnetic flux. In the pitting potential corrosion test, the Crutchfield clamp demonstrated good corrosion resistance with a pitting potential of 310 mV and no surface corrosion or pitting by scanning electron microscopy examination. The Selverstone clamp had lower pitting potentials and showed various degrees of corrosion and surface pitting by scanning electron microscopy. The Salibi pressure plate had a very low pitting potential of -525 mV and showed severe corrosion. By metallurgical criteria, only the Crutchfield clamp is suitable for long term implantation.

  7. Carbon nanotube-clamped metal atomic chain

    PubMed Central

    Tang, Dai-Ming; Yin, Li-Chang; Li, Feng; Liu, Chang; Yu, Wan-Jing; Hou, Peng-Xiang; Wu, Bo; Lee, Young-Hee; Ma, Xiu-Liang; Cheng, Hui-Ming

    2010-01-01

    Metal atomic chain (MAC) is an ultimate one-dimensional structure with unique physical properties, such as quantized conductance, colossal magnetic anisotropy, and quantized magnetoresistance. Therefore, MACs show great potential as possible components of nanoscale electronic and spintronic devices. However, MACs are usually suspended between two macroscale metallic electrodes; hence obvious technical barriers exist in the interconnection and integration of MACs. Here we report a carbon nanotube (CNT)-clamped MAC, where CNTs play the roles of both nanoconnector and electrodes. This nanostructure is prepared by in situ machining a metal-filled CNT, including peeling off carbon shells by spatially and elementally selective electron beam irradiation and further elongating the exposed metal nanorod. The microstructure and formation process of this CNT-clamped MAC are explored by both transmission electron microscopy observations and theoretical simulations. First-principles calculations indicate that strong covalent bonds are formed between the CNT and MAC. The electrical transport property of the CNT-clamped MAC was experimentally measured, and quantized conductance was observed. PMID:20427743

  8. π-Clamp Mediated Cysteine Conjugation

    PubMed Central

    Zhang, Chi; Welborn, Matthew; Zhu, Tianyu; Yang, Nicole J.; Santos, Michael S.; Van Voorhis, Troy; Pentelute, Bradley L.

    2016-01-01

    Site-selective functionalization of complex molecules is a grand challenge in chemistry. Protecting groups or catalysts must be used to selectively modify one site among many that are similarly reactive. General strategies are rare such the local chemical environment around the target site is tuned for selective transformation. Here we show a four amino acid sequence (Phe-Cys-Pro-Phe), which we call the “π-clamp”, tunes the reactivity of its cysteine thiol for the site-selective conjugation with perfluoroaromatic reagents. We used the π-clamp to selectively modify one cysteine site in proteins containing multiple endogenous cysteine residues (e.g. antibodies and cysteine-based enzymes), which was impossible with prior cysteine modification methods. The modified π-clamp antibodies retained binding affinity to their targets, enabling the synthesis of site-specific antibody-drug conjugates (ADCs) for selective killing of HER2-positive breast cancer cells. The π-clamp is an unexpected approach for site-selective chemistry and provides opportunities to modify biomolecules for research and therapeutics. PMID:26791894

  9. Temperature-Controlled Clamping and Releasing Mechanism

    NASA Technical Reports Server (NTRS)

    Rosing, David; Ford, Virginia

    2005-01-01

    A report describes the development of a mechanism that automatically clamps upon warming and releases upon cooling between temperature limits of approx. =180 K and approx. =293 K. The mechanism satisfied a need specific to a program that involved repeated excursions of a spectrometer between a room-temperature atmospheric environment and a cryogenic vacuum testing environment. The mechanism was also to be utilized in the intended application of the spectrometer, in which the spectrometer would be clamped for protection during launch of a spacecraft and released in the cold of outer space to allow it to assume its nominal configuration for scientific observations. The mechanism is passive in the sense that its operation does not depend on a control system and does not require any power other than that incidental to heating and cooling. The clamping and releasing action is effected by bolt-preloaded stacks of shape-memory-alloy (SMA) cylinders. In designing this mechanism, as in designing other, similar SMA mechanisms, it was necessary to account for the complex interplay among thermal expansion, elastic and inelastic deformation under load, and SMA thermomechanical properties.

  10. π-Clamp-mediated cysteine conjugation

    NASA Astrophysics Data System (ADS)

    Zhang, Chi; Welborn, Matthew; Zhu, Tianyu; Yang, Nicole J.; Santos, Michael S.; van Voorhis, Troy; Pentelute, Bradley L.

    2016-02-01

    Site-selective functionalization of complex molecules is one of the most significant challenges in chemistry. Typically, protecting groups or catalysts must be used to enable the selective modification of one site among many that are similarly reactive, and general strategies that selectively tune the local chemical environment around a target site are rare. Here, we show a four-amino-acid sequence (Phe-Cys-Pro-Phe), which we call the ‘π-clamp’, that tunes the reactivity of its cysteine thiol for site-selective conjugation with perfluoroaromatic reagents. We use the π-clamp to selectively modify one cysteine site in proteins containing multiple endogenous cysteine residues. These examples include antibodies and cysteine-based enzymes that would be difficult to modify selectively using standard cysteine-based methods. Antibodies modified using the π-clamp retained binding affinity to their targets, enabling the synthesis of site-specific antibody-drug conjugates for selective killing of HER2-positive breast cancer cells. The π-clamp is an unexpected approach to mediate site-selective chemistry and provides new avenues to modify biomolecules for research and therapeutics.

  11. Single molecule study of a processivity clamp sliding on DNA

    SciTech Connect

    Laurence, T A; Kwon, Y; Johnson, A; Hollars, C; O?Donnell, M; Camarero, J A; Barsky, D

    2007-07-05

    Using solution based single molecule spectroscopy, we study the motion of the polIII {beta}-subunit DNA sliding clamp ('{beta}-clamp') on DNA. Present in all cellular (and some viral) forms of life, DNA sliding clamps attach to polymerases and allow rapid, processive replication of DNA. In the absence of other proteins, the DNA sliding clamps are thought to 'freely slide' along the DNA; however, the abundance of positively charged residues along the inner surface may create favorable electrostatic contact with the highly negatively charged DNA. We have performed single-molecule measurements on a fluorescently labeled {beta}-clamp loaded onto freely diffusing plasmids annealed with fluorescently labeled primers of up to 90 bases. We find that the diffusion constant for 1D diffusion of the {beta}-clamp on DNA satisfies D {le} 10{sup -14} cm{sup 2}/s, much slower than the frictionless limit of D = 10{sup -10} cm{sup 2}/s. We find that the {beta} clamp remains at the 3-foot end in the presence of E. coli single-stranded binding protein (SSB), which would allow for a sliding clamp to wait for binding of the DNA polymerase. Replacement of SSB with Human RP-A eliminates this interaction; free movement of sliding clamp and poor binding of clamp loader to the junction allows sliding clamp to accumulate on DNA. This result implies that the clamp not only acts as a tether, but also a placeholder.

  12. 33 CFR 183.532 - Clips, straps, and hose clamps.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 33 Navigation and Navigable Waters 2 2011-07-01 2011-07-01 false Clips, straps, and hose clamps... (CONTINUED) BOATING SAFETY BOATS AND ASSOCIATED EQUIPMENT Fuel Systems Equipment Standards § 183.532 Clips, straps, and hose clamps. (a) Each clip, strap, and hose clamp must: (1) Be made from a...

  13. 33 CFR 183.532 - Clips, straps, and hose clamps.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 33 Navigation and Navigable Waters 2 2010-07-01 2010-07-01 false Clips, straps, and hose clamps... (CONTINUED) BOATING SAFETY BOATS AND ASSOCIATED EQUIPMENT Fuel Systems Equipment Standards § 183.532 Clips, straps, and hose clamps. (a) Each clip, strap, and hose clamp must: (1) Be made from a...

  14. Damping mechanisms of single-clamped and prestressed double-clamped resonant polymer microbeams

    NASA Astrophysics Data System (ADS)

    Schmid, S.; Hierold, C.

    2008-11-01

    In this article, an investigation of the damping mechanisms of resonant single- and double-clamped polymer microbeams for a frequency range from 10 kHz to 5 MHz is presented. The suspended structures are made of SU-8, an epoxy-type photoresist, by means of a sacrificial layer technique. The vibration was measured with a laser-Doppler vibrometer in high vacuum at different temperatures and at atmospheric pressure. The influence of air damping in rarefied air was investigated and the intrinsic damping mechanisms were determined in high vacuum (p <0.05 Pa). After excluding a variety of possible damping factors, the dominant intrinsic dissipation mechanism of the single-clamped microbeams was understood to be the material damping with maximum quality factors (Q) of around 70 at 20 °C. Quality factors of up to 720 at 20 °C were measured for stringlike double-clamped microbeams, which suggest a different intrinsic damping mechanism than material loss. It is shown that internal damping mechanisms due to flexure and elongation have a small impact on the damping of stretched strings. Modeling the clamping loss based on the wave transmission into the suspended anchor plates indicates that it is the dominant intrinsic dissipation in the prestressed double-clamped microresonators. At atmospheric pressure it was shown that at low frequencies the quality factors of single-clamped and stringlike double-clamped microbeams are limited by the squeeze-film air damping. At high frequencies the quality factors are limited by the specific intrinsic damping. In between the two particular regions with a specific dominant damping mechanism the quality factors show a maximum.

  15. Whole-cell patch clamp recordings from rhythmically active motoneurons in the isolated spinal cord of the chick embryo.

    PubMed

    Sernagor, E; O'Donovan, M J

    1991-07-22

    Whole-cell patch clamp recordings were obtained during motor activity from electrically identified motoneurons within the spinal cord of the chick embryo maintained in vitro. Most recordings were performed on E11-E13 motoneurons although it was also possible to record from younger cells (E7-E9). Voltage clamp recordings were used to characterize the synaptic currents expressed in femoro-tibialis (extensor) motoneurons during motor activity. These motoneurons exhibited rhythmic excitatory currents with reversal potentials near 0 mV. This powerful technique enables high resolution recordings from identified motoneurons in situ and allows investigation of the membrane and synaptic mechanisms involved in the development of embryonic motility.

  16. Robotic multi-well planar patch-clamp for native and primary mammalian cells

    PubMed Central

    Milligan, Carol J; Li, Jing; Sukumar, Piruthivi; Majeed, Yasser; Dallas, Mark L; English, Anne; Emery, Paul; Porter, Karen E; Smith, Andrew M; McFadzean, Ian; Beccano-Kelly, Dayne; Bahnasi, Yahya; Cheong, Alex; Naylor, Jacqueline; Zeng, Fanning; Liu, Xing; Gamper, Nikita; Jiang, Lin-Hua; Pearson, Hugh A; Peers, Chris; Robertson, Brian; Beech, David J

    2009-01-01

    Multi-well robotic planar patch-clamp has become common in drug development and safety programmes because it enables efficient and systematic testing of compounds against ion channels during voltage-clamp. It has not, however, been adopted significantly in other important areas of ion channel research, where conventional patch-clamp remains the favoured method. Here we show the wider potential of the multi-well approach with the capability for efficient intracellular solution exchange, describing protocols and success rates for recording from a range of native and primary mammalian cells derived from blood vessels, arthritic joints, and the immune and central nervous systems. The protocol involves preparing a suspension of single cells to be dispensed robotically into 4-8 microfluidic chambers each containing a glass chip with a small aperture. Under automated control, giga-seals and whole-cell access are achieved followed by pre-programmed routines of voltage paradigms and fast extracellular or intracellular solution exchange. Recording from 48 chambers usually takes 1-6 hr depending on the experimental design and yields 16-33 cell recordings. PMID:19197268

  17. A general modelling and control algorithm of a three-phase multilevel diode clamped inverter by means of a direct space vector control

    NASA Astrophysics Data System (ADS)

    Bouhali, O.; Francois, B.; Berkouk, E. M.; Saudemont, C.

    2005-07-01

    This paper presents a simple and general direct modulation strategy that enables to copy directly modulated waveforms onto output voltages of a multilevel three-phase Diode Clamped Inverter (DCI). A general modelling of this converter is presented. A space vector scheme is developed without using Park transforms. Based on this algorithm, the location of the reference voltage vector is determined and the voltage vectors for the modulation are deduced. Simultaneously, their durations are calculated. The proposed algorithm is general and can be directly applied to a (n+1) levels inverter independently on its topology (Diode Clamped Inverter, Neutral Point Clamped, Flying Capacitor Inverter...). To verify this algorithm, both control algorithms of a 5-level DCI and a 11-level DCI are considered and simulation results are given.

  18. Testing and evaluation of stretching strain in clamped-clamped beams for energy harvesting

    NASA Astrophysics Data System (ADS)

    Emad, Ahmed; Mahmoud, Mohamed A. E.; Ghoneima, Maged; Dessouky, Mohamed

    2016-11-01

    In this paper, evaluation of stretching strain capabilities to harvest energy from a piezoelectric clamped-clamped beam is theoretically modeled and experimentally tested. The utilization of stretching strain has many advantages as: elimination of any substrate material, and the simple electrode configuration. The doubly clamped structure exhibits a highly nonlinear frequency response (Hardening Duffing) that widens the bandwidth during the frequency up-ward sweep. The wide bandwidth makes it suitable for practical applications. A design of 53.5 {{mm}}3 (29.7 {{mm}}3 piezoelectric material + 23.8 {{mm}}3 proof mass) energy harvester was tested using PVDF (polyvinylidene fluoride) that can generate up to 15 μW from vibrations of 0.5{g} at 128 {Hz} and 2 MΩ load. The design can also generate up to 41 μ {{W}} from vibrations of 1{g} at 140 {Hz} and 2 MΩ load.

  19. Timing of umbilical cord clamping of term infants.

    PubMed

    Ceriani Cernadas, José María

    2017-04-01

    For at least over 200 years, multiple controversies have arisen around the timing of umbilical cord clamping. In the past decades, early cord clamping (within the first 15 seconds) had markedly prevailed. Only in the 21st century, randomized controlled trials have demonstrated the benefits of delayed cord clamping (at 2-3 minutes). Delayed cord clamping has been practiced in obstetrics based on the recommendations made by scientific societies and in systematic reviews, which have provided solid evidence to support this practice in term infants. This review describes the most relevant factors supporting the use of delayed cord clamping in term infants. It points out the essential role played by physiological mechanisms which, undoubtedly, allow us to understand the benefits of delayed cord clamping and advise us to wait for what nature has established. Other relevant aspects supporting delayed cord clamping are also described here.

  20. Patch Clamping Protoplasts from Vascular Plants 1

    PubMed Central

    Elzenga, J. Theo M.; Keller, Christopher P.; Van Volkenburgh, Elizabeth

    1991-01-01

    A method is described for the isolation of protoplasts (Pisum sativum, Phaseolus vulgaris, Avena sativa, Arabidopsis thaliana) in preparation for ion flux studies using patch clamp electrophysiology. Protoplasts that have been exposed to hydrolytic, cell wall degrading, enzymes for as little as 5 minutes form gigaseals (seal resistance higher than 10 giga Ohm) with the patch pipette with success rates greater than 40%. Sealing of these protoplasts is fast, averaging less than 2 minutes. This method yields high rates of gigaseal formation in a variety of tissues from both monocots and dicots and will enhance data collection in ion flux studies of plasma membranes of vascular plants. PMID:16668586

  1. The Clamp Loader Assembles the β Clamp onto Either a 3′ or 5′ Primer Terminus

    PubMed Central

    Park, Mee Sook; O'Donnell, Mike

    2009-01-01

    Clamp loaders assemble sliding clamps onto 3′ primed sites for DNA polymerases. Clamp loaders are thought to be specific for a 3′ primed site, and unable to bind a 5′ site. We demonstrate here that the Escherichia coli γ complex clamp loader can load the β clamp onto a 5′ primed site, although with at least 20-fold reduced efficiency relative to loading at a 3′ primed site. Preferential clamp loading at a 3′ site does not appear to be due to DNA binding, as the clamp loader forms an avid complex with β at a 5′ site. Preferential loading at a 3′ versus a 5′ site occurs at the ATP hydrolysis step, needed to close the ring around DNA. We also address DNA structural features that are recognized for preferential loading at a 3′ site. Although the single-stranded template strand extends in opposite directions from 3′ and 5′ primed sites, thus making it a favorite candidate for distinguishing between 3′ and 5′ sites, the single-strand polarity at a primed template junction does not determine 3′ site selection for clamp loading. Instead, we find that clamp loader recognition of a 3′ site lies in the duplex portion of the primed site, not the single-strand portion. We present evidence that the β clamp facilitates its own loading specificity for a 3′ primed site. Implications to eukaryotic clamp loader complexes are proposed. PMID:19759020

  2. Laser-assisted patch clamping: a methodology

    NASA Technical Reports Server (NTRS)

    Henriksen, G. H.; Assmann, S. M.; Evans, M. L. (Principal Investigator)

    1997-01-01

    Laser microsurgery can be used to perform both cell biological manipulations, such as targeted cell ablation, and molecular genetic manipulations, such as genetic transformation and chromosome dissection. In this report, we describe a laser microsurgical method that can be used either to ablate single cells or to ablate a small area (1-3 microns diameter) of the extracellular matrix. In plants and microorganisms, the extracellular matrix consists of the cell wall. While conventional patch clamping of these cells, as well as of many animal cells, requires enzymatic digestion of the extracellular matrix, we illustrate that laser microsurgery of a portion of the wall enables patch clamp access to the plasma membrane of higher plant cells remaining situated in their tissue environment. What follows is a detailed description of the construction and use of an economical laser microsurgery system, including procedures for single cell and targeted cell wall ablation. This methodology will be of interest to scientists wishing to perform cellular or subcellular ablation with a high degree of accuracy, or wishing to study how the extracellular matrix affects ion channel function.

  3. Negligible substrate clamping effect on piezoelectric response in (111)-epitaxial tetragonal Pb(Zr, Ti)O{sub 3} films

    SciTech Connect

    Yamada, Tomoaki; Yasumoto, Jun; Ito, Daisuke; Yoshino, Masahito; Nagasaki, Takanori; Sakata, Osami; Imai, Yasuhiko; Kiguchi, Takanori; Shiraishi, Takahisa; Shimizu, Takao; Funakubo, Hiroshi

    2015-08-21

    The converse piezoelectric responses of (111)- and (001)-epitaxial tetragonal Pb(Zr{sub 0.35}Ti{sub 0.65})O{sub 3} [PZT] films were compared to investigate the orientation dependence of the substrate clamping effect. Synchrotron X-ray diffraction (XRD) and piezoelectric force microscopy revealed that the as-grown (111)-PZT film has a polydomain structure with normal twin boundaries that are changed by the poling process to inclined boundaries, as predicted by Romanov et al. [Phys. Status Solidi A 172, 225 (1999)]. Time-resolved synchrotron XRD under bias voltage showed the negligible impact of substrate clamping on the piezoelectric response in the (111)-PZT film, unlike the case for (001)-PZT film. The origin of the negligible clamping effect in the (111)-PZT film is discussed from the viewpoint of the elastic properties and the compensation of lattice distortion between neighboring domains.

  4. Negligible substrate clamping effect on piezoelectric response in (111)-epitaxial tetragonal Pb(Zr, Ti)O3 films

    NASA Astrophysics Data System (ADS)

    Yamada, Tomoaki; Yasumoto, Jun; Ito, Daisuke; Sakata, Osami; Imai, Yasuhiko; Kiguchi, Takanori; Shiraishi, Takahisa; Shimizu, Takao; Funakubo, Hiroshi; Yoshino, Masahito; Nagasaki, Takanori

    2015-08-01

    The converse piezoelectric responses of (111)- and (001)-epitaxial tetragonal Pb(Zr0.35Ti0.65)O3 [PZT] films were compared to investigate the orientation dependence of the substrate clamping effect. Synchrotron X-ray diffraction (XRD) and piezoelectric force microscopy revealed that the as-grown (111)-PZT film has a polydomain structure with normal twin boundaries that are changed by the poling process to inclined boundaries, as predicted by Romanov et al. [Phys. Status Solidi A 172, 225 (1999)]. Time-resolved synchrotron XRD under bias voltage showed the negligible impact of substrate clamping on the piezoelectric response in the (111)-PZT film, unlike the case for (001)-PZT film. The origin of the negligible clamping effect in the (111)-PZT film is discussed from the viewpoint of the elastic properties and the compensation of lattice distortion between neighboring domains.

  5. Improving the electrochemical behavior of LiCoO 2 electrode by mixed Zr-Mg doping

    NASA Astrophysics Data System (ADS)

    Xu, H. Y.; Xie, S.; Zhang, C. P.; Chen, C. H.

    A new class of LiCo 1- xZr x/2 Mg x/2 O 2 ( x = 0, 0.02, 0.06, 0.10, 0.20) materials has been synthesized using a solution-combustion method with mixed acetates/nitrates as the starting materials. The structure of the synthesized oxides was analyzed using X-ray diffraction (XRD). Iodometry titration was used to measure the oxidation state of cobalt. The electrochemical performance of LiCo 1- xZr x/2 Mg x/2 O 2 electrodes was analyzed using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and galvanostatic charge-discharge cycling studies in the voltage range 2.7-4.2 V (versus Li metal). It is found that the maximum doping level ( x) is around 0.06, above which Li 2MgZrO 4 is formed as an impurity phase. The use of combined Zr-Mg doping has resulted in the decrease of the electrode impedance and increase of the specific capacity and the stability of 3.6 V plateau efficiency.

  6. Testing of Diode-Clamping in an Inductive Pulsed Plasma Thruster Circuit

    NASA Technical Reports Server (NTRS)

    Toftul, Alexandra; Polzin, Kurt A.; Martin, Adam K.; Hudgins, Jerry L.

    2014-01-01

    Testing of a 5.5 kV silicon (Si) diode and 5.8 kV prototype silicon carbide (SiC) diode in an inductive pulsed plasma thruster (IPPT) circuit was performed to obtain a comparison of the resulting circuit recapture efficiency,eta(sub r), defined as the percentage of the initial charge energy remaining on the capacitor bank after the diode interrupts the current. The diode was placed in a pulsed circuit in series with a silicon controlled rectifier (SCR) switch, and the voltages across different components and current waveforms were collected over a range of capacitor charge voltages. Reverse recovery parameters, including turn-off time and peak reverse recovery current, were measured and capacitor voltage waveforms were used to determine the recapture efficiency for each case. The Si fast recovery diode in the circuit was shown to yield a recapture efficiency of up to 20% for the conditions tested, while the SiC diode further increased recapture efficiency to nearly 30%. The data presented show that fast recovery diodes operate on a timescale that permits them to clamp the discharge quickly after the first half cycle, supporting the idea that diode-clamping in IPPT circuit reduces energy dissipation that occurs after the first half cycle

  7. High Performance ZVT with Bus Clamping Modulation Technique for Single Phase Full Bridge Inverters

    SciTech Connect

    Xia, Yinglai; Ayyanar, Raja

    2016-03-20

    This paper proposes a topology based on bus clamping modulation and zero-voltage-transition (ZVT) technique to realize zero-voltage-switching (ZVS) for all the main switches of the full bridge inverters, and inherent ZVS and/or ZCS for the auxiliary switches. The advantages of the strategy include significant reduction in the turn-on loss of the ZVT auxiliary switches which typically account for a major part of the total loss in other ZVT circuits, and reduction in the voltage ratings of auxiliary switches. The modulation scheme and the commutation stages are analyzed in detail. Finally, a 1kW, 500 kHz switching frequency inverter of the proposed topology using SiC MOSFETs has been built to validate the theoretical analysis. The ZVT with bus clamping modulation technique of fixed timing and adaptive timing schemes are implemented in DSP TMS320F28335 resulting in full ZVS for the main switches in the full bridge inverter. The proposed scheme can save up to 33 % of the switching loss compared with no ZVT case.

  8. Re-visiting the trans insertion model for complexin clamping

    PubMed Central

    Krishnakumar, Shyam S; Li, Feng; Coleman, Jeff; Schauder, Curtis M; Kümmel, Daniel; Pincet, Frederic; Rothman, James E; Reinisch, Karin M

    2015-01-01

    We have previously proposed that complexin cross-links multiple pre-fusion SNARE complexes via a trans interaction to function as a clamp on SNARE-mediated neurotransmitter release. A recent NMR study was unable to detect the trans clamping interaction of complexin and therefore questioned the previous interpretation of the fluorescence resonance energy transfer and isothermal titration calorimetry data on which the trans clamping model was originally based. Here we present new biochemical data that underscore the validity of our previous interpretation and the continued relevancy of the trans insertion model for complexin clamping. DOI: http://dx.doi.org/10.7554/eLife.04463.001 PMID:25831964

  9. An Optimal Cell Detection Technique for Automated Patch Clamping

    NASA Technical Reports Server (NTRS)

    McDowell, Mark; Gray, Elizabeth

    2004-01-01

    While there are several hardware techniques for the automated patch clamping of cells that describe the equipment apparatus used for patch clamping, very few explain the science behind the actual technique of locating the ideal cell for a patch clamping procedure. We present a machine vision approach to patch clamping cell selection by developing an intelligent algorithm technique that gives the user the ability to determine the good cell to patch clamp in an image within one second. This technique will aid the user in determining the best candidates for patch clamping and will ultimately save time, increase efficiency and reduce cost. The ultimate goal is to combine intelligent processing with instrumentation and controls in order to produce a complete turnkey automated patch clamping system capable of accurately and reliably patch clamping cells with a minimum amount of human intervention. We present a unique technique that identifies good patch clamping cell candidates based on feature metrics of a cell's (x, y) position, major axis length, minor axis length, area, elongation, roundness, smoothness, angle of orientation, thinness and whether or not the cell is only particularly in the field of view. A patent is pending for this research.

  10. Spectral infrared hemispherical reflectance measurements for LDEF tray clamps

    NASA Technical Reports Server (NTRS)

    Cromwell, B. K.; Shepherd, S. D.; Pender, C. W.; Wood, B. E.

    1993-01-01

    Infrared hemispherical reflectance measurements that were made on 58 chromic acid anodized tray clamps from LDEF are described. The measurements were made using a hemiellipsoidal mirror reflectometer with interferometer for wavelengths between 2-15 microns. The tray clamps investigated were from locations about the entire spacecraft and provided the opportunity for comparing the effects of atomic oxygen at each location. Results indicate there was essentially no dependence on atomic oxygen fluence for the surfaces studied, but there did appear to be a slight dependence on solar radiation exposure. The reflectances of the front sides of the tray clamps consistently were slightly higher than for the protected rear tray clamp surfaces.

  11. Screening fluorescent voltage indicators with spontaneously spiking HEK cells.

    PubMed

    Park, Jeehae; Werley, Christopher A; Venkatachalam, Veena; Kralj, Joel M; Dib-Hajj, Sulayman D; Waxman, Stephen G; Cohen, Adam E

    2013-01-01

    Development of improved fluorescent voltage indicators is a key challenge in neuroscience, but progress has been hampered by the low throughput of patch-clamp characterization. We introduce a line of non-fluorescent HEK cells that stably express NaV 1.3 and KIR 2.1 and generate spontaneous electrical action potentials. These cells enable rapid, electrode-free screening of speed and sensitivity of voltage sensitive dyes or fluorescent proteins on a standard fluorescence microscope. We screened a small library of mutants of archaerhodopsin 3 (Arch) in spiking HEK cells and identified two mutants with greater voltage-sensitivity than found in previously published Arch voltage indicators.

  12. Nanoindentation characteristics of clamped freestanding Cu membranes.

    PubMed

    Wang, Tong Hong; Fang, Te-Hua; Kang, Shao-Hui; Lin, Yu-Cheng

    2007-04-04

    This research employed instrumented nanoindentation to address the issue of bending to stretching-induced deformation of clamped freestanding Cu membranes. The experimental results show that indentation-induced plastic deformation only comes into effect at the centre and the indented edge of the Cu membrane when the indenter is applied, while the other locations remain undamaged. A step-by-step evolution was presumed for the time histories of the bending to stretching-induced deformation and for the timing of the significant change in slope of the load-deflection curve. Deformation was deliberately introduced at the transition from the single-point bending indentation to the surface stretching indentation at the impact location touched with the indenter. Good elastic recovery was found at locations away from the indenter. A similar finding can be arrived at by means of finite element analysis.

  13. Pipe lifting hook having clamp assembly

    SciTech Connect

    Codner, J.A.

    1984-06-12

    A pipe lifting hook is provided having a generally ''C'' shaped hook member having an elongated lower portion being insertable within the end of a joint of pipe and having an upper portion positionable above the pipe and provided with lifting connection means. The hook member is frictionally clamped to the pipe by grip shoe means that is movably supported by the upper portion of the hook member and is selectably movable from a released position out of contact with said pipe to a locked position in frictional locking engagement with the outer surface of the pipe. A ratchet mechanism couples said grip shoe means to the upper portion of the hook member and is manually positionable to lock said grip shoe means at said locked position or release said grip shoe means for movement toward said released position thereof.

  14. The PCNA-RFC families of DNA clamps and clamp loaders.

    PubMed

    Majka, Jerzy; Burgers, Peter M J

    2004-01-01

    The proliferating cell nuclear antigen PCNA functions at multiple levels in directing DNA metabolic pathways. Unbound to DNA, PCNA promotes localization of replication factors with a consensus PCNA-binding domain to replication factories. When bound to DNA, PCNA organizes various proteins involved in DNA replication, DNA repair, DNA modification, and chromatin modeling. Its modification by ubiquitin directs the cellular response to DNA damage. The ring-like PCNA homotrimer encircles double-stranded DNA and slides spontaneously across it. Loading of PCNA onto DNA at template-primer junctions is performed in an ATP-dependent process by replication factor C (RFC), a heteropentameric AAA+ protein complex consisting of the Rfc1, Rfc2, Rfc3, Rfc4, and Rfc5 subunits. Loading of yeast PCNA (POL30) is mechanistically distinct from analogous processes in E. coli (beta subunit by the gamma complex) and bacteriophage T4 (gp45 by gp44/62). Multiple stepwise ATP-binding events to RFC are required to load PCNA onto primed DNA. This stepwise mechanism should permit editing of this process at individual steps and allow for divergence of the default process into more specialized modes. Indeed, alternative RFC complexes consisting of the small RFC subunits together with an alternative Rfc1-like subunit have been identified. A complex required for the DNA damage checkpoint contains the Rad24 subunit, a complex required for sister chromatid cohesion contains the Ctf18 subunit, and a complex that aids in genome stability contains the Elg1 subunit. Only the RFC-Rad24 complex has a known associated clamp, a heterotrimeric complex consisting of Rad17, Mec3, and Ddc1. The other putative clamp loaders could either act on clamps yet to be identified or act on the two known clamps.

  15. OPTIMAL TIMING FOR CLAMPING THE UMBILICAL CORD AFTER BIRTH

    PubMed Central

    Raju, Tonse N. K.; Singal, Nalini

    2013-01-01

    Synopsis This paper provides a brief overview of pros and cons of clamping the cord too early (within seconds) after birth. It also highlights evolving data that suggests that delaying cord clamping for 30–60 seconds after birth is beneficial to the baby and the mother, with no measurable negative effects. PMID:23164185

  16. Combination Space Station Handrail Clamp and Pointing Device

    NASA Technical Reports Server (NTRS)

    Hughes, Stephen J. (Inventor)

    1999-01-01

    A device for attaching an experiment carrier to a space station handrail is provided. The device has two major components, a clamping mechanism for attachment to a space station handrail, and a pointing carrier on which an experiment package can be mounted and oriented. The handrail clamp uses an overcenter mechanism and the carrier mechanism uses an adjustable preload ball and socket for carrier positioning. The handrail clamp uses a stack of disk springs to provide a spring loaded button. This configuration provides consistent clamping force over a range of possible handrail thicknesses. Three load points are incorporated in the clamping mechanism thereby spreading the clamping load onto three separate points on the handrail. A four bar linkage is used to provide for a single actuation lever for all three load points. For additional safety, a secondary lock consisting of a capture plate and push lock keeps the clamp attached to the handrail in the event of main clamp failure. For the carrier positioning mechanism, a ball in a spring loaded socket uses friction to provide locking torque; however. the ball and socket are torque limited so that the ball ran slip under kick loads (125 pounds or greater). A lead screw attached to disk spring stacks is used to provide an adjustable spring force on the socket. A locking knob is attached to the lead screw to allow for hand manipulation of the lead screw.

  17. Theoretical Verification of Photoelectrochemical Water Oxidation Using Nanocrystalline TiO2 Electrodes.

    PubMed

    Yanagida, Shozo; Yanagisawa, Susumu; Yamashita, Koichi; Jono, Ryota; Segawa, Hiroshi

    2015-05-27

    Mesoscopic anatase nanocrystalline TiO2 (nc-TiO2) electrodes play effective and efficient catalytic roles in photoelectrochemical (PEC) H2O oxidation under short circuit energy gap excitation conditions. Interfacial molecular orbital structures of (H2O)3 &OH(TiO2)9H as a stationary model under neutral conditions and the radical-cation model of [(H2O)3&OH(TiO2)9H]+ as a working nc-TiO2 model are simulated employing a cluster model OH(TiO2)9H (Yamashita/Jono's model) and a H2O cluster model of (H2O)3 to examine excellent H2O oxidation on nc-TiO2 electrodes in PEC cells. The stationary model, (H2O)3&OH(TiO2)9H reveals that the model surface provides catalytic H2O binding sites through hydrogen bonding, van der Waals and Coulombic interactions. The working model, [(H2O)3&OH(TiO2)9H]+ discloses to have a very narrow energy gap (0.3 eV) between HOMO and LUMO potentials, proving that PEC nc-TiO2 electrodes become conductive at photo-irradiated working conditions. DFT-simulation of stepwise oxidation of a hydroxide ion cluster model of OH-(H2O)3, proves that successive two-electron oxidation leads to hydroxyl radical clusters, which should give hydrogen peroxide as a precursor of oxygen molecules. Under working bias conditions of PEC cells, nc-TiO2 electrodes are now verified to become conductive by energy gap photo-excitation and the electrode surface provides powerful oxidizing sites for successive H2O oxidation to oxygen via hydrogen peroxide.

  18. Photovoltaic performance of TiO2 electrode adsorbed with gardenia yellow purified by nonionic polymeric sorbent in dye-sensitized solar cells.

    PubMed

    Kwon, Oh Oun; Kim, Eui Jin; Lee, Jae Hyeok; Kim, Tae Young; Park, Kyung Hee; Kim, Sang Yook; Suh, Hwa Jin; Lee, Hyo Jung; Lee, Jae Wook

    2015-02-05

    To improve the photovoltaic conversion efficiency in dye-sensitized solar cells (DSSCs), TiO2 electrode adsorbed with gardenia yellow purified by nonionic polymeric sorbent was successfully formulated on nanoporous TiO2 surface. Adsorption and desorption properties of crude gardenia yellow solution on a macroporous resin, XAD-1600, were investigated to purify gardenia yellow because of its strong adsorption and desorption abilities as well as high selectivity. To this end, adsorption equilibrium and kinetic data were measured and fitted using adsorption isotherms and kinetic models. Adsorption and desorption breakthrough curves in a column packed with XAD-1600 resin was obtained to optimize the separation process of gardenia yellow. The photovoltaic performance of the photo-electrode adsorbed with the crude and purified gardenia yellow in DSSCs was compared from current-voltage measurements. The results showed that the photovoltaic conversion efficiency was highly dependent on how to separate and purify gardenia yellow as a photosensitizer.

  19. A novel measuring method of clamping force for electrostatic chuck in semiconductor devices

    NASA Astrophysics Data System (ADS)

    Kesheng, Wang; Jia, Cheng; Yin, Zhong; Linhong, Ji

    2016-04-01

    Electrostatic chucks are one of the core components of semiconductor devices. As a key index of electrostatic chucks, the clamping force must be controlled within a reasonable range. Therefore, it is essential to accurately measure the clamping force. To reduce the negative factors influencing measurement precision and repeatability, this article presents a novel method to measure the clamping force and we elaborate both the principle and the key procedure. A micro-force probe component is introduced to monitor, adjust, and eliminate the gap between the wafer and the electrostatic chuck. The contact force between the ruby probe and the wafer is selected as an important parameter to characterize de-chucking, and we have found that the moment of de-chucking can be exactly judged. Moreover, this article derives the formula calibrating equivalent action area of backside gas pressure under real working conditions, which can effectively connect the backside gas pressure at the moment of de-chucking and the clamping force. The experiments were then performed on a self-designed measuring platform. The de-chucking mechanism is discussed in light of our analysis of the experimental data. Determination criteria for de-chucking point are summed up. It is found that the relationship between de-chucking pressure and applied voltage conforms well to quadratic equation. Meanwhile, the result reveals that actual de-chucking behavior is much more complicated than the description given in the classical empirical formula. Project supported by No. 02 National Science and Technology Major Project of China (No. 2011ZX02403-004).

  20. Correlation of open cell-attached and excised patch clamp techniques.

    PubMed

    Filipovic, D; Hayslett, J P

    1995-11-01

    The excised patch clamp configuration provides a unique technique for some types of single channel analyses, but maintenance of stable, long-lasting preparations may be confounded by rundown and/or rapid loss of seal. Studies were performed on the amiloride-sensitive Na+ channel, located on the apical surface of A6 cells, to determine whether the nystatin-induced open cell-attached patch could serve as an alternative configuration. Compared to excised inside-out patches, stable preparations were achieved more readily with the open cell-attached patch (9% vs. 56% of attempts). In both preparations, the current voltage (I-V) relation was linear, current amplitudes were equal at opposite equivalent clamped voltages, and Erev was zero in symmetrical Na+ solutions, indicating similar Na+ activities on the cytosolic and external surfaces of the patch. Moreover, there was no evidence that nystatin altered channel activity in the patch because slope conductance (3-4pS) and Erev (75 mV), when the bath was perfused with a high K:low Na solution (ENa = 80 mV), were nearly equal in both patch configurations. Our results therefore indicate that the nystatin-induced open cell-attached patch can serve as an alternative approach to the excised inside-out patch when experiments require modulation of univalent ions in the cytosol.

  1. Cell-Detection Technique for Automated Patch Clamping

    NASA Technical Reports Server (NTRS)

    McDowell, Mark; Gray, Elizabeth

    2008-01-01

    A unique and customizable machinevision and image-data-processing technique has been developed for use in automated identification of cells that are optimal for patch clamping. [Patch clamping (in which patch electrodes are pressed against cell membranes) is an electrophysiological technique widely applied for the study of ion channels, and of membrane proteins that regulate the flow of ions across the membranes. Patch clamping is used in many biological research fields such as neurobiology, pharmacology, and molecular biology.] While there exist several hardware techniques for automated patch clamping of cells, very few of those techniques incorporate machine vision for locating cells that are ideal subjects for patch clamping. In contrast, the present technique is embodied in a machine-vision algorithm that, in practical application, enables the user to identify good and bad cells for patch clamping in an image captured by a charge-coupled-device (CCD) camera attached to a microscope, within a processing time of one second. Hence, the present technique can save time, thereby increasing efficiency and reducing cost. The present technique involves the utilization of cell-feature metrics to accurately make decisions on the degree to which individual cells are "good" or "bad" candidates for patch clamping. These metrics include position coordinates (x,y) in the image plane, major-axis length, minor-axis length, area, elongation, roundness, smoothness, angle of orientation, and degree of inclusion in the field of view. The present technique does not require any special hardware beyond commercially available, off-the-shelf patch-clamping hardware: A standard patchclamping microscope system with an attached CCD camera, a personal computer with an imagedata- processing board, and some experience in utilizing imagedata- processing software are all that are needed. A cell image is first captured by the microscope CCD camera and image-data-processing board, then the image

  2. Trends in Aortic Clamp Use During Coronary Artery Bypass Surgery: The Effect of Aortic Clamping Strategies on Neurologic Outcomes

    PubMed Central

    Daniel, William T.; Kilgo, Patrick; Puskas, John D.; Thourani, Vinod H.; Lattouf, Omar M.; Guyton, Robert A.; Halkos, Michael E.

    2013-01-01

    Objective The purpose of this study was to determine the impact of different clamping strategies during CABG on the incidence of postoperative stroke. Methods In this case-control study, all patients at Emory hospitals from 2002–2009 with postoperative stroke after isolated CABG (N=141) were matched 1:4 to a contemporaneous cohort of patients without postoperative stroke (N=565). Patients were matched according to the Society of Thoracic Surgeons (STS) Predicted Risk of Postoperative Stroke (PROPS), which is based on 26 variables. On- (ONCAB) and off-pump (OPCAB) patients were matched separately. Multiple logistic regression analysis with adjusted odds ratios (OR) was performed to identify operative variables associated with postoperative stroke. Results Among the ONCAB cohort, the single cross-clamp technique was associated with a decreased risk of stroke compared to the double clamp (cross clamp + partial clamp) technique (OR=0.385, p=0.044). Within the OPCAB cohort, there was no significant difference in stroke according to clamp use. Epiaortic ultrasound of the ascending aorta increased from 45.3% in 2002 to 89.4% in 2009. From 2002–2009, clamp use decreased from 97.7% of cases to 72.7%. Conclusions During ONCAB, the use of a single cross-clamp compared to the double clamp technique decreases the risk of postoperative stroke. The use of any aortic clamp has decreased and epiaortic ultrasound use has increased from 2002–2009, indicating a change in operative technique and surgeon awareness of the potential complications associated with manipulation of the aorta. PMID:23477689

  3. Avoiding aortic clamping during CABG reduces postoperative stroke

    PubMed Central

    Moss, Emmanuel; Puskas, John D; Thourani, Vinod H; Kilgo, Patrick; Chen, Edward P; Leshnower, Bradley G; Lattouf, Omar M; Guyton, Robert A.; Glas, Kathryn E; Halkos, Michael E.

    2014-01-01

    Objective The purpose of this study was to determine whether the incidence of postoperative stroke (PS) could be reduced by eliminating aortic clamping during CABG. Methods From 2002–2013, 12,079 patients underwent primary, isolated CABG at a single US academic institution. Aortic manipulation was completely avoided by using in-situ internal mammary arteries for inflow in 1,552 (12.9%) patients (no-touch), a clampless facilitating device (CFD) was used for proximal anastomoses in 1,548 (12.8%) patients, and aortic clamping was used in 8,979 (74.3%) patients. These strategies were assessed in a logistic regression model controlling for relevant variables. Results The overall incidence of PS was 1.4% (n=165), with an unadjusted incidence of 0.6% (n=10) in the no-touch group, 1.2% (n=18) in the CFD group, and 1.5% (n=137) in the clamp group (p<0.01 for no-touch vs clamp). The ratio of observed to expected stroke rate increased as the degree of aortic manipulation increased, from 0.48 in the no-touch group, to 0.61 in the CFD group, and 0.95 in the clamp group. Aortic clamping was independently associated with an increase in PS compared to a no-touch technique (AOR 2.50, p<0.01). When separated by CPB utilization, both the off-pump partial clamp and on-pump cross-clamp techniques increased the risk of PS compared to no-touch (AOR 2.52, p<0.01 and AOR 4.25, p<0.001, respectively). Conclusion A no-aortic touch technique has the lowest risk for postoperative stroke for patients undergoing CABG. Clamping the aorta during CABG increases the risk of PS, regardless of the severity of aortic disease. PMID:25293356

  4. Reciprocating clamp apparatus for thermoforming plastic containers

    SciTech Connect

    Beck, M.H.; Harry, I.L.; Krishnakumar, S.M.

    1984-03-06

    This relates to the forming of containers and like hollow articles from sheets or webs of thermoplastic material. Two webs or sheets are simultaneously acted upon by way of a forming apparatus which includes a reciprocating clamp first cooperable with one outer platen and then the other in sequence wherein, while a first web or sheet is being formed within a plurality of mold cavities to define a plurality of hollow articles such as containers, the other sheet or web may be stripped from its respective mold set and a new sheet or a new portion of a sheet or web may be advanced into position for molding. The forming apparatus may be constructed in a manner wherein the web portions which are to be formed may be billowed away from the mold cavities as an initial step in the stretching and orientation of the thermoplastic material. The thermoplastic material may be heated to the desired forming temperature using separate sets of rf electrodes so that only those portions of the thermoplastic material which are to be formed need be heated. Two sets of containers of different sizes may be formed from the webs or sheets, and then internested to form double wall containers wherein the walls may be formed of different materials and wherein the outer material may be a barrier material, or wherein the walls of the containers are spaced apart to form an insulated container. This abstract forms no part of the specification of this application and is not to be construed as limiting the claims of the application.

  5. High voltage load resistor array

    DOEpatents

    Lehmann, Monty Ray

    2005-01-18

    A high voltage resistor comprising an array of a plurality of parallel electrically connected resistor elements each containing a resistive solution, attached at each end thereof to an end plate, and about the circumference of each of the end plates, a corona reduction ring. Each of the resistor elements comprises an insulating tube having an electrode inserted into each end thereof and held in position by one or more hose clamps about the outer periphery of the insulating tube. According to a preferred embodiment, the electrode is fabricated from stainless steel and has a mushroom shape at one end, that inserted into the tube, and a flat end for engagement with the end plates that provides connection of the resistor array and with a load.

  6. A clamp-like biohybrid catalyst for DNA oxidation

    NASA Astrophysics Data System (ADS)

    van Dongen, Stijn F. M.; Clerx, Joost; Nørgaard, Kasper; Bloemberg, Tom G.; Cornelissen, Jeroen J. L. M.; Trakselis, Michael A.; Nelson, Scott W.; Benkovic, Stephen J.; Rowan, Alan E.; Nolte, Roeland J. M.

    2013-11-01

    In processive catalysis, a catalyst binds to a substrate and remains bound as it performs several consecutive reactions, as exemplified by DNA polymerases. Processivity is essential in nature and is often mediated by a clamp-like structure that physically tethers the catalyst to its (polymeric) template. In the case of the bacteriophage T4 replisome, a dedicated clamp protein acts as a processivity mediator by encircling DNA and subsequently recruiting its polymerase. Here we use this DNA-binding protein to construct a biohybrid catalyst. Conjugation of the clamp protein to a chemical catalyst with sequence-specific oxidation behaviour formed a catalytic clamp that can be loaded onto a DNA plasmid. The catalytic activity of the biohybrid catalyst was visualized using a procedure based on an atomic force microscopy method that detects and spatially locates oxidized sites in DNA. Varying the experimental conditions enabled switching between processive and distributive catalysis and influencing the sliding direction of this rotaxane-like catalyst.

  7. Studying mechanosensitive ion channels with an automated patch clamp.

    PubMed

    Barthmes, Maria; Jose, Mac Donald F; Birkner, Jan Peter; Brüggemann, Andrea; Wahl-Schott, Christian; Koçer, Armağan

    2014-03-01

    Patch clamp electrophysiology is the main technique to study mechanosensitive ion channels (MSCs), however, conventional patch clamping is laborious and success and output depends on the skills of the operator. Even though automated patch systems solve these problems for other ion channels, they could not be applied to MSCs. Here, we report on activation and single channel analysis of a bacterial mechanosensitive ion channel using an automated patch clamp system. With the automated system, we could patch not only giant unilamellar liposomes but also giant Escherichia coli (E. coli) spheroplasts. The tension sensitivity and channel kinetics data obtained in the automated system were in good agreement with that obtained from the conventional patch clamp. The findings will pave the way to high throughput fundamental and drug screening studies on mechanosensitive ion channels.

  8. Laser Transmission Welding of Thermoplastics With Dual Clamping Devices

    NASA Astrophysics Data System (ADS)

    Devrient, M.; Knoll, B.; Geiger, R.

    Beside well chosen process parameters a geometrical joining partner design suitable for laser transmission welding and adequate clamping pressure appliance is necessary to form high quality welds. Amongst other clamping techniques Dual Clamping Devices (DCD) are a promising approach to fulfill the process needs and to get to a robust and also fail safe clamping. When using DCD the laser beam has to pass thin non transmitting bars while a weld seam is formed. So the laser beam is partial refracted, reflected and absorbed. Here the influences of the bars onto the laser beam intensity distribution behind the bars rather in the interaction zone of the laser beam and the two joining partners are investigated. Welding experiments with DCD are carried out and discussed. Based on the experimental results thermal process simulations are performed, to get a deeper knowledge about the effect of the bars onto the spatially and temporally changing temperature field within the joining partners.

  9. The Application of Mechanical Clamps to Portsmouth Connectors.

    DTIC Science & Technology

    1981-11-23

    Resistance Readings ..... .............. .64 TABLE 12 - ALT SUMM!ARY - POLYURETHANE CONNECTORS Resis- ance Readings ..... .............. .65 v7 ft...34 FIGURE 17 Pressure Tank Pressure Fittings . ........ . 37 vi I THE APPLICATION OF MECHANICAL CLAMPS TO PORTSMOUTH CONNECTORS BACKGROUN )D This

  10. Mechanism of Electromagnetic Energy Effects on the Nervous System: Voltage-Clamp Study.

    DTIC Science & Technology

    1985-07-01

    Lin, Role of External Calcium in Microwave-Induced Snail Neuron Response, Abstract, 5th Annual Scientific Session Bioelectro - magnetics Society...Snail Nerve Cells to Noise Modulated Microwave Field, Abstract, 4th Annual Scientific Session of Bioelectro - magnetics Society, held in Los Angeles

  11. A loudspeaker-driven system for rapid and multiple solution exchanges in patch-clamp experiments.

    PubMed

    Méry, P F; Lechêne, P; Fischmeister, R

    1992-04-01

    A new and inexpensive system allowing rapid and synchronized changes of solutions around a membrane patch or a cell under voltage-clamp conditions is described. Four plastic capillary tubings (OD 640 microns; ID 430 microns) were glued together horizontally and attached to a coil of a commercially available loudspeaker. Servo-control of the position of the coil allowed the mouth of any of the capillaries to be positioned near the pipette tip within 6 ms. A high flow speed of the test solution was crucial to achieve rapid solution exchange. At a flow speed of 5 cm/s, complete exchange of the external environment of a frog ventricular cell was achieved within 20-30 ms. The time course of solution change was found to be 3-5 times faster at the tip of an open patch pipette. To preserve the physical integrity of the cell, the cell was usually perfused by a control capillary at a slow velocity (0.2-0.4 cm/s) and test solutions flowing out of adjacent capillaries at high velocity (4-5 cm/s) were applied to the cell only for short periods. Determination of the three-dimensional contamination profile around the mouth of the control capillary allowed the optimal conditions for the use of the system to be established and possible sources of contamination to be avoided between adjacent capillaries with unmatched flow speeds. Successive and multiple changes in external solutions could be easily synchronized with voltage-clamp depolarizations to examine the time course of the effect of drugs on voltage-operated ion channels.(ABSTRACT TRUNCATED AT 250 WORDS)

  12. Spatial dynamics, thermalization, and gain clamping in a photon condensate

    NASA Astrophysics Data System (ADS)

    Keeling, Jonathan; Kirton, Peter

    2016-01-01

    We study theoretically the effects of pump-spot size and location on photon condensates. By exploring the inhomogeneous molecular excitation fraction, we make clear the relation between spatial equilibration, gain clamping, and thermalization in a photon condensate. This provides a simple understanding of several recent experimental results. We find that as thermalization breaks down, gain clamping is imperfect, leading to "transverse spatial hole burning" and multimode condensation. This opens the possibility of engineering the gain profile to control the condensate structure.

  13. Analysis of the dynamic avalanche of carrier stored trench bipolar transistor (CSTBT) during clamped inductive turn-off transient

    NASA Astrophysics Data System (ADS)

    Xue, Peng; Fu, Guicui

    2017-03-01

    The dynamic avalanche has a huge impact on the switching robustness of carrier stored trench bipolar transistor (CSTBT). The purpose of this work is to investigate the CSTBT's dynamic avalanche mechanism during clamped inductive turn-off transient. At first, with a Mitsubishi 600 V/150 A CSTBT and a Infineon 600 V/200 A field stop insulated gate bipolar transistor (FS-IGBT) utilized, the clamped inductive turn-off characteristics are obtained by double pulse test. The unclamped inductive switching (UIS) test is also utilized to identify the CSTBT's clamping voltage under dynamic avalanche condition. After the test data analysis, it is found that the CSTBT's dynamic avalanche is abnormal and can be triggered under much looser condition than the conventional buffer layer IGBT. The comparison between the FS-IGBT and CSTBT's experimental results implies that the CSTBT's abnormal dynamic avalanche phenomenon may be induced by the carrier storage (CS) layer. Based on the semiconductor physics, the electric field distribution and dynamic avalanche generation in the depletion region are analyzed. The analysis confirms that the CS layer is the root cause of the CSTBT's abnormal dynamic avalanche mechanism. Moreover, the CSTBT's negative gate capacitance effect is also investigated to clarify the underlying mechanism of the gate voltage bump observed in the test. In the end, the mixed-mode numerical simulation is utilized to reproduce the CSTBT's dynamic avalanche behavior. The simulation results validate the proposed dynamic avalanche mechanisms.

  14. Fabrication and characterization of PbO2 electrode modified with polyvinylidene fluoride (PVDF)

    NASA Astrophysics Data System (ADS)

    Li, Xiaoliang; Xu, Hao; Yan, Wei

    2016-12-01

    A novel PbO2 electrode with a high oxygen evolution potential (OEP) and long service life was successfully fabricated by doping polyvinyl fluoride (PVDF) (marked as PbO2-PVDF) through co-deposition method. The morphology (SEM), elemental analysis (EDX), hydrophobic property (contact angle), crystalline structure (XRD), chemical state (XPS), electrochemical performances (Lsv and EIS) and stability (accelerated life test) were characterized. The results showed that PVDF doping could improve the film morphology, increase oxygen evolution potential (OEP) and reduce the electrode film impedance. In addition, the proportion of adsorbed hydroxyl oxygen (Oad) on the electrode also increased. During the electrochemical oxidation process, the PbO2-PVDF(2.0) electrode showed the best performance on degradation of phenol due to the highest removal rate, lowest energy consumption and minimum Pb dissolution, which could be attributed to its hydrophobic surface, high oxygen evolution potential (OEP) and strong capability of HOrad generation. Furthermore, the stability of the electrodes were greatly improved after PVDF modification. PbO2-PVDF(1.0) electrode showed the longest service life (501 h), which was more than 4 times longer than PbO2 electrode (118.5 h).

  15. Nonlinear response of a clamped-clamped beam with internal resonance under sinusoidal excitation

    NASA Astrophysics Data System (ADS)

    Afaneh, Abdul-Hafiz Ahmed

    1992-01-01

    The nonlinear response characteristics of a clamped-clamped beam is investigated analytically, numerically, and experimentally. The beam is under an initial static axial load and subjected to a harmonic excitation of its support. Two ranges of the axial load are considered. These are below (the beam is initially straight) and above Euler buckling load (the beam is initially buckled). Hamilton's principle is used to derive a fourth order partial differential equation of motion which is descritized and reduced to a set of second order ordinary differential equations by applying Galerkin's method. Under certain values of the static load, the normal modes are nonlinearly coupled and this coupling results in a fourth order internal resonance condition between the first three modes when the beam is initially straight. Second and third order internal resonance conditions occur between the first two modes for the case of initially buckled beam. The multiple scales method showed the significant effects of these internal resonance conditions on the system behavior. In the straight beam case, the third mode which is externally excited transfers energy to the first two modes within a small range of internal detuning. Outside this region, the response is governed by a unimodal response of the third mode. In the neighborhood of 1:1 internal resonance, it is found that within the region of two mode interaction, the solution is either stationary or nonstationary depending on the excitation level and system parameters. Saturation and jump phenomena are found to take place in the case of two mode interaction with 2:1 internal resonance. Numerical simulation and experimental testing confirmed these predictions and revealed the occurrence of multifurcation, snap-through (escaping from one well to the other in an irregular manner), and chaotic motion.

  16. A Transformerless Motor Drive with a Five-Level Diode-Clamped PWM Inverter for Fan/Blower Loads

    NASA Astrophysics Data System (ADS)

    Hasegawa, Kazunori; Hatti, Natchpong; Akagi, Hirofumi

    This paper describes a 6.6-kV adjustable-speed motor drive for use in fans, blowers, and pumps without a transformer. The power-conversion system consists of a diode rectifier, a five-level diode-clamped PWM inverter, and a voltage-balancing circuit. A 200-V 5.5-kW downscale model is developed, constructed, and tested. The five-level PWM inverter and the voltage-balancing circuit are studied in detail. Experimental results obtained from testing the 200-V downscale model confirm the viability and effectiveness of the 6.6-kV adjustable-speed motor drive, indicating that the dc mean voltages of the four split dc capacitors are well balanced under all the given operating conditions.

  17. Overproduction and purification of RFC-related clamp loaders and PCNA-related clamps from Saccharomyces cerevisiae.

    PubMed

    Bylund, Göran O; Majka, Jerzy; Burgers, Peter M J

    2006-01-01

    The replication clamp PCNA and its loader RFC (Replication Factor C) are central factors required for processive replication and coordinated DNA repair. Recently, several additional related clamp loaders have been identified. These alternative clamp loaders contain the small Rfc2-5 subunits of RFC, but replace the large Rfc1 subunit by a pathway-specific alternative large subunit, Rad24 for the DNA damage checkpoint, Ctf18 for the establishment of sister chromatid cohesion, and Elg1 for a general function in chromosome stability. In order to define biochemical functions for these loaders, the loaders were overproduced in yeast and purified at a milligram scale. To aid in purification, the large subunit of each clamp loader was fused to a GST-tag that, after purification could be easily removed by a rhinoviral protease. This methodology yielded all clamp loaders in high yield and with high enzymatic activity. The yeast 9-1-1 checkpoint clamp, consisting of Rad17, Mec3, and Ddc1, was overproduced and purified in a similar manner.

  18. Solid Electrolyte Interphase (SEI) at TiO2 Electrodes in Li-Ion Batteries: Defining Apparent and Effective SEI Based on Evidence from X-ray Photoemission Spectroscopy and Scanning Electrochemical Microscopy.

    PubMed

    Ventosa, Edgar; Madej, Edyta; Zampardi, Giorgia; Mei, Bastian; Weide, Philipp; Antoni, Hendrik; La Mantia, Fabio; Muhler, Martin; Schuhmann, Wolfgang

    2017-01-25

    The high (de)lithiation potential of TiO2 (ca. 1.7 V vs Li/Li(+) in 1 M Li(+)) decreases the voltage and, thus, the energy density of a corresponding Li-ion battery. On the other hand, it offers several advantages such as the (de)lithiation potential far from lithium deposition or absence of a solid electrolyte interphase (SEI). The latter is currently under controversial debate as several studies reported the presence of a SEI when operating TiO2 electrodes at potentials above 1.0 V vs Li/Li(+). We investigate the formation of a SEI at anatase TiO2 electrodes by means of X-ray photoemission spectroscopy (XPS) and scanning electrochemical microscopy (SECM). The investigations were performed in different potential ranges, namely, during storage (without external polarization), between 3.0-2.0 V and 3.0-1.0 V vs Li/Li(+), respectively. No SEI is formed when a completely dried and residues-free TiO2 electrode is cycled between 3.0 and 2.0 V vs Li/Li(+). A SEI is detected by XPS in the case of samples stored for 6 weeks or cycled between 3.0 and 1.0 V vs Li/Li(+). With use of SECM, it is verified that this SEI does not possess the electrically insulating character as expected for a "classic" SEI. Therefore, we propose the term apparent SEI for TiO2 electrodes to differentiate it from the protecting and effective SEI formed at graphite electrodes.

  19. Spectral infrared hemispherical reflectance measurements for LDEF tray clamps

    NASA Technical Reports Server (NTRS)

    Wood, Bobby E.; Cromwell, Brian K.; Pender, Charles W.; Shepherd, Seth D.

    1992-01-01

    This paper describes infrared hemispherical reflectance measurements (2-15 microns) that were made on 58 chromic acid anodized tray clamps retrieved from the LDEF spacecraft. These clamps were used for maintaining the experiments in place and were located at various locations about the spacecraft. Changes in reflectance of the tray clamps at these locations were compared with atomic oxygen fluxes at the same locations. A decrease in absorption band depth was seen for the surfaces exposed to space indicating that there was some surface layer erosion. In all of the surfaces measured, little evidence of contamination was observed and none of the samples showed evidence of the brown nicotine stain that was so prominent in other experiments. Total emissivity values were calculated for both exposed and unexposed tray clamp surfaces. Only small differences, usually less than 1 percent, were observed. The spectral reflectances were measured using a hemi-ellipsoidal mirror reflectometer matched with an interferometer spectrometer. The rapid scanning capability of the interferometer allowed the reflectance measurements to be made in a timely fashion. The ellipsoidal mirror has its two foci separated by 2 inches and located on the major axis. A blackbody source was located at one focus while the tray clamp samples were located at the conjugate focus. The blackbody radiation was modulated and then focused by the ellipsoid onto the tray clamps. Radiation reflected from the tray clamp was sampled by the interferometer by viewing through a hole in the ellipsoid. A gold mirror (reflectance approximately 98 percent) was used as the reference surface.

  20. Compartmental models of rat cerebellar Purkinje cells based on simultaneous somatic and dendritic patch-clamp recordings

    PubMed Central

    Roth, Arnd; Häusser, Michael

    2001-01-01

    Simultaneous dendritic and somatic patch-clamp recordings were made from Purkinje cells in cerebellar slices from 12- to 21-day-old rats. Voltage responses to current impulses injected via either the dendritic or the somatic pipette were obtained in the presence of the selective Ih blocker ZD 7288 and blockers of spontaneous synaptic input. Neurons were filled with biocytin for subsequent morphological reconstruction. Four neurons were reconstructed and converted into detailed compartmental models. The specific membrane capacitance (Cm), specific membrane resistance (Rm) and intracellular resistivity (Ri) were optimized by direct fitting of the model responses to the electrophysiological data from the same cell. Mean values were: Cm, 0.77 ± 0.17 μF cm−2 (mean ±s.d.; range, 0.64-1.00 μF cm−2), Rm, 122 ± 18 kΩ cm2 (98-141 kΩ cm2) and Ri, 115 ± 20 Ω cm (93-142 Ω cm). The steady-state electrotonic architecture of these cells was compact under the experimental conditions used. However, somatic voltage-clamp recordings of parallel fibre and climbing fibre synaptic currents were substantially filtered and attenuated. The detailed models were compared with a two-compartment model of Purkinje cells. The range of synaptic current kinetics that can be faithfully recorded using somatic voltage clamp is predicted fairly well by the two-compartment model, even though some of its underlying assumptions are violated. A model of Ih was constructed based on voltage-clamp data, and inserted into the passive compartmental models. Somatic EPSP amplitude was substantially attenuated compared to the amplitude of dendritic EPSPs at their site of generation. However, synaptic efficacy of the same quantal synaptic conductance, as measured by the somatic EPSP amplitude, was only weakly dependent on synaptic location on spiny branchlets. The passive electrotonic structure of Purkinje cells is unusual in that the steady-state architecture is very compact, while voltage transients

  1. Photoelectrochemical degradation of Methylene Blue with beta-PbO2 electrodes driven by visible light irradiation.

    PubMed

    Li, Guoting; Yip, Hoyin; Wong, Kin Hang; Hu, Chun; Qu, Jiuhui; Wong, Po Keung

    2011-01-01

    Beta-PbO2 electrodes were prepared by electro-deposition and characterized by scanning electron microscopy, X-ray diffraction, X-ray photoelectron spectroscopy, and linear sweep voltammetry. We confirmed pure (beta-PbO2 crystals were on the electrode and it had a high oxygen evolution potential. The photoactivity and photoelectrochemical (PEC) properties of the beta-PbO2 electrode were investigated under visible light irradiation (lambda > 420 nm) for the decolorization of Methylene Blue. Pseudo first-order kinetics parameter (K(app)) for dye decolorization using the beta-PbO2 electrode achieved 6.71 x 10(-4) min(-1) under visible light irradiation, which indicated its excellent visible light-induced photoactivity. The K(app) of the PEC process was as much as 1.41 x 10(-3) min(-1) and was 1.71 times that of visible light irradiation or electrolysis even in the presence of the beta-PbO2 electrode. A significant synergetic effect was observed in the PEC system. We also employed TiO2 modified beta-PbO2 electrodes in this test, which revealed that the TiO2 immobilized on the beta-PbO2 electrode inhibited the visible light-induced PEC efficiency despite the amount of TiO2 used for electrode preparation. The beta-PbO2 electrode was also superior to the dimensionally stable anode (Ti/Ru(0.3)Ti(0.7)O2) in visible light-induced photoactivity and PEC efficiency.

  2. Paired patch clamp recordings from motor-neuron and target skeletal muscle in zebrafish.

    PubMed

    Wen, Hua; Brehm, Paul

    2010-11-20

    Larval zebrafish represent the first vertebrate model system to allow simultaneous patch clamp recording from a spinal motor-neuron and target muscle. This is a direct consequence of the accessibility to both cell types and ability to visually distinguish the single segmental CaP motor-neuron on the basis of morphology and location. This video demonstrates the microscopic methods used to identify a CaP motor-neuron and target muscle cells as well as the methodologies for recording from each cell type. Identification of the CaP motor-neuron type is confirmed by either dye filling or by the biophysical features such as action potential waveform and cell input resistance. Motor-neuron recordings routinely last for one hour permitting long-term recordings from multiple different target muscle cells. Control over the motor-neuron firing pattern enables measurements of the frequency-dependence of synaptic transmission at the neuromuscular junction. Owing to a large quantal size and the low noise provided by whole cell voltage clamp, all of the unitary events can be resolved in muscle. This feature permits study of basic synaptic properties such as release properties, vesicle recycling, as well as synaptic depression and facilitation. The advantages offered by this in vivo preparation eclipse previous neuromuscular model systems studied wherein the motor-neurons are usually stimulated by extracellular electrodes and the muscles are too large for whole cell patch clamp. The zebrafish preparation is amenable to combining electrophysiological analysis with a wide range of approaches including transgenic lines, morpholino knockdown, pharmacological intervention and in vivo imaging. These approaches, coupled with the growing number of neuromuscular disease models provided by mutant lines of zebrafish, open the door for new understanding of human neuromuscular disorders.

  3. E-beam high voltage switching power supply

    DOEpatents

    Shimer, D.W.; Lange, A.C.

    1996-10-15

    A high-power power supply produces a controllable, constant high voltage output under varying and arcing loads. The power supply includes a voltage regulator, an inductor, an inverter for producing a high frequency square wave current of alternating polarity, an improved inverter voltage clamping circuit, a step up transformer, an output rectifier for producing a dc voltage at the output of each module, and a current sensor for sensing output current. The power supply also provides dynamic response to varying loads by controlling the voltage regulator duty cycle and circuitry is provided for sensing incipient arc currents at the output of the power supply to simultaneously decouple the power supply circuitry from the arcing load. The power supply includes a plurality of discrete switching type dc--dc converter modules. 5 figs.

  4. E-beam high voltage switching power supply

    DOEpatents

    Shimer, Daniel W.; Lange, Arnold C.

    1996-01-01

    A high-power power supply produces a controllable, constant high voltage put under varying and arcing loads. The power supply includes a voltage regulator, an inductor, an inverter for producing a high frequency square wave current of alternating polarity, an improved inverter voltage clamping circuit, a step up transformer, an output rectifier for producing a dc voltage at the output of each module, and a current sensor for sensing output current. The power supply also provides dynamic response to varying loads by controlling the voltage regulator duty cycle and circuitry is provided for sensing incipient arc currents at the output of the power supply to simultaneously decouple the power supply circuitry from the arcing load. The power supply includes a plurality of discrete switching type dc--dc converter modules.

  5. Whole-cell clamp of dissociated photoreceptors from the eye of Lima scabra.

    PubMed

    Nasi, E

    1991-01-01

    Voltage-dependent membrane currents were investigated in enzymatically dissociated photoreceptors of Lima scabra using the whole-cell clamp technique. Depolarizing steps to voltages more positive than -10 mV elicit a transient inward current followed by a delayed, sustained outward current. The outward current is insensitive to replacement of a large fraction of extracellular Cl- with the impermeant anion glucuronate. Superfusion with tetraethylammonium and 4-aminopyridine reversibly abolishes the outward current, and internal perfusion with cesium also suppresses it, indicating that it is mediated by potassium channels. Isolation of the inward current reveals a fast activation kinetics, the peak amplitude occurring as early as 4-5 ms after stimulus onset, and a relatively rapid, though incomplete inactivation. Within the range of voltages examined, spanning up to +90 mV, reversal was not observed. The inward current is not sensitive to tetrodotoxin at concentrations up to 10 microM, and survives replacement of extracellular Na with tetramethylammonium. On the other hand, it is completely eliminated by calcium removal from the perfusing solution, and it is partially blocked by submillimolar concentrations of cadmium, suggesting that it is entirely due to voltage-dependent calcium channels. Analysis of the kinetics and voltage dependence of the isolated calcium current indicates the presence of two components, possibly reflecting the existence of separate populations of channels. Barium and strontium can pass through these channels, though less easily than calcium. Both the activation and the inactivation become significantly more sluggish when these ions serve as the charge carrier. A large fraction of the outward current is activated by preceding calcium influx. Suppression of this calcium-dependent potassium current shows a small residual component resembling the delayed rectifier. In addition, a transient outward current sensitive to 4-aminopyridine (Ia) could

  6. Measuring beta-cell function relative to insulin sensitivity in youth: Does the hyperglycemic clamp suffice?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To compare beta-cell function relative to insulin sensitivity, disposition index (DI), calculated from two clamps (2cDI, insulin sensitivity from the hyperinsulinemic-euglycemic clamp and first-phase insulin from the hyperglycemic clamp) with the DI calculated from the hyperglycemic clamp alone (hcD...

  7. Subcellular Patch-clamp Recordings from the Somatodendritic Domain of Nigral Dopamine Neurons

    PubMed Central

    Engel, Dominique

    2016-01-01

    Dendrites of dopaminergic neurons receive and convey synaptic input, support action potential back-propagation and neurotransmitter release. Understanding these fundamental functions will shed light on the information transfer in these neurons. Dendritic patch-clamp recordings provide the possibility to directly examine the electrical properties of dendrites and underlying voltage-gated ion channels. However, these fine structures are not easily accessible to patch pipettes because of their small diameter. This report describes a step-by-step procedure to collect stable and reliable recordings from the dendrites of dopaminergic neurons in acute slices. Electrophysiological measurements are combined with post hoc recovery of cell morphology. Successful experiments rely on improved preparation of slices, solutions and pipettes, adequate adjustment of the optics and stability of the pipette in contact with the recorded structure. Standard principles of somatic patch-clamp recording are applied to dendrites but with a gentler approach of the pipette. These versatile techniques can be implemented to address various questions concerning the excitable properties of dendrites. PMID:27842379

  8. Polyacrylate bound TiSb2 electrodes for Li-ion batteries

    NASA Astrophysics Data System (ADS)

    Gómez-Cámer, Juan Luis; Novák, Petr

    2015-01-01

    Crystalline TiSb2 electrodes prepared using two different binders, PVDF and lithium polyacrylate (LiPAA), were examined as negative electrodes in Li-ion batteries. The cycle life of the electrodes is strongly influenced by the choice of the binder, reaching ca. 120 cycles with LiPAA vs. ca. 90 cycles achieved with the common binder PVDF. Moreover, rate capability is improved using LiPAA binder. The reduction in TiSb2 particle size is shown to influence the average practical specific charge at high charge/discharge rates. The reasons for this improvement are discussed and the optimized electrode was demonstrated in full Li-ion cells.

  9. High voltage dc-dc converter with dynamic voltage regulation and decoupling during load-generated arcs

    DOEpatents

    Shimer, Daniel W.; Lange, Arnold C.

    1995-01-01

    A high-power power supply produces a controllable, constant high voltage output under varying and arcing loads. The power supply includes a voltage regulator, an inductor, an inverter for producing a high frequency square wave current of alternating polarity, an improved inverter voltage clamping circuit, a step up transformer, an output rectifier for producing a dc voltage at the output of each module, and a current sensor for sensing output current. The power supply also provides dynamic response to varying loads by controlling the voltage regulator duty cycle and circuitry is provided for sensing incipient arc currents at the output of the power supply to simultaneously decouple the power supply circuitry from the arcing load. The power supply includes a plurality of discrete switching type dc--dc converter modules.

  10. High voltage dc--dc converter with dynamic voltage regulation and decoupling during load-generated arcs

    DOEpatents

    Shimer, D.W.; Lange, A.C.

    1995-05-23

    A high-power power supply produces a controllable, constant high voltage output under varying and arcing loads. The power supply includes a voltage regulator, an inductor, an inverter for producing a high frequency square wave current of alternating polarity, an improved inverter voltage clamping circuit, a step up transformer, an output rectifier for producing a dc voltage at the output of each module, and a current sensor for sensing output current. The power supply also provides dynamic response to varying loads by controlling the voltage regulator duty cycle and circuitry is provided for sensing incipient arc currents at the output of the power supply to simultaneously decouple the power supply circuitry from the arcing load. The power supply includes a plurality of discrete switching type dc--dc converter modules. 5 Figs.

  11. Measurement and analysis of postsynaptic potentials using a novel voltage-deconvolution method.

    PubMed

    Richardson, Magnus J E; Silberberg, Gilad

    2008-02-01

    Accurate measurement of postsynaptic potential amplitudes is a central requirement for the quantification of synaptic strength, dynamics of short-term and long-term plasticity, and vesicle-release statistics. However, the intracellular voltage is a filtered version of the underlying synaptic signal and so a method of accounting for the distortion caused by overlapping postsynaptic potentials must be used. Here a voltage-deconvolution technique is demonstrated that defilters the entire voltage trace to reveal an underlying signal of well-separated synaptic events. These isolated events can be cropped out and reconvolved to yield a set of isolated postsynaptic potentials from which voltage amplitudes may be measured directly-greatly simplifying this common task. The method also has the significant advantage of providing a higher temporal resolution of the dynamics of the underlying synaptic signal. The versatility of the method is demonstrated by a variety of experimental examples, including excitatory and inhibitory connections to neurons with passive membranes and those with activated voltage-gated currents. The deconvolved current-clamp voltage has many features in common with voltage-clamp current measurements. These similarities are analyzed using cable theory and a multicompartment cell reconstruction, as well as direct comparison to voltage-clamp experiments.

  12. The sliding-helix voltage sensor

    PubMed Central

    Peyser, Alexander; Nonner, Wolfgang

    2012-01-01

    The voltage sensor (VS) domain of voltage-gated ion channels underlies electrical excitability of living cells. We simulate a mesoscale model of the VS domain to determine the functional consequences of some of its physical elements. Our mesoscale model is based on VS charges, linear dielectrics and whole-body motion, applied to an S4 ‘sliding helix’. The electrostatics under voltage-clamped boundary conditions are solved consistently using a boundary element method. Based on electrostatic configurational energy, statistical-mechanical expectations of the experimentally observable relation between displaced charge and membrane voltage are predicted. Consequences of the model are investigated for variations of: S4 configuration (α- and 310-helical), countercharge alignment with S4 charges, protein polarizability, geometry of the gating canal, screening of S4 charges by the baths, and fixed charges located at the bath interfaces. The sliding helix VS domain has an inherent electrostatic stability in the explored parameter space: countercharges present in the region of weak dielectric always retain an equivalent S4 charge in that region but allow sliding movements displacing 3 to 4 e0. That movement is sensitive to small energy variations (< 2kT) along the path dependent on a number of electrostatic parameters tested in our simulations. These simulations show how the slope of the relation between displaced charge and voltage could be tuned in a channel. PMID:22907204

  13. Umbilical cord clamping. An analysis of a usual neonatological conduct.

    PubMed

    Papagno, L

    1998-01-01

    Here we described a critical analysis of the neonatological procedure of early cord clamping, meaning this, within 40 seconds after birth. Fifty three cases are here analysed, in which this practice was not performed, but instead a late umbilical cord clamping was done after birth or after the cord had stopped beating. Variations in hematocrito values within 24 to 36 hours after birth were studied. A transitory polycithemia, with a maximum peak 12 hours post-delivery was observed. These values returned to normal levels between 24 and 36 hours after birth. K vitamin was not administered to any of the newborns. No pathology appeared related to this transitory polycithemia. In can be concluded that the late umbilical cord clamping represents no risk to the new-born and that the pathological phenomena described under these circumstances may be attributed to the increase in K vitamin dependent coagulation factors that are induced by the routinary administration of phitonadione to all normal newborns.

  14. [Advantage of delayed umbilical cord clamping in the newborn infant].

    PubMed

    Menget, A; Mougey, C; Thiriez, G; Riethmuller, D

    2013-09-01

    The timing of umbilical cord clamping remains controversial. Although most maternity wards use the early clamping (5-15s), randomized studies and meta-analyses have demonstrated the benefit of delayed clamping for term and preterm newborn infants over the past 10 years. Indeed, placentofetal transfusion of 20-30 ml/kg in 2-3 min improves the iron status of term infants and prevents infant hypochromic anemia. Infant anemia is a public health problem in many developing countries. For preterm newborns, placental transfusion for 45 s or milking the cord for 15 s improves cardiovascular adaptation, with better hemodynamic stability, as well as decreased intraventricular hemorrhages, need for transfusion, and late-onset sepsis. A new look at this symbolic act is needed and professionals need to be persuaded of the importance of the "wait a minute" policy for a better physiological delivery.

  15. A self-strain feedback tuning-fork-shaped ionic polymer metal composite clamping actuator with soft matter elasticity-detecting capability for biomedical applications.

    PubMed

    Feng, Guo-Hua; Huang, Wei-Lun

    2014-12-01

    This paper presents a smart tuning-fork-shaped ionic polymer metal composite (IPMC) clamping actuator for biomedical applications. The two fingers of the actuator, which perform the clamping motion, can be electrically controlled through a unique electrode design on the IPMC material. The generated displacement or strain of the fingers can be sensed using an integrated soft strain-gage sensor. The IPMC actuator and associated soft strain gage were fabricated using a micromachining technique. A 13.5×4×2 mm(3) actuator was shaped from Nafion solution and a selectively grown metal electrode formed the active region. The strain gage consisted of patterned copper foil and polyethylene as a substrate. The relationship between the strain gage voltage output and the displacement at the front end of the actuator's fingers was characterized. The equivalent Young's modulus, 13.65 MPa, of the soft-strain-gage-integrated IPMC finger was analyzed. The produced clamping force exhibited a linear increasing rate of 1.07 mN/s, based on a dc driving voltage of 7 V. Using the developed actuator to clamp soft matter and simultaneously acquire its Young's modulus was achieved. This demonstrated the feasibility of the palpation function and the potential use of the actuator in minimally invasive surgery.

  16. Clamping compared to cauterization for subcutaneous hemostasis in Pfannenstiel incision.

    PubMed

    Ozkaya, Enis; Korkmaz, Vakkas; Kucukozkan, Tuncay

    2011-04-01

    We compared subcutaneous clamping and cauterization for hemostasis at laparotomy with Pfannenstiel incision with reference to surgical site infection, postoperative fever and time taken for incision. A total of 214 patients with consecutive hysterectomies were alternately assigned to incisional hemostasis by clamping (n= 107) or cauterization (n= 107). The groups were similar in terms of age, gravidity, parity, body mass index, uterine size and mean hemoglobin drop. Rates of surgical site infection, postoperative fever and time from skin incision to peritoneal cavity entry were significantly higher in the group with cauterization (p < 0.05).

  17. Plasma temperature clamping in filamentation laser induced breakdown spectroscopy

    SciTech Connect

    Harilal, Sivanandan S.; Yeak, J.; Phillips, Mark C.

    2015-10-19

    Ultrafast laser filament induced breakdown spectroscopy is a very promising method for remote material detection. We present characteristics of plasmas generated in a metal target by laser filaments in air. Our measurements show that the temperature of the ablation plasma is clamped along the filamentation channel due to intensity clamping in a filament. Nevertheless, significant changes in radiation intensity are noticeable, and this is essentially due to variation in the number density of emitting atoms. The present results also partly explains the reason for the occurrence of atomic plume during fs LIBS in air compared to long-pulse ns LIBS.

  18. Whole-Cell Patch-Clamp Recording of Mouse and Rat Inner Hair Cells in the Intact Organ of Corti.

    PubMed

    Goutman, Juan D; Pyott, Sonja J

    2016-01-01

    Whole-cell patch clamping is a widely applied method to record currents across the entire membrane of a cell. This protocol describes application of this method to record currents from the sensory inner hair cells in the intact auditory sensory epithelium, the organ of Corti, isolated from rats or mice. This protocol particularly outlines the basic equipment required, provides instructions for the preparation of solutions and small equipment items, and methodology for recording voltage-activated and evoked synaptic currents from the inner hair cells.

  19. Perforated patch-clamp analysis of the passive membrane properties of three classes of hippocampal neurons.

    PubMed

    Spruston, N; Johnston, D

    1992-03-01

    1. Perforated patch-clamp recordings were made from the three major classes of hippocampal neurons in conventional in vitro slices prepared from adult guinea pigs. This technique provided experimental estimates of passive membrane properties (input resistance, RN, and membrane time constant, tau m) determined in the absence of the leak conductance associated with microelectrode impalement or the washout of cytoplasmic constituents associated with conventional whole-cell recordings. 2. To facilitate comparison of our data with previous results and to determine the passive membrane properties under conditions as physiological as possible, recordings were made at the resting potential, in physiological saline, and without any added blockers of voltage-dependent conductances. 3. Membrane-potential responses to current steps were analyzed, and four criteria were used to identify voltage responses that were the least affected by activation of voltage-dependent conductances. tau m was estimated from the slowest component (tau 0) of multiexponential fits of responses deemed passive by these criteria. RN was estimated from the slope of the linear region in the hyperpolarizing direction of the voltage-current relation. 4. It was not possible to measure purely passive membrane properties that were completely independent of membrane potential in any of the three classes of hippocampal neurons. Changing the membrane potential by constant current injection resulted in changes in RN and tau 0; subthreshold depolarization produced an increase, and hyperpolarization a decrease, in both RN and tau 0 for all three classes of hippocampal neurons. 5. Each of the three classes of hippocampal neurons also displayed a depolarizing "sag" during larger hyperpolarizing voltage transients. To evaluate the effect of the conductances underlying this sag on passive membrane properties, 2-5 mM Cs+ was added to the physiological saline. Extracellular Cs+ effectively blocked the sag in all three

  20. 21 CFR 882.5175 - Carotid artery clamp.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Carotid artery clamp. 882.5175 Section 882.5175 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... (the principal artery in the neck that supplies blood to the brain) and has a removable...

  1. 21 CFR 882.5175 - Carotid artery clamp.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Carotid artery clamp. 882.5175 Section 882.5175 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... (the principal artery in the neck that supplies blood to the brain) and has a removable...

  2. 21 CFR 882.5175 - Carotid artery clamp.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Carotid artery clamp. 882.5175 Section 882.5175 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... (the principal artery in the neck that supplies blood to the brain) and has a removable...

  3. 21 CFR 882.5175 - Carotid artery clamp.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Carotid artery clamp. 882.5175 Section 882.5175 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... (the principal artery in the neck that supplies blood to the brain) and has a removable...

  4. 21 CFR 882.5175 - Carotid artery clamp.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Carotid artery clamp. 882.5175 Section 882.5175 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... (the principal artery in the neck that supplies blood to the brain) and has a removable...

  5. A band clamp with a spring toggle lever

    NASA Technical Reports Server (NTRS)

    Simmonds, M.

    1974-01-01

    Clamp could have several applications, as it provides tolerance for both expansion and contraction. It might be useful with firemen's breathing apparatus and luggage racks and other freight-carrying equipment. Also, using same piece as handle and spring reduces production costs by reducing number of parts.

  6. 30 CFR 18.40 - Cable clamps and grips.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 30 Mineral Resources 1 2011-07-01 2011-07-01 false Cable clamps and grips. 18.40 Section 18.40 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR TESTING, EVALUATION, AND APPROVAL OF MINING PRODUCTS ELECTRIC MOTOR-DRIVEN MINE EQUIPMENT AND ACCESSORIES Construction and...

  7. 30 CFR 18.40 - Cable clamps and grips.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Cable clamps and grips. 18.40 Section 18.40 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR TESTING, EVALUATION, AND APPROVAL OF MINING PRODUCTS ELECTRIC MOTOR-DRIVEN MINE EQUIPMENT AND ACCESSORIES Construction and...

  8. Membrane-pipette interactions underlie delayed voltage activation of mechanosensitive channels in Xenopus oocytes.

    PubMed Central

    Gil, Z; Magleby, K L; Silberberg, S D

    1999-01-01

    To investigate the mechanism for the delayed activation by voltage of the predominant mechanosensitive (MS) channel in Xenopus oocytes, currents were recorded from on-cell and excised patches of membrane with the patch clamp technique and from intact oocytes with the two-electrode voltage clamp technique. MS channels could be activated by stretch in inside-out, on-cell, and outside-out patch configurations, using pipettes formed of either borosilicate or soft glass. In inside-out patches formed with borosilicate glass pipettes, depolarizing voltage steps activated MS channels in a cooperative manner after delays of seconds. This voltage-dependent activation was not observed for outside-out patches. Voltage-dependent activation was also not observed when the borosilicate pipettes were either replaced with soft glass pipettes or coated with soft glass. When depolarizing voltage steps were applied to the whole oocyte with a two-electrode voltage clamp, currents that could be attributed to MS channels were not observed. Yet the same depolarizing steps activated MS channels in on-cell patches formed with borosilicate pipettes on the same oocyte. These observations suggest that the delayed cooperative activation of MS channels by depolarization is not an intrinsic property of the channels, but requires interaction between the membrane and patch pipette. PMID:10354436

  9. High-Voltage Pulse Voltage Generator,

    DTIC Science & Technology

    1979-12-21

    the invention: I. I. Kalyatskiy, V. I. Kurets, and V. I. Safronov Well-known are pulse voltage generators which employ the Arkad’yev- Marx principle of...P2, and hereafter the device operates like an ordinary GIN [pulse volt- age generator] according to the Arkad’yev- Marx principle. The Object of the...Invention The high-voltage pulse voltage generator, assembled according to the Arkad’yev- Marx arrangement, each stage of which incorporates reactive

  10. Polarization states and dielectric responses of elastically clamped ferroelectric nanocrystals

    NASA Astrophysics Data System (ADS)

    Azovtsev, A. V.; Pertsev, N. A.

    2016-12-01

    Polarization states and physical properties of ferroelectrics depend on the mechanical boundary conditions due to electrostrictive coupling between electric polarization and lattice strains. Here, we describe theoretically both equilibrium thermodynamic states and electric permittivities of ferroelectric nanocrystals subjected to the elastic three-dimensional (3D) clamping by a surrounding dielectric material. The problem is solved by the minimization of a special thermodynamic potential that describes the case of an ellipsoidal ferroelectric inclusion embedded into a linear elastic matrix. Numerical calculations are performed for BaTiO3, PbTiO3, and Pb(Zr0.5Ti0.5)O3 nanoparticles surrounded by silica glass. It is shown that, in the case of BaTiO3 and PbTiO3, elastic 3D clamping may change the order of a ferroelectric phase transition from first to second. Furthermore, the mechanical inclusion-matrix interaction shifts the temperatures of structural transitions between different ferroelectric states and even eliminates some ferroelectric phases existing in stress-free BaTiO3 and Pb(Zr0.5Ti0.5)O3 crystals. Another important effect of elastic clamping is the lowering of the symmetry of ferroelectric states in ellipsoidal inclusions, where orthorhombic and monoclinic phases may form instead of the tetragonal and rhombohedral bulk counterparts. Finally, our thermodynamic calculations show that the dielectric responses of studied perovskite ferroelectrics are sensitive to matrix-induced clamping as well. For instance, dielectric peaks occurring at structural transitions between different ferroelectric phases in BaTiO3 appear to be much higher in spherical inclusions than in the freestanding crystal. Predicted clamping-induced enhancement of certain dielectric responses at room temperature indicates that composite materials comprising nanocrystals of perovskite ferroelectrics are promising for device applications requiring the use of high-permittivity dielectrics.

  11. Clamped-filament elongation model for actin-based motors.

    PubMed Central

    Dickinson, Richard B; Purich, Daniel L

    2002-01-01

    Although actin-based motility drives cell crawling and intracellular locomotion of organelles and certain pathogens, the underlying mechanism of force generation remains a mystery. Recent experiments demonstrated that Listeria exhibit episodes of 5.4-nm stepwise motion corresponding to the periodicity of the actin filament subunits, and extremely small positional fluctuations during the intermittent pauses [S. C. Kuo and J. L. McGrath. 2000. Nature. 407:1026-1029]. These findings suggest that motile bacteria remain firmly bound to actin filament ends as they elongate, a behavior that appears to rule out previous models for actin-based motility. We propose and analyze a new mechanochemical model (called the "Lock, Load & Fire" mechanism) for force generation by means of affinity-modulated, clamped-filament elongation. During the locking step, the filament's terminal ATP-containing subunit binds tightly to a clamp situated on the surface of a motile object; in the loading step, actin.ATP monomer(s) bind to the filament end, an event that triggers the firing step, wherein ATP hydrolysis on the clamped subunit attenuates the filament's affinity for the clamp. This last step initiates translocation of the new ATP-containing terminus to the clamp, whereupon another cycle begins anew. This model explains how surface-tethered filaments can grow while exerting flexural or tensile force on the motile surface. Moreover, stochastic simulations of the model reproduce the signature motions of Listeria. This elongation motor, which we term actoclampin, exploits actin's intrinsic ATPase activity to provide a simple, high-fidelity enzymatic reaction cycle for force production that does not require elongating filaments to dissociate from the motile surface. This mechanism may operate whenever actin polymerization is called upon to generate the forces that drive cell crawling or intracellular organelle motility. PMID:11806905

  12. Voltage-dependent motion of the catalytic region of voltage-sensing phosphatase monitored by a fluorescent amino acid

    PubMed Central

    Sakata, Souhei; Jinno, Yuka; Kawanabe, Akira; Okamura, Yasushi

    2016-01-01

    The cytoplasmic region of voltage-sensing phosphatase (VSP) derives the voltage dependence of its catalytic activity from coupling to a voltage sensor homologous to that of voltage-gated ion channels. To assess the conformational changes in the cytoplasmic region upon activation of the voltage sensor, we genetically incorporated a fluorescent unnatural amino acid, 3-(6-acetylnaphthalen-2-ylamino)-2-aminopropanoic acid (Anap), into the catalytic region of Ciona intestinalis VSP (Ci-VSP). Measurements of Anap fluorescence under voltage clamp in Xenopus oocytes revealed that the catalytic region assumes distinct conformations dependent on the degree of voltage-sensor activation. FRET analysis showed that the catalytic region remains situated beneath the plasma membrane, irrespective of the voltage level. Moreover, Anap fluorescence from a membrane-facing loop in the C2 domain showed a pattern reflecting substrate turnover. These results indicate that the voltage sensor regulates Ci-VSP catalytic activity by causing conformational changes in the entire catalytic region, without changing their distance from the plasma membrane. PMID:27330112

  13. Fabrication and characterization of PbO2 electrode modified with [Fe(CN)6](3-) and its application on electrochemical degradation of alkali lignin.

    PubMed

    Hao, Xu; Quansheng, Yuan; Dan, Shao; Honghui, Yang; Jidong, Liang; Jiangtao, Feng; Wei, Yan

    2015-04-09

    PbO2 electrode modified by [Fe(CN)6](3-) (marked as FeCN-PbO2) was prepared by electro-deposition method and used for the electrochemical degradation of alkali lignin (AL). The surface morphology and the structure of the electrodes were characterized by scanning electronic microscopy (SEM) and X-ray diffraction (XRD), respectively. The stability and electrochemical activity of FeCN-PbO2 electrode were characterized by accelerated life test, linear sweep voltammetry, electrochemical impedance spectrum (EIS) and AL degradation. The results showed that [Fe(CN)6](3-) increased the average grain size of PbO2 and formed a compact surface coating. The service lifetime of FeCN-PbO2 electrode was 287.25 h, which was longer than that of the unmodified PbO2 electrode (100.5h). The FeCN-PbO2 electrode showed higher active surface area and higher oxygen evolution potential than that of the unmodified PbO2 electrode. In electrochemical degradation tests, the apparent kinetics coefficient of FeCN-PbO2 electrode was 0.00609 min(-1), which was higher than that of unmodified PbO2 electrode (0.00419 min(-1)). The effects of experimental parameters, such as applied current density, initial AL concentration, initial pH value and solution temperature, on electrochemical degradation of AL by FeCN-PbO2 electrode were evaluated.

  14. Studies on the physicochemical properties and performance characteristics of the PbO 2 electrode

    NASA Astrophysics Data System (ADS)

    Böttcher, F.; Panesar, H. S.

    At least 10 different parameters have been determined in situ as well as ex situ and statistically analyzed to study changes in the physicochemical properties of PbO 2 electrodes during the cycle life of traction batteries. The specific surface area, water content, microporosity and delta capacity are significant and strongly correlated. These variables, together with the high-frequency resistance, monitor the state of the positive active material as a function of the charge/discharge cycle-life. Insignificant variations in the resistance/capacitance and the Warburg term during the cycle life suggest that the changes in the properties of positive active material are not associated with the deviation in electrochemical activity. A negative correlation between the water content and specific surface area supports the assumption that the water originates from hydrated crystals, hydroxyl groups or protons present in PbO 2. A thermal decomposition mechanism has been proposed that suggests that the number of vacancies in the crystal lattice decreases with cycling and finally the positive active material attains an ideal structure. Furthermore, the area of intergrowth of crystals or necks (as proposed in the Kugelhaufen model) seems to exert a significant cause of the changes in the properties of PbO 2 during the cycle life.

  15. Patch clamp recordings of retinal bipolar cells in response to extracellular electrical stimulation in wholemount mouse retina.

    PubMed

    Walston, Steven T; Chow, Robert H; Weiland, James D

    2015-01-01

    Retinitis pigmentosa is a family of inherited retinal diseases identified by the degeneration of photoreceptors, which leads to blindness. In efforts to restore vision lost to retinitis pigmentosa, retinal prostheses have been developed to generate visual percepts by electrically stimulating the surviving retinal bipolar and ganglion cells. The response of retinal ganglion cells to electrical stimulation has been characterized through direct measurement. However, the response of bipolar cells has only been inferred by measuring retinal ganglion cell activity. This investigation reports on a novel tissue preparation technique facilitating bipolar cell patch clamp recordings in wholemount retina. We find that bipolar cells respond to extracellular electrical stimuli with time-locked voltage spike depolarizations, which are likely mediated by voltage-gated calcium channels.

  16. A patch-clamp ASIC for nanopore-based DNA analysis.

    PubMed

    Kim, Jungsuk; Maitra, Raj; Pedrotti, Kenneth D; Dunbar, William B

    2013-06-01

    In this paper, a fully integrated high-sensitivity patch-clamp system is proposed for single-molecule deoxyribonucleic acid (DNA) analysis using a nanopore sensor. This system is composed of two main blocks for amplification and compensation. The amplification block is composed of three stages: 1) a headstage, 2) a voltage-gain difference amplifier, and 3) a track-and-hold circuit, that amplify a minute ionic current variation sensed by the nanopore while the compensation block avoids the headstage saturation caused by the input parasitic capacitances during sensing. By employing design techniques novel for this application, such as an instrumentation--amplifier topology and a compensation switch, we minimize the deleterious effects of the input-offset voltage and the input parasitic capacitances while attaining hardware simplicity. This system is fabricated in a 0.35 μm 4M2P CMOS process and is demonstrated using an α-hemolysin protein nanopore for detection of individual molecules of single-stranded DNA that pass through the 1.5 nm-diameter pore. In future work, the refined system will functionalize single and multiple solid-state nanopores formed in integrated microfluidic devices for advanced DNA analysis, in scientific and diagnostic applications.

  17. High voltage bus and auxiliary heater control system for an electric or hybrid vehicle

    DOEpatents

    Murty, Balarama Vempaty

    2000-01-01

    A control system for an electric or hybrid electric vehicle includes a vehicle system controller and a control circuit having an electric immersion heater. The heater is electrically connected to the vehicle's high voltage bus and is thermally coupled to a coolant loop containing a heater core for the vehicle's climate control system. The system controller responds to cabin heat requests from the climate control system by generating a pulse width modulated signal that is used by the control circuit to operate the heater at a duty cycle appropriate for the amount of cabin heating requested. The control system also uses the heater to dissipate excess energy produced by an auxiliary power unit and to provide electric braking when regenerative braking is not desirable and manual braking is not necessary. The control system further utilizes the heater to provide a safe discharge of a bank of energy storage capacitors following disconnection of the battery or one of the high voltage connectors used to transmit high voltage operating power to the various vehicle systems. The control circuit includes a high voltage clamping circuit that monitors the voltage on the bus and operates the heater to clamp down the bus voltage when it exceeds a pre-selected maximum voltage. The control system can also be used to phase in operation of the heater when the bus voltage exceeds a lower threshold voltage and can be used to phase out the auxiliary power unit charging and regenerative braking when the battery becomes fully charged.

  18. High voltage bus and auxiliary heater control system for an electric or hybrid vehicle

    SciTech Connect

    Murty, B.V.

    2000-03-21

    A control system for an electric or hybrid electric vehicle includes a vehicle system controller and a control circuit having an electric immersion heater. The heater is electrically connected to the vehicle's high voltage bus and is thermally coupled to a coolant loop containing a heater core for the vehicle's climate control system. The system controller responds to cabin heat requests from the climate control system by generating a pulse width modulated signal that is used by the control circuit to operate the heater at a duty cycle appropriate for the amount of cabin heating requested. The control system also uses the heater to dissipate excess energy produced by an auxiliary power unit and to provide electric braking when regenerative braking is not desirable and manual braking is not necessary. The control system further utilizes the heater to provide a safe discharge of a bank of energy storage capacitors following disconnection of the battery or one of the high voltage connectors used to transmit high voltage operating power to the various vehicle systems. The control circuit includes a high voltage clamping circuit that monitors the voltage on the bus and operates the heater to clamp down the bus voltage when it exceeds a pre-selected maximum voltage. The control system can also be used to phase in operation of the heater when the bus voltage exceeds a lower threshold voltage and can be used to phase out the auxiliary power unit charging and regenerative braking when the battery becomes fully charged.

  19. Patch-clamp experiments under micro-gravity.

    PubMed

    Meissner, Klaus; Hanke, Wolfgang

    2002-07-01

    For human based space research it is of high importance to understand the influence of gravity on the properties of single ion channels in biological membranes, as these are involved in about all biological processes. The patch clamp technique is the best established method to investigate electrophysiological properties of single ion channels in detail. Consequently, a patch clamp set-up was designed for the drop tower in Bremen, Germany. Using this set-up among others, successfully leech neurons have been patched under micro-gravity, delivering data about ion channel behaviour, which were compared to results from bilayer experiments in the drop tower and to results from lab controls under 1 g and under higher gravity.

  20. An equivalent network representation of a clamped bimorph piezoelectric micromachined ultrasonic transducer with circular and annular electrodes using matrix manipulation techniques.

    PubMed

    Sammoura, Firas; Smyth, Katherine; Kim, Sang-Gook

    2013-09-01

    An electric circuit model for a clamped circular bimorph piezoelectric micromachined ultrasonic transducer (pMUT) was developed for the first time. The pMUT consisted of two piezoelectric layers sandwiched between three thin electrodes. The top and bottom electrodes were separated into central and annular electrodes by a small gap. While the middle electrode was grounded, the central and annular electrodes were biased with two independent voltage sources. The strain mismatch between the piezoelectric layers caused the plate to vibrate and transmit a pressure wave, whereas the received echo generated electric charges resulting from plate deformation. The clamped pMUT plate was separated into a circular and an annular plate, and the respective electromechanical transformation matrices were derived. The force and velocity vectors were properly selected using Hamilton's principle and the necessary boundary conditions were invoked. The electromechanical transformation matrix for the clamped circular pMUT was deduced using simple matrix manipulation techniques. The pMUT performance under three biasing schemes was elaborated: 1) central electrode only, 2) central and annular electrodes with voltages of the same magnitude and polarity, and 3) central and annular electrodes with voltages of the same magnitude and opposite polarity. The circuit parameters of the pMUT were extracted for each biasing scheme, including the transformer ratio, the clamped electric impedance, and the open-circuit mechanical impedance. Each pMUT scheme was characterized under different acoustic loadings using the theoretically developed model, which was verified with finite element modeling (FEM) simulation. The electrode size was optimized to maximize the electromechanical transformer ratio. As such, the developed model could provide more insight into the design, optimization, and characterization of pMUTs and allow for performance comparison with their cMUT counterparts.

  1. Batteries: Widening voltage windows

    NASA Astrophysics Data System (ADS)

    Xu, Kang; Wang, Chunsheng

    2016-10-01

    The energy output of aqueous batteries is largely limited by the narrow voltage window of their electrolytes. Now, a hydrate melt consisting of lithium salts is shown to expand such voltage windows, leading to a high-energy aqueous battery.

  2. Characterization of ryanodine receptor type 1 single channel activity using "on-nucleus" patch clamp.

    PubMed

    Wagner, Larry E; Groom, Linda A; Dirksen, Robert T; Yule, David I

    2014-08-01

    In this study, we provide the first description of the biophysical and pharmacological properties of ryanodine receptor type 1 (RyR1) expressed in a native membrane using the on-nucleus configuration of the patch clamp technique. A stable cell line expressing rabbit RyR1 was established (HEK-RyR1) using the FLP-in 293 cell system. In contrast to untransfected cells, RyR1 expression was readily demonstrated by immunoblotting and immunocytochemistry in HEK-RyR1 cells. In addition, the RyR1 agonists 4-CMC and caffeine activated Ca(2+) release that was inhibited by high concentrations of ryanodine. On nucleus patch clamp was performed in nuclei prepared from HEK-RyR1 cells. Raising the [Ca(2+)] in the patch pipette resulted in the appearance of a large conductance cation channel with well resolved kinetics and the absence of prominent subconductance states. Current versus voltage relationships were ohmic and revealed a chord conductance of ∼750pS or 450pS in symmetrical 250mM KCl or CsCl, respectively. The channel activity was markedly enhanced by caffeine and exposure to ryanodine resulted in the appearance of a subconductance state with a conductance ∼40% of the full channel opening with a Po near unity. In total, these properties are entirely consistent with RyR1 channel activity. Exposure of RyR1 channels to cyclic ADP ribose (cADPr), nicotinic acid adenine dinucleotide phosphate (NAADP) or dantrolene did not alter the single channel activity stimulated by Ca(2+), and thus, it is unlikely these molecules directly modulate RyR1 channel activity. In summary, we describe an experimental platform to monitor the single channel properties of RyR channels. We envision that this system will be influential in characterizing disease-associated RyR mutations and the molecular determinants of RyR channel modulation.

  3. Phosphatase activity of the voltage-sensing phosphatase, VSP, shows graded dependence on the extent of activation of the voltage sensor.

    PubMed

    Sakata, Souhei; Okamura, Yasushi

    2014-03-01

    The voltage-sensing phosphatase (VSP) consists of a voltage sensor and a cytoplasmic phosphatase region, and the movement of the voltage sensor is coupled to the phosphatase activity. However, its coupling mechanisms still remain unclear. One possible scenario is that the phosphatase is activated only when the voltage sensor is in a fully activated state. Alternatively, the enzymatic activity of single VSP proteins could be graded in distinct activated states of the voltage sensor, and partial activation of the voltage sensor could lead to partial activation of the phosphatase. To distinguish between these two possibilities, we studied a voltage sensor mutant of zebrafish VSP, where the voltage sensor moves in two steps as evidenced by analyses of charge movements of the voltage sensor and voltage clamp fluorometry. Measurements of the phosphatase activity toward phosphatidylinositol 4,5-bisphosphate revealed that both steps of voltage sensor activation are coupled to the tuning of phosphatase activities, consistent with the idea that the phosphatase activity is graded by the magnitude of the movement of the voltage sensor.

  4. Force-clamp laser trapping of rapidly interacting molecules

    NASA Astrophysics Data System (ADS)

    Capitanio, Marco; Monico, Carina; Vanzi, Francesco; Pavone, Francesco S.

    2013-06-01

    Forces play a fundamental role in a wide array of biological processes, regulating enzymatic activity, kinetics of molecular bonds, and molecular motors mechanics. Single molecule force spectroscopy techniques have enabled the investigation of such processes, but they are inadequate to probe short-lived (millisecond and sub-millisecond) molecular complexes. We developed an ultrafast force-clamp spectroscopy technique that uses a dual trap configuration to apply constant loads to a single intermittently interacting biological polymer and a binding protein. Our system displays a delay of only ˜10 μs between formation of the molecular bond and application of the force and is capable of detecting interactions as short as 100 μs. The force-clamp configuration in which our assay operates allows direct measurements of load-dependence of lifetimes of single molecular bonds. Moreover, conformational changes of single proteins and molecular motors can be recorded with sub-nanometer accuracy and few tens of microseconds of temporal resolution. We demonstrate our technique on molecular motors, using myosin II from fast skeletal muscle and on protein-DNA interaction, specifically on Lactose repressor (LacI). The apparatus is stabilized to less than 1 nm with both passive and active stabilization, allowing resolving specific binding regions along the actin filament and DNA molecule. Our technique extends single-molecule force-clamp spectroscopy to molecular complexes that have been inaccessible up to now, opening new perspectives for the investigation of the effects of forces on biological processes.

  5. A mathematical model of the euglycemic hyperinsulinemic clamp

    PubMed Central

    Picchini, Umberto; De Gaetano, Andrea; Panunzi, Simona; Ditlevsen, Susanne; Mingrone, Geltrude

    2005-01-01

    Background The Euglycemic Hyperinsulinemic Clamp (EHC) is the most widely used experimental procedure for the determination of insulin sensitivity, and in its usual form the patient is followed under insulinization for two hours. In the present study, sixteen subjects with BMI between 18.5 and 63.6 kg/m2 were studied by long-duration (five hours) EHC. Results From the results of this series and from similar reports in the literature it is clear that, in obese subjects, glucose uptake rates continue to increase if the clamp procedure is prolonged beyond the customary 2 hours. A mathematical model of the EHC, incorporating delays, was fitted to the recorded data, and the insulin resistance behaviour of obese subjects was assessed analytically. Obese subjects had significantly less effective suppression of hepatic glucose output and higher pancreatic insulin secretion than lean subjects. Tissue insulin resistance appeared to be higher in the obese group, but this difference did not reach statistical significance. Conclusion The use of a mathematical model allows a greater amount of information to be recovered from clamp data, making it easier to understand the components of insulin resistance in obese vs. normal subjects. PMID:16269082

  6. Cleaning patch-clamp pipettes for immediate reuse

    PubMed Central

    Kolb, I.; Stoy, W. A.; Rousseau, E. B.; Moody, O. A.; Jenkins, A.; Forest, C. R.

    2016-01-01

    Patch-clamp recording has enabled single-cell electrical, morphological and genetic studies at unparalleled resolution. Yet it remains a laborious and low-throughput technique, making it largely impractical for large-scale measurements such as cell type and connectivity characterization of neurons in the brain. Specifically, the technique is critically limited by the ubiquitous practice of manually replacing patch-clamp pipettes after each recording. To circumvent this limitation, we developed a simple, fast, and automated method for cleaning glass pipette electrodes that enables their reuse within one minute. By immersing pipette tips into Alconox, a commercially-available detergent, followed by rinsing, we were able to reuse pipettes 10 times with no degradation in signal fidelity, in experimental preparations ranging from human embryonic kidney cells to neurons in culture, slices, and in vivo. Undetectable trace amounts of Alconox remaining in the pipette after cleaning did not affect ion channel pharmacology. We demonstrate the utility of pipette cleaning by developing the first robot to perform sequential patch-clamp recordings in cell culture and in vivo without a human operator. PMID:27725751

  7. Cleaning patch-clamp pipettes for immediate reuse.

    PubMed

    Kolb, I; Stoy, W A; Rousseau, E B; Moody, O A; Jenkins, A; Forest, C R

    2016-10-11

    Patch-clamp recording has enabled single-cell electrical, morphological and genetic studies at unparalleled resolution. Yet it remains a laborious and low-throughput technique, making it largely impractical for large-scale measurements such as cell type and connectivity characterization of neurons in the brain. Specifically, the technique is critically limited by the ubiquitous practice of manually replacing patch-clamp pipettes after each recording. To circumvent this limitation, we developed a simple, fast, and automated method for cleaning glass pipette electrodes that enables their reuse within one minute. By immersing pipette tips into Alconox, a commercially-available detergent, followed by rinsing, we were able to reuse pipettes 10 times with no degradation in signal fidelity, in experimental preparations ranging from human embryonic kidney cells to neurons in culture, slices, and in vivo. Undetectable trace amounts of Alconox remaining in the pipette after cleaning did not affect ion channel pharmacology. We demonstrate the utility of pipette cleaning by developing the first robot to perform sequential patch-clamp recordings in cell culture and in vivo without a human operator.

  8. Automatic voltage imbalance detector

    DOEpatents

    Bobbett, Ronald E.; McCormick, J. Byron; Kerwin, William J.

    1984-01-01

    A device for indicating and preventing damage to voltage cells such as galvanic cells and fuel cells connected in series by detecting sequential voltages and comparing these voltages to adjacent voltage cells. The device is implemented by using operational amplifiers and switching circuitry is provided by transistors. The device can be utilized in battery powered electric vehicles to prevent galvanic cell damage and also in series connected fuel cells to prevent fuel cell damage.

  9. DNA Sliding Clamps: Just the Right Twist to Load onto DNA

    SciTech Connect

    Barsky, D; Venclovas, C

    2005-10-24

    Two recent papers illuminate a long sought step in DNA sliding clamp loading. One paper reveals the structure of the PCNA clamp wrapped around DNA--still open from being loaded--while a second paper discovers that the clamp may assist this process by forming a right-handed helix upon opening.

  10. Patch-clamp analysis in canine cardiac Purkinje cells of a novel sodium component in the pacemaker range.

    PubMed

    Rota, Marcello; Vassalle, Mario

    2003-04-01

    A putative Na+ component playing a role in the initiation and maintenance of spontaneous discharge in Purkinje fibres was studied by means of the whole-cell patch-clamp technique in canine cardiac single Purkinje cells. In 4 mM [K+]o, during depolarising clamp steps, a slowly inactivating current appeared at approximately -58 mV, negative to the threshold for the fast Na+ current (INa; approximately -50 mV). During depolarising ramps, the current underwent inward rectification with a negative slope region that began at approximately -60 mV. The current underlying the negative slope increased during faster ramps, decreased as a function of time when the initial depolarising ramp was over, decreased during depolarisations positive to approximately -35 mV and was much larger than the current during the symmetrical repolarising ramp. Increasing biphasic ('oscillatory') voltage ramps required much smaller currents at a holding potential (Vh) of -60 mV than at -80 mV and were associated with a marked decrease in slope conductance. At Vh -50/-40 mV, the oscillatory ramp currents and superimposed pulse currents reversed direction. The negative slope in the I-V relation as well as the change in current direction at -50/-40 mV were markedly reduced by tetrodotoxin (15 microM) and lidocaine (lignocaine, 100 microM) and therefore are due to a slowly inactivating Na+ current, labelled here INa3. Lower [K+]o (2.7 mM) reduced the steady state slope conductance as well as the current in the diastolic range, and increased as well as shifted INa3 in a negative direction. High [K+]o had the opposite effects. Cs+ (2 mM) and Ba2+ (2 mM) reduced the initial current during depolarising ramps but not INa3. In current-clamp mode, current-induced voltage oscillations elicited action potentials through a gradual transition between diastolic depolarisation and upstroke, consistent with the activation of INa3. Thus, the initiation and maintenance of spontaneous discharge in Purkinje strands

  11. Mixed voltage VLSI design

    NASA Technical Reports Server (NTRS)

    Panwar, Ramesh; Rennels, David; Alkalaj, Leon

    1993-01-01

    A technique for minimizing the power dissipated in a Very Large Scale Integration (VLSI) chip by lowering the operating voltage without any significant penalty in the chip throughput even though low voltage operation results in slower circuits. Since the overall throughput of a VLSI chip depends on the speed of the critical path(s) in the chip, it may be possible to sustain the throughput rates attained at higher voltages by operating the circuits in the critical path(s) with a high voltage while operating the other circuits with a lower voltage to minimize the power dissipation. The interface between the gates which operate at different voltages is crucial for low power dissipation since the interface may possibly have high static current dissipation thus negating the gains of the low voltage operation. The design of a voltage level translator which does the interface between the low voltage and high voltage circuits without any significant static dissipation is presented. Then, the results of the mixed voltage design using a greedy algorithm on three chips for various operating voltages are presented.

  12. Planar silicon patch-clamp electrodes integrated with polydimethylsiloxane microfluidics

    NASA Astrophysics Data System (ADS)

    Nagarah, John Michael

    The patch-clamp technique allows one to probe single ion channels and macroscopic ion channel activity in their native environment and resolve their activity as their physical and chemical surroundings are varied. The traditional method of patch-clamping cells involves bringing a clean, flame-polished glass pipette tip with a 1-2 mum diameter pore into contact with a cell membrane to form a high electrical resistance seal. This technique is the gold standard for cellular electrophysiology investigations because it allows the observation of single ion channel protein dynamics as well as activity from an ensemble ion channels from a single cell. Furthermore, any drug approved by federal drug agencies must be screened against particular ion channels with the patch-clamp technique. However, this technique by its nature is serial, time consuming, difficult when exchanging pipette solutions, and difficult to integrate with other technologies. These reasons have prompted several investigators to explore alternative approaches to traditional pipette patch-clamping to increase the throughput of measurements. Herein, I describe the development of a silicon-wafer based device platform that enables the measurement of ion channel activities. The electrical nature of the cell/wafer seal is characterized for several pore design variations. The majority of gigaohm seals obtained falls in the range of 10-20GO. The cell-attached and whole cell configurations are demonstrated. Whole cell ion channel activity originating from various cell fines is consistent with the more traditional micropipette patch-clamp recordings. The silicon fabrication methods developed, although novel, utilize established semiconductor technologies, making them amenable to batch fabrication techniques. I integrate these silicon devices with PDMS microfluidics with monolithic valves, allowing ultra-fast solution exchange as low as tens of milliseconds for the extracellular solution. Furthermore, I developed a

  13. Metal interactions with voltage- and receptor-activated ion channels.

    PubMed Central

    Vijverberg, H P; Oortgiesen, M; Leinders, T; van Kleef, R G

    1994-01-01

    Effects of Pb and several other metal ions on various distinct types of voltage-, receptor- and Ca-activated ion channels have been investigated in cultured N1E-115 mouse neuroblastoma cells. Experiments were performed using the whole-cell voltage clamp and single-channel patch clamp techniques. External superfusion of nanomolar to submillimolar concentrations of Pb causes multiple effects on ion channels. Barium current through voltage-activated Ca channels is blocked by micromolar concentrations of Pb, whereas voltage-activated Na current appears insensitive. Neuronal type nicotinic acetylcholine receptor-activated ion current is blocked by nanomolar concentrations of Pb and this block is reversed at micromolar concentrations. Serotonin 5-HT3 receptor-activated ion current is much less sensitive to Pb. In addition, external superfusion with micromolar concentrations of Pb as well as of Cd and aluminum induces inward current, associated with the direct activation of nonselective cation channels by these metal ions. In excised inside-out membrane patches of neuroblastoma cells, micromolar concentrations of Ca activate small (SK) and big (BK) Ca-activated K channels. Internally applied Pb activates SK and BK channels more potently than Ca, whereas Cd is approximately equipotent to Pb with respect to SK channel activation, but fails to activate BK channels. The results show that metal ions cause distinct, selective effects on the various types of ion channels and that metal ion interaction sites of ion channels may be highly selective for particular metal ions. PMID:7531139

  14. High Voltage SPT Performance

    NASA Technical Reports Server (NTRS)

    Manzella, David; Jacobson, David; Jankovsky, Robert

    2001-01-01

    A 2.3 kW stationary plasma thruster designed to operate at high voltage was tested at discharge voltages between 300 and 1250 V. Discharge specific impulses between 1600 and 3700 sec were demonstrated with thrust between 40 and 145 mN. Test data indicated that discharge voltage can be optimized for maximum discharge efficiency. The optimum discharge voltage was between 500 and 700 V for the various anode mass flow rates considered. The effect of operating voltage on optimal magnet field strength was investigated. The effect of cathode flow rate on thruster efficiency was considered for an 800 V discharge.

  15. Ion channelopathies in human induced pluripotent stem cell derived cardiomyocytes: a dynamic clamp study with virtual IK1

    PubMed Central

    Meijer van Putten, Rosalie M. E.; Mengarelli, Isabella; Guan, Kaomei; Zegers, Jan G.; van Ginneken, Antoni C. G.; Verkerk, Arie O.; Wilders, Ronald

    2015-01-01

    Human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) are widely used in studying basic mechanisms of cardiac arrhythmias that are caused by ion channelopathies. Unfortunately, the action potential profile of hiPSC-CMs—and consequently the profile of individual membrane currents active during that action potential—differs substantially from that of native human cardiomyocytes, largely due to almost negligible expression of the inward rectifier potassium current (IK1). In the present study, we attempted to “normalize” the action potential profile of our hiPSC-CMs by inserting a voltage dependent in silico IK1 into our hiPSC-CMs, using the dynamic clamp configuration of the patch clamp technique. Recordings were made from single hiPSC-CMs, using the perforated patch clamp technique at physiological temperature. We assessed three different models of IK1, with different degrees of inward rectification, and systematically varied the magnitude of the inserted IK1. Also, we modified the inserted IK1 in order to assess the effects of loss- and gain-of-function mutations in the KCNJ2 gene, which encodes the Kir2.1 protein that is primarily responsible for the IK1 channel in human ventricle. For our experiments, we selected spontaneously beating hiPSC-CMs, with negligible IK1 as demonstrated in separate voltage clamp experiments, which were paced at 1 Hz. Upon addition of in silico IK1 with a peak outward density of 4–6 pA/pF, these hiPSC-CMs showed a ventricular-like action potential morphology with a stable resting membrane potential near −80 mV and a maximum upstroke velocity >150 V/s (n = 9). Proarrhythmic action potential changes were observed upon injection of both loss-of-function and gain-of-function IK1, as associated with Andersen–Tawil syndrome type 1 and short QT syndrome type 3, respectively (n = 6). We conclude that injection of in silico IK1 makes the hiPSC-CM a more reliable model for investigating mechanisms underlying cardiac

  16. pH- and voltage-dependent conductances in toad skin.

    PubMed

    Lacaz-Vieira, F

    1995-11-01

    The present study focuses on two closely related topics on ion conductance in toad skins: (i) the interaction of apical protons with the apical voltage-dependent Cl(-)-activated channels of the mitochondria-rich cells, and (ii) the description and characterization of a novel subject, a voltage-dependent H(+)-activated conductance. The Cl- conductance (GCl) is activated by tissue hyperpolarization (which leads to apical membrane depolarization) and the presence of Cl- ions in the apical solution. Increasing apical proton concentration (from pH 8 to pH 4) impairs the process of activation of the Cl- conductive pathway, slowing the kinetics of It activation and reducing the steady-stage values of Gt and It. This effect is markedly voltage-dependent since no effect is seen at Vt = -100 mv and is fully present at -50 mV. The voltage-dependence of the pH effect suggests that the critical protonation sites of the apical Cl- channels are not freely exposed to the apical solution but dwell within the membrane electric field. An also coherent interpretation is that titration of apical proton binding sites affects the gating of the voltage-dependent Cl- channels, shifting the conductance-vs.-voltage curve to more negative clamping potentials. Tissue conductance in the absence of apical Cl- ions can be importantly affected by the pH of the apical solution (pHa), the effect being markedly dependent on the clamping potential. Generally speaking, the effect of rising apical proton concentration can be conspicuous at negative clamping potentials, while at positive potentials changes in tissue conductance were never observed. For a clamping potential of -100 mV, a turning point somewhere between pHa = 4 and pHa = 3 was observed. Apical acidification to pH 4 has no effect upon tissue conductance while apical acidification to pH 3 leads to a marked, slow and reversible increase of tissue conductance. A striking similitude exists between the voltage-dependent Cl(-)-gated conductance

  17. Clamping stiffness and its influence on load distribution between paired internal spinal fixation devices.

    PubMed

    Rohlmann, A; Calisse, J; Bergmann, G; Radvan, J; Mayer, H M

    1996-06-01

    The load distribution between two internal spinal fixation devices depends, besides other factors, on their stiffness. The stiffness ranges were determined experimentally for the clamps of the AO internal fixator with lateral nut and with posterior nut as well as for the clamps of the SOCON fixator. The stiffness of eight devices each differed by a factor of 3.1 for the clamp with lateral nut, by a factor of 1.5 for the clamp with posterior nut, and by a factor of 1.4 for the clamp of the SOCON fixator. For the AO clamp with lateral nut, the influence of the nut-tightening torque on the stiffness was determined. Using instrumented internal spinal fixation devices mounted to plastic vertebrae and simulating a corpectomy, the load distribution between the implants was measured for different tightening torques. It could be shown that, for the AO internal fixator whose clamps have a lateral nut, a nut-tightening torque > 5 Nm has only a negligible influence on load-sharing between the implants. Tooth damage occurs when the teeth of the clamp body and clamping jaw of the clamp with lateral nut do not gear together exactly, which leads to changes in the clamping stiffness and load-sharing between the two implants.

  18. M. tuberculosis Sliding β-Clamp Does Not Interact Directly with the NAD+ -Dependent DNA Ligase

    PubMed Central

    Kukshal, Vandna; Khanam, Taran; Chopra, Deepti; Singh, Nidhi; Sanyal, Sabyasachi; Ramachandran, Ravishankar

    2012-01-01

    The sliding β-clamp, an important component of the DNA replication and repair machinery, is drawing increasing attention as a therapeutic target. We report the crystal structure of the M. tuberculosis β-clamp (Mtbβ-clamp) to 3.0 Å resolution. The protein crystallized in the space group C2221 with cell-dimensions a = 72.7, b = 234.9 & c = 125.1 Å respectively. Mtbβ-clamp is a dimer, and exhibits head-to-tail association similar to other bacterial clamps. Each monomer folds into three domains with similar structures respectively and associates with its dimeric partner through 6 salt-bridges and about 21 polar interactions. Affinity experiments involving a blunt DNA duplex, primed-DNA and nicked DNA respectively show that Mtbβ-clamp binds specifically to primed DNA about 1.8 times stronger compared to the other two substrates and with an apparent Kd of 300 nM. In bacteria like E. coli, the β-clamp is known to interact with subunits of the clamp loader, NAD+ -dependent DNA ligase (LigA) and other partners. We tested the interactions of the Mtbβ-clamp with MtbLigA and the γ-clamp loader subunit through radioactive gel shift assays, size exclusion chromatography, yeast-two hybrid experiments and also functionally. Intriguingly while Mtbβ-clamp interacts in vitro with the γ-clamp loader, it does not interact with MtbLigA unlike in bacteria like E. coli where it does. Modeling studies involving earlier peptide complexes reveal that the peptide-binding site is largely conserved despite lower sequence identity between bacterial clamps. Overall the results suggest that other as-yet-unidentified factors may mediate interactions between the clamp, LigA and DNA in mycobacteria. PMID:22545130

  19. Analysis of motion during the breast clamping phase of mammography

    PubMed Central

    McEntee, Mark F; Mercer, Claire; Kelly, Judith; Millington, Sara; Hogg, Peter

    2016-01-01

    Objective: To measure paddle motion during the clamping phase of a breast phantom for a range of machine/paddle combinations. Methods: A deformable breast phantom was used to simulate a female breast. 12 mammography machines from three manufacturers with 22 flexible and 20 fixed paddles were evaluated. Vertical motion at the paddle was measured using two calibrated linear potentiometers. For each paddle, the motion in millimetres was recorded every 0.5 s for 40 s, while the phantom was compressed with 80 N. Independent t-tests were used to determine differences in paddle motion between flexible and fixed, small and large, GE Senographe Essential (General Electric Medical Systems, Milwaukee, WI) and Hologic Selenia Dimensions paddles (Hologic, Bedford, MA). Paddle tilt in the medial–lateral plane for each machine/paddle combination was calculated. Results: All machine/paddle combinations demonstrate highest levels of motion during the first 10 s of the clamping phase. The least motion is 0.17 ± 0.05 mm/10 s (n = 20) and the most motion is 0.51 ± 0.15 mm/10 s (n = 80). There is a statistical difference in paddle motion between fixed and flexible (p < 0.001), GE Senographe Essential and Hologic Selenia Dimensions paddles (p < 0.001). Paddle tilt in the medial–lateral plane is independent of time and varied from 0.04 ° to 0.69 °. Conclusion: All machine/paddle combinations exhibited motion and tilting, and the extent varied with machine and paddle sizes and types. Advances in knowledge: This research suggests that image blurring will likely be clinically insignificant 4 s or more after the clamping phase commences. PMID:26739577

  20. Relaxation of Isolated Ventricular Cardiomyocytes by a Voltage-Dependent Process

    NASA Astrophysics Data System (ADS)

    Bridge, John H. B.; Spitzer, Kenneth W.; Ershler, Philip R.

    1988-08-01

    Cell contraction and relaxation were measured in single voltage-clamped guinea pig cardiomyocytes to investigate the contribution of sarcolemmal Na+-Ca2+ exchange to mechanical relaxation. Cells clamped from -80 to 0 millivolts displayed initial phasic and subsequent tonic contractions; caffeine reduced or abolished the phasic and enlarged the tonic contraction. The rate of relaxation from tonic contractions was steeply voltage-dependent and was significantly slowed in the absence of a sarcolemmal Na+ gradient. Tonic contractions elicited in the absence of a Na+ gradient promptly relaxed when external Na+ was applied, reflecting activation of Na+-Ca2+ exchange. It appears that a voltage-dependent Na+-Ca2+ exchange can rapidly mechanically relax mammalian heart muscle.

  1. Contributions of counter-charge in a potassium channel voltage-sensor domain.

    PubMed

    Pless, Stephan A; Galpin, Jason D; Niciforovic, Ana P; Ahern, Christopher A

    2011-07-24

    Voltage-sensor domains couple membrane potential to conformational changes in voltage-gated ion channels and phosphatases. Highly coevolved acidic and aromatic side chains assist the transfer of cationic side chains across the transmembrane electric field during voltage sensing. We investigated the functional contribution of negative electrostatic potentials from these residues to channel gating and voltage sensing with unnatural amino acid mutagenesis, electrophysiology, voltage-clamp fluorometry and ab initio calculations. The data show that neutralization of two conserved acidic side chains in transmembrane segments S2 and S3, namely Glu293 and Asp316 in Shaker potassium channels, has little functional effect on conductance-voltage relationships, although Glu293 appears to catalyze S4 movement. Our results suggest that neither Glu293 nor Asp316 engages in electrostatic state-dependent charge-charge interactions with S4, likely because they occupy, and possibly help create, a water-filled vestibule.

  2. From unwinding to clamping - the DEAD box RNA helicase family.

    PubMed

    Linder, Patrick; Jankowsky, Eckhard

    2011-07-22

    RNA helicases of the DEAD box family are present in all eukaryotic cells and in many bacteria and Archaea. These highly conserved enzymes are required for RNA metabolism from transcription to degradation and are therefore important players in gene expression. DEAD box proteins use ATP to unwind short duplex RNA in an unusual fashion and remodel RNA-protein complexes, but they can also function as ATP-dependent RNA clamps to provide nucleation centres that establish larger RNA-protein complexes. Structural, mechanistic and molecular biological studies have started to reveal how these conserved proteins can perform such diverse functions and how accessory proteins have a central role in their regulation.

  3. Normal-pressure Tests of Circular Plates with Clamped Edges

    NASA Technical Reports Server (NTRS)

    Mcpherson, Albert E; Ramberg, Walter; Levy, Samuel

    1942-01-01

    A fixture is described for making normal-pressure tests of flat plates 5 inches in diameter in which particular care was taken to obtain rigid clamping at the edges. Results are given for 19 plates, ranging in thickness from 0.015 to 0.072 inch. The center deflections and the extreme-fiber stresses at low pressures were found to agree with theoretical values; the center deflections at high pressures were 4 to 12 percent greater than the theoretical values. Empirical curves are derived of the pressure for the beginning of permanent set as a function of the dimensions of the plate and the tensile properties of the material.

  4. Normal-Pressure Tests of Circular Plates with Clamped Edges

    NASA Technical Reports Server (NTRS)

    Mcpherson, Albert E; Ramberg, Walter; Levy, Samuel

    1942-01-01

    A fixture is described for making normal-pressure tests of flat plates 5 inches in diameter in which particular care was taken to obtain rigid clamping at the edges. Results are given for 19 plates, ranging in thickness form 0.015 to 0.072 inch. The center deflections and the extreme-fiber stresses at low pressures were found to agree with theoretical values; the center deflections at high pressures were 4 to 12 percent greater than the theoretical values. Empirical curves are derived of the pressure for the beginning of the permanent set as a function of the dimensions of the plate and the tensile properties of the material.

  5. Nanomechanics of PCNA: A protein-made DNA sliding clamp

    NASA Astrophysics Data System (ADS)

    Rydzewski, Jakub; Strzalka, Wojciech; Nowak, Wieslaw

    2015-08-01

    Proliferating Cell Nuclear Antigen (PCNA) is a homotrimeric DNA sliding clamp involved in DNA replication, repair and cell cycle control. Understanding determinants of mechanical stability of a PCNA monomer composed of two N- and C-terminal domains requires all-atom and coarse-grained steered molecular dynamics simulations. Three schemes of unfolding at a single molecule level indicate that the conserved curvature of the PCNA monomer is due to aggregation of side β-sheets. The force spectra suggest that rupturing an interface between the domains is less probable than opening an interface between the two monomers during PCNA loading onto DNA by Replication Factor C.

  6. Influence of TiCl4 post-treatment condition on TiO2 electrode for enhancement photovoltaic efficiency of dye-sensitized solar cells.

    PubMed

    Eom, Tae Sung; Kim, Kyung Hwan; Bark, Chung Wung; Choi, Hyung Wook

    2014-10-01

    Titanium tetrachloride (TiCl4) treatment processed by chemical bath deposition is usually adopted as pre- and post-treatment for nanocrystalline titanium dioxide (TiO2) film deposition in the dye-sensitized solar cells (DSSCs) technology. TiCl4 post-treatment is a widely known method capable of improving the performance of dye-sensitized solar cells. In this work, the effect of TiCl4 post-treatment on the TiO2 electrode is proposed and compared to the untreated film. A TiO2 passivating layer was deposited on FTO glass by RF magnetron sputtering. The TiO2 sol prepared sol-gel method, nanoporous TiO2 upper layer was deposited by screen printing method on the passivating layer. TiCl4 post-treatment was deposited on the substrate by hydrolysis of TiCl4 aqueous solution. Crystalline structure was adjusted by various TiCl4 concentration and dipping time: 20 mM-150 mM and 30 min-120 min. The conversion efficiency was measured by solar simulator (100 mW/cm2). The dye-sensitized solar cell using TiCl4 post-treatment was measured the maximum conversion efficiency of 5.04% due to electron transport effectively. As a result, the DSSCs based on TiCl4 post-treatment showed better photovoltaic performance than cells made purely of TiO2 nanoparticles. The relative DSSCs devices are characterized in terms of short circuit current density, open circuit voltage, fill factor, conversion efficiency.

  7. Binding inhibitors of the bacterial sliding clamp by design.

    PubMed

    Wijffels, Gene; Johnson, Wynona M; Oakley, Aaron J; Turner, Kathleen; Epa, V Chandana; Briscoe, Susan J; Polley, Mitchell; Liepa, Andris J; Hofmann, Albert; Buchardt, Jens; Christensen, Caspar; Prosselkov, Pavel; Dalrymple, Brian P; Alewood, Paul F; Jennings, Philip A; Dixon, Nicholas E; Winkler, David A

    2011-07-14

    The bacterial replisome is a target for the development of new antibiotics to combat drug resistant strains. The β(2) sliding clamp is an essential component of the replicative machinery, providing a platform for recruitment and function of other replisomal components and ensuring polymerase processivity during DNA replication and repair. A single binding region of the clamp is utilized by its binding partners, which all contain conserved binding motifs. The C-terminal Leu and Phe residues of these motifs are integral to the binding interaction. We acquired three-dimensional structural information on the binding site in β(2) by a study of the binding of modified peptides. Development of a three-dimensional pharmacophore based on the C-terminal dipeptide of the motif enabled identification of compounds that on further development inhibited α-β(2) interaction at low micromolar concentrations. We report the crystal structure of the complex containing one of these inhibitors, a biphenyl oxime, bound to β(2), as a starting point for further inhibitor design.

  8. Image processing techniques in computer-assisted patch clamping

    NASA Astrophysics Data System (ADS)

    Azizian, Mahdi; Patel, Rajni; Gavrilovici, Cezar; Poulter, Michael O.

    2010-02-01

    Patch clamping is used in electrophysiology to study single or multiple ion channels in cells. Multiple micropipettes are used as electrodes to collect data from several cells. Placement of these electrodes is a time consuming and complicated task due to the lack of depth perception, limited view through the microscope lens and the possibility of collisions between micro-pipettes. To aid in this process, a computer-assisted approach is developed using image processing techniques applied to images obtained through the microscope. Image processing algorithms are applied to perform autofocusing, relative depth estimation, distance estimation and tracking of the micro-pipettes in the images without making any major changes in the existing patch clamp equipment. An autofocusing algorithm with a micrometer precision is developed and the relative depth estimation is performed based on autofocusing. A micro-pipette tip detection algorithm is developed which can be used to initialize or reset the tracking algorithm and to calibrate the system by registering the relative image and micro-manipulator coordinates. An image-based tracking algorithm is also developed to track a micro-pipette tip in real time. The real-time tracking data is then used for visual servoing the micro-pipette tips and updating the calibration information.

  9. Clamping of fine Kirschner wires in external fixators.

    PubMed

    Zamani, Ahmad R; Oyadiji, S Olutunde

    2016-11-01

    In Ilizarov circular (ring) external fixators, fine Kirschner wires are used to fix the bone to the fixator. Clamping of the wires to the rings with different bolt torques has been studied. However, the relation between the bolt torque and the fixation load applied to the wire was not investigated. In this work, finite element method is used to address this problem. Here, a fully three-dimensional model of the wire fixation assembly was built, with geometric details like threads on the bolt to produce a realistic simulation of the clamping of the wire. Both cannulated and slotted bolt types were studied and values of 0.2, 0.25, 0.3 and 0.45 were used for coefficient of friction. A torque was applied to the nut while the ring section and bolt kept in place. The results for bolt load, nut rotation as well as axial and radial wire deformations were obtained. The results demonstrated a linear relation between the bolt load and the bolt torque. The coefficient of this relationship was shown to be inversely proportional to the coefficient of friction. For all results, the bolt load (N) was approximately 124 times the bolt torque (N m) divided by the friction coefficient. The results highlighted the difference between the cannulated and the slotted bolts in terms of their grip on the wire.

  10. Umbilical cord clamping and preterm infants: a randomised trial.

    PubMed Central

    Kinmond, S; Aitchison, T C; Holland, B M; Jones, J G; Turner, T L; Wardrop, C A

    1993-01-01

    OBJECTIVE--To investigate the clinical effects of regulating umbilical cord clamping in preterm infants. DESIGN--A prospective randomised study. SETTING--The Queen Mother's Hospital, Glasgow. SUBJECTS--36 vaginally delivered infants over 27 and under 33 weeks' gestation. INTERVENTION--Holding the infant 20 cm below the introitus for 30 seconds before clamping the umbilical cord ("regulated" group, 17 patients), or conventional management ("random" group, 19 patients). MAIN OUTCOME MEASURES--Initial packed cell volume, peak serum bilirubin concentrations, red cell transfusion requirements, and respiratory impairment (assessed by ventilatory requirements, arterial-alveolar oxygen tension ratio over the first day in ventilated infants, and duration of dependence on supplemental oxygen). RESULTS--There were statistically significant differences between the two groups in mean initial packed cell volume (regulated group 0.564, random group 0.509) and median red cell transfusion requirements (regulated group zero, random group 23 ml/kg). 13 infants from each group underwent mechanical ventilation and showed significant differences in mean minimum arterial-alveolar oxygen tension ratio on the first day (regulated group 0.42, random group 0.22) and in median duration of dependence on supplemental oxygen (regulated group three days, random group 10 days). Differences in final outcome measures such as duration of supplemental oxygen dependence and red cell transfusion requirements were mediated primarily through arterial-alveolar oxygen tension ratio and also packed cell volume. CONCLUSIONS--This intervention at preterm deliveries produces clinical and economic benefits. PMID:8443480

  11. Control of Local Intracellular Calcium Concentration with Dynamic-Clamp Controlled 2-Photon Uncaging

    PubMed Central

    Idoux, Erwin; Mertz, Jerome

    2011-01-01

    The variations of the intracellular concentration of calcium ion ([Ca2+]i) are at the heart of intracellular signaling, and their imaging is therefore of enormous interest. However, passive [Ca2+]i imaging provides no control over these variations, meaning that a full exploration of the functional consequences of [Ca2+]i changes is difficult to attain. The tools designed so far to modify [Ca2+]i, even qualitatively, suffer drawbacks that undermine their widespread use. Here, we describe an electro-optical technique to quantitatively set [Ca2+]i, in real time and with sub-cellular resolution, using two-photon Ca2+ uncaging and dynamic-clamp. We experimentally demonstrate, on neurons from acute olfactory bulb slices of Long Evans rats, various capabilities of this technique previously difficult to achieve, such as the independent control of the membrane potential and [Ca2+]i variations, the functional knocking-in of user-defined virtual voltage-dependent Ca2+ channels, and the standardization of [Ca2+]i patterns across different cells. Our goal is to lay the groundwork for this technique and establish it as a new and versatile tool for the study of cell signaling. PMID:22216105

  12. Ba2+ release from soda glass modifies single maxi K+ channel activity in patch clamp experiments.

    PubMed Central

    Copello, J; Simon, B; Segal, Y; Wehner, F; Ramanujam, V M; Alcock, N; Reuss, L

    1991-01-01

    Glasses used to fabricate patch pipettes may release components which affect ion channels (Cota, G., and C.M. Armstrong. 1988. Biophys. J. 53:107-109; Furman, R.E., and J.C. Tanaka. 1988. Biophys. J. 53:287-292; Rojas, L., and C. Zuazaga. 1988. Neurosci. Lett. 88:39-44). The gating properties of maxi K+ channels from Necturus gallbladder epithelium depend on whether borosilicate glass (BG) or blue tip hematocrit glass (SG) is used to construct the patch pipettes. The data are consistent with solubilization from SG of a component which exerts voltage-dependent, cytosolic-side specific block, closely resembling "slow block" by Ba2+ ions. Ringer's solution preincubated with SG, but not with BG, blocked inside-out maxi K+ channels when used as bathing solution. Mass spectrometry revealed that Ba2+ is released by the glass from fast and slow-release compartments (SG contains 3% wt/wt BaO), and is the only ion found in the solution at concentrations consistent with the observed channel block. Additionally, SG released O2-, Na+, Ca2+, and Mg2+, all to micromolar concentrations. These elements do not interfere with maxi K+ channels but they could in principle alter the properties of other ion channels. Thus, screening for channel-modifying substances released by the glass may be necessary for the adequate interpretation of patch-clamp results. PMID:1742460

  13. A study of Na(x)Pt3O4 as an O2 electrode bifunctional electrocatalyst

    NASA Technical Reports Server (NTRS)

    Fielder, William L.; Singer, Joseph

    1991-01-01

    The present study suggests that polytetrafluoroethylene (PTFE) bonded Na(X)Pt3O4 gas porous diffusion electrodes may be a viable candidate for bifunctional O2 reduction and evolution activity. The electrodes exhibited Tafel slopes of about 0.06 V/decade for both O2 reduction an evolution. For O2 reduction, the 0.06 slope doubled to 0.12 V/decade at larger current densities. Preliminary stability testing at 24 C suggest that the Na(x)Pt3O4 electrodes were relatively stable at reducing and oxidizing potentials typically encountered at the O2 electrodes in a regenerative fuel cell.

  14. Voltage correction power flow

    SciTech Connect

    Rajicic, D.; Ackovski, R.; Taleski, R. . Dept. of Electrical Engineering)

    1994-04-01

    A method for power flow solution of weakly meshed distribution and transmission networks is presented. It is based on oriented ordering of network elements. That allows an efficient construction of the loop impedance matrix and rational organization of the processes such as: power summation (backward sweep), current summation (backward sweep) and node voltage calculation (forward sweep). The first step of the algorithm is calculation of node voltages on the radial part of the network. The second step is calculation of the breakpoint currents. Then, the procedure continues with the first step, which is preceded by voltage correction. It is illustrated that using voltage correction approach, the iterative process of weakly meshed network voltage calculation is faster and more reliable.

  15. Voltage verification unit

    DOEpatents

    Martin, Edward J.

    2008-01-15

    A voltage verification unit and method for determining the absence of potentially dangerous potentials within a power supply enclosure without Mode 2 work is disclosed. With this device and method, a qualified worker, following a relatively simple protocol that involves a function test (hot, cold, hot) of the voltage verification unit before Lock Out/Tag Out and, and once the Lock Out/Tag Out is completed, testing or "trying" by simply reading a display on the voltage verification unit can be accomplished without exposure of the operator to the interior of the voltage supply enclosure. According to a preferred embodiment, the voltage verification unit includes test leads to allow diagnostics with other meters, without the necessity of accessing potentially dangerous bus bars or the like.

  16. Activation of Ih and TTX-sensitive sodium current at subthreshold voltages during CA1 pyramidal neuron firing.

    PubMed

    Yamada-Hanff, Jason; Bean, Bruce P

    2015-10-01

    We used dynamic clamp and action potential clamp techniques to explore how currents carried by tetrodotoxin-sensitive sodium channels and HCN channels (Ih) regulate the behavior of CA1 pyramidal neurons at resting and subthreshold voltages. Recording from rat CA1 pyramidal neurons in hippocampal slices, we found that the apparent input resistance and membrane time constant were strongly affected by both conductances, with Ih acting to decrease apparent input resistance and time constant and sodium current acting to increase both. We found that both Ih and sodium current were active during subthreshold summation of artificial excitatory postsynaptic potentials (EPSPs) generated by dynamic clamp, with Ih dominating at less depolarized voltages and sodium current at more depolarized voltages. Subthreshold sodium current-which amplifies EPSPs-was most effectively recruited by rapid voltage changes, while Ih-which blunts EPSPs-was maximal for slow voltage changes. The combined effect is to selectively amplify rapid EPSPs. We did similar experiments in mouse CA1 pyramidal neurons, doing voltage-clamp experiments using experimental records of action potential firing of CA1 neurons previously recorded in awake, behaving animals as command voltages to quantify flow of Ih and sodium current at subthreshold voltages. Subthreshold sodium current was larger and subthreshold Ih was smaller in mouse neurons than in rat neurons. Overall, the results show opposing effects of subthreshold sodium current and Ih in regulating subthreshold behavior of CA1 neurons, with subthreshold sodium current prominent in both rat and mouse CA1 pyramidal neurons and additional regulation by Ih in rat neurons.

  17. Activation of Ih and TTX-sensitive sodium current at subthreshold voltages during CA1 pyramidal neuron firing

    PubMed Central

    Yamada-Hanff, Jason

    2015-01-01

    We used dynamic clamp and action potential clamp techniques to explore how currents carried by tetrodotoxin-sensitive sodium channels and HCN channels (Ih) regulate the behavior of CA1 pyramidal neurons at resting and subthreshold voltages. Recording from rat CA1 pyramidal neurons in hippocampal slices, we found that the apparent input resistance and membrane time constant were strongly affected by both conductances, with Ih acting to decrease apparent input resistance and time constant and sodium current acting to increase both. We found that both Ih and sodium current were active during subthreshold summation of artificial excitatory postsynaptic potentials (EPSPs) generated by dynamic clamp, with Ih dominating at less depolarized voltages and sodium current at more depolarized voltages. Subthreshold sodium current—which amplifies EPSPs—was most effectively recruited by rapid voltage changes, while Ih—which blunts EPSPs—was maximal for slow voltage changes. The combined effect is to selectively amplify rapid EPSPs. We did similar experiments in mouse CA1 pyramidal neurons, doing voltage-clamp experiments using experimental records of action potential firing of CA1 neurons previously recorded in awake, behaving animals as command voltages to quantify flow of Ih and sodium current at subthreshold voltages. Subthreshold sodium current was larger and subthreshold Ih was smaller in mouse neurons than in rat neurons. Overall, the results show opposing effects of subthreshold sodium current and Ih in regulating subthreshold behavior of CA1 neurons, with subthreshold sodium current prominent in both rat and mouse CA1 pyramidal neurons and additional regulation by Ih in rat neurons. PMID:26289465

  18. Hyperinsulinemic-euglycemic clamp in the conscious rat.

    PubMed

    Hughey, Curtis C; Hittel, Dustin S; Johnsen, Virginia L; Shearer, Jane

    2011-02-07

    Type 2 diabetes (T2D) is rapidly rising in prevalence. Characterized by either inadequate insulin production or the inability to utilize insulin produced, T2D results in elevated blood glucose levels. The "gold-standard" in assessing insulin sensitivity is a hyperinsulinemic-euglycemic clamp or insulin clamp. In this procedure, insulin is infused at a constant rate resulting in a drop in blood glucose. To maintain blood glucose at a constant level, exogenous glucose (D50) is infused into the venous circulation. The amount of glucose infused to maintain homeostasis is indicative of insulin sensitivity. Here, we show the basic clamp procedure in the chronically catheterized, unrestrained, conscious rat. This model allows blood to be collected with minimal stress to the animal. Following the induction of anesthesia, a midline incision is made and the left common carotid artery and right jugular vein are catheterized. Inserted catheters are flushed with heparinized saline, then exteriorized and secured. Animals are allowed to recover for 4-5 days prior to experiments, with weight gain monitored daily. Only those animals who regain weight to pre-surgery levels are used for experiments. On the day of the experiment, rats are fasted and connected to pumps containing insulin and D50. Baseline glucose is assessed from the arterial line and used a benchmark throughout the experiment (euglycemia). Following this, insulin is infused at a constant rate into the venous circulation. To match the drop in blood glucose, D50 is infused. If the rate of D50 infusion is greater than the rate of uptake, a rise in glucose will occur. Similarly, if the rate is insufficient to match whole body glucose uptake, a drop will occur. Titration of glucose continues until stable glucose readings are achieved. Glucose levels and glucose infusion rates during this stable period are recorded and reported. Results provide an index of whole body insulin sensitivity. The technique can be refined to meet

  19. Use of vacuum tubes in test instrumentation for measuring characteristics of fast high-voltage semiconductor devices

    NASA Technical Reports Server (NTRS)

    Berning, D.

    1981-01-01

    Circuits are described that permit measurement of fast events occurring in power semiconductors. These circuits were developed for the dynamic characterization of transistors used in inductive-load switching applications. Fast voltage clamping using vacuum diodes is discussed, and reference is made to a unique circuit that was built for performing nondestructive, reverse-bias, second-breakdown tests on transistors.

  20. Optical electrophysiology for probing function and pharmacology of voltage-gated ion channels

    PubMed Central

    Zhang, Hongkang; Reichert, Elaine; Cohen, Adam E

    2016-01-01

    Voltage-gated ion channels mediate electrical dynamics in excitable tissues and are an important class of drug targets. Channels can gate in sub-millisecond timescales, show complex manifolds of conformational states, and often show state-dependent pharmacology. Mechanistic studies of ion channels typically involve sophisticated voltage-clamp protocols applied through manual or automated electrophysiology. Here, we develop all-optical electrophysiology techniques to study activity-dependent modulation of ion channels, in a format compatible with high-throughput screening. Using optical electrophysiology, we recapitulate many voltage-clamp protocols and apply to Nav1.7, a channel implicated in pain. Optical measurements reveal that a sustained depolarization strongly potentiates the inhibitory effect of PF-04856264, a Nav1.7-specific blocker. In a pilot screen, we stratify a library of 320 FDA-approved compounds by binding mechanism and kinetics, and find close concordance with patch clamp measurements. Optical electrophysiology provides a favorable tradeoff between throughput and information content for studies of NaV channels, and possibly other voltage-gated channels. DOI: http://dx.doi.org/10.7554/eLife.15202.001 PMID:27215841

  1. Modeling considerations for the analysis of LMFBR steam generator tube clamps

    SciTech Connect

    Lay, D.M.; Piper, R.M.

    1982-01-01

    In the design of the Babcock and Wilcox Helical Coil Liquid Metal Fast Breeder Reactor (LMFBR) Steam Generator, the tube bundle is connected to the feedwater and steam plenums via ''inlet/outlet tubes''. Of prime importance in the design of these tubes is the tube-to-tube and tube-to-shell clamps which are provided to prevent detrimental vibration. This paper presents a method of modeling the tube-to-tube clamps to accurately predict tube-to-clamp interaction in the finite element analysis. It also demonstrates the validity of specific modeling assumptions in determining stresses in the clamp assembly.

  2. Electrically integrated SU-8 clamped graphene drum resonators for strain engineering

    NASA Astrophysics Data System (ADS)

    Lee, Sunwoo; Chen, Changyao; Deshpande, Vikram V.; Lee, Gwan-Hyoung; Lee, Ilkyu; Lekas, Michael; Gondarenko, Alexander; Yu, Young-Jun; Shepard, Kenneth; Kim, Philip; Hone, James

    2013-04-01

    Graphene mechanical resonators are the ultimate two-dimensional nanoelectromechanical systems (NEMS) with applications in sensing and signal processing. While initial devices have shown promising results, an ideal graphene NEMS resonator should be strain engineered, clamped at the edge without trapping gas underneath, and electrically integratable. In this Letter, we demonstrate fabrication and direct electrical measurement of circular SU-8 polymer-clamped chemical vapor deposition graphene drum resonators. The clamping increases device yield and responsivity, while providing a cleaner resonance spectrum from eliminated edge modes. Furthermore, the clamping induces a large strain in the resonator, increasing its resonant frequency.

  3. Mechanism of ATP-driven PCNA clamp loading by S. cerevisiae RFC.

    PubMed

    Chen, Siying; Levin, Mikhail K; Sakato, Miho; Zhou, Yayan; Hingorani, Manju M

    2009-05-08

    Circular clamps tether polymerases to DNA, serving as essential processivity factors in genome replication, and function in other critical cellular processes as well. Clamp loaders catalyze clamp assembly onto DNA, and the question of how these proteins construct a topological link between a clamp and DNA, especially the mechanism by which ATP is utilized for the task, remains open. Here we describe pre-steady-state analysis of ATP hydrolysis, proliferating cell nuclear antigen (PCNA) clamp opening, and DNA binding by Saccharomyces cerevisiae replication factor C (RFC), and present the first kinetic model of a eukaryotic clamp-loading reaction validated by global data analysis. ATP binding to multiple RFC subunits initiates a slow conformational change in the clamp loader, enabling it to bind and open PCNA and to bind DNA as well. PCNA opening locks RFC into an active state, and the resulting RFC.ATP.PCNA((open)) intermediate is ready for the entry of DNA into the clamp. DNA binding commits RFC to ATP hydrolysis, which is followed by PCNA closure and PCNA.DNA release. This model enables quantitative understanding of the multistep mechanism of a eukaryotic clamp loader and furthermore facilitates comparative analysis of loaders from diverse organisms.

  4. Non-Linear Vibration Characteristics of Clamped Laminated Shallow Shells

    NASA Astrophysics Data System (ADS)

    ABE, A.; KOBAYASHI, Y.; YAMADA, G.

    2000-07-01

    This paper examines non-linear free vibration characteristics of first and second vibration modes of laminated shallow shells with rigidly clamped edges. Non-linear equations of motion for the shells based on the first order shear deformation and classical shell theories are derived by means of Hamilton's principle. We apply Galerkin's procedure to the equations of motion in which eigenvectors for first and second modes of linear vibration obtained by the Ritz method are employed as trial functions. Then simultaneous non-linear ordinary differential equations are derived in terms of amplitudes of the first and second vibration modes. Backbone curves for the first and second vibration modes are solved numerically by the Gauss-Legendre integration method and the shooting method respectively. The effects of lamination sequences and transverse shear deformation on the behavior are discussed. It is also shown that the motion of the first vibration mode affects the response for the second vibration mode.

  5. Self-clamping arc light reflector for welding torch

    NASA Technical Reports Server (NTRS)

    Gordon, Stephen S. (Inventor)

    1987-01-01

    This invention is directed to a coaxial extending metal mirror reflector attached to the electrode housing or gas cup on a welding torch. An electric welding torch with an internal viewing system for robotic welding is provded with an annular arc light reflector to reflect light from the arc back onto the workpiece. The reflector has a vertical split or gap in its surrounding wall to permit the adjacent wall ends forming the split to be sprung open slightly to permit the reflector to be removed or slipped onto the torch housing or gas cup. The upper opening of the reflector is slightly smaller than the torch housing or gas cup and therefore, when placed on the torch housing or gas cup has that springiness to cause it to clamp tightly on the housing or gas cup. The split or gap also serves to permit the feed of weld wire through to the weld area,

  6. Self-clamping arc light reflector for welding torch

    NASA Astrophysics Data System (ADS)

    Gordon, Stephen S.

    1987-07-01

    This invention is directed to a coaxial extending metal mirror reflector attached to the electrode housing or gas cup on a welding torch. An electric welding torch with an internal viewing system for robotic welding is provded with an annular arc light reflector to reflect light from the arc back onto the workpiece. The reflector has a vertical split or gap in its surrounding wall to permit the adjacent wall ends forming the split to be sprung open slightly to permit the reflector to be removed or slipped onto the torch housing or gas cup. The upper opening of the reflector is slightly smaller than the torch housing or gas cup and therefore, when placed on the torch housing or gas cup has that springiness to cause it to clamp tightly on the housing or gas cup. The split or gap also serves to permit the feed of weld wire through to the weld area,

  7. Priming and testing silicon patch-clamp neurochips.

    PubMed

    Py, Christophe; Denhoff, Michael W; Sabourin, Nicaulas; Weber, John; Shiu, Matthew; Zhao, Ping

    2014-09-25

    We report on the systematic and automated priming and testing of silicon planar patch-clamp chips after their assembly in Plexiglas packages and sterilization in an air plasma reactor. We find that almost 90% of the chips are successfully primed by our automated setup, and have a shunt capacitance of between 10 pF and 30 pF. Blocked chips are mostly due to glue invasion in the well, and variability in the manual assembly process is responsible for the distribution in shunt capacitance value. Priming and testing time with our automated setup is less than 5 min per chip, which is compatible with the production of large series for use in electrophysiology experiments.

  8. The human dynamic clamp as a paradigm for social interaction

    PubMed Central

    Dumas, Guillaume; de Guzman, Gonzalo C.; Tognoli, Emmanuelle; Kelso, J. A. Scott

    2014-01-01

    Social neuroscience has called for new experimental paradigms aimed toward real-time interactions. A distinctive feature of interactions is mutual information exchange: One member of a pair changes in response to the other while simultaneously producing actions that alter the other. Combining mathematical and neurophysiological methods, we introduce a paradigm called the human dynamic clamp (HDC), to directly manipulate the interaction or coupling between a human and a surrogate constructed to behave like a human. Inspired by the dynamic clamp used so productively in cellular neuroscience, the HDC allows a person to interact in real time with a virtual partner itself driven by well-established models of coordination dynamics. People coordinate hand movements with the visually observed movements of a virtual hand, the parameters of which depend on input from the subject’s own movements. We demonstrate that HDC can be extended to cover a broad repertoire of human behavior, including rhythmic and discrete movements, adaptation to changes of pacing, and behavioral skill learning as specified by a virtual “teacher.” We propose HDC as a general paradigm, best implemented when empirically verified theoretical or mathematical models have been developed in a particular scientific field. The HDC paradigm is powerful because it provides an opportunity to explore parameter ranges and perturbations that are not easily accessible in ordinary human interactions. The HDC not only enables to test the veracity of theoretical models, it also illuminates features that are not always apparent in real-time human social interactions and the brain correlates thereof. PMID:25114256

  9. Comparison of various organic compounds destruction on rare earths doped Ti/Sb-SnO2 electrodes.

    PubMed

    Cui, Yu-Hong; Feng, Yu-Jie; Liu, Junfeng; Ren, Nanqi

    2012-11-15

    Ti/Sb-SnO2 and three kinds of rare earths (REs), namely Ce, Gd, and Eu doped Ti/Sb-SnO2 electrodes were prepared and tested for their capacity on electrocatalytic degradation of three kinds of basal aromatic compounds (benzoquinone, hydroquinone and catechol) and six kinds of aliphatic acids (maleic acid, fumaric acid, succinic acid, malonic acid, oxalic acid and acetic acid). The elimination of selected organics as well as their TOC removal with different doped Ti/Sb-SnO2 electrodes was described by first-order kinetics. Compared with Ti/Sb-SnO2, the Gd and Eu doped electrodes show better performance on the degradation of most of the selected organics, while Ce doped electrode shows either closely or lower efficiency on the degradation of these selected organics. Besides electrode material, the molecular structure of organic compound has obvious effect on its degradation in the electrocatalytic process. Catechol is more resistant to the electrophilic attack by hydroxyl radicals than benzoquinone and hydroquinone. The compound with more complicate molecular structure or longer carbon chain is more difficult to be mineralized. The aliphatic acid with higher oxygen content or more double bonds is more readily to be oxidized in the electrocatalytic process.

  10. Voltage balanced multilevel voltage source converter system

    DOEpatents

    Peng, F.Z.; Lai, J.S.

    1997-07-01

    Disclosed is a voltage balanced multilevel converter for high power AC applications such as adjustable speed motor drives and back-to-back DC intertie of adjacent power systems. This converter provides a multilevel rectifier, a multilevel inverter, and a DC link between the rectifier and the inverter allowing voltage balancing between each of the voltage levels within the multilevel converter. The rectifier is equipped with at least one phase leg and a source input node for each of the phases. The rectifier is further equipped with a plurality of rectifier DC output nodes. The inverter is equipped with at least one phase leg and a load output node for each of the phases. The inverter is further equipped with a plurality of inverter DC input nodes. The DC link is equipped with a plurality of rectifier charging means and a plurality of inverter discharging means. The plurality of rectifier charging means are connected in series with one of the rectifier charging means disposed between and connected in an operable relationship with each adjacent pair of rectifier DC output nodes. The plurality of inverter discharging means are connected in series with one of the inverter discharging means disposed between and connected in an operable relationship with each adjacent pair of inverter DC input nodes. Each of said rectifier DC output nodes are individually electrically connected to the respective inverter DC input nodes. By this means, each of the rectifier DC output nodes and each of the inverter DC input nodes are voltage balanced by the respective charging and discharging of the rectifier charging means and the inverter discharging means. 15 figs.

  11. Voltage balanced multilevel voltage source converter system

    DOEpatents

    Peng, Fang Zheng; Lai, Jih-Sheng

    1997-01-01

    A voltage balanced multilevel converter for high power AC applications such as adjustable speed motor drives and back-to-back DC intertie of adjacent power systems. This converter provides a multilevel rectifier, a multilevel inverter, and a DC link between the rectifier and the inverter allowing voltage balancing between each of the voltage levels within the multilevel converter. The rectifier is equipped with at least one phase leg and a source input node for each of the phases. The rectifier is further equipped with a plurality of rectifier DC output nodes. The inverter is equipped with at least one phase leg and a load output node for each of the phases. The inverter is further equipped with a plurality of inverter DC input nodes. The DC link is equipped with a plurality of rectifier charging means and a plurality of inverter discharging means. The plurality of rectifier charging means are connected in series with one of the rectifier charging means disposed between and connected in an operable relationship with each adjacent pair of rectifier DC output nodes. The plurality of inverter discharging means are connected in series with one of the inverter discharging means disposed between and connected in an operable relationship with each adjacent pair of inverter DC input nodes. Each of said rectifier DC output nodes are individually electrically connected to the respective inverter DC input nodes. By this means, each of the rectifier DC output nodes and each of the inverter DC input nodes are voltage balanced by the respective charging and discharging of the rectifier charging means and the inverter discharging means.

  12. Low voltage to high voltage level shifter and related methods

    NASA Technical Reports Server (NTRS)

    Mentze, Erik J. (Inventor); Hess, Herbert L. (Inventor); Buck, Kevin M. (Inventor); Cox, David F. (Inventor)

    2006-01-01

    A shifter circuit comprises a high and low voltage buffer stages and an output buffer stage. The high voltage buffer stage comprises multiple transistors arranged in a transistor stack having a plurality of intermediate nodes connecting individual transistors along the stack. The transistor stack is connected between a voltage level being shifted to and an input voltage. An inverter of this stage comprises multiple inputs and an output. Inverter inputs are connected to a respective intermediate node of the transistor stack. The low voltage buffer stage has an input connected to the input voltage and an output, and is operably connected to the high voltage buffer stage. The low voltage buffer stage is connected between a voltage level being shifted away from and a lower voltage. The output buffer stage is driven by the outputs of the high voltage buffer stage inverter and the low voltage buffer stage.

  13. Single Event Transients in Low Voltage Dropout (LVDO) Voltage Regulators

    NASA Technical Reports Server (NTRS)

    LaBel, K.; Karsh, J.; Pursley, S.; Kleyner, I.; Katz, R.; Poivey, C.; Kim, H.; Seidleck, C.

    2006-01-01

    This viewgraph presentation reviews the use of Low Voltage Dropout (LVDO) Voltage Regulators in environments where heavy ion induced Single Event Transients are a concern to the designers.Included in the presentation are results of tests of voltage regulators.

  14. A non-linear voltage dependent charge movement in frog skeletal muscle.

    PubMed Central

    Chandler, W K; Rakowski, R F; Schneider, M F

    1976-01-01

    1. Voltage-clamp experiments were carried out using the three microelectrode technique. Using this method membrane current density at V1 is proportional to deltaV( = V2 - V1) where V1 and V2 are voltages at distances 1 and 21 from the end of a fibre. Voltage dependent sodium currents were blocked by tetrodotoxin, potassium by tetraethylammonium ions and rubidium. Contraction was blocked by adding sucrose, 467 mM. 2. The current deltaV (control) associated with a positive voltage step from a hyperpolarized conditioning voltage to the holding potential, -80 mV, showed two components, a capacitative transient which decayed rapidly and a maintained steady level... PMID:1082506

  15. Effects of voltage perturbation of the lingual receptive field on chorda tympani responses to Na+ and K+ salts in the rat: implications for gustatory transduction

    PubMed Central

    1994-01-01

    Taste sensory responses from the chorda tympani nerve of the rat were recorded with the lingual receptive field under current or voltage clamp. Consistent with previous results (Ye, Q., G. L. Heck, and J. A. DeSimone. 1993. Journal of Neurophysiology. 70:167-178), responses to NaCl were highly sensitive to lingual voltage clamp condition. This can be attributed to changes in the electrochemical driving force for Na+ ions through apical membrane transducer channels in taste cells. In contrast, responses to KCl over the concentration range 50-500 mM were insensitive to the voltage clamp condition of the receptive field. These results indicate the absence of K+ conductances comparable to those for Na+ in the apical membranes of taste cells. This was supported by the strong anion dependence of K salt responses. At zero current clamp, the potassium gluconate (KGlu) threshold was > 250 mM, and onset kinetics were slow (12 s to reach half-maximal response). Faster onset kinetics and larger responses to KGlu occurred at negative voltage clamp (-50 mV). This indicates that when K+ ion is transported as a current, and thereby uncoupled from gluconate mobility, its rate of delivery to the K+ taste transducer increases. Analysis of conductances shows that the paracellular pathway in the lingual epithelium is 28 times more permeable to KCl than to KGlu. Responses to KGlu under negative voltage clamp were not affected by agents that are K+ channel blockers in other systems. The results indicate that K salt taste transduction is under paracellular diffusion control, which limits chemoreception efficiency. We conclude that rat K salt taste occurs by means of a subtight junctional transducer for K+ ions with access limited by anion mobility. The data suggest that this transducer is not cation selective which also accounts for the voltage and amiloride insensitive part of the response to NaCl. PMID:7876827

  16. Zero Voltage Soft Switching Duty Cycle Pulse Modulated High Frequency Inverter-Fed

    NASA Astrophysics Data System (ADS)

    Ishitobi, Manabu; Matsushige, Takayuki; Nakaoka, Mutsuo; Bessyo, Daisuke; Omori, Hideki; Terai, Haruo

    The utility grid voltage of commercial AC power source in Japan and USA is 100V, but in other Asian and European countries, it is 220V. In recent years, in Japan 200V outputted single-phase three-wire system begins to be used for high power applications. In 100V utility AC power applications and systems, an active voltage clamped quasi-resonant inverter circuit topology sing IGBTs has been effectively used so far for the consumer microwave oven. In this paper, presented is a half bridge type voltage-clamped asymmetrical soft switching PWM high-frequency inverter type AC-DC converter using IGBTs which is designed for consumer magnetron drive used as the consumer microwave oven in 200V utility AC power system. The zero voltage soft switching inverter treated here can use the same power rated switching semiconductor devices and three-winding high frequency transformer as those of the active voltage clamped quasi-resonant inverter using the IGBTs that has already been used for 100V utility AC power source. The operating performances of the voltage source single ended push pull (SEPP) type soft switching PWM inverter are evaluated and discussed for 100V and 200V common use consumer microwave oven. The harmonic line current components in the utility AC power side of the AC-DC power converter with ZVS-PWM SEPP inverter are reduced and improved on the basis of sine wave like pulse frequency modulation and sine wave like pulse width modulation for the utility AC voltage source.

  17. Analysis of impactor residues in tray clamps from the Long Duration Exposure Facility. Part 1: Clamps from Bay A of the satellite

    NASA Technical Reports Server (NTRS)

    Zolensky, Michael E.; Bernhard, Ronald P.

    1993-01-01

    The Long Duration Exposure Facility (LDEF) was placed in low Earth orbit (LEO) in 1984 and was recovered 5.7 years later. The LDEF was host to several individual experiments that were specifically designed to characterize critical aspects of meteoroid and debris environment in LEO. It was realized from the beginning, however, that the most efficient use of the satellite would be to examine the entire surface of the Earth for impact features. In this regard, particular interest has centered on common exposed materials that faced in all LDEF pointing directions. Among the most important of these materials is the tray clamps. Therefore, in an effort to understand the nature of particulates in LEO and their effects on spacecraft hardware better, we are analyzing residues found in impact features on LDEF tray clamp surfaces. This catalog presents all data from clamps from Bay A of the LDEF. Subsequent catalogs will include clamps from succeeding bays of the satellite.

  18. High voltage DC power supply

    DOEpatents

    Droege, T.F.

    1989-12-19

    A high voltage DC power supply having a first series resistor at the output for limiting current in the event of a short-circuited output, a second series resistor for sensing the magnitude of output current, and a voltage divider circuit for providing a source of feedback voltage for use in voltage regulation is disclosed. The voltage divider circuit is coupled to the second series resistor so as to compensate the feedback voltage for a voltage drop across the first series resistor. The power supply also includes a pulse-width modulated control circuit, having dual clock signals, which is responsive to both the feedback voltage and a command voltage, and also includes voltage and current measuring circuits responsive to the feedback voltage and the voltage developed across the second series resistor respectively. 7 figs.

  19. High voltage DC power supply

    DOEpatents

    Droege, Thomas F.

    1989-01-01

    A high voltage DC power supply having a first series resistor at the output for limiting current in the event of a short-circuited output, a second series resistor for sensing the magnitude of output current, and a voltage divider circuit for providing a source of feedback voltage for use in voltage regulation is disclosed. The voltage divider circuit is coupled to the second series resistor so as to compensate the feedback voltage for a voltage drop across the first series resistor. The power supply also includes a pulse-width modulated control circuit, having dual clock signals, which is responsive to both the feedback voltage and a command voltage, and also includes voltage and current measuring circuits responsive to the feedback voltage and the voltage developed across the second series resistor respectively.

  20. High voltage power supply

    NASA Technical Reports Server (NTRS)

    Ruitberg, A. P.; Young, K. M. (Inventor)

    1985-01-01

    A high voltage power supply is formed by three discrete circuits energized by a battery to provide a plurality of concurrent output signals floating at a high output voltage on the order of several tens of kilovolts. In the first two circuits, the regulator stages are pulse width modulated and include adjustable ressistances for varying the duty cycles of pulse trains provided to corresponding oscillator stages while the third regulator stage includes an adjustable resistance for varying the amplitude of a steady signal provided to a third oscillator stage. In the first circuit, the oscillator, formed by a constant current drive network and a tuned resonant network included a step up transformer, is coupled to a second step up transformer which, in turn, supplies an amplified sinusoidal signal to a parallel pair of complementary poled rectifying, voltage multiplier stages to generate the high output voltage.

  1. High Voltage Distribution

    NASA Astrophysics Data System (ADS)

    Norbeck, Edwin; Miller, Michael; Onel, Yasar

    2010-11-01

    For detector arrays that require 5 to 10 kV at a few microamps each for hundreds of detectors, using hundreds of HV power supplies is unreasonable. Bundles of hundreds of HV cables take up space that should be filled with detectors. A typical HV module can supply 1 ma, enough current for hundreds of detectors. It is better to use a single HV module and distribute the current as needed. We show a circuit that, for each detector, measures the current, cuts off the voltage if the current exceeds a set maximum, and allows the HV to be turned on or off from a control computer. The entire array requires a single HV cable and 2 or 3 control lines. This design provides the same voltage to all of the detectors, the voltage set by the single HV module. Some additional circuitry would allow a computer controlled voltage drop between the HV and each individual detector.

  2. High-voltage distributors

    NASA Technical Reports Server (NTRS)

    Mcchesney, J. F., Jr.

    1974-01-01

    Two distributors reduce high-voltage breakdowns and corona discharges. Both distributors are constructed to prevent air traps and facilitate servicing without soldering. Occurrence of coronas is also minimized due to smooth surfaces of device.

  3. The effect of internal sodium and caesium on phasic contraction of patch-clamped rabbit ventricular myocytes.

    PubMed

    Levi, A J; Mitcheson, J S; Hancox, J C

    1996-04-01

    1. The voltage dependence of phasic contraction was assessed in rabbit ventricular myocytes. Phasic contraction at all potentials was abolished by exposure to ryanodine-thapsigargin, showing that it was due primarily to Ca2+ release from the sarcoplasmic reticulum (SR). Experiments were performed at 35 degrees C, cells were whole-cell patch clamped and contraction was measured optically as unloaded shortening. Cells were held at -40 mV to inactivate the Na+ current (INa) and T-type Ca2+ current. A standard cellular Ca2+ load was established by applying a train of conditioning pulses at 0.5 Hz before each test pulse. The effect of replacing K+ with Cs+ in the dialysing pipette solution, and the effect of altering dialysing [Na+] between 0 and 20 mM, was assessed on contraction. 2. Cells dialysed with a K(+)-based, Na(+)-free solution exhibited a 'bell-shaped' voltage dependence of the L-type Ca2+ channel current (ICa,L), with a maximum ICa,L at +10 mV. Replacing internal K+ with Cs+, or altering pipette [Na+], did not affect the voltage dependence of ICa,L. 3. The voltage dependence of phasic contraction in cells dialysed with a K(+)-based solution was modulated by pipette [Na+]. The voltage dependence of phasic contraction was bell-shaped with 0 Na+, became much loss bell-shaped with 10 mM Na+ and with 20 mM Na+ the phasic contraction elicited at +100 mV was 1.6-fold larger than that at +10 mV. 4. Replacing 80% of K+ with Cs+ in the pipette dialysis solution led to a significant reduction in contraction amplitude and a more rapid decline in contraction amplitude after beginning the dialysis of the cell. 5. Cells dialysed with a Cs(+)-based solution displayed a voltage dependence of phasic contraction which was more bell-shaped (i.e. more similar to that of ICa,L) than that obtained with the corresponding K(+)-based dialysis solution. The level of pipette [Na+] still modulated the voltage dependence of phasic contraction in cells dialysed with a Cs(+)-based solution

  4. Allosteric substrate switching in a voltage-sensing lipid phosphatase.

    PubMed

    Grimm, Sasha S; Isacoff, Ehud Y

    2016-04-01

    Allostery provides a critical control over enzyme activity, biasing the catalytic site between inactive and active states. We found that the Ciona intestinalis voltage-sensing phosphatase (Ci-VSP), which modifies phosphoinositide signaling lipids (PIPs), has not one but two sequential active states with distinct substrate specificities, whose occupancy is allosterically controlled by sequential conformations of the voltage-sensing domain (VSD). Using fast fluorescence resonance energy transfer (FRET) reporters of PIPs to monitor enzyme activity and voltage-clamp fluorometry to monitor conformational changes in the VSD, we found that Ci-VSP switches from inactive to a PIP3-preferring active state when the VSD undergoes an initial voltage-sensing motion and then into a second PIP2-preferring active state when the VSD activates fully. This two-step allosteric control over a dual-specificity enzyme enables voltage to shape PIP concentrations in time, and provides a mechanism for the complex modulation of PIP-regulated ion channels, transporters, cell motility, endocytosis and exocytosis.

  5. Allosteric substrate switching in a voltage sensing lipid phosphatase

    PubMed Central

    Grimm, Sasha S.; Isacoff, Ehud Y.

    2016-01-01

    Allostery provides a critical control over enzyme activity, biasing the catalytic site between inactive and active states. We find the Ciona intestinalis voltage-sensing phosphatase (Ci-VSP), which modifies phosphoinositide signaling lipids (PIPs), to have not one but two sequential active states with distinct substrate specificities, whose occupancy is allosterically controlled by sequential conformations of the voltage sensing domain (VSD). Using fast FRET reporters of PIPs to monitor enzyme activity and voltage clamp fluorometry to monitor conformational changes in the VSD, we find that Ci-VSP switches from inactive to a PIP3-preferring active state when the VSD undergoes an initial voltage sensing motion and then into a second PIP2-preferring active state when the VSD activates fully. This novel 2-step allosteric control over a dual specificity enzyme enables voltage to shape PIP concentrations in time, and provides a mechanism for the complex modulation of PIP-regulated ion channels, transporters, cell motility and endo/exocytosis. PMID:26878552

  6. Calibration of Voltage Transformers and High- Voltage Capacitors at NIST

    PubMed Central

    Anderson, William E.

    1989-01-01

    The National Institute of Standards and Technology (NIST) calibration service for voltage transformers and high-voltage capacitors is described. The service for voltage transformers provides measurements of ratio correction factors and phase angles at primary voltages up to 170 kV and secondary voltages as low as 10 V at 60 Hz. Calibrations at frequencies from 50–400 Hz are available over a more limited voltage range. The service for high-voltage capacitors provides measurements of capacitance and dissipation factor at applied voltages ranging from 100 V to 170 kV at 60 Hz depending on the nominal capacitance. Calibrations over a reduced voltage range at other frequencies are also available. As in the case with voltage transformers, these voltage constraints are determined by the facilities at NIST. PMID:28053409

  7. Structural insight into β-Clamp and its interaction with DNA Ligase in Helicobacter pylori.

    PubMed

    Pandey, Preeti; Tarique, Khaja Faisal; Mazumder, Mohit; Rehman, Syed Arif Abdul; Kumari, Nilima; Gourinath, Samudrala

    2016-08-08

    Helicobacter pylori, a gram-negative and microaerophilic bacterium, is the major cause of chronic gastritis, gastric ulcers and gastric cancer. Owing to its central role, DNA replication machinery has emerged as a prime target for the development of antimicrobial drugs. Here, we report 2Å structure of β-clamp from H. pylori (Hpβ-clamp), which is one of the critical components of DNA polymerase III. Despite of similarity in the overall fold of eubacterial β-clamp structures, some distinct features in DNA interacting loops exists that have not been reported previously. The in silico prediction identified the potential binders of β-clamp such as alpha subunit of DNA pol III and DNA ligase with identification of β-clamp binding regions in them and validated by SPR studies. Hpβ-clamp interacts with DNA ligase in micromolar binding affinity. Moreover, we have successfully determined the co-crystal structure of β-clamp with peptide from DNA ligase (not reported earlier in prokaryotes) revealing the region from ligase that interacts with β-clamp.

  8. A Comparison of the Performance and Application Differences Between Manual and Automated Patch-Clamp Techniques

    PubMed Central

    Yajuan, Xiao; Xin, Liang; Zhiyuan, Li

    2012-01-01

    The patch clamp technique is commonly used in electrophysiological experiments and offers direct insight into ion channel properties through the characterization of ion channel activity. This technique can be used to elucidate the interaction between a drug and a specific ion channel at different conformational states to understand the ion channel modulators’ mechanisms. The patch clamp technique is regarded as a gold standard for ion channel research; however, it suffers from low throughput and high personnel costs. In the last decade, the development of several automated electrophysiology platforms has greatly increased the screen throughput of whole cell electrophysiological recordings. New advancements in the automated patch clamp systems have aimed to provide high data quality, high content, and high throughput. However, due to the limitations noted above, automated patch clamp systems are not capable of replacing manual patch clamp systems in ion channel research. While automated patch clamp systems are useful for screening large amounts of compounds in cell lines that stably express high levels of ion channels, the manual patch clamp technique is still necessary for studying ion channel properties in some research areas and for specific cell types, including primary cells that have mixed cell types and differentiated cells that derive from induced pluripotent stem cells (iPSCs) or embryonic stem cells (ESCs). Therefore, further improvements in flexibility with regard to cell types and data quality will broaden the applications of the automated patch clamp systems in both academia and industry. PMID:23346269

  9. Assessing clamp-related vascular injuries by measurement of associated vascular dysfunction.

    PubMed

    Barone, G W; Conerly, J M; Farley, P C; Flanagan, T L; Kron, I L

    1989-04-01

    The development of vascular clamps requires a reliable method to quantitate clamp-related vascular injuries. The degree of vessel damage usually is estimated subjectively from photomicrographs made with scanning electron microscopy. In order to test whether the use of vascular rings to assess residual vascular function may be a better method, rabbit thoracic aortas were occluded by five types of clamps: a Fogarty softjaw bulldog, a Fogarty Hydragrip, a 6-inch Satinsky clamp, an Edslab bulldog, and a silicone vessel band. Each area of clamp injury was sectioned into a vascular ring and suspended in a tissue bath. Residual vascular function was determined by contraction in response to phenylephrine and by relaxation in response to methacholine chloride and sodium nitroprusside. Morphologic studies with use of Evans blue dye and scanning electron microscopy complemented the vascular-function studies. The Fogarty bulldog clamp was the best at preserving vascular contraction and relaxation; the more crushing Satinsky clamp was the worst. The testing of vascular rings for residual function appears to be a useful technique for objectively quantitating vascular clamp-related vascular damage.

  10. Structural insight into β-Clamp and its interaction with DNA Ligase in Helicobacter pylori

    PubMed Central

    Pandey, Preeti; Tarique, Khaja Faisal; Mazumder, Mohit; Rehman, Syed Arif Abdul; kumari, Nilima; Gourinath, Samudrala

    2016-01-01

    Helicobacter pylori, a gram-negative and microaerophilic bacterium, is the major cause of chronic gastritis, gastric ulcers and gastric cancer. Owing to its central role, DNA replication machinery has emerged as a prime target for the development of antimicrobial drugs. Here, we report 2Å structure of β-clamp from H. pylori (Hpβ-clamp), which is one of the critical components of DNA polymerase III. Despite of similarity in the overall fold of eubacterial β-clamp structures, some distinct features in DNA interacting loops exists that have not been reported previously. The in silico prediction identified the potential binders of β-clamp such as alpha subunit of DNA pol III and DNA ligase with identification of β-clamp binding regions in them and validated by SPR studies. Hpβ-clamp interacts with DNA ligase in micromolar binding affinity. Moreover, we have successfully determined the co-crystal structure of β-clamp with peptide from DNA ligase (not reported earlier in prokaryotes) revealing the region from ligase that interacts with β-clamp. PMID:27499105

  11. Voltage- and calcium-dependent motility of saccular hair bundles

    NASA Astrophysics Data System (ADS)

    Quiñones, Patricia M.; Meenderink, Sebastiaan W. F.; Bozovic, Dolores

    2015-12-01

    Active bundle motility, which is hypothesized to supply feedback for mechanical amplification of signals, is thought to enhance sensitivity and sharpen tuning in vestibular and auditory organs. To study active hair bundle motility, we combined high-speed camera recordings of bullfrog sacculi, which were mounted in a two-compartment chamber, and voltage-clamp of the hair cell membrane potential. Using this paradigm, we measured three types of bundle motions: 1) spontaneous oscillations which can be analyzed to measure the physiological operating range of the transduction channel; 2) a sustained quasi-static movement of the bundle that depends on membrane potential; and 3) a fast, transient and asymmetric movement that resets the bundle position and depends on changes in the membrane potential. These data support a role for both calcium and voltage in the transduction-channel function.

  12. Control method for peak power delivery with limited DC-bus voltage

    DOEpatents

    Edwards, John; Xu, Longya; Bhargava, Brij B.

    2006-09-05

    A method for driving a neutral point-clamped multi-level voltage source inverter supplying a synchronous motor is provided. A DC current is received at a neutral point-clamped multi-level voltage source inverter. The inverter has first, second, and third output nodes. The inverter also has a plurality of switches. A desired speed of a synchronous motor connected to the inverter by the first second and third nodes is received by the inverter. The synchronous motor has a rotor and the speed of the motor is defined by the rotational rate of the rotor. A position of the rotor is sensed, current flowing to the motor out of at least two of the first, second, and third output nodes is sensed, and predetermined switches are automatically activated by the inverter responsive to the sensed rotor position, the sensed current, and the desired speed.

  13. Intracellular calcium affects prestin's voltage operating point indirectly via turgor-induced membrane tension

    NASA Astrophysics Data System (ADS)

    Song, Lei; Santos-Sacchi, Joseph

    2015-12-01

    Recent identification of a calmodulin binding site within prestin's C-terminus indicates that calcium can significantly alter prestin's operating voltage range as gauged by the Boltzmann parameter Vh (Keller et al., J. Neuroscience, 2014). We reasoned that those experiments may have identified the molecular substrate for the protein's tension sensitivity. In an effort to understand how this may happen, we evaluated the effects of turgor pressure on such shifts produced by calcium. We find that the shifts are induced by calcium's ability to reduce turgor pressure during whole cell voltage clamp recording. Clamping turgor pressure to 1kPa, the cell's normal intracellular pressure, completely counters the calcium effect. Furthermore, following unrestrained shifts, collapsing the cells abolishes induced shifts. We conclude that calcium does not work by direct action on prestin's conformational state. The possibility remains that calcium interaction with prestin alters water movements within the cell, possibly via its anion transport function.

  14. On the mechanism of loading the PCNA sliding clamp by RFC.

    PubMed

    Dionne, Isabelle; Brown, Nicola J; Woodgate, Roger; Bell, Stephen D

    2008-04-01

    Sliding clamps play central roles in a broad range of DNA replication and repair processes. The clamps form circular molecules that must be opened and resealed around DNA by the clamp loader complex to fulfil their function. While most eukaryotes and many archea possess a homo-trimeric PCNA, the PCNA of Sulfolobus solfataricus is a heterotrimer. Here, we exploit the asymmetry of S. solfataricus PCNA to create a series of circularly permuted PCNA subunit fusions, thereby covalently closing defined interfaces within the heterotrimer. Using these concatamers, we investigate the requirements for loading the clamp onto DNA and reveal that a single defined interface within the heterotrimer is opened during the loading process. Subunit-specific interactions between S. solfataricus RFC clamp loader and PCNA permit us to superimpose our data upon the structure of yeast RFC-PCNA complex, thereby presenting a general model for PCNA loading by RFC in archaea and eukaryotes.

  15. Device for monitoring cell voltage

    DOEpatents

    Doepke, Matthias [Garbsen, DE; Eisermann, Henning [Edermissen, DE

    2012-08-21

    A device for monitoring a rechargeable battery having a number of electrically connected cells includes at least one current interruption switch for interrupting current flowing through at least one associated cell and a plurality of monitoring units for detecting cell voltage. Each monitoring unit is associated with a single cell and includes a reference voltage unit for producing a defined reference threshold voltage and a voltage comparison unit for comparing the reference threshold voltage with a partial cell voltage of the associated cell. The reference voltage unit is electrically supplied from the cell voltage of the associated cell. The voltage comparison unit is coupled to the at least one current interruption switch for interrupting the current of at least the current flowing through the associated cell, with a defined minimum difference between the reference threshold voltage and the partial cell voltage.

  16. High voltage pulse generator

    DOEpatents

    Fasching, George E.

    1977-03-08

    An improved high-voltage pulse generator has been provided which is especially useful in ultrasonic testing of rock core samples. An N number of capacitors are charged in parallel to V volts and at the proper instance are coupled in series to produce a high-voltage pulse of N times V volts. Rapid switching of the capacitors from the paralleled charging configuration to the series discharging configuration is accomplished by using silicon-controlled rectifiers which are chain self-triggered following the initial triggering of a first one of the rectifiers connected between the first and second of the plurality of charging capacitors. A timing and triggering circuit is provided to properly synchronize triggering pulses to the first SCR at a time when the charging voltage is not being applied to the parallel-connected charging capacitors. Alternate circuits are provided for controlling the application of the charging voltage from a charging circuit to be applied to the parallel capacitors which provides a selection of at least two different intervals in which the charging voltage is turned "off" to allow the SCR's connecting the capacitors in series to turn "off" before recharging begins. The high-voltage pulse-generating circuit including the N capacitors and corresponding SCR's which connect the capacitors in series when triggered "on" further includes diodes and series-connected inductors between the parallel-connected charging capacitors which allow sufficiently fast charging of the capacitors for a high pulse repetition rate and yet allow considerable control of the decay time of the high-voltage pulses from the pulse-generating circuit.

  17. The role of K+ currents in frequency-dependent spike broadening in Aplysia R20 neurons: a dynamic-clamp analysis.

    PubMed

    Ma, M; Koester, J

    1996-07-01

    The R20 neurons of Aplysia exhibit frequency-dependent spike broadening. Previously, we had used two-electrode voltage clamp to examine the mechanisms of this spike broadening (Ma and Koester, 1995). We identified three K+ currents that mediate action-potential repolarization: a transient A-type K+ current (I(Adepol)), a delayed rectifier current (IK-V), and a Ca(2+)-sensitive K+ current(IK-CA). A major constraint in that study was the lack of completely selective blockers for I(Adepol) and I(K-V), resulting in an inability to assess directly the effects of their activation and inactivation on spike broadening. In the present study, the dynamic-clamp technique, which employs computer simulation to inject biologically realistic currents into a cell under current-clamp conditions (Sharp et al., 1993a,b), was used either to block I(Adepol) or I(K-V) or to modify their inactivation properties. The data in this paper, together with earlier results, lead to the following hypothesis for the mechanism of spike broadening in the R20 cells. As the spike train progresses, the primary responsibility for spike repolarization gradually shifts from I(Adepol) to I(K-V) to I(K-Ca). This sequence can be explained on the basis of the relative rates of activation and inactivation of each current with respect to the constantly changing spike durations, the cumulative inactivation of I(Adepol) and I(K-V), and the progressive potentiation of I(K-Ca). Positive feedback interactions between spike broadening and inactivation contribute to the cumulative inactivation of both I(Adepol) and I(K-V). The data also illustrate that when two or more currents have similar driving forces and partially overlapping activation characteristics, selectively blocking one current under current-clamp conditions can lead to a significant underestimate of its normal physiological importance.

  18. Neuroprotective effect of interleukin-6 regulation of voltage-gated Na(+) channels of cortical neurons is time- and dose-dependent.

    PubMed

    Xia, Wei; Peng, Guo-Yi; Sheng, Jiang-Tao; Zhu, Fang-Fang; Guo, Jing-Fang; Chen, Wei-Qiang

    2015-04-01

    Interleukin-6 has been shown to be involved in nerve injury and nerve regeneration, but the effects of long-term administration of high concentrations of interleukin-6 on neurons in the central nervous system is poorly understood. This study investigated the effects of 24 hour exposure of interleukin-6 on cortical neurons at various concentrations (0.1, 1, 5 and 10 ng/mL) and the effects of 10 ng/mL interleukin-6 exposure to cortical neurons for various durations (2, 4, 8, 24 and 48 hours) by studying voltage-gated Na(+) channels using a patch-clamp technique. Voltage-clamp recording results demonstrated that interleukin-6 suppressed Na(+) currents through its receptor in a time- and dose-dependent manner, but did not alter voltage-dependent activation and inactivation. Current-clamp recording results were consistent with voltage-clamp recording results. Interleukin-6 reduced the action potential amplitude of cortical neurons, but did not change the action potential threshold. The regulation of voltage-gated Na(+) channels in rat cortical neurons by interleukin-6 is time- and dose-dependent.

  19. Non-Hookean statistical mechanics of clamped graphene ribbons

    NASA Astrophysics Data System (ADS)

    Bowick, Mark J.; Košmrlj, Andrej; Nelson, David R.; Sknepnek, Rastko

    2017-03-01

    Thermally fluctuating sheets and ribbons provide an intriguing forum in which to investigate strong violations of Hooke's Law: Large distance elastic parameters are in fact not constant but instead depend on the macroscopic dimensions. Inspired by recent experiments on free-standing graphene cantilevers, we combine the statistical mechanics of thin elastic plates and large-scale numerical simulations to investigate the thermal renormalization of the bending rigidity of graphene ribbons clamped at one end. For ribbons of dimensions W ×L (with L ≥W ), the macroscopic bending rigidity κR determined from cantilever deformations is independent of the width when W <ℓth , where ℓth is a thermal length scale, as expected. When W >ℓth , however, this thermally renormalized bending rigidity begins to systematically increase, in agreement with the scaling theory, although in our simulations we were not quite able to reach the system sizes necessary to determine the fully developed power law dependence on W . When the ribbon length L >ℓp , where ℓp is the W -dependent thermally renormalized ribbon persistence length, we observe a scaling collapse and the beginnings of large scale random walk behavior.

  20. One-channel Cell-attached Patch-clamp Recording

    PubMed Central

    Maki, Bruce A.; Cummings, Kirstie A.; Paganelli, Meaghan A.; Murthy, Swetha E.; Popescu, Gabriela K.

    2014-01-01

    Ion channel proteins are universal devices for fast communication across biological membranes. The temporal signature of the ionic flux they generate depends on properties intrinsic to each channel protein as well as the mechanism by which it is generated and controlled and represents an important area of current research. Information about the operational dynamics of ion channel proteins can be obtained by observing long stretches of current produced by a single molecule. Described here is a protocol for obtaining one-channel cell-attached patch-clamp current recordings for a ligand gated ion channel, the NMDA receptor, expressed heterologously in HEK293 cells or natively in cortical neurons. Also provided are instructions on how to adapt the method to other ion channels of interest by presenting the example of the mechano-sensitive channel PIEZO1. This method can provide data regarding the channel’s conductance properties and the temporal sequence of open-closed conformations that make up the channel’s activation mechanism, thus helping to understand their functions in health and disease. PMID:24961614

  1. Indentation metrology of clamped, ultra-thin elastic sheets.

    PubMed

    Vella, Dominic; Davidovitch, Benny

    2017-03-06

    We study the indentation of ultrathin elastic sheets clamped to the edge of a circular hole. This classical setup has received considerable attention lately, being used by various experimental groups as a probe to measure the surface properties and stretching modulus of thin solid films. Despite the apparent simplicity of this method, the geometric nonlinearity inherent in the mechanical response of thin solid objects renders the analysis of the resulting data a nontrivial task. Importantly, the essence of this difficulty is in the geometric coupling between in-plane stress and out-of-plane deformations, and hence is present in the behaviour of Hookean solids even when the slope of the deformed membrane remains small. Here we take a systematic approach to address this problem, using the membrane limit of the Föppl-von-Kármán equations. This approach highlights some of the dangers in the use of approximate formulae in the metrology of solid films, which can introduce large errors; we suggest how such errors may be avoided in performing experiments and analyzing the resulting data.

  2. Simultaneous patch-clamping and calcium imaging in developing dendrites.

    PubMed

    Kleindienst, Thomas; Lohmann, Christian

    2014-03-01

    Calcium imaging has been used extensively to explore the role of action potential (AP) firing in the development of neuronal structure and synaptic function because increases in intracellular calcium ([Ca(2+)]i) reliably and, within a certain range, linearly reflect neuronal spiking activity. Patterns of APs in individual cells can be deduced from calcium recordings, which have typically been performed at the level of cell bodies. However, neurons are particularly susceptible to phototoxicity when they are illuminated at the soma. Furthermore, for some imaging experiments (e.g., those that address the interactions between dendrites and axons during synapse formation), the cell body of a given neuron may simply not be in the field of view. In these situations, it would be helpful to determine the spiking patterns of a neuron from the calcium activity in its subcellular compartments such as stretches of dendrites or axons. Here, we describe an approach for determining the relationship between AP firing and dendritic calcium transients by simultaneously imaging calcium transients in small dendritic stretches of hippocampal pyramidal neurons in slice cultures from neonatal rats and recording spiking activity with whole-cell patch-clamp recordings in these neurons. These experiments allow us to correlate the electrophysiological spiking pattern with the accompanying changes in the calcium concentration in individual dendritic segments.

  3. Performance of Li-Ion Cells Under Battery Voltage Charge Control

    NASA Technical Reports Server (NTRS)

    Rao, Gopalakrishna M.; Vaidyanathan, Hari; Day, John H. (Technical Monitor)

    2001-01-01

    A study consisting of electrochemical characterization and Low-Earth-Orbit (LEO) cycling of Li-Ion cells from three vendors was initiated in 1999 to determine the cycling performance and to infuse the new technology in the future NASA missions. The 8-cell batteries included in this evaluation are prismatic cells manufactured by Mine Safety Appliances Company (MSA), cylindrical cells manufactured by SAFT and prismatic cells manufactured by Yardney Technical Products, Inc. (YTP). The three batteries were cycle tested in the LEO regime at 40% depth of discharge, and under a charge control technique that consists of battery voltage clamp with a current taper. The initial testing was conducted at 20 C; however, the batteries were cycled also intermittently at low temperatures. YTP 20 Ah cells consisted of mixed-oxide (Co and Ni) positive, graphitic carbon negative, LIPF6 salt mixed with organic carbonate solvents. The battery voltage clamp was 32 V. The low temperature cycling tests started after 4575 cycles at 20 C. The cells were not capable of cycling. at low temperature since the charge acceptance at battery level was poor. There was a cell in the battery that showed too high an end-of-charge (EOC) voltage thereby limiting the ability to charge the rest of the cells in the battery. The battery has completed 6714 cycles. SAFT 12 Ah cells consisted of mixed-oxide (Co and NO positive, graphitic carbon negative, LiPF6 salt mixed with organic carbonate solvents. The battery voltage clamp was for 30.8 V. The low temperature cycling tests started after 4594 cycles at 20 C. A cell that showed low end of discharge (EOD) and EOC voltages and three other cells that showed higher EOC voltages limited the charge acceptance at the selected voltage limit during charge. The cells were capable of cycling at 10 C and 0 C but the charge voltage limit had to be increased to 34.3 V (4.3 V per cell). The low temperature cycling may have induced poor chargeability since the voltage had to

  4. E-beam high voltage switching power supply

    DOEpatents

    Shimer, Daniel W.; Lange, Arnold C.

    1997-01-01

    A high power, solid state power supply is described for producing a controllable, constant high voltage output under varying and arcing loads suitable for powering an electron beam gun or other ion source. The present power supply is most useful for outputs in a range of about 100-400 kW or more. The power supply is comprised of a plurality of discrete switching type dc-dc converter modules, each comprising a voltage regulator, an inductor, an inverter for producing a high frequency square wave current of alternating polarity, an improved inverter voltage clamping circuit, a step up transformer, and an output rectifier for producing a dc voltage at the output of each module. The inputs to the converter modules are fed from a common dc rectifier/filter and are linked together in parallel through decoupling networks to suppress high frequency input interactions. The outputs of the converter modules are linked together in series and connected to the input of the transmission line to the load through a decoupling and line matching network. The dc-dc converter modules are phase activated such that for n modules, each module is activated equally 360.degree./n out of phase with respect to a successive module. The phased activation of the converter modules, combined with the square current waveforms out of the step up transformers, allows the power supply to operate with greatly reduced output capacitance values which minimizes the stored energy available for discharge into an electron beam gun or the like during arcing. The present power supply also provides dynamic response to varying loads by controlling the voltage regulator duty cycle using simulated voltage feedback signals and voltage feedback loops. Circuitry is also provided for sensing incipient arc currents reflected at the output of the power supply and for simultaneously decoupling the power supply circuitry from the arcing load.

  5. E-beam high voltage switching power supply

    DOEpatents

    Shimer, D.W.; Lange, A.C.

    1997-03-11

    A high power, solid state power supply is described for producing a controllable, constant high voltage output under varying and arcing loads suitable for powering an electron beam gun or other ion source. The present power supply is most useful for outputs in a range of about 100-400 kW or more. The power supply is comprised of a plurality of discrete switching type dc-dc converter modules, each comprising a voltage regulator, an inductor, an inverter for producing a high frequency square wave current of alternating polarity, an improved inverter voltage clamping circuit, a step up transformer, and an output rectifier for producing a dc voltage at the output of each module. The inputs to the converter modules are fed from a common dc rectifier/filter and are linked together in parallel through decoupling networks to suppress high frequency input interactions. The outputs of the converter modules are linked together in series and connected to the input of the transmission line to the load through a decoupling and line matching network. The dc-dc converter modules are phase activated such that for n modules, each module is activated equally 360{degree}/n out of phase with respect to a successive module. The phased activation of the converter modules, combined with the square current waveforms out of the step up transformers, allows the power supply to operate with greatly reduced output capacitance values which minimizes the stored energy available for discharge into an electron beam gun or the like during arcing. The present power supply also provides dynamic response to varying loads by controlling the voltage regulator duty cycle using simulated voltage feedback signals and voltage feedback loops. Circuitry is also provided for sensing incipient arc currents reflected at the output of the power supply and for simultaneously decoupling the power supply circuitry from the arcing load. 7 figs.

  6. High voltage variable diameter insulator

    DOEpatents

    Vanacek, D.L.; Pike, C.D.

    1982-07-13

    A high voltage feedthrough assembly having a tubular insulator extending between the ground plane ring and the high voltage ring. The insulator is made of Pyrex and decreases in diameter from the ground plane ring to the high voltage ring, producing equipotential lines almost perpendicular to the wall of the insulator to optimize the voltage-holding capability of the feedthrough assembly.

  7. Voltage controlled current source

    DOEpatents

    Casne, Gregory M.

    1992-01-01

    A seven decade, voltage controlled current source is described for use in testing intermediate range nuclear instruments that covers the entire test current range of from 10 picoamperes to 100 microamperes. High accuracy is obtained throughout the entire seven decades of output current with circuitry that includes a coordinated switching scheme responsive to the input signal from a hybrid computer to control the input voltage to an antilog amplifier, and to selectively connect a resistance to the antilog amplifier output to provide a continuous output current source as a function of a preset range of input voltage. An operator controlled switch provides current adjustment for operation in either a real-time simulation test mode or a time response test mode.

  8. Electron launching voltage monitor

    DOEpatents

    Mendel, Clifford W.; Savage, Mark E.

    1992-01-01

    An electron launching voltage monitor measures MITL voltage using a relationship between anode electric field and electron current launched from a cathode-mounted perturbation. An electron launching probe extends through and is spaced from the edge of an opening in a first MITL conductor, one end of the launching probe being in the gap between the MITL conductor, the other end being adjacent a first side of the first conductor away from the second conductor. A housing surrounds the launching probe and electrically connects the first side of the first conductor to the other end of the launching probe. A detector detects the current passing through the housing to the launching probe, the detected current being representative of the voltage between the conductors.

  9. Electron launching voltage monitor

    DOEpatents

    Mendel, C.W.; Savage, M.E.

    1992-03-17

    An electron launching voltage monitor measures MITL voltage using a relationship between anode electric field and electron current launched from a cathode-mounted perturbation. An electron launching probe extends through and is spaced from the edge of an opening in a first MITL conductor, one end of the launching probe being in the gap between the MITL conductor, the other end being adjacent a first side of the first conductor away from the second conductor. A housing surrounds the launching probe and electrically connects the first side of the first conductor to the other end of the launching probe. A detector detects the current passing through the housing to the launching probe, the detected current being representative of the voltage between the conductors. 5 figs.

  10. High voltage coaxial switch

    DOEpatents

    Rink, John P.

    1983-07-19

    A coaxial high voltage, high current switch having a solid cylindrical cold cathode coaxially surrounded by a thin hollow cylindrical inner electrode and a larger hollow cylindrical outer electrode. A high voltage trigger between the cathode and the inner electrode causes electrons to be emitted from the cathode and flow to the inner electrode preferably through a vacuum. Some of the electrons penetrate the inner electrode and cause a volumetric discharge in the gas (which may be merely air) between the inner and outer electrodes. The discharge provides a low impedance path between a high voltage charge placed on the outer electrode and a load (which may be a high power laser) coupled to the inner electrode. For high repetition rate the gas between the inner and outer electrodes may be continuously exchanged or refreshed under pressure.

  11. High voltage coaxial switch

    DOEpatents

    Rink, J.P.

    1983-07-19

    A coaxial high voltage, high current switch having a solid cylindrical cold cathode coaxially surrounded by a thin hollow cylindrical inner electrode and a larger hollow cylindrical outer electrode. A high voltage trigger between the cathode and the inner electrode causes electrons to be emitted from the cathode and flow to the inner electrode preferably through a vacuum. Some of the electrons penetrate the inner electrode and cause a volumetric discharge in the gas (which may be merely air) between the inner and outer electrodes. The discharge provides a low impedance path between a high voltage charge placed on the outer electrode and a load (which may be a high power laser) coupled to the inner electrode. For high repetition rate the gas between the inner and outer electrodes may be continuously exchanged or refreshed under pressure. 3 figs.

  12. Voltage Regulators for Photovoltaic Systems

    NASA Technical Reports Server (NTRS)

    Delombard, R.

    1986-01-01

    Two simple circuits developed to provide voltage regulation for highvoltage (i.e., is greater than 75 volts) and low-voltage (i.e., is less than 36 volts) photovoltaic/battery power systems. Use of these circuits results in voltage regulator small, low-cost, and reliable, with very low power dissipation. Simple oscillator circuit controls photovoltaic-array current to regulate system voltage and control battery charging. Circuit senses battery (and system) voltage and adjusts array current to keep battery voltage from exceeding maximum voltage.

  13. Geomagnetism and induced voltage

    NASA Astrophysics Data System (ADS)

    Abdul-Razzaq, W.; Biller, R. D.

    2010-07-01

    Introductory physics laboratories have seen an influx of conceptual integrated science over time in their classrooms with elements of other sciences such as chemistry, biology, Earth science, and astronomy. We describe a laboratory to introduce this development, as it attracts attention to the voltage induced in the human brain as it is initiated by the change in the magnetic flux due to the Earth's magnetic field and movement. This simple and enjoyable experiment will demonstrate how basic concepts in physics and geology can help us think about possible health effects due to the induced voltage.

  14. Peptide Pores in Lipid Bilayers: Voltage Facilitation Pleads for a Revised Model

    NASA Astrophysics Data System (ADS)

    Fadda, G. C.; Lairez, D.; Guennouni, Z.; Koutsioubas, A.

    2013-07-01

    We address the problem of antimicrobial peptides that create pores in lipid bilayers, focusing on voltage-temperature dependence of pore opening. Two novel experiments (voltage clamp with alamethicin as an emblematic representative of these peptides and neutron reflectivity of lipid monolayer at solid-water interface under electric field) serve to revise the only current theoretical model. We introduce a general contribution of peptide adsorption and electric field as being responsible for an unbalanced tension of the two bilayer leaflets and we claim that the main entropy cost of one pore opening is due to the corresponding excluded area for lipid translation.

  15. Motion of a DNA Sliding Clamp Observed by Single Molecule Fluorescence Spectroscopy*S⃞

    PubMed Central

    Laurence, Ted A.; Kwon, Youngeun; Johnson, Aaron; Hollars, Christopher W.; O'Donnell, Mike; Camarero, Julio A.; Barsky, Daniel

    2008-01-01

    DNA sliding clamps attach to polymerases and slide along DNA to allow rapid, processive replication of DNA. These clamps contain many positively charged residues that could curtail the sliding due to attractive interactions with the negatively charged DNA. By single-molecule spectroscopy we have observed a fluorescently labeled sliding clamp (polymerase III β subunit or β clamp) loaded onto freely diffusing, single-stranded M13 circular DNA annealed with fluorescently labeled DNA oligomers of up to 90 bases. We find that the diffusion constant for the β clamp diffusing along DNA is on the order of 10–14 m2/s, at least 3 orders of magnitude less than that for diffusion through water alone. We also find evidence that the β clamp remains at the 3′ end in the presence of Escherichia coli single-stranded-binding protein. These results may imply that the clamp not only acts to hold the polymerase on the DNA but also prevents excessive drifting along the DNA. PMID:18556658

  16. Endovascular balloon versus transthoracic aortic clamping for minimally invasive mitral valve surgery: impact on cerebral microemboli.

    PubMed

    Maselli, Daniele; Pizio, Raffaella; Borelli, Gabriele; Musumeci, Francesco

    2006-04-01

    To evaluate micro embolic events occurrence during minimally invasive mitral valve procedures, comparing balloon endovascular aortic occlusion (Group I) and transthoracic aortic clamping technique (Group II), 36 patients (20 in Group I and 16 in Group II) undergoing minimally invasive mitral valve surgery were selected by CT scan and Doppler studies for absence of atherosclerotic disease at aortic, coronary or peripheral level. Assignment to one of the two groups was made on the basis of surgeon's preference. Continuous automated intra-operative transcranial Doppler was used to monitor micro embolic events during five operative steps: cardiopulmonary bypass (CPB) setup, time interval from CPB start until aortic clamp positioning, first minute after clamp-on, first minute after clamp-off, first ten minutes after CPB weaning start. More embolic events were observed in Group II than in Group I (total 143.4+/-30.6 per patient vs. 78.9+/-28.6 per patient). A large amount of embolic events occurring mainly when the aortic clamp was positioned and released accounted for the observed differences. In a low risk population for embolic events occurrence, endovascular balloon aortic clamping determined less embolic signals than transthoracic aortic clamping.

  17. Generation of sodium hypochlorite (NaOCl) from sodium chloride solution using C/PbO2 and Pb/PbO2 electrodes

    NASA Astrophysics Data System (ADS)

    Ghalwa, Nasser Abu; Tamos, Hassan; ElAskalni, Mohamed; El Agha, Abed Rhman

    2012-06-01

    Two modified electrodes (Pb/PbO2 and C/PbO2) were prepared by electrodepositing a lead oxide layer on lead and carbon substrates. These modified electrodes were used as anodes for the generation of sodium hypochlorite (NaOCl) from sodium chloride solution. Different operating conditions and factors affecting the treatment process of NaOCl generation, including current density, pH values, conductive electrolytes, and electrolysis time, were studied and optimized. By comparison the C/PbO2 electrode shows a higher efficiency than the Pb/PbO2 electrode for the generation of NaOCl.

  18. Performance of Li-Ion Cells Under Battery Voltage Charge Control

    NASA Technical Reports Server (NTRS)

    Vaidyanathan, Hari; Rao, Gopalakrishna M.

    2002-01-01

    Li-ion cells manufactured by YTP, SAFT, and MSA have completed 6714, 6226, and 3441 cycles, respectively. An increase in the charge voltage limit was required in all cases to maintain the discharge voltage. SAFT and MSA cells were capable of cycling at -10 C and 0 C with an increase in the charge voltage limit, whereas Yardney cells could not be cycled. Reconditioning improved the discharge voltage of SAFT and MSA cells; it is important to note that the effect has been temporary as in Ni-H and Ni-Cd batteries. It was demonstrated that the charge operation with VT clamp at battery rather than at cell level is feasible. Continuation of testing depends on the health of the cells and on the funding situation.

  19. Geomagnetism and Induced Voltage

    ERIC Educational Resources Information Center

    Abdul-Razzaq, W.; Biller, R. D.

    2010-01-01

    Introductory physics laboratories have seen an influx of "conceptual integrated science" over time in their classrooms with elements of other sciences such as chemistry, biology, Earth science, and astronomy. We describe a laboratory to introduce this development, as it attracts attention to the voltage induced in the human brain as it…

  20. Measuring Breakdown Voltage.

    ERIC Educational Resources Information Center

    Auer, Herbert J.

    1978-01-01

    The article discusses an aspect of conductivity, one of the electrical properties subdivisions, and describes a tester that can be shop-built. Breakdown voltage of an insulation material is specifically examined. Test procedures, parts lists, diagrams, and test data form are included. (MF)

  1. High Voltage Insulation Technology

    NASA Astrophysics Data System (ADS)

    Scherb, V.; Rogalla, K.; Gollor, M.

    2008-09-01

    In preparation of new Electronic Power Conditioners (EPC's) for Travelling Wave Tub Amplifiers (TWTA's) on telecom satellites a study for the development of new high voltage insulation technology is performed. The initiative is mandatory to allow compact designs and to enable higher operating voltages. In a first task a market analysis was performed, comparing different materials with respect to their properties and processes. A hierarchy of selection criteria was established and finally five material candidates (4 Epoxy resins and 1 Polyurethane resin) were selected to be further investigated in the test program. Samples for the test program were designed to represent core elements of an EPC, the high voltage transformer and Printed Circuit Boards of the high voltage section. All five materials were assessed in the practical work flow of the potting process and electrical, mechanical, thermal and lifetime testing was performed. Although the lifetime tests results were overlayed by a larges scatter, finally two candidates have been identified for use in a subsequent qualification program. This activity forms part of element 5 of the ESA ARTES Programme.

  2. Voltage-Controlled Oscillator

    NASA Technical Reports Server (NTRS)

    1995-01-01

    Integrated Component Systems, Inc. incorporated information from a NASA Tech Briefs article into a voltage-controlled oscillator it designed for a customer. The company then applied the technology to its series of phase-locked loop synthesizers, which offer superior phase noise performance.

  3. [A data interface based on USB bus technology for full auto patch-clamp system].

    PubMed

    Liu, Youlin; Hu, Yang; Qu, Anlian

    2006-04-01

    A USB bus based data interface technology for full auto Patch-Clamp system is discussed in the article. The main controller is CY2131QC (Cypress) and the logic controller is EPM3256A (Altera). Optocouplers are used to get rid of the noise from the interface. It makes the installation of the Patch-Clamp system easier by using the USB bus, and is suitable for the new generation of the Patch-Clamp system with a high speed of 1M bytes/s.

  4. Voltage-Independent Calcium Release in Heart Muscle

    NASA Astrophysics Data System (ADS)

    Niggli, Ernst; Lederer, W. Jonathan

    1990-10-01

    The Ca2+ that activates contraction in heart muscle is regulated as in skeletal muscle by processes that depend on voltage and intracellular Ca2+ and involve a positive feedback system. How the initial electrical signal is amplified in heart muscle has remained controversial, however. Analogous protein structures from skeletal muscle and heart muscle have been identified physiologically and sequenced; these include the Ca2+ channel of the sarcolemma and the Ca2+ release channel of the sarcoplasmic reticulum. Although the parallels found in cardiac and skeletal muscles have provoked valuable experiments in both tissues, separation of the effects of voltage and intracellular Ca2+ on sarcoplasmic reticulum Ca2+ release in heart muscle has been imperfect. With the use of caged Ca2+ and flash photolysis in voltage-clamped heart myocytes, effects of membrane potential in heart muscle cells on Ca2+ release from intracellular stores have been studied. Unlike the response in skeletal muscle, voltage across the sarcolemma of heart muscle does not affect the release of Ca2+ from the sarcoplasmic reticulum, suggesting that other regulatory processes are needed to control Ca2+-induced Ca2+ release.

  5. Resurgent current of voltage-gated Na+ channels

    PubMed Central

    Lewis, Amanda H; Raman, Indira M

    2014-01-01

    Resurgent Na+ current results from a distinctive form of Na+ channel gating, originally identified in cerebellar Purkinje neurons. In these neurons, the tetrodotoxin-sensitive voltage-gated Na+ channels responsible for action potential firing have specialized mechanisms that reduce the likelihood that they accumulate in fast inactivated states, thereby shortening refractory periods and permitting rapid, repetitive, and/or burst firing. Under voltage clamp, step depolarizations evoke transient Na+ currents that rapidly activate and quickly decay, and step repolarizations elicit slower channel reopening, or a ‘resurgent’ current. The generation of resurgent current depends on a factor in the Na+ channel complex, probably a subunit such as NaVβ4 (Scn4b), which blocks open Na+ channels at positive voltages, competing with the fast inactivation gate, and unblocks at negative voltages, permitting recovery from an open channel block along with a flow of current. Following its initial discovery, resurgent Na+ current has been found in nearly 20 types of neurons. Emerging research suggests that resurgent current is preferentially increased in a variety of clinical conditions associated with altered cellular excitability. Here we review the biophysical, molecular and structural mechanisms of resurgent current and their relation to the normal functions of excitable cells as well as pathophysiology. PMID:25172941

  6. The voltage-dependent proton pumping in bacteriorhodopsin is characterized by optoelectric behavior.

    PubMed

    Geibel, S; Friedrich, T; Ormos, P; Wood, P G; Nagel, G; Bamberg, E

    2001-10-01

    The light-driven proton pump bacteriorhodopsin (bR) was functionally expressed in Xenopus laevis oocytes and in HEK-293 cells. The latter expression system allowed high time resolution of light-induced current signals. A detailed voltage clamp and patch clamp study was performed to investigate the DeltapH versus Deltapsi dependence of the pump current. The following results were obtained. The current voltage behavior of bR is linear in the measurable range between -160 mV and +60 mV. The pH dependence is less than expected from thermodynamic principles, i.e., one DeltapH unit produces a shift of the apparent reversal potential of 34 mV (and not 58 mV). The M(2)-BR decay shows a significant voltage dependence with time constants changing from 20 ms at +60 mV to 80 ms at -160 mV. The linear I-V curve can be reconstructed by this behavior. However, the slope of the decay rate shows a weaker voltage dependence than the stationary photocurrent, indicating that an additional process must be involved in the voltage dependence of the pump. A slowly decaying M intermediate (decay time > 100 ms) could already be detected at zero voltage by electrical and spectroscopic means. In effect, bR shows optoelectric behavior. The long-lived M can be transferred into the active photocycle by depolarizing voltage pulses. This is experimentally demonstrated by a distinct charge displacement. From the results we conclude that the transport cycle of bR branches via a long-lived M(1)* in a voltage-dependent manner into a nontransporting cycle, where the proton release and uptake occur on the extracellular side.

  7. Interaction of the β sliding clamp with MutS, ligase, and DNA polymerase I

    PubMed Central

    López de Saro, Francisco J.; O'Donnell, Mike

    2001-01-01

    The β and proliferating cell nuclear antigen (PCNA) sliding clamps were first identified as components of their respective replicases, and thus were assigned a role in chromosome replication. Further studies have shown that the eukaryotic clamp, PCNA, interacts with several other proteins that are involved in excision repair, mismatch repair, cellular regulation, and DNA processing, indicating a much wider role than replication alone. Indeed, the Escherichia coli β clamp is known to function with DNA polymerases II and V, indicating that β also interacts with more than just the chromosomal replicase, DNA polymerase III. This report demonstrates three previously undetected protein–protein interactions with the β clamp. Thus, β interacts with MutS, DNA ligase, and DNA polymerase I. Given the diverse use of these proteins in repair and other DNA transactions, this expanded list of β interactive proteins suggests that the prokaryotic β ring participates in a wide variety of reactions beyond its role in chromosomal replication. PMID:11459978

  8. Shock Response of the Clamped Disk in Small Form Factor Hard Disk Drive

    NASA Astrophysics Data System (ADS)

    Gu, Bin; Shu, Dongwei; Shi, Baojun; Lu, Guoxing

    As small form factor (one-inch and smaller) hard disk drives are widely used in portable consumer appliances and gadgets, their mechanical robustness is of greater concern. In the previous work, it is found that when the disk is more tightly clamped, it helps to decrease the shock response of the disk and then avoid the head slap. In this paper, the real boundary condition of the disk for a small form factor hard disk drive from Seagate is investigated numerically. The disk is clamped between the clamp and the hub. The shock response of the disk under a half-sine acceleration pulse is simulated by using the finite element method. In the finite element model, both contact between disk and clamp and contact between disk and hub are considered. According to the simulation results, how to decrease the shock response of the disk is suggested.

  9. Modeling considerations for the analysis of LMFBR steam generator tube clamps

    SciTech Connect

    Lay, D.M.; Piper, R.M.

    1983-10-01

    In the design of the Babcock and Wilcox Helical Coil Liquid Metal Fast Breeder Reactor (LMFBR) Steam Generator, the tube bundle is connected to the feedwater and steam plenums via ''inlet/outlet tubes.'' Of prime importance in the design of these tubes is the tube-to-tube and tube-to-shell clamps which are provided to prevent detrimental vibration. This paper presents a method of modeling the tubeto-tube clamps to accurately predict tube-to-clamp interaction in the finite element analysis. It is also the objective of this paper to demonstrate the validity of specific modeling assumptions in determining stresses in the clamp assembly. As this paper deals only with the analytical approach taken, no detailed results are presented.

  10. Acoustic plane waves incident on an oblique clamped panel in a rectangular duct

    NASA Technical Reports Server (NTRS)

    Unz, H.; Roskam, J.

    1980-01-01

    The theory of acoustic plane waves incident on an oblique clamped panel in a rectangular duct was developed from basic theoretical concepts. The coupling theory between the elastic vibrations of the panel (plate) and the oblique incident acoustic plane wave in infinite space was considered in detail, and was used for the oblique clamped panel in the rectangular duct. The partial differential equation which governs the vibrations of the clamped panel (plate) was modified by adding to it stiffness (spring) forces and damping forces. The Transmission Loss coefficient and the Noise Reduction coefficient for oblique incidence were defined and derived in detail. The resonance frequencies excited by the free vibrations of the oblique finite clamped panel (plate) were derived and calculated in detail for the present case.

  11. Nanofibrous TiO2 improving performance of mesoporous TiO2 electrode in dye-sensitized solar cell

    NASA Astrophysics Data System (ADS)

    Zukalová, Markéta; Kavan, Ladislav; Procházka, Jan; Zukal, Arnošt; Yum, Jun-Ho; Graetzel, Michael

    2013-05-01

    A method of direct coating of conducting glass by electrospinning was developed. Electrospun fibrous TiO2 consisting of closely packed anatase nanocrystals of 40-50 nm in size was incorporated into mesoporous TiO2 thin film stabilized by phosphorus. The mesoporous framework formed by walls with 5-6 nm TiO2 nanocrystals surrounding 20 nm mesopores exhibits extreme porosity and consequently limited number of necking points. TiO2 with fibrous morphology was found to solidify mesoporous titania and to be beneficial for the performance of corresponding photoanode in dye-sensitized solar cell (DSC). Obviously, its wire-like structure suitably interconnects mesoporous network and thus increases the electron collection efficiency from the TiO2 layer to the F-doped SnO2 electrode. The solar conversion efficiency of a DSC employing optimized photoanode consisting of nanocrystalline fibrous bottom layer, four mesoporous layers, and one nanocrystalline anatase scattering top layer sensitized with the N945 dye reached 5.35 %. This represents an improvement of about 9 % compared to the solar conversion efficiency of a DSC employing purely mesoporous TiO2 layer prepared by means of phosphorus doping (5.05 %).

  12. Pseudocapacitive performance of a solution-processed β-Co(OH)2 electrode monitored through its surface morphology and area.

    PubMed

    Gaikar, P S; Navale, S T; Gaikwad, S L; Al-Osta, Ahmed; Jadhav, V V; Arjunwadkar, P R; Naushad, Mu; Mane, Rajaram S

    2017-03-07

    In the present study, beta-cobalt hydroxide (β-Co(OH)2) electrodes of various nanostructures and surface areas, viz. nano-rhombuses (NRs), nano-plates (NPs), and nano-grass (NGs), have been synthesized directly onto a stainless-steel (SS) substrate using a simple, economical and binder-free chemical solution-process, utilizing three cobalt precursor salts, i.e. cobalt acetate, cobalt chloride, and cobalt nitrate, respectively. Structural elucidation proves the crystallite size, type and phase-purity of β-Co(OH)2, whereas the surface morphology analysis supports the evolution of the above mentioned nanostructures of various surface areas. The electrochemical pseudocapacitor performance investigation demonstrates a specific capacitance (Sc) of 367 F g(-1) at 1 mA cm(-2) for the NP-type morphology, which is higher than that that displayed by the other morphologies. This change in Sc value is attributed to different charge transfer resistance values, which have been obtained from electrochemical impedance spectroscopy spectra. Finally, we attempt to correlate the relationship between the surface morphology, i.e. surface area, and the charge transfer resistance with the obtained specific capacitance value of the respective electrode.

  13. One Step Synthesis of Uniform SnO2 Electrode by UV Curing Technology toward Enhanced Lithium-Ion Storage.

    PubMed

    Wei, Hang; Xia, Zhonghong; Xia, Dingguo

    2017-03-01

    A uniform anode material composed of ultrasmall tin oxide (SnO2) nanoparticles with an excellent lithium-ion (Li-ion) storage performance is obtained for the first time through one step UV curing technology. The diameter of ∼3 nm-sized SnO2 particles is uniformly dispersed in the styrylpyridinium (SbQ) polymer because of its photo-cross-linking property. The in situ cross-linking of SbQ polymer not only assist synthesis of uniform ultrasmall SnO2, but act as a strong adhesion binder on SnO2 nanoparticles, thereby effectively accommodating the volume expansion of SnO2 anodes during cycling process. The uniform electrode exhibits substantially higher specific capacity and longer cycling stability compared with the SnO2 nanoparticles electrodes treated by traditional PVDF-mixing method. A stable specific capacity of 572.5 mA h g(-1) of the SnO2 electrode derived from UV curing technology is obtained at a current density of 0.2 C (156.2 mA g(-1)) after 150 cycles. Even at high rate of 5 C (3905 mA g(-1)), the electrode still demonstrates specific capacity of 440.2 mA h g(-1). Therefore, the scalable and low-cost synthetic approach described herein can readily be extended to other nanomaterials electrodes to improve their lithium-storage properties.

  14. Highly efficient electrochemical degradation of perfluorooctanoic acid (PFOA) by F-doped Ti/SnO2 electrode.

    PubMed

    Yang, Bo; Jiang, Chaojin; Yu, Gang; Zhuo, Qiongfang; Deng, Shubo; Wu, Jinhua; Zhang, Hong

    2015-12-15

    The novel F-doped Ti/SnO2 electrode prepared by SnF4 as the single-source precursor was used for electrochemical degradation of aqueous perfluorooctanoic acid (PFOA). Higher oxidation reactivity and significantly longer service life were achieved for Ti/SnO2-F electrode than Ti/SnO2-X (X=Cl, Br, I, or Sb) electrode, which could decomposed over 99% of PFOA (50 mL of 100 mg L(-1)) within 30-min electrolysis. The property of Ti/SnO2-F electrode and its electrooxidation mechanism were investigated by XRD, SEM-EDX, EIS, LSV, and interfacial resistance measurements. We propose that the similar ionic radii of F and O as well as strong electronegativity of F caused its electrochemical stability with high oxygen evolution potential (OEP) and smooth surface to generate weakly adsorbed OH. The preparation conditions of electrode were also optimized including F doping amount, calcination temperature, and dip coating times, which revealed the formation process of electrode. Additionally, the major mineralization product, F(-), and low concentration of shorter chain perfluorocarboxylic acids (PFCAs) were detected in solution. So the reaction pathway of PFOA electrooxidation was proposed by intermediate analysis. These results demonstrate that Ti/SnO2-F electrode is promising for highly efficient treatment of PFOA in wastewater.

  15. Tenderization of hot-boned broiler breast meat by clamping during chilling.

    PubMed

    Cason, J A; Lyon, C E; Dickens, J A

    2002-01-01

    Hot-boned broiler breast fillets were tightly clamped between rigid aluminum plates during chilling to determine whether tenderness is increased if breast fillets are not allowed to shorten during rigor. In two experiments, 6-wk-old broilers were processed in a pilot plant. Approximately 5 min after evisceration, the breast fillets (pectoralis major) were deboned, and each fillet was subjected to one of two treatments while chilling for 2 h in ice slush. Fillets were placed in perforated plastic bags (hot-boned control) or clamped between rigid aluminum plates that compressed the meat to a uniform thickness of 7.2 mm during chilling. In Experiment 2, chilling time in ice slush was 1 h, and a third treatment was added to make an incomplete block design in which one breast half was left intact on the carcass and was deboned immediately after chilling. All breast fillets were sealed in plastic bags after the chilling period, held overnight at 4 C, and then cooked at 85 C for 30 min in a steam kettle. In Experiment 1, clamping for 2 h reduced Warner-Bratzler shear values of hot-boned fillets from 11.4 to 2.7 kg. In Experiment 2, shear values for the treatments were 13.0, 9.2, and 5.1 kg for the hot-boned, cold-boned, and hot-boned clamped treatments, respectively, with significantly lower shear values for the clamped fillets. Clamped fillets were significantly thinner than the control fillets in both experiments. Cooked yield as a percentage of postchill weight was significantly higher for the clamped compared to the hot-boned control pieces, 81.1 versus 77.3%, with cold-boned pieces being intermediate and not different from the other treatments. Shear values were reduced, and cooked yield was increased by clamping hot-boned fillets during chilling.

  16. Patch Clamp: A Powerful Technique for Studying the Mechanism of Acupuncture

    PubMed Central

    Zhang, D.

    2012-01-01

    Cellular and molecular events can be investigated using electrophysiological techniques. In particular, the patch-clamp method provides detailed information. In addition, the patch-clamp technique has become a powerful method for investigating the mechanisms underlying the effects of acupuncture. In this paper, recent researches on how acupuncture might modulate electrophysiological responses in the central nervous system (CNS) and affect peripheral structures are reviewed. PMID:23133497

  17. The TCF C-clamp DNA binding domain expands the Wnt transcriptome via alternative target recognition

    PubMed Central

    Hoverter, Nate P.; Zeller, Michael D.; McQuade, Miriam M.; Garibaldi, Angela; Busch, Anke; Selwan, Elizabeth M.; Hertel, Klemens J.; Baldi, Pierre; Waterman, Marian L.

    2014-01-01

    LEF/TCFs direct the final step in Wnt/β-catenin signalling by recruiting β-catenin to genes for activation of transcription. Ancient, non-vertebrate TCFs contain two DNA binding domains, a High Mobility Group box for recognition of the Wnt Response Element (WRE; 5′-CTTTGWWS-3′) and the C-clamp domain for recognition of the GC-rich Helper motif (5′-RCCGCC-3′). Two vertebrate TCFs (TCF-1/TCF7 and TCF-4/TCF7L2) use the C-clamp as an alternatively spliced domain to regulate cell-cycle progression, but how the C-clamp influences TCF binding and activity genome-wide is not known. Here, we used a doxycycline inducible system with ChIP-seq to assess how the C-clamp influences human TCF1 binding genome-wide. Metabolic pulse-labeling of nascent RNA with 4′Thiouridine was used with RNA-seq to connect binding to the Wnt transcriptome. We find that the C-clamp enables targeting to a greater number of gene loci for stronger occupancy and transcription regulation. The C-clamp uses Helper sites concurrently with WREs for gene targeting, but it also targets TCF1 to sites that do not have readily identifiable canonical WREs. The coupled ChIP-seq/4′Thiouridine-seq analysis identified new Wnt target genes, including additional regulators of cell proliferation. Thus, C-clamp containing isoforms of TCFs are potent transcriptional regulators with an expanded transcriptome directed by C-clamp-Helper site interactions. PMID:25414359

  18. Clamping of the Linewidth Enhancement Factor in Narrow Quantum-Well GRINSCH Semiconductor Lasers

    SciTech Connect

    Bossert, D.; Chow, W.W.; Hader, J.; Koch, S.W.; Moloney, J.V.; Stohls, J.

    1999-01-20

    The linewidth enhancement factor in single quantum-well GRINSCH semiconductor lasers is investigated theoretically and experimentally. For thin wells a small linewidth enhancement factor is obtained which clamps with increasing carrier density, in contrast to the monotonous increase observed for thicker wells. Microscopic many-body calculations reproduce the experimental observations attributing the clamping to a subtle interplay between excitation dependent gain shifts and carrier population distributions.

  19. Molecular Analyses of a Three-Subunit Euryarchaeal Clamp Loader Complex from Methanosarcina acetivorans▿ †

    PubMed Central

    Chen, Yi-Hsing; Lin, Yuyen; Yoshinaga, Aya; Chhotani, Benazir; Lorenzini, Jenna L.; Crofts, Alexander A.; Mei, Shou; Mackie, Roderick I.; Ishino, Yoshizumi; Cann, Isaac K. O.

    2009-01-01

    Chromosomal DNA replication is dependent on processive DNA synthesis. Across the three domains of life and in certain viruses, a toroidal sliding clamp confers processivity to replicative DNA polymerases by encircling the DNA and engaging the polymerase in protein/protein interactions. Sliding clamps are ring-shaped; therefore, they have cognate clamp loaders that open and load them onto DNA. Here we use biochemical and mutational analyses to study the structure/function of the Methanosarcina acetivorans clamp loader or replication factor C (RFC) homolog. M. acetivorans RFC (RFCMa), which represents an intermediate between the common archaeal RFC and the eukaryotic RFC, comprises two different small subunits (RFCS1 and RFCS2) and a large subunit (RFCL). Size exclusion chromatography suggested that RFCS1 exists in oligomeric states depending on protein concentration, while RFCS2 exists as a monomer. Protein complexes of RFCS1/RFCS2 formed in solution; however, they failed to stimulate DNA synthesis by a cognate DNA polymerase in the presence of its clamp. Determination of the subunit composition and previous mutational analysis allowed the prediction of the spatial distribution of subunits in this new member of the clamp loader family. Three RFCS1 subunits are flanked by an RFCS2 and an RFCL. The spatial distribution is, therefore, reminiscent of the minimal Escherichia coli clamp loader that exists in space as three γ-subunits (motor) flanked by the δ′ (stator) and the δ (wrench) subunits. Mutational analysis, however, suggested that the similarity between the two clamp loaders does not translate into the complete conservation of the functions of individual subunits within the RFCMa complex. PMID:19717601

  20. Molecular modeling-based analysis of interactions in the RFC-dependent clamp-loading process.

    PubMed

    Venclovas, Ceslovas; Colvin, Michael E; Thelen, Michael P

    2002-10-01

    Replication and related processes in eukaryotic cells require replication factor C (RFC) to load a molecular clamp for DNA polymerase in an ATP-driven process, involving multiple molecular interactions. The detailed understanding of this mechanism is hindered by the lack of data regarding structure, mutual arrangement, and dynamics of the players involved. In this study, we analyzed interactions that take place during loading onto DNA of either the PCNA clamp or the Rad9-Rad1-Hus1 checkpoint complex, using computationally derived molecular models. Combining the modeled structures for each RFC subunit with known structural, biochemical, and genetic data, we propose detailed models of how two of the RFC subunits, RFC1 and RFC3, interact with the C-terminal regions of PCNA. RFC1 is predicted to bind PCNA similarly to the p21-PCNA interaction, while the RFC3-PCNA binding is proposed to be similar to the E. coli delta-beta interaction. Additional sequence and structure analysis, supported by experimental data, suggests that RFC5 might be the third clamp loader subunit to bind the equivalent PCNA region. We discuss functional implications stemming from the proposed model of the RFC1-PCNA interaction and compare putative clamp-interacting regions in RFC1 and its paralogs, Rad17 and Ctf18. Based on the individual intermolecular interactions, we propose RFC and PCNA arrangement that places three RFC subunits in association with each of the three C-terminal regions in PCNA. The two other RFC subunits are positioned at the two PCNA interfaces, with the third PCNA interface left unobstructed. In addition, we map interactions at the level of individual subunits between the alternative clamp loader/clamp system, Rad17-RFC(2-5)/Rad9-Rad1-Hus1. The proposed models of interaction between two clamp/clamp loader pairs provide both structural framework for interpretation of existing experimental data and a number of specific findings that can be subjected to direct experimental

  1. High Voltage Seismic Generator

    NASA Astrophysics Data System (ADS)

    Bogacz, Adrian; Pala, Damian; Knafel, Marcin

    2015-04-01

    This contribution describes the preliminary result of annual cooperation of three student research groups from AGH UST in Krakow, Poland. The aim of this cooperation was to develop and construct a high voltage seismic wave generator. Constructed device uses a high-energy electrical discharge to generate seismic wave in ground. This type of device can be applied in several different methods of seismic measurement, but because of its limited power it is mainly dedicated for engineering geophysics. The source operates on a basic physical principles. The energy is stored in capacitor bank, which is charged by two stage low to high voltage converter. Stored energy is then released in very short time through high voltage thyristor in spark gap. The whole appliance is powered from li-ion battery and controlled by ATmega microcontroller. It is possible to construct larger and more powerful device. In this contribution the structure of device with technical specifications is resented. As a part of the investigation the prototype was built and series of experiments conducted. System parameter was measured, on this basis specification of elements for the final device were chosen. First stage of the project was successful. It was possible to efficiently generate seismic waves with constructed device. Then the field test was conducted. Spark gap wasplaced in shallowborehole(0.5 m) filled with salt water. Geophones were placed on the ground in straight line. The comparison of signal registered with hammer source and sparker source was made. The results of the test measurements are presented and discussed. Analysis of the collected data shows that characteristic of generated seismic signal is very promising, thus confirms possibility of practical application of the new high voltage generator. The biggest advantage of presented device after signal characteristics is its size which is 0.5 x 0.25 x 0.2 m and weight approximately 7 kg. This features with small li-ion battery makes

  2. Increased voltage photovoltaic cell

    NASA Technical Reports Server (NTRS)

    Ross, B.; Bickler, D. B.; Gallagher, B. D. (Inventor)

    1985-01-01

    A photovoltaic cell, such as a solar cell, is provided which has a higher output voltage than prior cells. The improved cell includes a substrate of doped silicon, a first layer of silicon disposed on the substrate and having opposite doping, and a second layer of silicon carbide disposed on the first layer. The silicon carbide preferably has the same type of doping as the first layer.

  3. Insulators for high voltages

    SciTech Connect

    Looms, J.S.T.

    1987-01-01

    This book describes electrical insulators for high voltage applications. Topics considered include the insulating materials, the manufacture of wet process porcelain, the manufacture of tempered glass, the glass-fibre core, the polymeric housing, the common problem - terminating an insulator, mechanical constraints, the physics of pollution flashover, the physics of contamination, testing of insulators, conclusions from testing, remedies for flashover, insulators for special cases, interference and noise, and the insulator of the future.

  4. High voltage generator

    DOEpatents

    Schwemin, A. J.

    1959-03-17

    A generator for producing relatively large currents at high voltages is described. In general, the invention comprises a plurality of capacitors connected in series by a plurality of switches alternately disposed with the capacitors. The above-noted circuit is mounted for movement with respect to contact members and switch closure means so that a load device and power supply are connected across successive numbers of capacitors, while the other capacitors are successively charged with the same power supply.

  5. High voltage pulse conditioning

    DOEpatents

    Springfield, Ray M.; Wheat, Jr., Robert M.

    1990-01-01

    Apparatus for conditioning high voltage pulses from particle accelerators in order to shorten the rise times of the pulses. Flashover switches in the cathode stalk of the transmission line hold off conduction for a determinable period of time, reflecting the early portion of the pulses. Diodes upstream of the switches divert energy into the magnetic and electrostatic storage of the capacitance and inductance inherent to the transmission line until the switches close.

  6. HIGH VOLTAGE GENERATOR

    DOEpatents

    Schwemin, A.J.

    1959-03-17

    A generator is presented for producing relatively large currents at high voltages. In general, the invention comprises a plurality of capacitors connected in series by a plurality of switches alternately disposed with the capacitors. The circuit is mounted for movement with respect to contact members and switch closure means so that a load device and power supply are connected across successive numbers of capacitors, while the other capacitors are successively charged with the same power supply.

  7. HIGH VOLTAGE ION SOURCE

    DOEpatents

    Luce, J.S.

    1960-04-19

    A device is described for providing a source of molecular ions having a large output current and with an accelerated energy of the order of 600 kv. Ions are produced in an ion source which is provided with a water-cooled source grid of metal to effect maximum recombination of atomic ions to molecular ions. A very high accelerating voltage is applied to withdraw and accelerate the molecular ions from the source, and means are provided for dumping the excess electrons at the lowest possible potentials. An accelerating grid is placed adjacent to the source grid and a slotted, grounded accelerating electrode is placed adjacent to the accelerating grid. A potential of about 35 kv is maintained between the source grid and accelerating grid, and a potential of about 600 kv is maintained between the accelerating grid and accelerating electrode. In order to keep at a minimum the large number of oscillating electrons which are created when such high voltages are employed in the vicinity of a strong magnetic field, a plurality of high voltage cascaded shields are employed with a conventional electron dumping system being employed between each shield so as to dump the electrons at the lowest possible potential rather than at 600 kv.

  8. Effect of non-ideal clamping shape on the resonance frequencies of silicon nanocantilevers.

    PubMed

    Guillon, Samuel; Saya, Daisuke; Mazenq, Laurent; Perisanu, Sorin; Vincent, Pascal; Lazarus, Arnaud; Thomas, Olivier; Nicu, Liviu

    2011-06-17

    In this paper, we investigate the effects of non-ideal clamping shapes on the dynamic behavior of silicon nanocantilevers. We fabricated silicon nanocantilevers using silicon on insulator (SOI) wafers by employing stepper ultraviolet (UV) lithography, which permits a resolution of under 100 nm. The nanocantilevers were driven by electrostatic force inside a scanning electron microscope (SEM). Both lateral and out-of-plane resonance frequencies were visually detected with the SEM. Next, we discuss overhanging of the cantilever support and curvature at the clamping point in the silicon nanocantilevers, which generally arises in the fabrication process. We found that the fundamental out-of-plane frequency of a realistically clamped cantilever is always lower than that for a perfectly clamped cantilever, and depends on the cantilever width and the geometry of the clamping point structure. Using simulation with the finite-elements method, we demonstrate that this discrepancy is attributed to the particular geometry of the clamping point (non-zero joining curvatures and a flexible overhanging) that is obtained in the fabrication process. The influence of the material orthotropy is also investigated and is shown to be negligible.

  9. A novel monolithic piezoelectric actuated flexure-mechanism based wire clamp for microelectronic device packaging

    NASA Astrophysics Data System (ADS)

    Liang, Cunman; Wang, Fujun; Tian, Yanling; Zhao, Xingyu; Zhang, Hongjie; Cui, Liangyu; Zhang, Dawei; Ferreira, Placid

    2015-04-01

    A novel monolithic piezoelectric actuated wire clamp is presented in this paper to achieve fast, accurate, and robust microelectronic device packaging. The wire clamp has compact, flexure-based mechanical structure and light weight. To obtain large and robust jaw displacements and ensure parallel jaw grasping, a two-stage amplification composed of a homothetic bridge type mechanism and a parallelogram leverage mechanism was designed. Pseudo-rigid-body model and Lagrange approaches were employed to conduct the kinematic, static, and dynamic modeling of the wire clamp and optimization design was carried out. The displacement amplification ratio, maximum allowable stress, and natural frequency were calculated. Finite element analysis (FEA) was conducted to evaluate the characteristics of the wire clamp and wire electro discharge machining technique was utilized to fabricate the monolithic structure. Experimental tests were carried out to investigate the performance and the experimental results match well with the theoretical calculation and FEA. The amplification ratio of the clamp is 20.96 and the working mode frequency is 895 Hz. Step response test shows that the wire clamp has fast response and high accuracy and the motion resolution is 0.2 μm. High speed precision grasping operations of gold and copper wires were realized using the wire clamper.

  10. APPARATUS FOR REGULATING HIGH VOLTAGE

    DOEpatents

    Morrison, K.G.

    1951-03-20

    This patent describes a high-voltage regulator of the r-f type wherein the modulation of the r-f voltage is accomplished at a high level, resulting in good stabilization over a large range of load conditions.

  11. Analysis of impactor residues in tray clamps from the Long Duration Exposure Facility. Part 2: Clamps from Bay B of the satellite

    NASA Technical Reports Server (NTRS)

    Bernhard, Ronald P.; Zolensky, Michael E.

    1994-01-01

    The Long Duration Exposure Facility (LDEF) was placed in low-Earth orbit (LEO) in 1984 and recovered 5.7 years later. The LDEF was host to several individual experiments specifically designed to characterize critical aspects of meteoroid and debris environment in LEO. However, it was realized from the beginning that the most efficient use of the satellite would be to examine the entire surface for impact features. In this regard, particular interest centered on common exposed materials that faced in all LDEF pointing directions. Among the most important of these materials was the tray clamps. Therefore, in an effort to better understand the nature of particulates in LEO and their effects on spacecraft hardware, residues found in impact features on LDEF tray clamp surfaces are being analyzed. This catalog presents all data from clamps from Bay B of the LDEF. NASA Technical Memorandum 104759 has cataloged impacts that occurred on Bay B (published March 1993). Subsequent catalogs will include clamps from succeeding bays of the satellite.

  12. High voltage variable diameter insulator

    DOEpatents

    Vanecek, David L.; Pike, Chester D.

    1984-01-01

    A high voltage feedthrough assembly (10) having a tubular insulator (15) extending between the ground plane ring (16) and the high voltage ring (30). The insulator (15) is made of Pyrex and decreases in diameter from the ground plane ring (16) to the high voltage ring (30), producing equipotential lines almost perpendicular to the wall (27) of the insulator (15) to optimize the voltage-holding capability of the feedthrough assembly (10).

  13. Modulation of voltage-gated sodium channels hyperpolarizes the voltage threshold for activation in spinal motoneurones.

    PubMed

    Power, Kevin E; Carlin, Kevin P; Fedirchuk, Brent

    2012-03-01

    Previous work has shown that motoneurone excitability is enhanced by a hyperpolarization of the membrane potential at which an action potential is initiated (V(th)) at the onset, and throughout brainstem-evoked fictive locomotion in the adult decerebrate cat and neonatal rat. Modeling work has suggested the modulation of Na(+) conductance as a putative mechanism underlying this state-dependent change in excitability. This study sought to determine whether modulation of voltage-gated sodium channels could induce V(th) hyperpolarization. Whole-cell patch-clamp recordings were made from antidromically identified lumbar spinal motoneurones in an isolated neonatal rat spinal cord preparation. Recordings were made with and without the bath application of veratridine, a plant alkaloid neurotoxin that acts as a sodium channel modulator. As seen in HEK 293 cells expressing Nav1.2 channels, veratridine-modified channels demonstrated a hyperpolarizing shift in their voltage-dependence of activation and a slowing of inactivation that resulted in an enhanced inward current in response to voltage ramp stimulations. In the native rat motoneurones, veratridine-modified sodium channels induced a hyperpolarization of V(th) in all 29 neonatal rat motoneurones examined (mean hyperpolarization: -6.6 ± 4.3 mV). V(th) hyperpolarization was not due to the effects on Ca(2+) and/or K(+) channels as blockade of these currents did not alter V(th). Veratridine also significantly increased the amplitude of persistent inward currents (PICs; mean increase: 72.5 ± 98.5 pA) evoked in response to slow depolarizing current ramps. However, the enhancement of the PIC amplitude had a slower time course than the hyperpolarization of V(th), and the PIC onset voltage could be either depolarized or hyperpolarized, suggesting that PIC facilitation did not mediate the V(th) hyperpolarization. We therefore suggest that central neuronal circuitry in mammals could affect V(th) in a mechanism similar to that of

  14. Charge-pump voltage converter

    DOEpatents

    Brainard, John P.; Christenson, Todd R.

    2009-11-03

    A charge-pump voltage converter for converting a low voltage provided by a low-voltage source to a higher voltage. Charge is inductively generated on a transfer rotor electrode during its transit past an inductor stator electrode and subsequently transferred by the rotating rotor to a collector stator electrode for storage or use. Repetition of the charge transfer process leads to a build-up of voltage on a charge-receiving device. Connection of multiple charge-pump voltage converters in series can generate higher voltages, and connection of multiple charge-pump voltage converters in parallel can generate higher currents. Microelectromechanical (MEMS) embodiments of this invention provide a small and compact high-voltage (several hundred V) voltage source starting with a few-V initial voltage source. The microscale size of many embodiments of this invention make it ideally suited for MEMS- and other micro-applications where integration of the voltage or charge source in a small package is highly desirable.

  15. Late cardiac sodium current can be assessed using automated patch-clamp

    PubMed Central

    Gawali, Vaibhavkumar; Todt, Hannes; Knott, Thomas; Scheel, Olaf; Abriel, Hugues

    2014-01-01

    The cardiac late Na + current is generated by a small fraction of voltage-dependent Na + channels that undergo a conformational change to a burst-gating mode, with repeated openings and closures during the action potential (AP) plateau. Its magnitude can be augmented by inactivation-defective mutations, myocardial ischemia, or prolonged exposure to chemical compounds leading to drug-induced (di)-long QT syndrome, and results in an increased susceptibility to cardiac arrhythmias. Using CytoPatch™ 2 automated patch-clamp equipment, we performed whole-cell recordings in HEK293 cells stably expressing human Nav1.5, and measured the late Na + component as average current over the last 100 ms of 300 ms depolarizing pulses to -10 mV from a holding potential of -100 mV, with a repetition frequency of 0.33 Hz. Averaged values in different steady-state experimental conditions were further corrected by the subtraction of current average during the application of tetrodotoxin (TTX) 30 μM. We show that ranolazine at 10 and 30 μM in 3 min applications reduced the late Na + current to 75.0 ± 2.7% (mean ± SEM, n = 17) and 58.4 ± 3.5% ( n = 18) of initial levels, respectively, while a 5 min application of veratridine 1 μM resulted in a reversible current increase to 269.1 ± 16.1% ( n = 28) of initial values. Using fluctuation analysis, we observed that ranolazine 30 μM decreased mean open probability p from 0.6 to 0.38 without modifying the number of active channels n, while veratridine 1 μM increased n 2.5-fold without changing p. In human iPSC-derived cardiomyocytes, veratridine 1 μM reversibly increased APD90 2.12 ± 0.41-fold (mean ± SEM, n = 6). This effect is attributable to inactivation removal in Nav1.5 channels, since significant inhibitory effects on hERG current were detected at higher concentrations in hERG-expressing HEK293 cells, with a 28.9 ± 6.0% inhibition (mean ± SD, n = 10) with 50 μM veratridine.        PMID:25383189

  16. Thin Film Nanocrystalline TiO2 Electrodes: Dependence of Flat Band Potential on pH and Anion Adsorption.

    PubMed

    Minella, M; Maurino, V; Minero, C; Pelizzetti, E

    2015-05-01

    Thin nanocrystalline TiO2 films were produced on ITO conductive glass by dip-coating of a sol-gel TiO2 precursor. The transparent films were characterized from the optical and structural point of view with UV-Vis, Spectroscopic Ellipsometry, Raman and X-ray photoelectron spectroscopies, the roughness of the coating by AFM. The changes in the electrochemical properties features of ITO/TiO2 electrodes were evaluated in the presence of different electrolytes (KCI, Na2SO4 and phosphate buffer) with the aim to clarify the role of the ion adsorption on the structure of the electrical double layer. Electrochemical tests (Cyclic Voltammetry, CV, and Impedance Electrochemical Spectroscopy, EIS) showed a strong influence of the electrolyte properties on the semiconductor band edge position in the electrochemical scale and on band bending. The CV profiles recorded can be explained by considering that the interface capacity is due to the charging of surface states (e.g., Ti(IV) surface sites coordinated by oxygen atoms, ≡Ti-OH or Ti-O-Ti). The surface charge is strongly affected also by the density and nature of adsorbed ions and by dissociation of surficial OH. Of interest the fact that for the produced nanocrystalline electrodes the flat band potential, measured from the Mott-Schottky analysis of the space charge layer capacity obtained with EIS, showed a non Nernstian behavior with the pH probably caused by a change in the surface acidity as a consequence of specific anion adsorption. The modulation of flat band potential with adsorbed ions is of interest for many applications, in particular for photocatalysis (change in the redox potential of photogenerated carriers) and for photovoltaic applications like DSSC (change in the photopotentials).

  17. Photovoltaic conversion using Zn chlorophyll derivative assembled in hydrophobic domain onto nanocrystalline TiO2 electrode.

    PubMed

    Amao, Yutaka; Yamada, Yuriko

    2007-02-15

    Photovoltaic conversion using zinc chlorin-e6 (ZnChl-e6), which is zinc chlorophyll-a derivative, and fatty acid (myristic acid or cholic acid) co-adsorbed nanocrystalline TiO2 layer onto ITO glass (OTE) electrode is developed. The maximum peaks of photocurrent action spectrum of the ZnChl-e6 adsorbed TiO2 layer onto OTE (ZnChl-e6/TiO2) are 400, 660 and 800 nm, respectively. Especially the IPCE value at 800 nm (7.5%) is larger than that of 660 nm (6.9%). This result indicates that ZnChl-e6 molecules is aggregated or formed dimer on a nanocrystalline TiO2 layer onto OTE and the absorption band is shifted to near IR region. The photocurrent action spectrum of ZnChl-e6 and cholic acid adsorbed TiO2 layer onto OTE (ZnChl-e6-Cho/TiO2 is similar to that of the UV-vis absorption spectrum in methanol solution, and IPCE values at 400 and 660 nm (8.1%) increase and the IPCE value at 800 nm (4.1%) decreases, indicating that the aggregation of ZnChl-e6 molecules on the TiO2 is suppressed by cholic acid. By using ZnChl-e6-Cho/TiO2, the short-circuit photocurrent density and open-circuit photovoltage also increase compared with that of ZnChl-e6 adsorbed nanocrystalline TiO2 electrode.

  18. Improvement in cardioplegic perfusion technique in single aortic clamping - initial results

    PubMed Central

    Sobral, Marcelo Luiz Peixoto; dos Santos Júnior, Sérgio Francisco; de Sá, Juliano Cavalcante; Terrazas, Anderson da Silva; Trompieri, Daniel Francisco de Mendonça; de Sousa, Thierry Araújo Nunes; dos Santos, Gilmar Geraldo; Stolf, Noedir Antonio Groppo

    2014-01-01

    Introduction The most common method used for myocardial protection is administering cardioplegic solution in the coronary circulation. Nevertheless, protection may be achieved by intermittent perfusion of the coronary system with patient's own blood. The intermittent perfusion may be performed by multiple sequences of clamping and opening of the aortic clamp or due single clamping and accessory cannulation of the aortic root as in the improved technique proposed in this study, reperfusion without the need for multiple clamping of the aorta. Objective To evaluate the clinical outcome and the occurrence of neurological events in in-hospital patients submitted to myocardial revascularization surgery with the "improved technique" of intermittent perfusion of the aortic root with single clamping. Methods This is a prospective, cross-sectional, observational study that describes a myocardial management technique that consists of intermittent perfusion of the aortic root with single clamping in which 50 patients (mean age 58.5±7.19 years old) have been submitted to the myocardial revasculrization surgery under the proposed technique. Clinical and laboratory variables, pre- and post-surgery, have been assessed. Results The mean peak level of post-surgery CKMB was 51.64±27.10 U/L in the second post-surgery and of troponin I was 3.35±4.39 ng/ml in the fourth post-surgery, within normal limits. No deaths have occurred and one patient presented mild neurological disorder. Hemodynamic monitoring has not indicated any changes. Conclusion The myocardial revascularization surgery by perfusion with the improved technique with intermittent aortic root with single clamping proved to be safe, enabling satisfactory clinical results. PMID:25140473

  19. A central swivel point in the RFC clamp loader controls PCNA opening and loading on DNA.

    PubMed

    Sakato, Miho; O'Donnell, Mike; Hingorani, Manju M

    2012-02-17

    Replication factor C (RFC) is a five-subunit complex that loads proliferating cell nuclear antigen (PCNA) clamps onto primer-template DNA (ptDNA) during replication. RFC subunits belong to the AAA(+) superfamily, and their ATPase activity drives interactions between the clamp loader, the clamp, and the ptDNA, leading to topologically linked PCNA·ptDNA. We report the kinetics of transient events in Saccharomyces cerevisiae RFC-catalyzed PCNA loading, including ATP-induced RFC activation, PCNA opening, ptDNA binding, ATP hydrolysis, PCNA closing, and PCNA·ptDNA release. This detailed perspective enables assessment of individual RFC-A, RFC-B, RFC-C, RFC-D, and RFC-E subunit functions in the reaction mechanism. Functions have been ascribed to RFC subunits previously based on a steady-state analysis of 'arginine-finger' ATPase mutants; however, pre-steady-state analysis provides a different view. The central subunit RFC-C serves as a critical swivel point in the clamp loader. ATP binding to this subunit initiates RFC activation, and the clamp loader adopts a spiral conformation that stabilizes PCNA in a corresponding open spiral. The importance of RFC subunit response to ATP binding decreases as RFC-C>RFC-D>RFC-B, with RFC-A being unnecessary. RFC-C-dependent activation of RFC also enables ptDNA binding, leading to the formation of the RFC·ATP·PCNA(open)·ptDNA complex. Subsequent ATP hydrolysis leads to complex dissociation, with RFC-D activity contributing the most to rapid ptDNA release. The pivotal role of the RFC-B/C/D subunit ATPase core in clamp loading is consistent with the similar central location of all three ATPase active subunits of the Escherichia coli clamp loader.

  20. Effects of baclofen on mechanical noxious and innocuous transmission in the spinal dorsal horn of the adult rat: in vivo patch-clamp analysis.

    PubMed

    Fukuhara, Kaori; Katafuchi, Toshihiko; Yoshimura, Megumu

    2013-11-01

    The effects of a GABAB agonist, baclofen, on mechanical noxious and innocuous synaptic transmission in the substantia gelatinosa (SG) were investigated in adult rats with the in vivo patch-clamp technique. Under current-clamp conditions, perfusion with baclofen (10 μm) on the surface of the spinal cord caused hyperpolarisation of SG neurons and a decrease in the number of action potentials elicited by pinch and touch stimuli applied to the receptive field of the ipsilateral hindlimb. The suppression of action potentials was preserved under blockade of postsynaptic G-proteins, although baclofen-induced hyperpolarisation was completely blocked. These findings suggest presynaptic effects of baclofen on the induced action potentials. Under voltage-clamp conditions, application of baclofen reduced the frequency, but not the amplitude, of miniature excitatory postsynaptic currents (mEPSCs), whereas the GABAB receptor antagonist CGP55845 increased the frequency of mEPSCs without affecting the amplitude. Furthermore, application of a GABA uptake inhibitor, nipecotic acid, decreased the frequency of mEPSCs; this effect was blocked by CGP55845, but not by the GABAA antagonist bicuculline. Both the frequency and the amplitude of the pinch-evoked barrage of excitatory postsynaptic currents (EPSCs) were suppressed by baclofen in a dose-dependent manner. The frequency and amplitude of touch-evoked EPSCs was also suppressed by baclofen, but the suppression was significantly smaller than that of pinch-evoked EPSCs. We conclude that mechanical noxious transmission is presynaptically blocked through GABAB receptors in the SG, and is more effectively suppressed than innocuous transmission, which may account for a part of the mechanism of the efficient analgesic effects of baclofen.

  1. Direct GABAergic and glycinergic inhibition of the substantia gelatinosa from the rostral ventromedial medulla revealed by in vivo patch-clamp analysis in rats.

    PubMed

    Kato, Go; Yasaka, Toshiharu; Katafuchi, Toshihiko; Furue, Hidemasa; Mizuno, Masaharu; Iwamoto, Yukihide; Yoshimura, Megumu

    2006-02-08

    Stimulation of the rostral ventromedial medulla (RVM) is believed to exert analgesic effects through the activation of the serotonergic system descending to the spinal dorsal horn; however, how nociceptive transmission is modulated by the descending system has not been fully clarified. To investigate the inhibitory mechanisms affected by the RVM, an in vivo patch-clamp technique was used to record IPSCs from the substantia gelatinosa (SG) of the spinal cord evoked by chemical (glutamate injection) and electrical stimulation (ES) of the RVM in adult rats. In the voltage-clamp mode, the RVM glutamate injection and RVM-ES produced an increase in both the frequency and amplitude of IPSCs in SG neurons that was not blocked by glutamate receptor antagonists. Serotonin receptor antagonists were unexpectedly without effect, but a GABAA receptor antagonist, bicuculline, or a glycine receptor antagonist, strychnine, completely suppressed the RVM stimulation-induced increase in IPSCs. The RVM-ES-evoked IPSCs showed fixed latency and no failure at 20 Hz stimuli with a conduction velocity of >3 m/s (3.1-20.7 m/s), suggesting descending monosynaptic GABAergic and/or glycinergic inputs from the RVM to the SG through myelinated fibers. In the current-clamp mode, action potentials elicited by noxious mechanical stimuli applied to the receptive field of the ipsilateral hindlimb were suppressed by the RVM-ES in more than half of the neurons tested (63%; 10 of 16). These findings suggest that the RVM-mediated antinociceptive effects on noxious inputs to the SG may be exerted preferentially by the direct GABAergic and glycinergic pathways to the SG.

  2. Ion channel drug discovery and research: the automated Nano-Patch-Clamp technology.

    PubMed

    Brueggemann, A; George, M; Klau, M; Beckler, M; Steindl, J; Behrends, J C; Fertig, N

    2004-01-01

    Unlike the genomics revolution, which was largely enabled by a single technological advance (high throughput sequencing), rapid advancement in proteomics will require a broader effort to increase the throughput of a number of key tools for functional analysis of different types of proteins. In the case of ion channels -a class of (membrane) proteins of great physiological importance and potential as drug targets- the lack of adequate assay technologies is felt particularly strongly. The available, indirect, high throughput screening methods for ion channels clearly generate insufficient information. The best technology to study ion channel function and screen for compound interaction is the patch clamp technique, but patch clamping suffers from low throughput, which is not acceptable for drug screening. A first step towards a solution is presented here. The nano patch clamp technology, which is based on a planar, microstructured glass chip, enables automatic whole cell patch clamp measurements. The Port-a-Patch is an automated electrophysiology workstation, which uses planar patch clamp chips. This approach enables high quality and high content ion channel and compound evaluation on a one-cell-at-a-time basis. The presented automation of the patch process and its scalability to an array format are the prerequisites for any higher throughput electrophysiology instruments.

  3. Transistor voltage comparator performs own sensing

    NASA Technical Reports Server (NTRS)

    Cliff, R. A.

    1965-01-01

    Detection of the highest voltage input among a group of varying voltage inputs is accomplished by a transistorized voltage comparison circuit. The collector circuits of the transistors perform the sensing function. Input voltage levels are governed by the transistors.

  4. A Tight-Seal Whole Cell Study of the Voltage-Dependent Gating Mechanism of K+-Channels of Protoplasmic Droplets of Chara corallina1

    PubMed Central

    Homblé, Fabrice

    1987-01-01

    The biophysical properties of voltage-dependent K+-channels of protoplasmic droplets of Chara corallina Klein ex Willd., em, R.D.W. were investigated using the tight-seal whole cell method. Two potassium currents were observed in voltage-clamp mode and they can be used to explain the transient membrane potential time course observed in current-clamp mode. The K+-channels are identified by the effect of tetraethylammonium chloride which blocks both currents. A two-state, constant dipole moment model is used to fit the voltage-conductance curve. From this model the minimum equivalent gating charge involved in the gating mechanism of K+-channels of Chara can be estimated. PMID:16665457

  5. An Induction Motor Drive System Based on the Back-To-Back Connection of Two Five-Level Diode-Clamped PWM Converters

    NASA Astrophysics Data System (ADS)

    Kondo, Yosuke; Natchpong, Hatti; Akagi, Hirofumi

    This paper discusses a 6.6-kV transformerless motor drive system based on the back-to-back connection of two five-level diode-clamped PWM converters. In addition, this paper designs, constructs and experimentally tests a down-scaled model rated at 200V and 5.5kW. This system consist of the two five-level converters, a common dc-link, and voltage-balancing circuits using two bi-directional buck-boost choppers. Experimental results obtained from the down-scaled model verify the validity of the system design, giving promise of the viability of the 6.6-kV transformerless motor drive system.

  6. High voltage feedthrough bushing

    DOEpatents

    Brucker, John P.

    1993-01-01

    A feedthrough bushing for a high voltage diode provides for using compression sealing for all sealing surfaces. A diode assembly includes a central conductor extending through the bushing and a grading ring assembly circumferentially surrounding and coaxial with the central conductor. A flexible conductive plate extends between and compressively seals against the central conductor and the grading ring assembly, wherein the flexibility of the plate allows inner and outer portions of the plate to axially translate for compression sealing against the central conductor and the grading ring assembly, respectively. The inner portion of the plate is bolted to the central conductor for affecting sealing. A compression beam is also bolted to the central conductor and engages the outer portion of the plate to urge the outer portion toward the grading ring assembly to obtain compression sealing therebetween.

  7. High voltage isolation transformer

    NASA Technical Reports Server (NTRS)

    Clatterbuck, C. H.; Ruitberg, A. P. (Inventor)

    1985-01-01

    A high voltage isolation transformer is provided with primary and secondary coils separated by discrete electrostatic shields from the surfaces of insulating spools on which the coils are wound. The electrostatic shields are formed by coatings of a compound with a low electrical conductivity which completely encase the coils and adhere to the surfaces of the insulating spools adjacent to the coils. Coatings of the compound also line axial bores of the spools, thereby forming electrostatic shields separating the spools from legs of a ferromagnetic core extending through the bores. The transformer is able to isolate a high constant potential applied to one of its coils, without the occurrence of sparking or corona, by coupling the coatings, lining the axial bores to the ferromagnetic core and by coupling one terminal of each coil to the respective coating encasing the coil.

  8. The Mechanism of Voltage Dependent Gating of the NaChBac Prokaryotic Sodium Channel

    NASA Astrophysics Data System (ADS)

    Decaen, Paul G.

    Electrical signaling in cells depends on selective conductance of ions through membrane proteins called 'voltage gated ion channels'. These channels are characterized by their ability turn on and off the flow of ionic current by opening and closing their conductive pore in response to changes in membrane potential. The opening and closing of the pore is a mechanically linked to conformational movement of the positively charged fourth transmembrane segment (S4) in 'the voltage sensor' region. How the S4 moves in response to membrane potential is a controversial subject. In this thesis, we used the prokaryotic sodium channel NaChBac as our model sodium channel to study voltage dependent movement of the S4 in the voltage sensor. We use a disulfide-locking method where we introduced pairs of cysteines in the voltage sensor that crosslink and trap the S4 in its path after depolarization. We screened over one hundred mutations of the NaChBac channel in the whole cell patch clamp assay and demonstrated discrete and sequential voltage dependent ion pair interactions that occur in at least three states between the positively charged residues of the S4 segment and the acidic residues in the S1, S2 and S3 segments. In conjunction with structural modeling of the voltage sensor and our disulfide locking data, we propose that the S4 moves in and out of the plane of the membrane 8-13 A, forming distinct gating charge interactions with counter charges of the voltage sensor and adopts a 310 helix over a portion of its structure during activation. These findings are compatible with the sliding helix model and refine our understanding of the structural determinates of voltage sensor function in voltage gated ion channels.

  9. Comparative High Voltage Impulse Measurement

    PubMed Central

    FitzPatrick, Gerald J.; Kelley, Edward F.

    1996-01-01

    A facility has been developed for the determination of the ratio of pulse high voltage dividers over the range from 10 kV to 300 kV using comparative techniques with Kerr electro-optic voltage measurement systems and reference resistive voltage dividers. Pulse voltage ratios of test dividers can be determined with relative expanded uncertainties of 0.4 % (coverage factor k = 2 and thus a two standard deviation estimate) or less using the complementary resistive divider/Kerr cell reference systems. This paper describes the facility and specialized procedures used at NIST for the determination of test voltage divider ratios through comparative techniques. The error sources and special considerations in the construction and use of reference voltage dividers to minimize errors are discussed, and estimates of the measurement uncertainties are presented. PMID:27805083

  10. Large-deflection theory for end compression of long rectangular plates rigidly clamped along two edges

    NASA Technical Reports Server (NTRS)

    Levy, Samuel; Krupen, Philip

    1943-01-01

    The von Karman equations for flat plates are solved beyond the buckling load up to edge strains equal to eight time the buckling strain, for the extreme case of rigid clamping along the edges parallel to the load. Deflections, bending stresses, and membrane stresses are given as a function of end compressive load. The theoretical values of effective width are compared with the values derived for simple support along the edges parallel to the load. The increases in effective width due to rigid clamping drops from about 20 percent near the buckling strain to about 8 percent at an edge strain equal to eight times the buckling strain. Experimental values of effective width in the elastic range reported in NACA Technical Note No. 684 are between the theoretical curves for the extremes of simple support and rigid clamping.

  11. The Role of the C-Clamp in Wnt-Related Colorectal Cancers

    PubMed Central

    Ravindranath, Aditi J.; Cadigan, Ken M.

    2016-01-01

    T-cell Factor/Lymphoid Enhancer Factor (TCF/LEF) transcription factors are major regulators of Wnt targets, and the products of the TCF7 and TCF7L2 genes have both been implicated in the progression of colorectal cancer in animal models and humans. TCFs recognize specific DNA sequences through their high mobility group (HMG) domains, but invertebrate TCFs and some isoforms of vertebrate TCF7 and TCF7L2 contain a second DNA binding domain known as the C-clamp. This review will cover the basic properties of C-clamps and their importance in Wnt signaling, using data from Drosophila, C. elegans, and mammalian cell culture. The connection between C-clamp containing TCFs and colorectal cancer will also be discussed. PMID:27527215

  12. Sequential dissection of multiple ionic currents in single cardiac myocytes under action potential-clamp

    PubMed Central

    Banyasz, Tamas; Horvath, Balazs; Jian, Zhong; Izu, Leighton T.; Chen-Izu, Ye

    2011-01-01

    The cardiac action potential (AP) is shaped by myriad ionic currents. In this study we develop an innovative AP-clamp Sequential Dissection technique to enable recording of multiple ionic currents in the single cell under AP-clamp. This new technique presents a significant step beyond the traditional way of recording only one current in any one cell. The ability to measure many currents in a single cell has revealed two hitherto unknown characteristics of the ionic currents in cardiac cells: coordination of currents within a cell and large variation of currents between cells. Hence, the AP-clamp Sequential Dissection method provides a unique and powerful tool for studying individual cell electrophysiology. PMID:21215755

  13. Sequential dissection of multiple ionic currents in single cardiac myocytes under action potential-clamp.

    PubMed

    Banyasz, Tamas; Horvath, Balazs; Jian, Zhong; Izu, Leighton T; Chen-Izu, Ye

    2011-03-01

    The cardiac action potential (AP) is shaped by myriad ionic currents. In this study, we develop an innovative AP-clamp Sequential Dissection technique to enable the recording of multiple ionic currents in the single cell under AP-clamp. This new technique presents a significant step beyond the traditional way of recording only one current in any one cell. The ability to measure many currents in a single cell has revealed two hitherto unknown characteristics of the ionic currents in cardiac cells: coordination of currents within a cell and large variation of currents between cells. Hence, the AP-clamp Sequential Dissection method provides a unique and powerful tool for studying individual cell electrophysiology.

  14. Delayed cord clamping in red blood cell alloimmunization: safe, effective, and free?

    PubMed Central

    2016-01-01

    Hemolytic disease of the newborn (HDN), an alloimmune disorder due to maternal and fetal blood type incompatibility, is associated with fetal and neonatal complications related to red blood cell (RBC) hemolysis. After delivery, without placental clearance, neonatal hyperbilirubinemia may develop from ongoing maternal antibody-mediated RBC hemolysis. In cases refractory to intensive phototherapy treatment, exchange transfusions (ET) may be performed to prevent central nervous system damage by reducing circulating bilirubin levels and to replace antibody-coated red blood cells with antigen-negative RBCs. The risks and costs of treating HDN are significant, but appear to be decreased by delayed umbilical cord clamping at birth, a strategy that promotes placental transfusion to the newborn. Compared to immediate cord clamping (ICC), safe and beneficial short-term outcomes have been demonstrated in preterm and term neonates receiving delayed cord clamping (DCC), a practice that may potentially be effective in cases RBC alloimmunization. PMID:27186530

  15. Faster voltage-dependent activation of Na+ channels in growth cones versus somata of neuroblastoma N1E-115 cells.

    PubMed Central

    Zhang, J; Loew, L M; Davidson, R M

    1996-01-01

    Kinetics of voltage-gated ionic channels fundamentally reflect the response of the channels to local electric fields. In this report cell-attached patch-clamp studies reveal that the voltage-dependent activation rate of sodium channels residing in the growth cone membrane differs from that of soma sodium channels in differentiating N1E-115 neuroblastoma cells. Because other electrophysiological properties of these channels do not differ, this finding may be a reflection of the difference in intramembrane electric field in these two regions of the cell. This represents a new mechanism for channels to attain a range of activities both within and between cells. PMID:8913589

  16. A Randomized Trial of Mogen Clamp versus Plastibell for Neonatal Male Circumcision in Botswana

    PubMed Central

    Plank, Rebeca M; Ndubuka, Nnamdi O; Wirth, Kathleen E; Mwambona, Janet T; Kebaabetswe, Poloko; Bassil, Barbara; Lesetedi, Chiapo; Hussein, Fatma M; Magetse, Jane; Nkgau, Maggie; Makhema, Joseph; Mmalane, Mompati; Creek, Tracy; Powis, Kathleen M; Shapiro, Roger; Lockman, Shahin

    2013-01-01

    Background Male circumcision can reduce the risk of heterosexually-acquired HIV-1 infection in men. Neonatal male circumcision (NMC) has many potential advantages over circumcision at older ages but little is known about its feasibility and safety in resource-limited settings. Methods We performed a randomized trial in southeastern Botswana of Mogen clamp and Plastibell, two commonly used devices for NMC. Follow-up visits occurred at six weeks and four months postpartum. Adverse events, parental satisfaction and staff impressions were recorded. Results Of 302 male neonates randomized, 300 (99%) underwent circumcision, 153 (51%) with Mogen clamp and 147 (49%) with Plastibell. There were no major adverse events in the Mogen clamp arm but there were two major adverse events in the Plastibell arm (both were a proximally migrated ring that had to be removed by study staff). Minor adverse events were more common with the Mogen clamp compared with the Plastibell, specifically removal of too little skin and formation of skin bridges or adhesions (12 vs. 1 and 11 vs. 3, respectively, all P<0.05). Five (3%) infants in the Mogen clamp arm and none in the Plastibell arm had minor bleeding (P=0.03). More than 94% of mothers reported being highly or completely satisfied with the procedure. Conclusions NMC can be performed in Botswana with a low rate of adverse events and high parental satisfaction. Although the risk of migration and retention of the Plastibell is small, the Mogen clamp may be safer for NMC in regions where immediate emergent medical attention is not available. PMID:23314413

  17. Analysis and testing of the DIII-D ohmic heating coil lead repair clamp

    SciTech Connect

    Reis, E.E.; Anderson, P.M.; Chin, E.; Robinson, J.I.

    1997-11-01

    DIII-D has been operating for the last year with limited volt-second capabilities due to structural failure of a conductor lead to one of the ohmic heating (OH) solenoids. The conductor failure was due to poor epoxy impregnation of the overwrap of the lead pack, resulting in copper fatigue and a water leak. A number of structural analyses were performed to assist in determining the failure scenario and to evaluate various repair options. A fatigue stress analysis of the leads with a failed epoxy overwrap indicated crack initiation after 1,000 cycles at the maximum operating conditions. The failure occurred in a very inaccessible area which restricted design repair options to concepts which could be implemented remotely. Several design options were considered for repairing the lead so that it can sustain the loads for 7.5 Vs conditions at full toroidal field. A clamp, along with preloaded banding straps and shim bags, provides a system that guarantees that the stress at the crack location is always compressive and prevents further crack growth in the conductor. Due to the limited space available for the repair, it was necessary to design the clamp system to operate at the material yield stress. The primary components of the clamp system were verified by load tests prior to installation. The main body of the clamp contains a load cell and potentiometer for monitoring the load-deflection characteristics of the clamp and conductors during plasma operation. Strain gages provides redundant instrumentation. If required, the preload on the conductors can be increased remotely by a special wrench attached to the clamp assembly.

  18. Growth hormone response to GHRH + GHRP-6 in type 2 diabetes during euglycemic and hyperglycemic clamp.

    PubMed

    Micic, Dragan; Kendereski, Aleksandra; Sumarac-Dumanovic, Mirjana; Cvijovic, Goran; Popovic, Vera; Dieguez, Carlos; Casanueva, Felipe

    2004-01-01

    The aim of this study was to investigate the effect of two different glucose levels on GH response to the combined administration of GHRH+GHRP-6 in patients with type 2 diabetes. GH response to i.v. bolus of GHRH+GHRP-6 (100 mcg, each) was measured in 12 male patients with type 2 diabetes (mean age: 53.9+/-1.59 years; BMI: 25.58+/-0.39 kg/m(2); mean HbA(1c): 8.7+/-0.42%), during a euglycemic (mean glucose: 4.92+/-0.08 mmol) hyperinsulinemic clamp (insulin infusion rate of 100 mU/kg/h) and a hyperglycemic clamp (mean glucose: 12.19+/-0.11 mmol/l). There was no difference in basal GH levels between the hyperglycemic and euglycemic clamps (2.9+/-0.99 mU/l versus 1.48+/-0.44 mU/l; P>0.05). Peak GH response to GHRH+GHRP-6 during the hyperglycemic clamp was lower than in the englycemic clamp (112.45+/-14.45 mU/l versus 151.06+/-16.87 mU/l; P<0.05). Area under the GH curve was lower in the hyperglycemic than in the euglycemic clamp (6974.49+/-1001.95 mU/l/min versus 9560.75+/-1140.65 mU/l/min; P<0.05). It is concluded that hyperglycemia significantly reduces GH response to combined administration of GHRH+GHRP-6 in normal weight patients with type 2 diabetes. It is suggested that ambient glucose levels should be taken into account during interpretation of GH response to combined administration of GHRH+GHRP-6 in patients with type 2 diabetes.

  19. Design and fabrication of a planar patch-clamp substrate using a silicon-on-insulator wafer

    NASA Astrophysics Data System (ADS)

    Zhenlong, Zhang; Xiangyang, Liu; Yanli, Mao

    2009-09-01

    The planar patch-clamp technique has been applied to high throughput screening in drug discovery. The key feature of this technique is the fabrication of a planar patch-clamp substrate using appropriate materials. In this study, a planar patch-clamp substrate was designed and fabricated using a silicon-on-insulator (SOI) wafer. The access resistance and capacitance of SOI-based planar patch-clamp substrates are smaller than those of bulk silicon-based planar substrates, which will reduce the distributed RC noise.

  20. The interplay of primer-template DNA phosphorylation status and single-stranded DNA binding proteins in directing clamp loaders to the appropriate polarity of DNA

    PubMed Central

    Hayner, Jaclyn N.; Douma, Lauren G.; Bloom, Linda B.

    2014-01-01

    Sliding clamps are loaded onto DNA by clamp loaders to serve the critical role of coordinating various enzymes on DNA. Clamp loaders must quickly and efficiently load clamps at primer/template (p/t) junctions containing a duplex region with a free 3′OH (3′DNA), but it is unclear how clamp loaders target these sites. To measure the Escherichia coli and Saccharomyces cerevisiae clamp loader specificity toward 3′DNA, fluorescent β and PCNA clamps were used to measure clamp closing triggered by DNA substrates of differing polarity, testing the role of both the 5′phosphate (5′P) and the presence of single-stranded binding proteins (SSBs). SSBs inhibit clamp loading by both clamp loaders on the incorrect polarity of DNA (5′DNA). The 5′P groups contribute selectivity to differing degrees for the two clamp loaders, suggesting variations in the mechanism by which clamp loaders target 3′DNA. Interestingly, the χ subunit of the E. coli clamp loader is not required for SSB to inhibit clamp loading on phosphorylated 5′DNA, showing that χ·SSB interactions are dispensable. These studies highlight a common role for SSBs in directing clamp loaders to 3′DNA, as well as uncover nuances in the mechanisms by which SSBs perform this vital role. PMID:25159615

  1. Patch-clamp array with on-chip electronics, optics, flow control and mechanical actuation.

    SciTech Connect

    James, Conrad D.; Okandan, Murat; Draper, Bruce Leroy; Mani, Seethambal S.

    2003-07-01

    Fast and quantitative analysis of cellular activity, signaling and responses to external stimuli is a crucial capability and it has been the goal of several projects focusing on patch clamp measurements. To provide the maximum functionality and measurement options, we have developed a patch clamp array device that incorporates on-chip electronics, mechanical, optical and microfluidic coupling as well as cell localization through fluid flow. The preliminary design, which integrated microfluidics, electrodes and optical access, was fabricated and tested. In addition, new designs which further combine mechanical actuation, on-chip electronics and various electrode materials with the previous designs are currently being fabricated.

  2. Ultrafast spectroscopy of super high frequency mechanical modes of doubly clamped beams

    NASA Astrophysics Data System (ADS)

    Ristow, Oliver; Merklein, Moritz; Grossmann, Martin; Hettich, Mike; Schubert, Martin; Bruchhausen, Axel; Grebing, Jochen; Erbe, Artur; Mounier, Denis; Gusev, Vitalyi; Scheer, Elke; Dekorsy, Thomas; Barretto, Elaine C. S.

    2013-12-01

    We use ultrafast pump-probe spectroscopy to study the mechanical vibrations in the time domain of doubly clamped silicon nitride beams. Beams with two different clamping conditions are investigated. Finite element method calculations are performed to analyse the mode spectra of both structures. By calculating the strain integral on the surface of the resonators, we are able to reproduce the effect of the detection mechanism and identify all the measured modes. We show that our spectroscopy technique combined with our modelling tools allow the investigation of several different modes in the super high frequency range (3-30 GHz) and above, bringing more information about the vibration modes of nanomechanical resonators.

  3. Clamping drainage is unnecessary after minimally invasive total knee arthroplasty in patients with tranexamic acid

    PubMed Central

    Wu, Yuangang; Yang, Timin; Zeng, Yi; Li, Canfeng; Shen, Bin; Pei, Fuxing

    2017-01-01

    Abstract Background: Drainage and tranexamic acid (TXA) have been widely used in total knee arthroplasty (TKA). However, it remains unclear whether it is necessary to clamp the drain after minimally invasive TKA (MIS-TKA) when TXA is used. We therefore conducted a randomized controlled trial to compare the effects of clamping versus not clamping drainage following MIS-TKA in patients in whom TXA was used. Methods: From January 2015 to December 2015, 121 patients undergoing unilateral primary MIS-TKA were enrolled and randomly divided into 2 groups. In the clamping group (N = 60), drainage was clamped for the 1st 4 postoperative hours. In the nonclamping group (N = 61), drainage was not clamped. All patients underwent a minimidvastus approach and received 10 mg/kg TXA intravenously before tourniquet deflation. We recorded the total blood loss, drainage volume, and transfusion requirements in the postoperative period. We also measured the hemoglobin (Hb) and hematocrit (Hct) levels on postoperative days 1, 3, and 5. Other factors, including range of motion (ROM), visual analog scale (VAS), and occurrence of wound-related complications, deep vein thrombosis (DVT), and pulmonary embolism (PE) were recorded at the time of discharge and 1 and 6 months postoperatively. No statistically significant differences were found between the 2 groups with regard to age, gender, weight, BMI, preoperative Hb and Hct levels, preoperative ROM, VAS, duration of surgery, anesthesia method, and the American Society of Anesthesiologists classification. Results: The clamping group experienced better drainage volume results than the nonclamping group (P < 0.001). There were no statistically significant differences in TBL and transfusion requirements (P = 0.105 and 0.276, respectively); Hb and Hct levels on postoperative days 1, 3, and 5 were similar between the 2 groups. No significant differences were found for ROM, VAS, DVT, PE, wound-related complications, and hospital

  4. Electrochemical properties of the erbium-chitosan-fluorine-modified PbO2 electrode for the degradation of 2,4-dichlorophenol in aqueous solution.

    PubMed

    Wang, Ying; Shen, Zhenyao; Li, Yang; Niu, Junfeng

    2010-05-01

    The erbium (Er)-chitosan-fluorine (F) modified PbO(2) electrode was prepared by electrodeposition method, and its use for adsorption and electrochemical degradation of 2,4-dichlorophenol (2,4-DCP) in aqueous solution was compared with F-PbO(2) and Er-F-PbO(2) electrodes in a batch experiment. The electrodes were characterized by scanning electron microscopy, X-ray diffraction and cyclic voltammetry. Degradation of 2,4-DCP depending on Er and chitosan contents was discussed. The results showed that Er(2)O(3) and chitosan were scattered between the prevailing crystal structure of beta-PbO(2) and thus decreased the internal stress of PbO(2) film. Prior to each electrolysis, the modified PbO(2) anode was first pre-saturated with 2,4-DCP solution for 360 min to preclude the 2,4-DCP decrease due to adsorption. Among the electrodes examined in our study, the highest adsorption and electrochemical degradation for 2,4-DCP and TOC removals that are due to oxidation and adsorption of the organic products onto the chitosan was observed on Er-chitosan-F-PbO(2) electrode. At an applied current density of 5 mAcm(-2), the removal percentages of 2,4-DCP and TOC (solution volume: 180 mL, initial 2,4-DCP concentration: 90 mgL(-1)) were 95% after 120 min and 53% after 360 min, respectively. At Er amount of 10mM in the precursor coating solution, the degradation and mineralization removal for 2,4-DCP on the Er-F-PbO(2) electrode reached a maximum. At chitosan amount of 5 gL(-1), the highest TOC removal on the Er-chitosan-F-PbO(2) electrode was observed. Intermediates mainly including aliphatic carboxylic acids were examined and a possible degradation pathway for 2,4-DCP in aqueous solution involving dechlorination and hydroxylation reactions was proposed.

  5. Effects of acidic pH on voltage-gated ion channels in rat trigeminal mesencephalic nucleus neurons

    PubMed Central

    Han, Jin-Eon; Cho, Jin-Hwa; Choi, In-Sun; Kim, Do-Yeon

    2017-01-01

    The effects of acidic pH on several voltage-dependent ion channels, such as voltage-dependent K+ and Ca2+ channels, and hyperpolarization-gated and cyclic nucleotide-activated cation (HCN) channels, were examined using a whole-cell patch clamp technique on mechanically isolated rat mesencephalic trigeminal nucleus neurons. The application of a pH 6.5 solution had no effect on the peak amplitude of voltage-dependent K+ currents. A pH 6.0 solution slightly, but significantly inhibited the peak amplitude of voltage-dependent K+ currents. The pH 6.0 also shifted both the current-voltage and conductance-voltage relationships to the depolarization range. The application of a pH 6.5 solution scarcely affected the peak amplitude of membrane currents mediated by HCN channels, which were profoundly inhibited by the general HCN channel blocker Cs+ (1 mM). However, the pH 6.0 solution slightly, but significantly inhibited the peak amplitude of HCN-mediated currents. Although the pH 6.0 solution showed complex modulation of the current-voltage and conductance-voltage relationships, the midpoint voltages for the activation of HCN channels were not changed by acidic pH. On the other hand, voltage-dependent Ca2+ channels were significantly inhibited by an acidic pH. The application of an acidic pH solution significantly shifted the current-voltage and conductance-voltage relationships to the depolarization range. The modulation of several voltage-dependent ion channels by an acidic pH might affect the excitability of mesencephalic trigeminal nucleus neurons, and thus physiological functions mediated by the mesencephalic trigeminal nucleus could be affected in acidic pH conditions. PMID:28280415

  6. Voltage-activated proton current in eosinophils from human blood.

    PubMed Central

    Gordienko, D V; Tare, M; Parveen, S; Fenech, C J; Robinson, C; Bolton, T B

    1996-01-01

    1. The resting membrane potential of freshly purified normodense human eosinophils bathed in and dialysed with quasi-physiological solutions was -63 +/- 2 mV (n = 100). 2. In voltage-clamp mode with quasi-physiological internal and external solutions, voltage steps from the holding potential of -60 mV to levels positive to +20 mV resulted in development of a quasi-instantaneous outward current and a slowly developing outward current. The instantaneous current was absent when the cells were bathed in and dialysed with K(+)-free solution. 3. The slow outward current persisted following simultaneous replacement of K+, Na+ and most of the Cl- with largely impermeant ions (tetraethylammonium, N-methyl-D-glucamine and methanesulphonate) and was augmented when the cell was dialysed with a solution of increased buffering capacity for protons. The observed reversal potential of the current closely followed the hydrogen equilibrium potential over a wide range of internal-external pH combinations, indicating that the conductance underlying the slow outward current was highly selective for H+ ions. 4. Acidification of the pipette solution (increasing [H+]i) augmented the outward H+ current and shifted its activation range negatively, whilst acidification of the external solution had the opposite effect. The voltage dependence of the current is modulated by the transmembrane pH gradient so the only outward current could be activated. However, when the outward current was activated by a voltage step, rapid acidification of external solution produced an inward H+ current which rapidly deactivated. 5. The proton current was reversibly inhibited in a voltage-dependent manner by extracellular application of Zn2+. The apparent dissociation constants were 8 nM (at +40 mV), 36 nM (at +70 mV) and 200 nM (at +100 mV). 6. The proton current was augmented by exposure to 10 microM arachidonic acid. This augmentation consisted of a shift of the voltage dependence of activation to more

  7. Voltage-gated potassium currents within the dorsal vagal nucleus: inhibition by BDS toxin.

    PubMed

    Dallas, Mark L; Morris, Neil P; Lewis, David I; Deuchars, Susan A; Deuchars, Jim

    2008-01-16

    Voltage-gated potassium (Kv) channels are essential components of neuronal excitability. The Kv3.4 channel protein is widely distributed throughout the central nervous system (CNS), where it can form heteromeric or homomeric Kv3 channels. Electrophysiological studies reported here highlight a functional role for this channel protein within neurons of the dorsal vagal nucleus (DVN). Current clamp experiments revealed that blood depressing substance (BDS) and intracellular dialysis of an anti-Kv3.4 antibody prolonged the action potential duration. In addition, a BDS sensitive, voltage-dependent, slowly inactivating outward current was observed in voltage clamp recordings from DVN neurons. Electrical stimulation of the solitary tract evoked EPSPs and IPSPs in DVN neurons and BDS increased the average amplitude and decreased the paired pulse ratio, consistent with a presynaptic site of action. This presynaptic modulation was action potential dependent as revealed by ongoing synaptic activity. Given the role of the Kv3 proteins in shaping neuronal excitability, these data highlight a role for homomeric Kv3.4 channels in spike timing and neurotransmitter release in low frequency firing neurons of the DVN.

  8. Mapping of voltage sensor positions in resting and inactivated mammalian sodium channels by LRET.

    PubMed

    Kubota, Tomoya; Durek, Thomas; Dang, Bobo; Finol-Urdaneta, Rocio K; Craik, David J; Kent, Stephen B H; French, Robert J; Bezanilla, Francisco; Correa, Ana M

    2017-03-07

    Voltage-gated sodium channels (Navs) play crucial roles in excitable cells. Although vertebrate Nav function has been extensively studied, the detailed structural basis for voltage-dependent gating mechanisms remain obscure. We have assessed the structural changes of the Nav voltage sensor domain using lanthanide-based resonance energy transfer (LRET) between the rat skeletal muscle voltage-gated sodium channel (Nav1.4) and fluorescently labeled Nav1.4-targeting toxins. We generated donor constructs with genetically encoded lanthanide-binding tags (LBTs) inserted at the extracellular end of the S4 segment of each domain (with a single LBT per construct). Three different Bodipy-labeled, Nav1.4-targeting toxins were synthesized as acceptors: β-scorpion toxin (Ts1)-Bodipy, KIIIA-Bodipy, and GIIIA-Bodipy analogs. Functional Nav-LBT channels expressed in Xenopus oocytes were voltage-clamped, and distinct LRET signals were obtained in the resting and slow inactivated states. Intramolecular distances computed from the LRET signals define a geometrical map of Nav1.4 with the bound toxins, and reveal voltage-dependent structural changes related to channel gating.

  9. Mapping of voltage sensor positions in resting and inactivated mammalian sodium channels by LRET

    PubMed Central

    Kubota, Tomoya; Durek, Thomas; Dang, Bobo; Finol-Urdaneta, Rocio K.; Craik, David J.; Kent, Stephen B. H.; French, Robert J.; Bezanilla, Francisco; Correa, Ana M.

    2017-01-01

    Voltage-gated sodium channels (Navs) play crucial roles in excitable cells. Although vertebrate Nav function has been extensively studied, the detailed structural basis for voltage-dependent gating mechanisms remain obscure. We have assessed the structural changes of the Nav voltage sensor domain using lanthanide-based resonance energy transfer (LRET) between the rat skeletal muscle voltage-gated sodium channel (Nav1.4) and fluorescently labeled Nav1.4-targeting toxins. We generated donor constructs with genetically encoded lanthanide-binding tags (LBTs) inserted at the extracellular end of the S4 segment of each domain (with a single LBT per construct). Three different Bodipy-labeled, Nav1.4-targeting toxins were synthesized as acceptors: β-scorpion toxin (Ts1)-Bodipy, KIIIA-Bodipy, and GIIIA-Bodipy analogs. Functional Nav-LBT channels expressed in Xenopus oocytes were voltage-clamped, and distinct LRET signals were obtained in the resting and slow inactivated states. Intramolecular distances computed from the LRET signals define a geometrical map of Nav1.4 with the bound toxins, and reveal voltage-dependent structural changes related to channel gating. PMID:28202723

  10. Pacemaking in dopaminergic ventral tegmental area neurons: depolarizing drive from background and voltage-dependent sodium conductances

    PubMed Central

    Khaliq, Zayd M.; Bean, Bruce P.

    2010-01-01

    Dopaminergic neurons in the ventral tegmental area (VTA) fire spontaneously in a pacemaker-like manner. We analyzed the ionic currents that drive pacemaking in dopaminergic VTA neurons, studied in mouse brain slices. Pacemaking was not inhibited by blocking hyperpolarization-activated cation current (Ih) or blocking all calcium current by Mg2+ replacement of Ca2+. Tetrodotoxin (TTX) stopped spontaneous activity and usually resulted in stable resting potentials near −60 mV to −55 mV, 10–15 mV below the action potential threshold. When external sodium was replaced by N-methyl-D-glucamine (NMDG) with TTX present, cells hyperpolarized by an average of −11 mV, suggesting a significant resting sodium conductance not sensitive to TTX. Voltage-clamp experiments using slow (10 mV/s) ramps showed a steady-state, steeply voltage-dependent current blocked by TTX that activates near −60 mV, as well as a sodium “background” current with little voltage-sensitivity, revealed by NMDG replacement for sodium with TTX present. We quantified these two components of sodium current during the pacemaking trajectory using action potential clamp. The initial phase of depolarization, up to about −55 mV, is driven mainly by non-voltage-dependent sodium background current. Above −55 mV, TTX-sensitive voltage-dependent “persistent” Na current helps drive the final phase of depolarization to the spike threshold. Voltage-dependent calcium current is small at all subthreshold voltages. The pacemaking mechanism in VTA neurons differs from that in substantia nigra pars compacta (SNc) neurons, where subthreshold calcium current plays a dominant role. In addition, we found that non-voltage-dependent background sodium current is much smaller in SNc neurons than VTA neurons. PMID:20505107

  11. Voltage sensor and dielectric material

    DOEpatents

    Yakymyshyn, Christopher Paul; Yakymyshyn, Pamela Jane; Brubaker, Michael Allen

    2006-10-17

    A voltage sensor is described that consists of an arrangement of impedance elements. The sensor is optimized to provide an output ratio that is substantially immune to changes in voltage, temperature variations or aging. Also disclosed is a material with a large and stable dielectric constant. The dielectric constant can be tailored to vary with position or direction in the material.

  12. Smaller insulators handle higher voltage

    SciTech Connect

    Wilt, G.

    1997-10-01

    Researcher at Lawrence Livermore have designed the Ultra High Gradient Insulator, a device that can reliably withstand electrical voltages four times greater than before. The Ultra-HGI is designed with alternating layers which divide voltages so finely that the chances of failure are small, and when they do occur, they are confined to a very small portion of the insulator.

  13. Transient voltage oscillations in coils

    SciTech Connect

    Chowdhuri, P.

    1985-01-01

    Magnet coils may be excited into internal voltage oscillations by transient voltages. Such oscillations may electrically stress the magnet's dielectric components to many times its normal stress. This may precipitate a dielectric failure, and the attendant prolonged loss of service and costly repair work. Therefore, it is important to know the natural frequencies of oscillations of a magnet during the design stage, and to determine whether the expected switching transient voltages can excite the magnet into high-voltage internal oscillations. The series capacitance of a winding significantly affects its natural frequencies. However, the series capacitance is difficult to calculate, because it may comprise complex capacitance network, consisting of intra- and inter-coil turn-to-turn capacitances of the coil sections. A method of calculating the series capacitance of a winding is proposed. This method is rigorous but simple to execute. The time-varying transient voltages along the winding are also calculated.

  14. Construction, Calibration, and Validation of a Simple Patch-Clamp Amplifier for Physiology Education

    ERIC Educational Resources Information Center

    Rouzrokh, Ali; Ebrahimi, Soltan Ahmed; Mahmoudian, Massoud

    2009-01-01

    A modular patch-clamp amplifier was constructed based on the Strickholm design, which was initially published in 1995. Various parts of the amplifier such as the power supply, input circuit, headstage, feedback circuit, output and nulling circuits were redesigned to use recent software advances and fabricated using the common lithographic printed…

  15. A Single-Strand Annealing Protein Clamps DNA to Detect and Secure Homology

    PubMed Central

    Ander, Marcel; Subramaniam, Sivaraman; Fahmy, Karim; Stewart, A. Francis; Schäffer, Erik

    2015-01-01

    Repair of DNA breaks by single-strand annealing (SSA) is a major mechanism for the maintenance of genomic integrity. SSA is promoted by proteins (single-strand-annealing proteins [SSAPs]), such as eukaryotic RAD52 and λ phage Redβ. These proteins use a short single-stranded region to find sequence identity and initiate homologous recombination. However, it is unclear how SSAPs detect homology and catalyze annealing. Using single-molecule experiments, we provide evidence that homology is recognized by Redβ monomers that weakly hold single DNA strands together. Once annealing begins, dimerization of Redβ clamps the double-stranded region and nucleates nucleoprotein filament growth. In this manner, DNA clamping ensures and secures a successful detection for DNA sequence homology. The clamp is characterized by a structural change of Redβ and a remarkable stability against force up to 200 pN. Our findings not only present a detailed explanation for SSAP action but also identify the DNA clamp as a very stable, noncovalent, DNA–protein interaction. PMID:26271032

  16. Experiments on vertical slender flexible cylinders clamped at both ends and subjected to axial flow.

    PubMed

    Modarres-Sadeghi, Y; Païdoussis, M P; Semler, C; Grinevich, E

    2008-04-13

    Three series of experiments were conducted on vertical clamped-clamped cylinders in order to observe experimentally the dynamical behaviour of the system, and the results are compared with theoretical predictions. In the first series of experiments, the downstream end of the clamped-clamped cylinder was free to slide axially, while in the second, the downstream end was fixed; the influence of externally applied axial compression was also studied in this series of experiments. The third series of experiments was similar to the second, except that a considerably more slender, hollow cylinder was used. In these experiments, the cylinder lost stability by divergence at a sufficiently high flow velocity and the amplitude of buckling increased thereafter. At higher flow velocities, the cylinder lost stability by flutter (attainable only in the third series of experiments), confirming experimentally the existence of a post-divergence oscillatory instability, which was previously predicted by both linear and nonlinear theory. Good quantitative agreement is obtained between theory and experiment for the amplitude of buckling, and for the critical flow velocities.

  17. Enhancement of heat transfer by clamped flags in a Poiseuille channel flow

    NASA Astrophysics Data System (ADS)

    Lee, Jae Bok; Park, Sung Goon; Kim, Boyoung; Sung, Hyung Jin

    2016-11-01

    A pair of flexible flags clamped vertically in a heated channel was numerically modeled to study an enhancement of heat transfer by the clamped flags in a Poiseuille channel flow. The penalty immersed boundary method was adopted to analyze the fluid-structure-thermal interaction between the surrounding fluid and the clamped flags. The dynamics of the clamped flags was categorized into three distinctive modes: a flapping mode, a fully deflected mode, and an irregular mode. The distinctive modes that depended on the relationship between the hydrodynamic force and the restoring force displayed different movement patterns. The flapping mode provided superior thermal performance to the other modes. Vortices generated from the flapping flags swept out the thermal boundary layer and entrained the fluid near channel walls into the channel core flow while passing through the wake periodically. Compared to rigid flags, the flapping flags significantly improved the thermal efficiency. In addition, the effects of channel height and Reynolds number on the thermal efficiency were explored to obtain an optimal parameter set, which presented the highest thermal performance in present study. The flexible flags regarding the optimal parameter set showed an increase of up to 230% in net heat flux, compared to the baseline flow. Dynamic modes decomposition (DMD) method was adopted to examine the correlation between the vorticity and temperature fields.

  18. Influence of clamp-up force on the strength of bolted composite joints

    NASA Astrophysics Data System (ADS)

    Horn, Walter J.; Schmitt, Ron R.

    1994-03-01

    Composite materials offer the potential for a reduction in the number of individual parts and joints in a structure because large one-piece components can replace multipart assemblies. Nevertheless, there are many situations where composite parts must be joined and often mechanical fasteners provide the only practical method of joining those parts. The long-term strength of mechanically fastened joints of composite members can be directly affected by the clamp-up force of the fastener and thus perhaps by the relaxation of this force due to the viscoelastic character of the composite materials of the joint. Methods for predicting the effect of bolt clamp-up force relaxation on the strength of mechanically fastened joints of thermoplastic composite materials were investigated during the present study. A test program, using two thermoplastic composite materials, was conducted to determine the influence of clamp-up force on joint strength, to measure the relaxation of the joint clamp-up force with time, and to measure the change of joint strength as a function of time.

  19. Chromosomal Replication Dynamics and Interaction with the β Sliding Clamp Determine Orientation of Bacterial Transposable Elements

    PubMed Central

    Gómez, Manuel J.; Díaz-Maldonado, Héctor; González-Tortuero, Enrique; López de Saro, Francisco J.

    2014-01-01

    Insertion sequences (ISs) are small transposable elements widespread in bacterial genomes, where they play an essential role in chromosome evolution by stimulating recombination and genetic flow. Despite their ubiquity, it is unclear how ISs interact with the host. Here, we report a survey of the orientation patterns of ISs in bacterial chromosomes with the objective of gaining insight into the interplay between ISs and host chromosomal functions. We find that a significant fraction of IS families present a consistent and family-specific orientation bias with respect to chromosomal DNA replication, especially in Firmicutes. Additionally, we find that the transposases of up to nine different IS families with different transposition pathways interact with the β sliding clamp, an essential replication factor, suggesting that this is a widespread mechanism of interaction with the host. Although we find evidence that the interaction with the β sliding clamp is common to all bacterial phyla, it also could explain the observed strong orientation bias found in Firmicutes, because in this group β is asymmetrically distributed during synthesis of the leading or lagging strands. Besides the interaction with the β sliding clamp, other asymmetries also play a role in the biased orientation of some IS families. The utilization of the highly conserved replication sliding clamps suggests a mechanism for host regulation of IS proliferation and also a universal platform for IS dispersal and transmission within bacterial populations and among phylogenetically distant species. PMID:24614824

  20. Alpha 1-adrenergic agonists selectively suppress voltage-dependent K+ current in rat ventricular myocytes.

    PubMed Central

    Apkon, M; Nerbonne, J M

    1988-01-01

    The effects of alpha 1-adrenergic agonists on the waveforms of action potentials and voltage-gated ionic currents were examined in isolated adult rat ventricular myocytes by the whole-cell patch-clamp recording technique. After "puffer" applications of either of two alpha 1 agonists, phenylephrine and methoxamine, action-potential durations were increased. In voltage-clamped cells, phenylephrine (5-20 microM) or methoxamine (5-10 microM) reduced the amplitudes of Ca2+-independent voltage-activated outward K+ currents (Iout); neither the kinetics nor the voltage-dependent properties of Iout were significantly affected. The effects of phenylephrine or methoxamine on Iout were larger and longer-lasting at higher concentrations and after prolonged or repeated exposures; in all experiments, however, Iout recovered completely when puffer applications were discontinued. The suppression of Iout is attributed to the activation of alpha 1-adrenergic receptors, as neither beta- nor alpha 2-adrenergic agonists had measurable effects on Iout; in addition, the effect of phenylephrine was attenuated in the presence of the alpha antagonist phentolamine (10 microM), but not in the presence of the beta antagonist propranolol (10 microM). Voltage-gated Ca2+ currents, in contrast, were not altered measurably by phenylephrine or methoxamine and no currents were activated directly by these agents. Suppression of Iout was also observed during puffer applications of either of two protein kinase C activators, phorbol 12-myristate 13-acetate (10 nM-1 microM) and 1-oleoyl-2-acetylglycerol (60 microM). We conclude that the activation of alpha 1-adrenergic receptors in adult rat ventricular myocytes leads to action-potential prolongation as a result of the specific suppression of Iout and that this effect may be mediated by activation of protein kinase C. PMID:2903506

  1. Non-selective voltage-activated cation channel in the human red blood cell membrane.

    PubMed

    Kaestner, L; Bollensdorff, C; Bernhardt, I

    1999-02-04

    Using the patch-clamp technique, a non-selective voltage-activated Na+ and K+ channel in the human red blood cell membrane was found. The channel operates only at positive membrane potentials from about +30 mV (inside positive) onwards. For sodium and potassium ions, similar conductances of about 21 pS were determined. Together with the recently described K+(Na+)/H+ exchanger, this channel is responsible for the increase of residual K+ and Na+ fluxes across the human red blood cell membrane when the cells are suspended in low ionic strength medium.

  2. Exaggerated natriuresis during clamping of systemic NO supply in healthy young men.

    PubMed

    Simonsen, Jane A; Rasmussen, Mona S; Vach, Werner; Høilund-Carlsen, Poul F; Bie, Peter

    2012-01-01

    NO (nitric oxide) may be involved in fluid homoeostasis. We hypothesized that increases in NO synthesis contribute to acute, saline-induced natriuresis, which, therefore, should be blunted when NO availability is stabilized. Young men were studied during simultaneous infusions of L-NAME [NG-nitro-L-arginine methyl ester; bolus of 750 μg·kg⁻¹ of body weight and 8.3 μg·min⁻¹·kg⁻¹ of body weight] and SNP (sodium nitroprusside), the latter at a rate preventing L-NAME from increasing total peripheral resistance ('NO-clamping'). Slow volume expansion (saline, 20 μmol of NaCl·min⁻¹·kg⁻¹ of body weight for 3 h) was performed with and without concomitant NO-clamping. NO-clamping itself decreased RPF (renal plasma flow; P~0.02) and tended to decrease arterial blood pressure [MABP (mean arterial blood pressure)]. Volume expansion markedly decreased the plasma levels of renin, AngII (angiotensin II) and aldosterone (all P<0.001), while MABP (oscillometry), heart rate, cardiac output (impedance cardiography), RPF (by p-aminohippurate), GFR [glomerular filtration rate; by using 51Cr-labelled EDTA] and plasma [Na+] and [K+] remained constant. Volume expansion increased sodium excretion (P<0.02) at constant filtered load, but more so during NO-clamping than during control (+184% compared with 52%; P<0.0001). Urinary nitrate/nitrite excretion increased during volume expansion; plasma cGMP and plasma vasopressin were unchanged. The results demonstrate that NO-clamping augments sodium excretion in response to volume expansion at constant MABP and GFR, reduced RPF and decreased renin system activity, a response termed hypernatriuresis. The results indicate that mediator(s) other than MABP, RPF, GFR and renin system activity contribute significantly to the homoeostatic response to saline loading, but the specific mechanisms of hypernatriuresis remain obscure.

  3. Scanning electron microscope/energy dispersive x ray analysis of impact residues on LDEF tray clamps

    NASA Technical Reports Server (NTRS)

    Bernhard, Ronald P.; Durin, Christian; Zolensky, Michael E.

    1992-01-01

    To better understand the nature of particulates in low-Earth orbit (LEO), and their effects on spacecraft hardware, we are analyzing residues found in impacts on the Long Duration Exposure Facility (LDEF) tray clamps. LDEF experiment trays were held in place by 6 to 8 chromic-anodized aluminum (6061-T6) clamps that were fastened to the spacecraft frame using three stainless steel hex bolts. Each clamp exposed an area of approximately 58 sq cm (4.8 cm x 12.7 cm x .45 cm, minus the bolt coverage). Some 337 out of 774 LDEF tray clamps were archived at JSC and are available through the Meteoroid & Debris Special Investigation Group (M&D SIG). Optical scanning of clamps, starting with Bay/Row A01 and working toward H25, is being conducted at JSC to locate and document impacts as small as 40 microns. These impacts are then inspected by Scanning Electron Microscopy/Energy Dispersive X-ray Analysis (SEM/EDXA) to select those features which contain appreciable impact residue material. Based upon the composition of projectile remnants, and using criteria developed at JSC, we have made a preliminary discrimination between micrometeoroid and space debris residue-containing impact features. Presently, 13 impacts containing significant amounts of unmelted and semi-melted micrometeoritic residues were forwarded to Centre National d'Etudes Spatiales (CNES) in France. At the CNES facilities, the upgraded impacts were analyzed using a JEOL T330A SEM equipped with a NORAN Instruments, Voyager X-ray Analyzer. All residues were quantitatively characterized by composition (including oxygen and carbon) to help understand interplanetary dust as possibly being derived from comets and asteroids.

  4. Ventilation before Umbilical Cord Clamping Improves the Physiological Transition at Birth

    PubMed Central

    Bhatt, Sasmira; Polglase, Graeme R.; Wallace, Euan M.; te Pas, Arjan B.; Hooper, Stuart B.

    2014-01-01

    The transition from a fetus to a neonate at birth represents a critical phase in our life. Most infants make this transition without complications, but preterm infants usually require some form of assistance due to immature cardiopulmonary systems that predispose them to lifelong sequelae. As the incidence of preterm birth is increasing, there is now an urgent need for the development of management strategies that facilitate this transition, which will likely include improved strategies for the management of the maternal third stage of labor. For instance, recent studies on the physiological transition at birth have led to the discovery that establishing ventilation in the infant before the umbilical cord is clamped greatly stabilizes the cardiovascular transition at birth. While most benefits of delayed clamping previously have been attributed to an increase in placenta to infant blood transfusion, clearly there are other significant benefits for the infant, which are not well understood. Nevertheless, if ventilation can be established before cord clamping in a preterm infant, the large adverse changes in cardiac function that normally accompanies umbilical cord clamping can be avoided. As preterm infants have an immature cerebral vascular bed, large swings in cardiovascular function places them at high risk of cerebral vascular rupture and the associated increased risk of mortality and morbidity. In view of the impact that cord clamping has on the cardiovascular transition at birth, it is also time to re-examine some of the strategies used in the management of the third stage of labor. These include the appropriate timing of uterotonic administration in relation to delivery of the infant and placenta. As there is a lack of evidence on the effects these individual practices have on the infant, there is a necessity to improve our understanding of their impact in order to develop strategies that facilitate the transition to newborn life. PMID:25368858

  5. Improved Programmable High-Voltage Power Supply

    NASA Technical Reports Server (NTRS)

    Castell, Karen; Rutberg, Arthur

    1994-01-01

    Improved dc-to-dc converter functions as programmable high-voltage power supply with low-power-dissipation voltage regulator on high-voltage side. Design of power supply overcomes deficiencies of older designs. Voltage regulation with low power dissipation provided on high-voltage side.

  6. Slow unloading leads to DNA-bound β2-sliding clamp accumulation in live Escherichia coli cells

    PubMed Central

    Moolman, M. Charl; Krishnan, Sriram Tiruvadi; Kerssemakers, Jacob W. J.; van den Berg, Aafke; Tulinski, Pawel; Depken, Martin; Reyes-Lamothe, Rodrigo; Sherratt, David J.; Dekker, Nynke H.

    2014-01-01

    The ubiquitous sliding clamp facilitates processivity of the replicative polymerase and acts as a platform to recruit proteins involved in replication, recombination and repair. While the dynamics of the E. coli β2-sliding clamp have been characterized in vitro, its in vivo stoichiometry and dynamics remain unclear. To probe both β2-clamp dynamics and stoichiometry in live E. coli cells, we use custom-built microfluidics in combination with single-molecule fluorescence microscopy and photoactivated fluorescence microscopy. We quantify the recruitment, binding and turnover of β2-sliding clamps on DNA during replication. These quantitative in vivo results demonstrate that numerous β2-clamps in E. coli remain on the DNA behind the replication fork for a protracted period of time, allowing them to form a docking platform for other enzymes involved in DNA metabolism. PMID:25520215

  7. Slow unloading leads to DNA-bound β2-sliding clamp accumulation in live Escherichia coli cells

    NASA Astrophysics Data System (ADS)

    Moolman, M. Charl; Krishnan, Sriram Tiruvadi; Kerssemakers, Jacob W. J.; van den Berg, Aafke; Tulinski, Pawel; Depken, Martin; Reyes-Lamothe, Rodrigo; Sherratt, David J.; Dekker, Nynke H.

    2014-12-01

    The ubiquitous sliding clamp facilitates processivity of the replicative polymerase and acts as a platform to recruit proteins involved in replication, recombination and repair. While the dynamics of the E. coli β2-sliding clamp have been characterized in vitro, its in vivo stoichiometry and dynamics remain unclear. To probe both β2-clamp dynamics and stoichiometry in live E. coli cells, we use custom-built microfluidics in combination with single-molecule fluorescence microscopy and photoactivated fluorescence microscopy. We quantify the recruitment, binding and turnover of β2-sliding clamps on DNA during replication. These quantitative in vivo results demonstrate that numerous β2-clamps in E. coli remain on the DNA behind the replication fork for a protracted period of time, allowing them to form a docking platform for other enzymes involved in DNA metabolism.

  8. An accurate continuous calibration system for high voltage current transformer

    SciTech Connect

    Tong Yue; Li Binhong

    2011-02-15

    A continuous calibration system for high voltage current transformers is presented in this paper. The sensor of this system is based on a kind of electronic instrument current transformer, which is a clamp-shape air core coil. This system uses an optical fiber transmission system for its signal transmission and power supply. Finally the digital integrator and fourth-order convolution window algorithm as error calculation methods are realized by the virtual instrument with a personal computer. It is found that this system can calibrate a high voltage current transformer while energized, which means avoiding a long calibrating period in the power system and the loss of power metering expense. At the same time, it has a wide dynamic range and frequency band, and it can achieve a high accuracy measurement in a complex electromagnetic field environment. The experimental results and the on-site operation results presented in the last part of the paper, prove that it can reach the 0.05 accuracy class and is easy to operate on site.

  9. A New Control Method of a Resonant Switched-Capacitor Converter and the Application for Balancing of the Split DC Voltages in a Multilevel Inverter

    NASA Astrophysics Data System (ADS)

    Sano, Kenichiro; Fujita, Hideaki

    This paper proposes a new voltage-balancing circuit for the split dc voltages in a diode-clamped five-level inverter. The proposed circuit is based on a resonant switched-capacitor converter (RSCC), which consists of two half-bridge inverters, a resonant inductor and a resonant capacitor. A new phase-shift control of the RSCC is proposed to improve voltage balancing performance. Theoretical analysis reveals the rating of the RSCC and stored energy in the resonant inductor. Experimental results confirm the reduction of the inductor to one tenth in volume as compared to a conventional voltage-balancing circuit based on buck-boost topology. Moreover, the proposed phase-shift control has demonstrated that it is possible to eliminate the voltage deviation between the dc capacitors.

  10. Low voltage nonprimary explosive detonator

    DOEpatents

    Dinegar, Robert H.; Kirkham, John

    1982-01-01

    A low voltage, electrically actuated, nonprimary explosive detonator is disclosed wherein said detonation is achieved by means of an explosive train in which a deflagration-to-detonation transition is made to occur. The explosive train is confined within a cylindrical body and positioned adjacent to low voltage ignition means have electrical leads extending outwardly from the cylindrical confining body. Application of a low voltage current to the electrical leads ignites a self-sustained deflagration in a donor portion of the explosive train which then is made to undergo a transition to detonation further down the train.

  11. Voltage Sensors Monitor Harmful Static

    NASA Technical Reports Server (NTRS)

    2009-01-01

    A tiny sensor, small enough to be worn on clothing, now monitors voltage changes near sensitive instruments after being created to alert Agency workers to dangerous static buildup near fuel operations and avionics. San Diego s Quasar Federal Systems received a Small Business Innovation Research (SBIR) contract from Kennedy Space Center to develop its remote voltage sensor (RVS), a dime-sized electrometer designed to measure triboelectric changes in the environment. One of the unique qualities of the RVS is that it can detect static at greater distances than previous devices, measuring voltage changes from a few centimeters to a few meters away, due to its much-improved sensitivity.

  12. Power-MOSFET Voltage Regulator

    NASA Technical Reports Server (NTRS)

    Miller, W. N.; Gray, O. E.

    1982-01-01

    Ninety-six parallel MOSFET devices with two-stage feedback circuit form a high-current dc voltage regulator that also acts as fully-on solid-state switch when fuel-cell out-put falls below regulated voltage. Ripple voltage is less than 20 mV, transient recovery time is less than 50 ms. Parallel MOSFET's act as high-current dc regulator and switch. Regulator can be used wherever large direct currents must be controlled. Can be applied to inverters, industrial furnaces photovoltaic solar generators, dc motors, and electric autos.

  13. Diacylglycerols induce both ion pumping in patch-clamped guard-cell protoplasts and opening of intact stomata.

    PubMed Central

    Lee, Y; Assmann, S M

    1991-01-01

    Stomatal guard cells in leaves regulate the apertures of microscopic pores through which photosynthetic gas exchange and water vapor loss occur. Environmental signals, including light, high humidity, and low CO2 concentrations, open stomata by increasing the volume of guard cells. Activation of a plasma membrane H+ pump initiates K+ and Cl- influx, accompanied by malate synthesis, resulting in osmotic water flow into the guard cells, a bowing apart of the guard-cell pair, and consequent stomatal opening. Physiological and electrophysiological techniques were employed to investigate the possibility that a second-messenger lipid, 1,2-diacylglycerol, is involved in the transduction of opening stimuli. The synthetic diacylglycerols 1,2-dihexanoylglycerol and 1,2-dioctanoylglycerol enhanced light-induced stomatal opening in Commelina communis and induced stomatal opening under darkness, whereas an isomer with no known second-messenger role, 1,3-dioctanoylglycerol, did not affect stomatal responses. 1-(5-Isoquinolinylsulfonyl)-2-methylpiperazine (H-7), an inhibitor of protein kinase C, the enzyme typically activated by 1,2-diacylglycerol in animal cells, inhibited light-stimulated stomatal opening and enhanced dark-induced stomatal closure. N-[(2-Methylamino)ethyl]-5-isoquinolinesulfonamide (H-8), which inhibits cyclic nucleotide-dependent protein kinases preferentially over lipid-dependent protein kinases such as protein kinase C, had little effect on stomatal apertures. Whole-cell patch clamping of guard-cell protoplasts of Vicia faba revealed that 1,2-dihexanoylglycerol and 1-oleoyl-2-acetylglycerol activated an ATP-dependent, voltage-independent current, suggesting activation of an electrogenic ion pump such as the H+ pump. Diacylglycerol or functionally similar lipids may act through protein phosphorylation to provide the intracellular signals that mediate H+-ATPase activation and stomatal opening in response to light or other opening stimuli. PMID:11607161

  14. Estimating use-dependent synaptic gain in autonomic ganglia by computational simulation and dynamic-clamp analysis.

    PubMed

    Wheeler, Diek W; Kullmann, Paul H M; Horn, John P

    2004-11-01

    Biological gain mechanisms regulate the sensitivity and dynamics of signaling pathways at the systemic, cellular, and molecular levels. In the sympathetic nervous system, gain in sensory-motor feedback loops is essential for homeostatic regulation of blood pressure and body temperature. This study shows how synaptic convergence and plasticity can interact to generate synaptic gain in autonomic ganglia and thereby enhance homeostatic control. Using a conductance-based computational model of an idealized sympathetic neuron, we simulated the postganglionic response to noisy patterns of presynaptic activity and found that a threefold amplification in postsynaptic spike output can arise in ganglia, depending on the number and strength of nicotinic synapses, the presynaptic firing rate, the extent of presynaptic facilitation, and the expression of muscarinic and peptidergic excitation. The simulations also showed that postsynaptic refractory periods serve to limit synaptic gain and alter postsynaptic spike timing. Synaptic gain was measured by stimulating dissociated bullfrog sympathetic neurons with 1-10 virtual synapses using a dynamic clamp. As in simulations, the threshold synaptic conductance for nicotinic excitation of firing was typically 10-15 nS, and synaptic gain increased with higher levels of nicotinic convergence. Unlike the model, gain in neurons sometimes declined during stimulation. This postsynaptic effect was partially blocked by 10 microM Cd2+, which inhibits voltage-dependent calcium currents. These results support a general model in which the circuit variations observed in parasympathetic and sympathetic ganglia, as well as other neural relays, can enable functional subsets of neurons to behave either as 1:1 relays, variable amplifiers, or switches.

  15. A study of the bovine adrenal chromaffin nicotinic receptor using patch clamp and concentration-jump techniques.

    PubMed Central

    Maconochie, D J; Knight, D E

    1992-01-01

    1. Voltage clamp records have been obtained from bovine adrenal chromaffin cells in the outside-out and whole-cell configurations, in response to step changes of acetylcholine (ACh) concentration. The concentrations used ranged from 50 nM to 20 mM. 2. At high acetylcholine concentrations, the activation and desensitization kinetics of the nicotinic receptor, as observed in outside-out patches, may be described by a model incorporating a single, fast agonist binding step, and relatively slow isomerization to the open state. The affinity of the closed receptor for ACh is 310 microM, the channel opening rate constant is 460 s-1, and the closing rate constant is 29 s-1. 3. Single channel events, observed when nanomolar ACh concentrations are applied to whole cells, have two distinct channel lifetimes: 0.6 ms and 11-15 ms. The variation of the frequencies of the events with ACh concentration, suggests that the short lifetimes are openings of a singly liganded receptor and the longer lifetimes are openings of a doubly liganded receptor. 4. Only a single exponential associated with receptor desensitization is seen with outside-out patches, but two are seen with whole cells. It is postulated that there are two nicotinic receptor types present on adrenal chromaffin cells. 5. The rate of desensitization (9 s-1 and 26 s-1, whole cells; 24 s-1, patches), is fast enough to be significant in determining the open channel lifetime. 6. A sudden increase in current (rebound) is observed when a high concentration of ACh is abruptly removed from outside-out patches. This is evidence for a blocked state. The affinity of the blocking site for ACh is 1400 microM (outside-out patches). 7. The total number of activatable nicotinic channels per whole cell is estimated to be 2600. PMID:1282154

  16. Two distinct voltage-sensing domains control voltage sensitivity and kinetics of current activation in CaV1.1 calcium channels.

    PubMed

    Tuluc, Petronel; Benedetti, Bruno; Coste de Bagneaux, Pierre; Grabner, Manfred; Flucher, Bernhard E

    2016-06-01

    Alternative splicing of the skeletal muscle CaV1.1 voltage-gated calcium channel gives rise to two channel variants with very different gating properties. The currents of both channels activate slowly; however, insertion of exon 29 in the adult splice variant CaV1.1a causes an ∼30-mV right shift in the voltage dependence of activation. Existing evidence suggests that the S3-S4 linker in repeat IV (containing exon 29) regulates voltage sensitivity in this voltage-sensing domain (VSD) by modulating interactions between the adjacent transmembrane segments IVS3 and IVS4. However, activation kinetics are thought to be determined by corresponding structures in repeat I. Here, we use patch-clamp analysis of dysgenic (CaV1.1 null) myotubes reconstituted with CaV1.1 mutants and chimeras to identify the specific roles of these regions in regulating channel gating properties. Using site-directed mutagenesis, we demonstrate that the structure and/or hydrophobicity of the IVS3-S4 linker is critical for regulating voltage sensitivity in the IV VSD, but by itself cannot modulate voltage sensitivity in the I VSD. Swapping sequence domains between the I and the IV VSDs reveals that IVS4 plus the IVS3-S4 linker is sufficient to confer CaV1.1a-like voltage dependence to the I VSD and that the IS3-S4 linker plus IS4 is sufficient to transfer CaV1.1e-like voltage dependence to the IV VSD. Any mismatch of transmembrane helices S3 and S4 from the I and IV VSDs causes a right shift of voltage sensitivity, indicating that regulation of voltage sensitivity by the IVS3-S4 linker requires specific interaction of IVS4 with its corresponding IVS3 segment. In contrast, slow current kinetics are perturbed by any heterologous sequences inserted into the I VSD and cannot be transferred by moving VSD I sequences to VSD IV. Thus, CaV1.1 calcium channels are organized in a modular manner, and control of voltage sensitivity and activation kinetics is accomplished by specific molecular mechanisms

  17. High voltage solar array experiments

    NASA Technical Reports Server (NTRS)

    Kennerud, K. L.

    1974-01-01

    The interaction between the components of a high voltage solar array and a simulated space plasma is studied to obtain data for the design of a high voltage solar array capable of 15kW at 2 to 16kV. Testing was conducted in a vacuum chamber 1.5-m long by 1.5-m diameter having a plasma source which simulated the plasma conditions existing in earth orbit between 400 nautical miles and synchronous altitude. Test samples included solar array segments pinholes in insulation covering high voltage electrodes, and plain dielectric samples. Quantitative data are presented in the areas of plasma power losses, plasma and high voltage induced damage, and dielectric properties. Limitations of the investigation are described.

  18. Induction of DNA DSB and its rejoining in clamped and non-clamped tumours after exposure to carbon ion beams in comparison to X rays.

    PubMed

    Hirayama, R; Uzawa, A; Matsumoto, Y; Noguchi, M; Kase, Y; Takase, N; Ito, A; Koike, S; Ando, K; Okayasu, R; Furusawa, Y

    2011-02-01

    We studied double-strand breaks (DSB) induction and rejoining in clamped and non-clamped transplanted tumours in mice leg after exposure to 80 keV µm(-1) carbon ions and X rays. The yields of DSB in the tumours were analysed by a static-field gel electrophoresis. The OER of DSB after X rays was 1.68±0.31, and this value was not changed after 1 h rejoining time (1.40±0.26). These damages in oxygenated conditions were rejoined 60-70% within 1 h in situ. No difference was found between the exposure to X rays and carbon ions for the induction and rejoining of DSB. Thus, the values of OER and rejoined fraction after exposure to carbon ions were similar to those after X rays, and the calculated relative biological effectivenesses of carbon ion were around 1 under both oxygen conditions. The yields of DSB in vivo depend on exposure doses, oxygen conditions and rejoining time, but not on the types of radiation quality.

  19. Propagation in the transverse tubular system and voltage dependence of calcium release in normal and mdx mouse muscle fibres

    PubMed Central

    Woods, Christopher E; Novo, David; DiFranco, Marino; Capote, Joana; Vergara, Julio L

    2005-01-01

    Using a two-microelectrode voltage clamp technique, we investigated possible mechanisms underlying the impaired excitation–contraction coupling in skeletal muscle fibres of the mdx mouse, a model of the human disease Duchenne muscular dystrophy. We evaluated the role of the transverse tubular system (T-system) by using the potentiometric indicator di-8 ANEPPS, and that of the sarcoplasmic reticulum (SR) Ca2+ release by measuring Ca2+ transients with a low affinity indicator in the presence of high EGTA concentrations under voltage clamp conditions. We observed minimal differences in the T-system structure and the T-system electrical propagation was not different between normal and mdx mice. Whereas the maximum Ca2+ release elicited by voltage pulses was reduced by ∼67% in mdx fibres, in agreement with previous results obtained using AP stimulation, the voltage dependence of SR Ca2+ release was identical to that seen in normal fibres. Taken together, our data suggest that the intrinsic ability of the sarcoplasmic reticulum to release Ca2+ may be altered in the mdx mouse. PMID:16123111

  20. High voltage power transistor development

    NASA Technical Reports Server (NTRS)

    Hower, P. L.

    1981-01-01

    Design considerations, fabrication procedures, and methods of evaluation for high-voltage power-transistor development are discussed. Technique improvements such as controlling the electric field at the surface and perserving lifetimes in the collector region which have advanced the state of the art in high-voltage transistors are discussed. These improvements can be applied directly to the development of 1200 volt, 200 ampere transistors.

  1. Switched-Capacitor Voltage Multiplier

    NASA Technical Reports Server (NTRS)

    Sridharan, Govind

    1991-01-01

    Dc-to-dc power converter multiplies input supply potential by factor of nearly 40. Design does not make use of transformers or inductors but effects voltage boost-up by capacitive energy transfer. Circuit primarily made up of banks of capacitors, connected by network of integrated-circuit relays. Converter functionally linear voltage amplifier with fixed gain figure. Bipolar in operation. Output fully floating, and excellent dc isolation between input and output terminals.

  2. Thyratron Marx High Voltage Generator.

    DTIC Science & Technology

    This invention relates to a high voltage pulse generator of the Marx type, in which capacitors are charged in parallel and discharged in series...Amongst the many techniques for producing high voltage pulses, the Marx generator is probably the best known and most widely used. For the combination of...short risetime and low output impendance (i.e. high power), large energy, high efficiency and waveform flexibility -- the Marx principle is peerless

  3. Voltage-Boosting Driver For Switching Regulator

    NASA Technical Reports Server (NTRS)

    Trump, Ronald C.

    1990-01-01

    Driver circuit assures availability of 10- to 15-V gate-to-source voltage needed to turn on n-channel metal oxide/semiconductor field-effect transistor (MOSFET) acting as switch in switching voltage regulator. Includes voltage-boosting circuit efficiently providing gate voltage 10 to 15 V above supply voltage. Contains no exotic parts and does not require additional power supply. Consists of NAND gate and dual voltage booster operating in conjunction with pulse-width modulator part of regulator.

  4. A Matter of Quantum Voltages

    SciTech Connect

    Sellner, Bernhard; Kathmann, Shawn M.

    2014-11-14

    Voltages inside matter are relevant to crystallization, materials science, biology, catalysis, and aqueous chemistry. Electron holography is able to measure the variation of voltages in matter and modern supercomputers allow the calculation of quantum voltages with practically unlimited spatial and temporal resolution of bulk systems. Of particular interest is the Mean Inner Potential (Vo) - the spatial average of these voltages. Voltages are very sensitive to the distribution of electrons and provide metrics to understand interactions in condensed phases. In the present study, we find excellent agreement with measurements of Vo for vitrified water and salt crystals and demonstrate the impact of covalent and ionic bonding as well as intermolecular/atomic interactions. Furthermore, we predict Vo as well as the fluctuations of these voltages in aqueous NaCl electrolytes and characterize the changes in their behavior as the resolution increases below the size of atoms. This work was supported by the U.S. Department of Energy, Office of Basic Energy Sciences, Division of Chemical Sciences, Geosciences & Biosciences. Pacific Northwest National Laboratory (PNNL) is a multiprogram national laboratory operated for DOE by Battelle. This research used resources of the National Energy Research Scientific Computing Center, which is supported by the Office of Science of the U.S. Department of Energy under Contract No. DE-AC02-05CH11231.

  5. A matter of quantum voltages

    SciTech Connect

    Sellner, Bernhard; Kathmann, Shawn M.

    2014-11-14

    Voltages inside matter are relevant to crystallization, materials science, biology, catalysis, and aqueous chemistry. The variation of voltages in matter can be measured by experiment, however, modern supercomputers allow the calculation of accurate quantum voltages with spatial resolutions of bulk systems well beyond what can currently be measured provided a sufficient level of theory is employed. Of particular interest is the Mean Inner Potential (V{sub o}) – the spatial average of these quantum voltages referenced to the vacuum. Here we establish a protocol to reliably evaluate V{sub o} from quantum calculations. Voltages are very sensitive to the distribution of electrons and provide metrics to understand interactions in condensed phases. In the present study, we find excellent agreement with measurements of V{sub o} for vitrified water and salt crystals and demonstrate the impact of covalent and ionic bonding as well as intermolecular/atomic interactions. Certain aspects in this regard are highlighted making use of simple model systems/approximations. Furthermore, we predict V{sub o} as well as the fluctuations of these voltages in aqueous NaCl electrolytes and characterize the changes in their behavior as the resolution increases below the size of atoms.

  6. A matter of quantum voltages.

    PubMed

    Sellner, Bernhard; Kathmann, Shawn M

    2014-11-14

    Voltages inside matter are relevant to crystallization, materials science, biology, catalysis, and aqueous chemistry. The variation of voltages in matter can be measured by experiment, however, modern supercomputers allow the calculation of accurate quantum voltages with spatial resolutions of bulk systems well beyond what can currently be measured provided a sufficient level of theory is employed. Of particular interest is the Mean Inner Potential (V(o))--the spatial average of these quantum voltages referenced to the vacuum. Here we establish a protocol to reliably evaluate V(o) from quantum calculations. Voltages are very sensitive to the distribution of electrons and provide metrics to understand interactions in condensed phases. In the present study, we find excellent agreement with measurements of V(o) for vitrified water and salt crystals and demonstrate the impact of covalent and ionic bonding as well as intermolecular/atomic interactions. Certain aspects in this regard are highlighted making use of simple model systems/approximations. Furthermore, we predict V(o) as well as the fluctuations of these voltages in aqueous NaCl electrolytes and characterize the changes in their behavior as the resolution increases below the size of atoms.

  7. The voltage dependence of NADPH oxidase reveals why phagocytes need proton channels

    NASA Astrophysics Data System (ADS)

    DeCoursey, Thomas E.; Morgan, Deri; Cherny, Vladimir V.

    2003-04-01

    The enzyme NADPH oxidase in phagocytes is important in the body's defence against microbes: it produces superoxide anions (O2-, precursors to bactericidal reactive oxygen species). Electrons move from intracellular NADPH, across a chain comprising FAD (flavin adenine dinucleotide) and two haems, to reduce extracellular O2 to O2-. NADPH oxidase is electrogenic, generating electron current (Ie) that is measurable under voltage-clamp conditions. Here we report the complete current-voltage relationship of NADPH oxidase, the first such measurement of a plasma membrane electron transporter. We find that Ie is voltage-independent from -100mV to >0mV, but is steeply inhibited by further depolarization, and is abolished at about +190mV. It was proposed that H+ efflux mediated by voltage-gated proton channels compensates Ie, because Zn2+ and Cd2+ inhibit both H+ currents and O2- production. Here we show that COS-7 cells transfected with four NADPH oxidase components, but lacking H+ channels, produce O2- in the presence of Zn2+ concentrations that inhibit O2- production in neutrophils and eosinophils. Zn2+ does not inhibit NADPH oxidase directly, but through effects on H+ channels. H+ channels optimize NADPH oxidase function by preventing membrane depolarization to inhibitory voltages.

  8. Development of voltage-dependent calcium, sodium, and potassium currents in Xenopus spinal neurons.

    PubMed

    O'Dowd, D K; Ribera, A B; Spitzer, N C

    1988-03-01

    Action potentials of embryonic nerve and muscle cells often have a different ionic dependence and longer duration than those of mature cells. The action potential of spinal cord neurons from Xenopus laevis exhibits a prominent calcium component at early stages of development that diminishes with age as the impulse becomes principally sodium dependent. Whole-cell voltage-clamp analysis has been undertaken to characterize the changes in membrane currents during development of these neurons in culture. Four voltage-dependent currents of cells were identified and examined during the first day in vitro, when most of the change in the action potential occurs. There are no changes in the peak density of the calcium current (ICa), its voltage dependence, or time to half-maximal activation; a small increase in inactivation is apparent. The major change in sodium current (INa) is a 2-fold increase in its density. In addition, more subtle changes in the kinetics of the macroscopic sodium current were noted. The peak density of voltage-dependent potassium current (IKv) increases 3-fold, and this current becomes activated almost twice as fast. No changes were noted in the extent of its inactivation. The calcium-dependent potassium current (IKc) consists of an inactivating and a sustained component. The former increases 2-fold in peak current density, and the latter increases similarly at less depolarized voltages. The changes in these currents contribute to the decrease in duration and the change in ionic dependence of the impulse.

  9. Highly efficient and stable Zr-doped nanocrystalline PbO2 electrode for mineralization of perfluorooctanoic acid in a sequential treatment system.

    PubMed

    Xu, Zesheng; Yu, Yanxin; Liu, Han; Niu, Junfeng

    2017-02-01

    Zr-doped nanocrystalline PbO2 (Zr-PbO2) film electrodes were prepared at different bath temperatures. The Zr-PbO2 electrode doped at 75°C (75-Zr-PbO2) featured high oxygen evolution overpotential, large effective area and good electrocatalytic performance. The oxygen evolution potential and the effective area of 75-Zr-PbO2 achieved 1.91V (vs. SCE) and 9.1cm(2), respectively. The removal efficiency and the defluorination ratio of PFOA reached 97.0% and 88.1% after 90min electrolysis. The primary mineralization products (i.e., F(-) and intermediates) and their change trends were determined. The 75-Zr-PbO2 electrode was introduced to sequentially treat the PFOA wastewater. In an 116h of 75-Zr-PbO2 electrocatalysis sequential process, the PFOA, PFHpA, PFHxA, PFPeA, PFBA, PFPrA, TFA, and TOC concentrations were reduced to below 30, 2.5, 1.3, 1.0, 0.5, 0.2, 0.1, and 9mgL(-1), respectively, demonstrating the promising application of the sequential treatment system for the treatment of PFOA wastewater.

  10. Degradation effects on the surface of commercial LiNi0.5Co0.2Mn0.3O2 electrodes

    NASA Astrophysics Data System (ADS)

    Börner, M.; Horsthemke, F.; Kollmer, F.; Haseloff, S.; Friesen, A.; Niehoff, P.; Nowak, S.; Winter, M.; Schappacher, F. M.

    2016-12-01

    A comprehensive analysis of the degradation mechanisms on the surface of commercial LiNi0.5Co0.2Mn0.3O2 electrodes is presented. Irregularly distributed particle cracking and the formation of a cathode electrolyte interphase on the surface of the active material were identified to be the main degradation mechanisms. The particle cracking originates from inhomogeneity of the composite electrode, leading to deviations in the local current density and the state of charge which results in overcharge conditions for particular LiNi0.5Co0.2Mn0.3O2 particles. Therein, the highly delithiated structure suffers from anisotropic stress due to repulsive interactions between adjacent layers and the formation of new phases which eventually cause particle cracking. The structural changes were confirmed by the presence of a spinel phase on the surface of the cracked particles. Furthermore, the migration of transition metal ions in the highly delithiated structure can facilitate their dissolution into the electrolyte. The investigation of the re-deposited transition metals reveals a predominant dissolution of manganese from the overcharged particles. In addition, electrochemical cycling of the LiNi0.5Co0.2Mn0.3O2 electrodes in laboratory cells show an increasing severity of the particle cracking at higher C-rates which can influence the thermal stability of the active material. Moreover, an increased electrolyte decomposition was observed for higher cut-off potentials.

  11. Design and experimental research of a novel inchworm type piezo-driven rotary actuator with the changeable clamping radius.

    PubMed

    Zhao, Hongwei; Fu, Lu; Ren, Luquan; Huang, Hu; Fan, Zunqiang; Li, Jianping; Qu, Han

    2013-01-01

    In this paper, a novel piezo-driven rotary actuator with the changeable clamping radius is developed based on the inchworm principle. This actuator mainly utilizes three piezoelectric actuators, a flexible gripper, a clamping block, and a rotor to achieve large stroke rotation with high resolution. The design process of the flexible gripper consisting of the driving unit and the clamping unit is described. Lever-type mechanisms were used to amplify the micro clamping displacements. The amplifying factor and parasitic displacement of the lever-type mechanism in the clamping unit was analyzed theoretically and experimentally. In order to investigate the rotation characteristics of the actuator, a series of experiments was carried out. Experimental results indicate that the actuator can rotate at a speed of 77,488 μrad/s with a driving frequency of 167 Hz. The rotation resolution and maximum load torque of the actuator are 0.25 μrad and 37 N mm, respectively. The gripper is movable along the z direction based on an elevating platform, and the clamping radius can change from 10.6 mm to 25 mm. Experimental results confirm that the actuator can achieve different rotation speeds by changing the clamping radius.

  12. Voltage-dependent K channels in protoplasts of trap-lobe cells of Dionaea muscipula.

    PubMed

    Iijima, T; Hagiwara, S

    1987-01-01

    The outward rectification of the K+ current in mesophyll cell protoplasts from trap-lobes of Dionaea muscipula was studied with the patch-clamp technique. The rectification had instantaneous and time-dependent components. Changes in [K+]i strongly affected the conductance voltage relation of the plasma membrane while changes in [K+]o had little effect on the relation. Thus, the outward rectification depends on the membrane voltage and the concentration of intracellular K+. Corresponding single-channel activities were observed both in the intact membrane (cell-attached recording) and in excised patches. The single-channel conductance was about 3.3 pS with symmetrical solutions containing 30 mM K+.

  13. Suppression of voltage-gated Na(+) channels and neuronal excitability by imperatorin.

    PubMed

    Wu, King-Chuen; Chen, Yi-Hung; Cheng, Ka-Shun; Kuo, Yueh-Hsiung; Yang, Chin-Tsang; Wong, Kar-Lok; Tu, Yuan-Kun; Chan, Paul; Leung, Yuk-Man

    2013-12-05

    Imperatorin is a naturally occurring furocoumarin compound isolated from plants such as Angelica archangelica and Cnidium monnieri. It has multiple pharmacological effects including anticonvulsant effects. Here we determined the effects of imperatorin on voltage-gated Na(+) channels (VGSC) using whole-cell patch clamp techniques in differentiated neuronal NG108-15 cells. We showed that imperatorin inhibited VGSC; such inhibition did not show state-dependence. Imperatorin caused a left shift in the steady-state inactivation curve without affecting activation gating. The inhibition of VGSC by imperatorin displayed a mild frequency-dependence. Imperatorin was also shown to inhibit VGSC and action potential amplitude without affecting voltage-gated K(+) channels in rat hippocampal CA1 neurons. In conclusion, our results suggest that imperatorin dampens neuronal excitability by inhibiting VGSC.

  14. Estimates on the eigenvalues of the clamped plate problem on domains in Euclidean spaces

    NASA Astrophysics Data System (ADS)

    Yolcu, Selma Yıldırım; Yolcu, Türkay

    2013-04-01

    The purpose of this article is two-fold. First, we obtain some certain bounds for the sums of (positive and negative) powers of the eigenvalues of the clamped plate problem of the Dirichlet bi-Laplacian operator Δ 2|_{{D}}, restricted to a bounded domain {D}subset {{R}}^d with d ⩾ 2. Second, we establish lower bounds for the sums of eigenvalues of Δ ^2|_{{D}} sharper than the bounds recently obtained by Cheng and Wei ["A lower bound for eigenvalues of a clamped plate problem," Calculus Var. Partial Differ. Equ. 42(3-4), 579-590 (2011)], 10.1007/s00526-011-0399-6. All these estimates are sharp in the sense of Weyl asymptotics.

  15. Robotic surgery and hemostatic agents in partial nephrectomy: a high rate of success without vascular clamping.

    PubMed

    Morelli, Luca; Morelli, John; Palmeri, Matteo; D'Isidoro, Cristiano; Kauffmann, Emanuele Federico; Tartaglia, Dario; Caprili, Giovanni; Pisano, Roberta; Guadagni, Simone; Di Franco, Gregorio; Di Candio, Giulio; Mosca, Franco

    2015-09-01

    Robot-assisted partial nephrectomy has been proposed as a technique to overcome technical challenges of laparoscopic partial nephrectomy. We prospectively collected and analyzed data from 31 patients who underwent robotic partial nephrectomy with systematic use of hemostatic agents, between February 2009 and October 2014. Thirty-three renal tumors were treated in 31 patients. There were no conversions to open surgery, intraoperative complications, or blood transfusions. The mean size of the resected tumors was 27 mm (median 20 mm, range 5-40 mm). Twenty-seven of 33 lesions (82%) did not require vascular clamping and therefore were treated in the absence of ischemia. All margins were negative. The high partial nephrectomy success rate without vascular clamping suggests that robotic nephron-sparing surgery with systematic use of hemostatic agents may be a safe, effective method to completely avoid ischemia in the treatment of selected renal masses.

  16. [Traumatic section of the thoracic aorta: its repair by direct aortic clamping].

    PubMed

    Cairols, M A; Sieyro, F; Miralles, M; Blanes, I; Lozano, P

    1991-01-01

    Traumatic section of descendent aorta, severe complication of a thoracic traumatism, requires an early recognition and restoration because its high mortality rate. Between 1988 and 1990, three patients underwent surgical approach, by direct aortic clamping [correction of clamplaje], at our Service. In two cases, a heart stoppage appeared during the clamping/unclamping [correction of clampaje/desclampaje] maneuvers. One of them had previously electrocardiographic abnormalities, suggestive of heart contusion. Two patients died, one of them during surgical procedure because an irreversible heart stoppage, and the second patient died after 7 days because of a brain hemorrhage. One case presented postoperative paraplegia. Respective rates and literature about the main factors implicated in the diagnosis and treatment of such pathology are reviewed.

  17. Fatal Delayed Esophageal Rupture Following Aortic Clamping for Treatment of Stanford Type B Dissection

    SciTech Connect

    Ito, Hisao; Yamada, Takayuki; Ishibashi, Tadashi; Akiyama, Masatoshi; Nakame, Takahiko; Ito, Yasuhiro; Konnai, Toshiaki

    2003-11-15

    A 65-year-old man underwent a thromboexclusionoperation for management of chronic Stanford type B dissecting aneurysmin 1991. However, long-term follow-up CT scans after the operation revealed that the ascending aorta gradually enlarged and was eventually complicated by recurrent aortic dissection. The patient complained of frequent bloody sputum, whereas chest roentogenography showed no pulmonary abnormalities. Subsequent swallow esophagogram demonstrated that the upper esophagus was deviated to the right and the middle esophagus was greatly compressed by the aortic clamp. Esophageal endoscopy showed a bloody inner surface and marked swelling of the middle esophagus. The patient eventually died of massive hematemesis in 2001. We describe the imaging features of unanticipated complications such as recurrent dissecting aneurysm or impending esophageal rupture.Furthermore, we discuss the cause of hematemesis and document that the aortic clamp migrated and resulted in development of a recurrent aneurysmal dissection, which in turn resulted in esophageal rupture with aneurysmal disruption.

  18. Simplified Estimating Method for Shock Response Spectrum Envelope of V-Band Clamp Separation Shock

    NASA Astrophysics Data System (ADS)

    Iwasa, Takashi; Shi, Qinzhong

    A simplified estimating method for the Shock Response Spectrum (SRS) envelope at the spacecraft interface near the V-band clamp separation device has been established. This simplified method is based on the pyroshock analysis method with a single degree of freedom (D.O.F) model proposed in our previous paper. The parameters required in the estimating method are only geometrical information of the interface and a tension of the V-band clamp. According to the use of these parameters, a simplified calculation of the SRS magnitude at the knee frequency is newly proposed. By comparing the estimation results with actual pyroshock test results, it was verified that the SRS envelope estimated with the simplified method appropriately covered the pyroshock test data of the actual space satellite systems except some specific high frequency responses.

  19. An investigation of the behavior of the clamp-induced bending stresses

    NASA Astrophysics Data System (ADS)

    Huang, S. N.

    1991-12-01

    The Fast Flux Test Facility is a demonstration and test facility for the sodium cooled fast breeder reactor. Insulated pipe clamps are used in the heat transport and safety related systems. This investigation determines whether the clamp induced pipe stresses should be classified as primary or secondary stresses. Three finite element models were developed using the ANSYS computer program. Inelastic analyses were performed to investigate the behavior of meridional bending stress and hoop bending stress. The double exponential creep law of 316 stainless steel was used in the creep analysis. Results indicate that pipe bending stresses do not completely relax with time. Therefore, a portion of the meridional bending stress and the hoop bending stress should be classified as primary stress.

  20. Whole-cell patch-clamp recordings in freely moving animals.

    PubMed

    Lee, Albert K; Epsztein, Jérôme; Brecht, Michael

    2014-01-01

    The patch-clamp technique and the whole-cell measurements derived from it have greatly advanced our understanding of the coding properties of individual neurons by allowing for a detailed analysis of their excitatory/inhibitory synaptic inputs, intrinsic electrical properties, and morphology. Because such measurements require a high level of mechanical stability they have for a long time been limited to in vitro and anesthetized preparations. Recently, however, a considerable amount of effort has been devoted to extending these techniques to awake restrained/head-fixed preparations allowing for the study of the input-output functions of neurons during behavior. In this chapter we describe a technique extending patch-clamp recordings to awake animals free to explore their environments.

  1. Clamped Hybridization Chain Reactions for the Self-Assembly of Patterned DNA Hydrogels.

    PubMed

    Wang, Jianbang; Chao, Jie; Liu, Huajie; Su, Shao; Wang, Lianhui; Huang, Wei; Willner, Itamar; Fan, Chunhai

    2017-02-13

    DNA hydrogels hold great potential for biological and biomedical applications owing to their programmable nature and macroscopic sizes. However, most previous studies involve spontaneous and homogenous gelation procedures in solution, which often lack precise control. A clamped hybridization chain reaction (C-HCR)-based strategy has been developed to guide DNA self-assembly to form macroscopic hydrogels. Analogous to catalysts in chemical synthesis or seeds in crystal growth, we introduced DNA initiators to induce the gelation process, including crosslinked self-assembly and clamped hybridization in three dimensions with spatial and temporal control. The formed hydrogels show superior mechanical properties. The use of printed, surface-confined DNA initiators was also demonstrated for fabricating 2D hydrogel patterns without relying on external confinements. This simple method can be used to construct DNA hydrogels with defined geometry, composition, and order for various bioapplications.

  2. Implementation and Tuning of an Optical Tweezers Force-Clamp Feedback System.

    PubMed

    Bugiel, Michael; Jannasch, Anita; Schäffer, Erik

    2017-01-01

    Feedback systems can be used to control the value of a system variable. In optical tweezers, active feedback is often implemented to either keep the position or tension applied to a single biomolecule constant. Here, we describe the implementation of the latter: an optical force-clamp setup that can be used to study the motion of processive molecular motors under a constant load. We describe the basics of a software-implemented proportional-integral-derivative (PID) controller, how to tune it, and how to determine its optimal feedback rate. Limitations, possible feed-forward applications, and extensions into two- and three-dimensional optical force clamps are discussed. The feedback is ultimately limited by thermal fluctuations and the compliance of the involved molecules. To investigate a particular mechanical process, understanding the basics and limitations of the feedback system will be helpful for choosing the proper feedback hardware, for optimizing the system parameters, and for the design of the experiment.

  3. Scanning electron microscope/energy dispersive x ray analysis of impact residues in LDEF tray clamps

    NASA Technical Reports Server (NTRS)

    Bernhard, Ronald P.; Durin, Christian; Zolensky, Michael E.

    1993-01-01

    Detailed optical scanning of tray clamps is being conducted in the Facility for the Optical Inspection of Large Surfaces at JSC to locate and document impacts as small as 40 microns in diameter. Residues from selected impacts are then being characterized by Scanning Electron Microscopy/Energy Dispersive X-ray Analysis at CNES. Results from this analysis will be the initial step to classifying projectile residues into specific sources.

  4. A comparative in vitro study of fixation of mandibular fractures with paraskeletal clamps or screw plates.

    PubMed

    Crofts, C E; Trowbridge, A; Maung Aung, T; Brook, I M

    1990-05-01

    The use of porcine rib pairs as an in vitro analog for the edentulous mandible is described. Using this model, the relative degree of fixation achieved with a screw plate (Champy) and a paraskeletal clamp plate (Mennen) has been evaluated. The fractured plated ribs failed at significantly lower bending forces than the non-sectioned controls. However, no statistically significant difference in force at failure between the two different methods of plate fixation could be demonstrated.

  5. False aneurysm of aorta secondary to partial occlusion clamp injury: diagnosis by nuclear flow study

    SciTech Connect

    Becker, R.M.; Wexler, J.; Frater, R.W.

    1981-09-01

    A 72-year-old woman presented 12 months postaortic valve replacement with a false aneurysm near the aortic cannulation site. The diagnosis was suspected from clinical findings and confirmed by a nuclear flow study. The patient refused surgery and died shortly afterward. At autopsy, a smooth-walled 1 cm defect adjacent to the cannulation site (presumably related to injury from a partial occlusion clamp) was found; this would have been easily reparable with surgery.

  6. Adverse impact of intermittent portal clamping on long-term postoperative outcomes in hepatocellular carcinoma.

    PubMed

    Hao, S; Chen, S; Yang, X; Wan, C

    2017-01-01

    Introduction To evaluate the impact of intermittent portal clamping (IPC) on long-term postoperative outcomes in patients with hepatocellular carcinoma (HCC). Methods Clinical records of 355 patients underwent curative liver resection for HCC in January 2007 to December 2010 were retrospectively reviewed. According to how portal clamping was performed, patients were grouped as: IPC, n=113; other portal clamping (OPC), n=190; and no portal clamping (NPC), n=52. Results Median recurrence-free survival (RFS) was statistically significantly shorter in the IPC (39.4 months) than OPC (47.3 months, p=0.010) and NPC groups (51.4 months, p=0.008). Median overall survival (OS) was also significantly shorter with IPC (46.3 months), versus 52.9 months with OPC (p=0.022) and 56.2 months with NPC (p=0.015). Kaplan-Meier survival analysis revealed that 5-year cumulative RFS was much lower in the IPC (42.5%) than OPC (50.9%, p=0.014) and NPC groups (49.6%, p=0.013). Five-year cumulative OS was also much lower in the IPC (44.9%) than OPC (58.0%, p=0.020) and NPC groups (57.7%, p=0.025). On univariate analysis, tumour grade, size and number, TNM stage, blood transfusion, vascular invasion and IPC were significantly inversely correlated with RFS and OS. On multivariate analysis, tumour size and number, blood transfusion, vascular invasion and IPC remained significant. Conclusions Our study suggests that IPC is an independent risk factor for poor long-term postoperative outcomes in patients with HCC.

  7. Small deflection of a class of clamped thin plates using collocation

    NASA Technical Reports Server (NTRS)

    Worley, W. J.

    1977-01-01

    Equations are given for the optimization of a class of two-and three-dimensional structures. The application of existing analytical techniques to the response of thin clamped plates is described. The ratios of deflections to plate thickness are given for uniform transverse loads as well as for uniform plus linearly varying transverse loads. Deflections are presented at angular increments of 5 degrees and at radial increments of 0.1 of the radius.

  8. A Nerve Clamp Electrode Design for Indirect Stimulation of Skeletal Muscle

    DTIC Science & Technology

    2010-10-01

    Reports www.BioTechniques.com739Vol. 49 | No. 4 | 2010 Ex vivo assays to measure muscle paralysis induced by botulinum neurotoxin (BoNT) have been...Keywords: stimulating electrode; botulinum neurotoxin; skeletal muscle; paralysis A nerve clamp electrode was developed to indirectly stimulate skeletal...attached nerve. Indirect muscle stimulation is critical for studying the para- lytic actions of presynaptic-acting toxins such as botulinum neurotoxins

  9. On the vibration of stretched strings with clamped ends and non-zero flexural rigidity

    NASA Astrophysics Data System (ADS)

    Wittrick, W. H.

    1986-10-01

    A simple iterative procedure, suitable for a programmable calculator, is described for computing correction factors to the natural frequencies of a stretched string with clamped ends due to its flexural rigidity, which may be moderately large. Also an accurate power series approximation, suitable for small values of the flexural rigidity, is derived. Detailed information about the convergence of the iterative procedure and the accuracy of the series approximation is presented.

  10. Modeling hepatic insulin sensitivity during a meal: validation against the euglycemic hyperinsulinemic clamp

    PubMed Central

    Dalla Man, Chiara; Piccinini, Francesca; Basu, Rita; Basu, Ananda; Rizza, Robert A.

    2013-01-01

    Recently, we proposed a model describing the suppression of endogenous glucose production (EGP) during a meal. It assumes that EGP suppression depends on glucose concentration and its rate of change and on delayed insulin action. Hepatic insulin sensitivity (SILmeal) can be derived from EGP model parameters. This model was shown to adequately describe EGP profiles measured with multiple tracer techniques; however, SILmeal has never been compared directly with its euglycemic hyperinsulinemic clamp counterpart (SILclamp). To do so, 62 subjects with different degrees of glucose tolerance underwent a triple-tracer mixed meal. Fifty-seven subjects also underwent a labeled ([3-3H]glucose) euglycemic hyperinsulinemic clamp. From the triple-tracer meal data, virtually model-independent estimates of EGP were obtained using the tracer-to-tracee clamp technique, and the EGP model was identified in each subject. Model fit was satisfactory, and SILmeal was estimated with good precision. Correlation between SILclamp and SILmeal was good (r = 0.72, P < 0.001); however, SILmeal was lower than SILclamp (4.60 ± 0.64 vs. 8.73 ± 1.07 10−4 dl·kg−1·min−1 per μU/ml, P < 0.01). This difference may be due to different ranges of insulin explored during the two tests (ΔIclamp = 15.60 ± 1.61 vs. ΔImeal= 83.37 ± 10.71 μU/ml) as well as steady- vs. non-steady-state glucose and insulin profiles. In conclusion, the new EGP model provides an estimate of hepatic insulin sensitivity during a meal that is in good agreement with that derived in the same individuals with a hyperinsulinemic clamp. When used in conjunction with the minimal model, the approach potentially enables estimation of hepatic insulin sensitivity from a single-tracer labeled meal or oral glucose tolerance test. PMID:23443923

  11. Ultra-fast force-clamp laser trapping of single molecular motors and DNA binding proteins

    NASA Astrophysics Data System (ADS)

    Capitanio, Marco; Monico, Carina; Vanzi, Francesco; Pavone, Francesco S.

    2013-09-01

    Forces play a fundamental role in a wide array of biological processes, regulating enzymatic activity, kinetics of molecular bonds, and molecular motors mechanics. Single molecule force spectroscopy techniques have enabled the investigation of such processes, but they are inadequate to probe short-lived (millisecond and sub-millisecond) molecular complexes. We developed an ultrafast force-clamp spectroscopy technique that uses a dual trap configuration to apply constant loads to a single intermittently interacting biological polymer and a binding protein. Our system displays a delay of only ˜10 μs between formation of the molecular bond and application of the force and is capable of detecting interactions as short as 100 μs. The force-clamp configuration in which our assay operates allows direct measurements of load-dependence of lifetimes of single molecular bonds. Moreover, conformational changes of single proteins and molecular motors can be recorded with sub-nanometer accuracy and few tens of microseconds of temporal resolution. We demonstrate our technique on molecular motors, using myosin II from fast skeletal muscle and on protein-DNA interaction, specifically on Lactose repressor (LacI). The apparatus is stabilized to less than 1 nm with both passive and active stabilization, allowing resolving specific binding regions along the actin filament and DNA molecule. Our technique extends single-molecule force-clamp spectroscopy to molecular complexes that have been inaccessible up to now, opening new perspectives for the investigation of the effects of forces on biological processes.

  12. Molecular jamming--the cystine slipknot mechanical clamp in all-atom simulations.

    PubMed

    Pepłowski, Lukasz; Sikora, Mateusz; Nowak, Wiesław; Cieplak, Marek

    2011-02-28

    A recent survey of 17 134 proteins has identified a new class of proteins which are expected to yield stretching induced force peaks in the range of 1 nN. Such high force peaks should be due to forcing of a slip-loop through a cystine ring, i.e., by generating a cystine slipknot. The survey has been performed in a simple coarse grained model. Here, we perform all-atom steered molecular dynamics simulations on 15 cystine knot proteins and determine their resistance to stretching. In agreement with previous studies within a coarse grained structure based model, the level of resistance is found to be substantially higher than in proteins in which the mechanical clamp operates through shear. The large stretching forces arise through formation of the cystine slipknot mechanical clamp and the resulting steric jamming. We elucidate the workings of such a clamp in an atomic detail. We also study the behavior of five top strength proteins with the shear-based mechanostability in which no jamming is involved. We show that in the atomic model, the jamming state is relieved by moving one amino acid at a time and there is a choice in the selection of the amino acid that advances the first. In contrast, the coarse grained model also allows for a simultaneous passage of two amino acids.

  13. Transposase interaction with the β sliding clamp: effects on insertion sequence proliferation and transposition rate

    PubMed Central

    Díaz-Maldonado, Héctor; Gómez, Manuel J.; Moreno-Paz, Mercedes; San Martín-Úriz, Patxi; Amils, Ricardo; Parro, Víctor; López de Saro, Francisco J.

    2015-01-01

    Insertion sequences (ISs) are ubiquitous and abundant mobile genetic elements in prokaryotic genomes. ISs often encode only one protein, the transposase, which catalyzes their transposition. Recent studies have shown that transposases of many different IS families interact with the β sliding clamp, a DNA replication factor of the host. However, it was unclear to what extent this interaction limits or favors the ability of ISs to colonize a chromosome from a phylogenetically-distant organism, or if the strength of this interaction affects the transposition rate. Here we describe the proliferation of a member of the IS1634 family in Acidiphilium over ~600 generations of cultured growth. We demonstrate that the purified transposase binds to the β sliding clamp of Acidiphilium, Leptospirillum and E. coli. Further, we also demonstrate that the Acidiphilium IS1634 transposase binds to the archaeal sliding clamp (PCNA) from Methanosarcina, and that the transposase encoded by Methanosarcina IS1634 binds to Acidiphilium β. Finally, we demonstrate that increasing the strength of the interaction between β and transposase results in a higher transposition rate in vivo. Our results suggest that the interaction could determine the potential of ISs to be mobilized in bacterial populations and also their ability to proliferate within chromosomes. PMID:26306550

  14. Low noise patch-clamp current amplification by nanoparticles plasmonic-photonic coupling (analysis and modelling).

    PubMed

    Haberal, E O; SalmanOgli, A; Nasseri, B

    2016-10-01

    In this article, a patch-clamp low noise current amplification based on nanoparticles plasmonic radiation is analyzed. It is well-known, a very small current is flowing from different membrane channels and so, for extra processing the current amplification is necessary. It is notable that there are some problems in traditional electronic amplifier due to its noise and bandwidth problem. Because of the important role of the patch-clamp current in cancer research and especially its small amplitude, it is vital to intensify it without adding any noises. In this study, the current amplification is performed firstly: from the excitement of nanoparticles by the patch-clamp pico-ampere current and then, the effect of nanoparticles plasmonic far-field radiation on conductor's carriers, which will cause the current amplification. This relates to the plasmonic-photonic coupling and their effect on conductor carriers as the current perturbation agent. In the steady state, the current amplification can reach to 1000 times of initial level. Furthermore, we investigated the nanoparticles morphology changing effect such as size, nanoparticles inter-distance, and nanoparticles distance from the conductor on the amplifier parameters. Finally, it should note that the original aim is to use nanoparticles plasmonic engineering and their coupling to photonics for output current manipulating.

  15. Ion Channels in Native Chloroplast Membranes: Challenges and Potential for Direct Patch-Clamp Studies

    PubMed Central

    Pottosin, Igor; Dobrovinskaya, Oxana

    2015-01-01

    Photosynthesis without any doubt depends on the activity of the chloroplast ion channels. The thylakoid ion channels participate in the fine partitioning of the light-generated proton-motive force (p.m.f.). By regulating, therefore, luminal pH, they affect the linear electron flow and non-photochemical quenching. Stromal ion homeostasis and signaling, on the other hand, depend on the activity of both thylakoid and envelope ion channels. Experimentally, intact chloroplasts and swollen thylakoids were proven to be suitable for direct measurements of the ion channels activity via conventional patch-clamp technique; yet, such studies became infrequent, although their potential is far from being exhausted. In this paper we wish to summarize existing challenges for direct patch-clamping of native chloroplast membranes as well as present available results on the activity of thylakoid Cl− (ClC?) and divalent cation-permeable channels, along with their tentative roles in the p.m.f. partitioning, volume regulation, and stromal Ca2+ and Mg2+ dynamics. Patch-clamping of the intact envelope revealed both large-conductance porin-like channels, likely located in the outer envelope membrane and smaller conductance channels, more compatible with the inner envelope location. Possible equivalent model for the sandwich-like arrangement of the two envelope membranes within the patch electrode will be discussed, along with peculiar properties of the fast-activated cation channel in the context of the stromal pH control. PMID:26733887

  16. A stepwise aortic clamp procedure to treat porcelain aorta associated with aortic valve stenosis and hemodialysis.

    PubMed

    Isoda, Susumu; Osako, Motohiko; Kimura, Tamizo; Nishimura, Kenji; Yamanaka, Nozomu; Nakamura, Shingo; Maehara, Tadaaki

    2014-01-01

    A 62-year-old man was referred for an aortic-valve surgery because of severe aortic stenosis. Thirty years ago, he had undergone a mitral valve commissurotomy and after 9 years, the valve had been replaced by a mechanical valve. He had been undergoing hemodialysis for the past 8 years. A computed tomographic (CT) scan of the chest and abdomen showed a dense circumferential calcification in the wall of the entire thoracic and abdominal aorta, pulmonary artery, and left and right atrium. A conventional aortic-valve replacement was performed. To avoid an embolic event, a "stepwise aortic clamp" procedure was attempted and involved the following: (1) brief circulatory arrest and aortotomy during moderate hypothermia; (2) balloon occlusion at the ascending aorta during low-flow cardiopulmonary bypass (CPB); (3) endoarterectomy by using an ultrasonic surgical aspirator to enable aortic cross-clamping; and (4) a cross-clamp reinforced with felt and full-flow CPB. The patient recovered without any thromboembolic events. Using this procedure to treat a porcelain aorta seemed to reduce the time limit and reduced the risk of brain injury during cardiac surgery.

  17. Dwell-time distribution analysis of polyprotein unfolding using force-clamp spectroscopy.

    PubMed

    Brujic, Jasna; Hermans, Rodolfo I Z; Garcia-Manyes, Sergi; Walther, Kirstin A; Fernandez, Julio M

    2007-04-15

    Using the recently developed single molecule force-clamp technique we quantitatively measure the kinetics of conformational changes of polyprotein molecules at a constant force. In response to an applied force of 110 pN, we measure the dwell times of 1647 unfolding events of individual ubiquitin modules within each protein chain. We then establish a rigorous method for analyzing force-clamp data using order statistics. This allows us to test the success of a history-independent, two-state model in describing the kinetics of the unfolding process. We find that the average unfolding trajectory is independent of the number of protein modules N in each trajectory, which varies between 3 and 12 (the engineered protein length), suggesting that the unfolding events in each chain are uncorrelated. We then derive a binomial distribution of dwell times to describe the stochastic dynamics of protein unfolding. This distribution successfully describes 81% of the data with a single rate constant of alpha = 0.6 s(-1) for all N. The remainder of the data that cannot be accounted for suggests alternative unfolding barriers in the energy landscape of the protein. This method investigates the statistical features of unfolding beyond the average measurement of a single rate constant, thus providing an attractive alternative for measuring kinetics by force-clamp spectroscopy.

  18. The metabolic response to hyperglycaemic clamping in insulin-dependent diabetes.

    PubMed

    Nosadini, R; Noy, G A; Alberti, K G; Hodson, A; Orskoy, H

    1981-02-01

    The metabolic and hormonal effects of stable hyperglycaemia (10-12 mmol/l) have been examined in five insulin-dependent diabetics and compared with the results of 8 h (1200 to 2000 h) normoglycaemic (5-6 mmol/l) clamping. Glucose levels were maintained using a glucose controlled insulin infusion system. Mean blood lactate, pyruvate, total ketone bodies, glycerol and plasma non-esterified fatty acids were similar during the period of stable glycaemia at the two glucose levels. In contrast mean blood alanine was markedly elevated during hyperglycaemic clamping (0.384 +/- 0.008 vs 0.298 +/- 0.021 mmol/l) and 3-hydroxybutyrate was slightly decreased (0.068 +/- 0.007 vs 0.084 +/- 0.008 mmol/l). Plasma glucagon levels were raised during hyperglycaemic clamping and growth hormone slightly decreased. There was a close positive correlation between mean blood alanine and mean blood glucose (r = 0.79, p less than 0.01), and a negative correlation of alanine with the amount of insulin infused (r = -0.72, p less than 0.01). It is suggested that the raised alanine results from increased peripheral glucose utilisation. In general a short period of stable hyperglycaemia is not associated with a worsening of metabolic abnormalities in insulin-dependent diabetic subjects.

  19. PIP2 in pancreatic β-cells regulates voltage-gated calcium channels by a voltage-independent pathway.

    PubMed

    de la Cruz, Lizbeth; Puente, Erika I; Reyes-Vaca, Arturo; Arenas, Isabel; Garduño, Julieta; Bravo-Martínez, Jorge; Garcia, David E

    2016-10-01

    Phosphatidylinositol-4,5-bisphosphate (PIP2) is a membrane phosphoinositide that regulates the activity of many ion channels. Influx of calcium primarily through voltage-gated calcium (CaV) channels promotes insulin secretion in pancreatic β-cells. However, whether CaV channels are regulated by PIP2, as is the case for some non-insulin-secreting cells, is unknown. The purpose of this study was to investigate whether CaV channels are regulated by PIP2 depletion in pancreatic β-cells through activation of a muscarinic pathway induced by oxotremorine methiodide (Oxo-M). CaV channel currents were recorded by the patch-clamp technique. The CaV current amplitude was reduced by activation of the muscarinic receptor 1 (M1R) in the absence of kinetic changes. The Oxo-M-induced inhibition exhibited the hallmarks of voltage-independent regulation and did not involve PKC activation. A small fraction of the Oxo-M-induced CaV inhibition was diminished by a high concentration of Ca(2+) chelator, whereas ≥50% of this inhibition was prevented by diC8-PIP2 dialysis. Localization of PIP2 in the plasma membrane was examined by transfecting INS-1 cells with PH-PLCδ1, which revealed a close temporal association between PIP2 hydrolysis and CaV channel inhibition. Furthermore, the depletion of PIP2 by a voltage-sensitive phosphatase reduced CaV currents in a way similar to that observed following M1R activation. These results indicate that activation of the M1R pathway inhibits the CaV channel via PIP2 depletion by a Ca(2+)-dependent mechanism in pancreatic β- and INS-1 cells and thereby support the hypothesis that membrane phospholipids regulate ion channel activity by interacting with ion channels.

  20. Voltage-dependent gating mechanism for single fast chloride channels from rat skeletal muscle.

    PubMed Central

    Weiss, D S; Magleby, K L

    1992-01-01

    1. A voltage-dependent gating mechanism for the fast Cl- channel was developed from the analysis of single-channel current records obtained with the patch clamp technique from primary cultures of rat skeletal muscle. Up to 10(6) open and shut intervals were analysed from each of five different excised patches of membrane containing a single fast Cl- channel. 2. Rate constants for a kinetic scheme with six closed and two open states (scheme I) were estimated at a given voltage by maximum likelihood fitting of open and closed dwell-time distributions obtained at that voltage. This procedure was then repeated for data obtained at each of three to eight different membrane potentials for each channel. 3. Plots of the estimated rate constants against membrane potential typically appeared linear on semilogarithmic co-ordinates, consistent with rate constants that are exponentially dependent on voltage. 4. Regression analysis of these plots yielded two parameters for each rate constant: the value of the rate constant at -50 mV (B) and its voltage sensitivity (A). The dwell-time distributions predicted with these parameters and scheme I gave a good description of the experimental dwell-time distributions at all the studied voltages, lending further support for an exponential dependence of rate constants on membrane potential. 5. Estimates of A and B were also obtained by simultaneously fitting dwell-time distributions obtained at three to eight different voltages, in order to better define these parameters. Predicted dwell-time distributions obtained with these estimates and scheme I could approach the theoretical best description of the data for discrete-state Markov models. 6. Eight to twelve of the fourteen rate constants in scheme I appeared voltage sensitive, with effective gating charges ranging from about -1.5 to +1.0 units of electronic charge. 7. The estimated rate constants and their voltage sensitivities for the five analysed channels were generally similar, but

  1. NMR resonance assignments for the N-terminal domain of the δ subunit of the E. coli γ clamp loader complex.

    PubMed

    Alyami, Esmael M; Rizzo, Alessandro A; Beuning, Penny J; Korzhnev, Dmitry M

    2017-03-06

    The β-clamp protein and the γ clamp loader complex are essential components of bacterial DNA replication machinery. The β-clamp is a ring-shaped homodimer that encircles DNA and increases the efficiency of replication by providing a binding platform for DNA polymerases and other replication-related proteins. The β-clamp is loaded onto DNA by the five-subunit γ clamp loader complex in a multi-step ATP-dependent process. The initial steps of this process involve the cooperative binding of the β-clamp by the five subunits of ATP-bound clamp loader, which induces or traps an open conformation of the clamp. Remarkably, the δ subunit of the E. coli clamp loader, or even its 140 residue N-terminal domain (called mini-δ), alone can shift conformational equilibrium of the β-clamp towards the open state. Here we report nearly complete backbone and side-chain (1)H, (13)C and (15)N NMR resonance assignments of mini-δ that will facilitate NMR studies of the mechanisms of β-clamp opening and its loading on DNA by the clamp loader.

  2. Electrode voltage fall and total voltage of a transient arc

    NASA Astrophysics Data System (ADS)

    Valensi, F.; Ratovoson, L.; Razafinimanana, M.; Masquère, M.; Freton, P.; Gleizes, A.

    2016-06-01

    This paper deals with an experimental study of the components of a transient arc total voltage with duration of a few tens of ms and a current peak close to 1000 A. The cathode tip is made of graphite whereas the flat anode is made either of copper or of graphite; the electrodes gap is a few mm. The analysis of the electrical parameters is supported and validated by fast imaging and by two models: the first one is a 2D physical model of the arc allowing to calculate both the plasma temperature field and the arc voltage; the second model is able to estimate the transient heating of the graphite electrode. The main aim of the study was to detect the possible change of the cathode voltage fall (CVF) during the first instants of the arc. Indeed it is expected that during the first ms the graphite cathode is rather cool and the main mechanism of the electron emission should be the field effect emission, whereas after several tens of ms the cathode is strongly heated and thermionic emission should be predominant. We have observed some change in the apparent CVF but we have shown that this apparent change can be attributed to the variation of the solid cathode resistance. On the other hand, the possible change of CVF corresponding to the transition between a ‘cold’ and a ‘hot’ cathode should be weak and could not be characterized considering our measurement uncertainty of about 2 V. The arc column voltage (ACV) was estimated by subtracting the electrode voltage fall from the total arc voltage. The experimental transient evolution of the ACV is in very good agreement with the theoretical variation predicted by the model, showing the good ability of the model to study this kind of transient arc.

  3. Modeling hepatic insulin sensitivity during a meal: validation against the euglycemic hyperinsulinemic clamp.

    PubMed

    Dalla Man, Chiara; Piccinini, Francesca; Basu, Rita; Basu, Ananda; Rizza, Robert A; Cobelli, Claudio

    2013-04-15

    Recently, we proposed a model describing the suppression of endogenous glucose production (EGP) during a meal. It assumes that EGP suppression depends on glucose concentration and its rate of change and on delayed insulin action. Hepatic insulin sensitivity (S(I)(Lmeal)) can be derived from EGP model parameters. This model was shown to adequately describe EGP profiles measured with multiple tracer techniques; however, S(I)(Lmeal) has never been compared directly with its euglycemic hyperinsulinemic clamp counterpart (S(I)(Lclamp)). To do so, 62 subjects with different degrees of glucose tolerance underwent a triple-tracer mixed meal. Fifty-seven subjects also underwent a labeled ([3-(3)H]glucose) euglycemic hyperinsulinemic clamp. From the triple-tracer meal data, virtually model-independent estimates of EGP were obtained using the tracer-to-tracee clamp technique, and the EGP model was identified in each subject. Model fit was satisfactory, and S(I)(Lmeal) was estimated with good precision. Correlation between S(I)(Lclamp) and S(I)(Lmeal) was good (r = 0.72, P < 0.001); however, S(I)(Lmeal) was lower than S(I)(Lclamp) (4.60 ± 0.64 vs. 8.73 ± 1.07 10(-4) dl·kg(-1)·min(-1) per μU/ml, P < 0.01). This difference may be due to different ranges of insulin explored during the two tests (ΔI(clamp) = 15.60 ± 1.61 vs. ΔI(meal)= 83.37 ± 10.71 μU/ml) as well as steady- vs. non-steady-state glucose and insulin profiles. In conclusion, the new EGP model provides an estimate of hepatic insulin sensitivity during a meal that is in good agreement with that derived in the same individuals with a hyperinsulinemic clamp. When used in conjunction with the minimal model, the approach potentially enables estimation of hepatic insulin sensitivity from a single-tracer labeled meal or oral glucose tolerance test.

  4. Timing of clamping and factors associated with iron stores in full-term newborns

    PubMed Central

    Oliveira, Fabiana de Cássia Carvalho; Assis, Karine Franklin; Martins, Mariana Campos; do Prado, Mara Rúbia Maciel Cardoso; Ribeiro, Andréia Queiroz; Sant’Ana, Luciana Ferreira da Rocha; Priore, Silvia Eloiza; Franceschini, Sylvia do Carmo Castro

    2014-01-01

    OBJECTIVE To analyze the impact of timing of clamping and obstetric, biological and socioeconomic factors on the iron stores of full-term newborns. METHODS Cross-sectional study between October 2011 and July 2012 in which hematological parameters were evaluated for newborns in Viçosa, MG, Southeastern Brazil. It involved collecting 7 mL of umbilical cord blood from 144 full-term not underweight newborns. The parameters investigated were complete blood count, serum iron, ferritin and C-reactive protein. The time of umbilical cord clamping was measured using a digital timer without interfering in the procedures of childbirth. The birth data were collected from Live Birth Certificates and other information was obtained from the mother through a questionnaire applied in the first month postpartum. Analysis of multiple linear regression was then used to estimate the influence of biological, obstetrics and socioeconomic factors on the ferritin levels at birth. RESULTS The median ferritin was 130.3 µg/L (n = 129, minimum = 16.4; maximum = 420.5 µg/L), the mean serum iron was 137.9 μg/dL (n = 144, SD = 39.29) and mean hemoglobin was 14.7 g/dL (n = 144, SD = 1.47). The median time of cord clamping was 36 seconds, ranging between 7 and 100. The bivariate analysis detected an association between ferritin levels and color of the child, timing clamping of 60 seconds, type of delivery, the presence of gestational diabetes and per capita family income. In multivariate analysis, the variables per capita income, number of antenatal visits and length at birth accounted for 22.0% of variation in ferritin levels. CONCLUSIONS Iron stores at birth were influenced by biological, obstetric and social characteristics. Tackling anemia should involve creating policies aimed at reducing social inequalities, improving the quality of antenatal care, as well as implementing a criterion of delayed clamping of the umbilical cord within the guidelines of labor. PMID:24789632

  5. Comparative Effects of Halogenated Inhaled Anesthetics on Voltage-gated Na+ Channel Function

    PubMed Central

    OuYang, Wei; Herold, Karl F.; Hemmings, Hugh C.

    2009-01-01

    Background Inhibition of voltage-gated Na+ channels (Nav) is implicated in the synaptic actions of volatile anesthetics. We studied the effects of the major halogenated inhaled anesthetics (halothane, isoflurane, sevoflurane, enflurane and desflurane) on Nav1.4, a well characterized pharmacological model for Nav effects. Methods Na+ currents (INa) from rat Nav1.4 α-subunits heterologously expressed in Chinese hamster ovary cells were analyzed by whole cell voltage-clamp electrophysiological recording. Results Halogenated inhaled anesthetics reversibly inhibited Nav1.4 in a concentration- and voltage-dependent manner at clinical concentrations. At equi-anesthetic concentrations, peak INa was inhibited with a rank order of desflurane > halothane ≈ enflurane > isoflurane ≈ sevoflurane from a physiological holding potential (−80 mV). This suggests that the contribution of Na+ channel block to anesthesia might vary in an agent-specific manner. From a hyperpolarized holding potential that minimizes inactivation (−120 mV), peak INa was inhibited with a rank order of potency for tonic inhibition of peak INa of halothane > isoflurane ≈ sevoflurane > enflurane > desflurane. Desflurane produced the largest negative shift in voltage-dependence of fast inactivation consistent with its more prominent voltage-dependent effects. A comparison between isoflurane and halothane showed that halothane produced greater facilitation of current decay, slowing of recovery from fast inactivation, and use-dependent block than isoflurane. Conclusions Five halogenated inhaled anesthetics all inhibit a voltage-gated Na+ channel by voltage- and use-dependent mechanisms. Agent-specific differences in efficacy for Na+ channel inhibition due to differential state-dependent mechanisms creates pharmacologic diversity that could underlie subtle differences in anesthetic and nonanesthetic actions. PMID:19225394

  6. A Novel NaV1.5 Voltage Sensor Mutation Associated with Severe Atrial and Ventricular Arrhythmias

    PubMed Central

    Wang, Hong-Gang; Zhu, Wandi; Kanter, Ronald J.; Silva, Jonathan R.; Honeywell, Christina; Gow, Robert M.; Pitt, Geoffrey S.

    2016-01-01

    Background Inherited autosomal dominant mutations in cardiac sodium channels (NaV1.5) cause various arrhythmias, such as long QT syndrome and Brugada syndrome. Although dozens of mutations throughout the protein have been reported, there are few reported mutations within a voltage sensor S4 transmembrane segment and few that are homozygous. Here we report analysis of a novel lidocaine-sensitive recessive mutation, p.R1309H, in the NaV1.5 DIII/S4 voltage sensor in a patient with a complex arrhythmia syndrome. Methods and Results We expressed the wild type or mutant NaV1.5 heterologously for analysis with the patch-clamp and voltage clamp fluorometry (VCF) techniques. p.R1309H depolarized the voltage-dependence of activation, hyperpolarized the voltage-dependence of inactivation, and slowed recovery from inactivation, thereby reducing the channel availability at physiologic membrane potentials. Additionally, p.R1309H increased the “late” Na+ current. The location of the mutation in DIIIS4 prompted testing for a gating pore current. We observed an inward current at hyperpolarizing voltages that likely exacerbates the loss-of-function defects at resting membrane potentials. Lidocaine reduced the gating pore current. Conclusions The p.R1309H homozygous NaV1.5 mutation conferred both gain-of-function and loss-of-function effects on NaV1.5 channel activity. Reduction of a mutation-induced gating pore current by lidocaine suggested a therapeutic mechanism. PMID:26801742

  7. Modulated voltage metastable ionization detector

    NASA Technical Reports Server (NTRS)

    Carle, G. C.; Kojiro, D. R.; Humphrey, D. E. (Inventor)

    1985-01-01

    The output current from a metastable ionization detector (MID) is applied to a modulation voltage circuit. An adjustment is made to balance out the background current, and an output current, above background, is applied to an input of a strip chart recorder. For low level concentrations, i.e., low detected output current, the ionization potential will be at a maximum and the metastable ionization detector will operate at its most sensitive level. When the detected current from the metastable ionization detector increases above a predetermined threshold level, a voltage control circuit is activated which turns on a high voltage transistor which acts to reduce the ionization potential. The ionization potential applied to the metastable ionization detector is then varied so as to maintain the detected signal level constant. The variation in ionization potential is now related to the concentration of the constituent and a representative amplitude is applied to another input of said strip chart recorder.

  8. Voltage-Gated Hydrophobic Nanopores

    SciTech Connect

    Lavrik, Nickolay V

    2011-01-01

    Hydrophobicity is a fundamental property that is responsible for numerous physical and biophysical aspects of molecular interactions in water. Peculiar behavior is expected for water in the vicinity of hydrophobic structures, such as nanopores. Indeed, hydrophobic nanopores can be found in two distinct states, dry and wet, even though the latter is thermodynamically unstable. Transitions between these two states are kinetically hindered in long pores but can be much faster in shorter pores. As it is demonstrated for the first time in this paper, these transitions can be induced by applying a voltage across a membrane with a single hydrophobic nanopore. Such voltage-induced gating in single nanopores can be realized in a reversible manner through electrowetting of inner walls of the nanopores. The resulting I-V curves of such artificial hydrophobic nanopores mimic biological voltage-gated channels.

  9. Fiber-optic voltage sensor

    NASA Astrophysics Data System (ADS)

    Wood, C. B.

    1990-07-01

    A fiber-optic voltage sensor is described which includes a source of light, a reference fiber for receiving a known percentage of the light and an electrostrictive element having terminals across which is applied, and a voltage to be measured. The electrostrictive element is responsive to the applied voltage to assume an altered physical state. A measuring fiber also receives a known percentage of light from the light source and is secured about the electrostrictive element. The measuring fiber is provided with a cladding and exhibits an evanescent wave in the cladding. The measuring fiber has a known length which is altered when the electrostrictive element assumes its altered physical state. A differential sensor is provided which senses the intensity of light in both the reference fiber and the measuring fiber and provides an output indicative of the difference between the intensities.

  10. A low voltage CMOS low drop-out voltage regulator

    NASA Astrophysics Data System (ADS)

    Bakr, Salma Ali; Abbasi, Tanvir Ahmad; Abbasi, Mohammas Suhaib; Aldessouky, Mohamed Samir; Abbasi, Mohammad Usaid

    2009-05-01

    A low voltage implementation of a CMOS Low Drop-Out voltage regulator (LDO) is presented. The requirement of low voltage devices is crucial for portable devices that require extensive computations in a low power environment. The LDO is implemented in 90nm generic CMOS technology. It generates a fixed 0.8V from a 2.5V supply which on discharging goes to 1V. The buffer stage used is unity gain configured unbuffered OpAmp with rail-to-rail swing input stage. The simulation result shows that the implemented circuit provides load regulation of 0.004%/mA and line regulation of -11.09mV/V. The LDO provides full load transient response with a settling time of 5.2μs. Further, the dropout voltage is 200mV and the quiescent current through the pass transistor (Iload=0) is 20μA. The total power consumption of this LDO (excluding bandgap reference) is only 80μW.

  11. Conformational analysis of processivity clamps in solution demonstrates that tertiary structure does not correlate with protein dynamics.

    PubMed

    Fang, Jing; Nevin, Philip; Kairys, Visvaldas; Venclovas, Ceslovas; Engen, John R; Beuning, Penny J

    2014-04-08

    The relationship between protein sequence, structure, and dynamics has been elusive. Here, we report a comprehensive analysis using an in-solution experimental approach to study how the conservation of tertiary structure correlates with protein dynamics. Hydrogen exchange measurements of eight processivity clamp proteins from different species revealed that, despite highly similar three-dimensional structures, clamp proteins display a wide range of dynamic behavior. Differences were apparent both for structurally similar domains within proteins and for corresponding domains of different proteins. Several of the clamps contained regions that underwent local unfolding with different half-lives. We also observed a conserved pattern of alternating dynamics of the α helices lining the inner pore of the clamps as well as a correlation between dynamics and the number of salt bridges in these α helices. Our observations reveal that tertiary structure and dynamics are not directly correlated and that primary structure plays an important role in dynamics.

  12. L-band all-optical gain-clamped EDFA by utilizing C-band backward ASE

    NASA Astrophysics Data System (ADS)

    Zhang, Hao; Jin, Yanli; Dou, Qingying; Liu, Yange; Yuan, Shuzhong; Dong, Xiaoyi

    2006-04-01

    By using an optical circulator and C/L-band wavelength division multiplexer to recycle the C-band backward ASE, an L-band gain-clamped erbium-doped fiber amplifier is presented. We have experimentally studied the static gain clamping property of this amplifier. As the ASE feedback attenuation is set to 0, the gain at 1585 nm can be clamped at 18.84 ± 0.26 dB within dynamic range of 25 dB and the critical power reaches about -15.09 dBm. The gain variation and saturated output power at 1585 nm for 0 dB attenuation are 1 dB lower and 2.17 dB higher than those for 30 dB attenuation, which indicates that the L-band EDFA gain can be effectively clamped via the ASE injection technique.

  13. Conformational analysis of processivity clamps in solution demonstrates that tertiary structure does not correlate with protein dynamics

    PubMed Central

    Kairys, Visvaldas; Venclovas, Česlovas; Engen, John R.; Beuning, Penny J.

    2014-01-01

    Summary The relationship between protein sequence, structure, and dynamics has been elusive. We report one of the first comprehensive analyses using an in-solution experimental approach to study how the conservation of tertiary structure correlates with protein dynamics. Hydrogen exchange measurements of eight processivity clamp proteins from different species revealed that, despite highly similar three-dimensional structures, clamp proteins display a wide range of dynamic behavior. Differences were apparent both for structurally similar domains within proteins and for corresponding domains of different proteins. Several of the clamps contained regions that underwent local unfolding with different half-lives. We also observed a conserved pattern of alternating dynamics of the α-helices lining the inner pore of the clamps as well as a correlation between dynamics and the number of salt bridges in these α-helices. Our observations reveal that tertiary structure and dynamics are not directly correlated and that primary structure plays an important role in dynamics. PMID:24613485

  14. First clinical experience with the air purge control and electrical remote-controlled tubing clamp in mini bypass.

    PubMed

    Huybregts, Rien M A J M; Veerman, Derk P; Vonk, Alexander B A; Nesselaar, Alfred F; Paulus, Reggie C E; Thone-Passchier, Deirdre H; Smith, Annette L; de Vroege, Roel

    2006-09-01

    Most mini bypass systems do not contain a venous and cardiotomy reservoir in the cardiopulmonary bypass (CPB) circuit and lack the capability to remove venous air. In conjunction with the manufacturer the air purge control system, a system which automatically removes air that is captured in a venous bubble trap, has been developed. This system is combined with an electrical remote clamp, which automatically clamps the arterial line in case air leaves the bubble trap. Twenty consecutive patients undergoing surgery with CPB were included in this clinical validation. Venous air was removed by the air purge control during bypass. The electrical remote clamp was never activated by the system, confirming that the air purge control adequately removed venous air during these cases. The air purge control, in conjunction with the electrical remote clamp, is a valuable safety feature in mini bypass, enhancing patient safety and user friendliness while providing a level of safety equivalent to those of conventional bypass systems.

  15. A new balancing three level three dimensional space vector modulation strategy for three level neutral point clamped four leg inverter based shunt active power filter controlling by nonlinear back stepping controllers.

    PubMed

    Chebabhi, Ali; Fellah, Mohammed Karim; Kessal, Abdelhalim; Benkhoris, Mohamed F

    2016-07-01

    In this paper is proposed a new balancing three-level three dimensional space vector modulation (B3L-3DSVM) strategy which uses a redundant voltage vectors to realize precise control and high-performance for a three phase three-level four-leg neutral point clamped (NPC) inverter based Shunt Active Power Filter (SAPF) for eliminate the source currents harmonics, reduce the magnitude of neutral wire current (eliminate the zero-sequence current produced by single-phase nonlinear loads), and to compensate the reactive power in the three-phase four-wire electrical networks. This strategy is proposed in order to gate switching pulses generation, dc bus voltage capacitors balancing (conserve equal voltage of the two dc bus capacitors), and to switching frequency reduced and fixed of inverter switches in same times. A Nonlinear Back Stepping Controllers (NBSC) are used for regulated the dc bus voltage capacitors and the SAPF injected currents to robustness, stabilizing the system and to improve the response and to eliminate the overshoot and undershoot of traditional PI (Proportional-Integral). Conventional three-level three dimensional space vector modulation (C3L-3DSVM) and B3L-3DSVM are calculated and compared in terms of error between the two dc bus voltage capacitors, SAPF output voltages and THDv, THDi of source currents, magnitude of source neutral wire current, and the reactive power compensation under unbalanced single phase nonlinear loads. The success, robustness, and the effectiveness of the proposed control strategies are demonstrated through simulation using Sim Power Systems and S-Function of MATLAB/SIMULINK.

  16. Direct Observation of a Cytosine Analogue that Forms Five Hydrogen Bonds to Guanosine: Guanyl G-Clamp

    SciTech Connect

    Wilds, C.J.; Maier, M.A.; Tereshko, V.; Manoharan, M.; Egli, M.

    2010-03-08

    A novel heterocyclic base modification, the guanidino G-clamp, is designed to allow two Hoogsteen-type hydrogen bonds to form between the amino and imino nitrogen atoms of a tethered guanidinium group to O6 and N7 of guanosine, which results in a total of five hydrogen bonds (broken lines, see picture). Details of a crystal structure at 1.0-{angstrom} resolution of a modified DNA decamer containing this guanidino G-clamp analogue demonstrate its mechanism of binding.

  17. Structure-function analysis of the C-clamp of TCF/Pangolin in Wnt/ß-catenin signaling.

    PubMed

    Ravindranath, Aditi J; Ravindranath, Aditi; Cadigan, Ken M

    2014-01-01

    The evolutionarily conserved Wnt/ß-catenin (Wnt/ß-cat) pathway plays an important role in animal development in metazoans. Many Wnt targets are regulated by members of the TCF/LEF1 (TCF) family of transcription factors. All TCFs contain a High Mobility Group (HMG) domain that bind specific DNA sequences. Invertebrate TCFs and some vertebrate TCF isoforms also contain another domain, called the C-clamp, which allows TCFs to recognize an additional DNA motif known as the Helper site. While the C-clamp has been shown to be important for regulating several Wnt reporter genes in cell culture, its physiological role in regulating Wnt targets is less clear. In addition, little is known about this domain, except that two of the four conserved cysteines are functionally important. Here, we carried out a systematic mutagenesis and functional analysis of the C-clamp from the Drosophila TCF/Pangolin (TCF/Pan) protein. We found that the C-clamp is a zinc-binding domain that is sufficient for binding to the Helper site. In addition to this DNA-binding activity, the C-clamp also inhibits the HMG domain from binding its cognate DNA site. Point mutations were identified that specifically affected DNA-binding or reduced the inhibitory effect. These mutants were characterized in TCF/Pan rescue assays. The specific DNA-binding activity of the C-clamp was essential for TCF/Pan function in cell culture and in patterning the embryonic epidermis of Drosophila, demonstrating the importance of this C-clamp activity in regulating Wnt target gene expression. In contrast, the inhibitory mutation had a subtle effect in cell culture and no effect on TCF/Pan activity in embryos. These results provide important information about the functional domains of the C-clamp, and highlight its importance for Wnt/ß-cat signaling in Drosophila.

  18. In situ electrochemical activation of Ni-based colloids from an NiCl2 electrode and their advanced energy storage performance.

    PubMed

    Chen, Kunfeng; Xue, Dongfeng

    2016-10-06

    The formation of electrochemical activated cations in electrode materials to induce multiple-electron transfer reactions is a challenge for high-energy storage systems. Herein, highly electroactive Ni-based colloidal electrode materials have been synthesized by in situ electrochemical activation of a NiCl2 electrode. The highest specific capacitance of the activated Ni-based electrodes was 10 286 F g(-1) at a current density of 3 A g(-1), indicating that a three-electron Faradaic redox reaction (Ni(3+) ↔ Ni) occurred. Upon potential cycling and constant potential activation, a decrease in the charge transfer resistance can be found. Activation and utilization of multiple-electron reactions is an efficient route to increase the energy density of supercapacitors. This newly designed colloidal pseudocapacitor is compatible with inorganic pseudocapacitor chemistry, which enables us to use metal cations directly via their commercial salts rather than their oxide/hydroxide compounds.

  19. Effects of lithium content on the electrochemical lithium intercalation reaction into LiNiO 2 and LiCoO 2 electrodes

    NASA Astrophysics Data System (ADS)

    Choi, Young-Min; Pyun, Su-Il; Bae, Joon-Sung; Moon, Seong-In

    The electrochemical lithium intercalation reaction into LiNiO 2 and LiCoO 2 electrodes in 1 M LiClO 4—propylene carbonate solution is investigated as a function of lithium content in the oxide electrodes by using X-ray diffractometry (XRD), electrochemical impedance spectroscopy (EIS), and a galvanostatic intermittent titration technique (GITT). Li 1-δNiO 2 shows a greater loss in capacity during the first intermittent discharge, as well as a higher resistance for the electrochemical intercalation reaction, in comparison with Li 1-δCoO 2. This is attributed to a partial cation mixing in Li 1-δNiO 2 which is substantiated by XRD studies. The electrochemical impedance spectra of the Li 1-δNiO 2 electrode reveals that the magnitude of the intermediate frequency arc that is associated with the absorption reaction decreases with increasing lithium content, (1 — δ), in the range from 0.5 to 0.7. By contrast, Li 1-δCoO 2 exhibits the reverse behaviour.—The component diffusivities of lithium ions display a nearly constant value, in the order of 10 -11 cm 2 s -1, for both electrodes at room temperature, irrespective of the value of (1 — δ) over the range 0.5-0.7. It is suggested that lithium-ion diffusion through both the layered oxides is affected by the number of empty sites within the lithium-ion layer, and not by the lattice parameter.

  20. A conserved physical and functional interaction between the cell cycle checkpoint clamp loader and DNA ligase I of eukaryotes.

    PubMed

    Song, Wei; Levin, David S; Varkey, Johnson; Post, Sean; Bermudez, Vladimir P; Hurwitz, Jerard; Tomkinson, Alan E

    2007-08-03

    DNA ligase I joins Okazaki fragments during DNA replication and completes certain excision repair pathways. The participation of DNA ligase I in these transactions is directed by physical and functional interactions with proliferating cell nuclear antigen, a DNA sliding clamp, and, replication factor C (RFC), the clamp loader. Here we show that DNA ligase I also interacts with the hRad17 subunit of the hRad17-RFC cell cycle checkpoint clamp loader, and with each of the subunits of its DNA sliding clamp, the heterotrimeric hRad9-hRad1-hHus1 complex. In contrast to the inhibitory effect of RFC, hRad17-RFC stimulates joining by DNA ligase I. Similar results were obtained with the homologous Saccharomyces cerevisiae proteins indicating that the interaction between the replicative DNA ligase and checkpoint clamp is conserved in eukaryotes. Notably, we show that hRad17 preferentially interacts with and specifically stimulates dephosphorylated DNA ligase I. Moreover, there is an increased association between DNA ligase I and hRad17 in S phase following DNA damage and replication blockage that occurs concomitantly with DNA damage-induced dephosphorylation of chromatin-associated DNA ligase I. Thus, our results suggest that the in vivo interaction between DNA ligase I and the checkpoint clamp loader is regulated by post-translational modification of DNA ligase I.

  1. Catch and Patch: A Pipette-Based Approach for Automating Patch Clamp That Enables Cell Selection and Fast Compound Application.

    PubMed

    Danker, Timm; Braun, Franziska; Silbernagl, Nikole; Guenther, Elke

    2016-03-01

    Manual patch clamp, the gold standard of electrophysiology, represents a powerful and versatile toolbox to stimulate, modulate, and record ion channel activity from membrane fragments and whole cells. The electrophysiological readout can be combined with fluorescent or optogenetic methods and allows for ultrafast solution exchanges using specialized microfluidic tools. A hallmark of manual patch clamp is the intentional selection of individual cells for recording, often an essential prerequisite to generate meaningful data. So far, available automation solutions rely on random cell usage in the closed environment of a chip and thus sacrifice much of this versatility by design. To parallelize and automate the traditional patch clamp technique while perpetuating the full versatility of the method, we developed an approach to automation, which is based on active cell handling and targeted electrode placement rather than on random processes. This is achieved through an automated pipette positioning system, which guides the tips of recording pipettes with micrometer precision to a microfluidic cell handling device. Using a patch pipette array mounted on a conventional micromanipulator, our automated patch clamp process mimics the original manual patch clamp as closely as possible, yet achieving a configuration where recordings are obtained from many patch electrodes in parallel. In addition, our implementation is extensible by design to allow the easy integration of specialized equipment such as ultrafast compound application tools. The resulting system offers fully automated patch clamp on purposely selected cells and combines high-quality gigaseal recordings with solution switching in the millisecond timescale.

  2. Cyclosporin A pharmacokinetics in liver transplant recipients in relation to biliary T-tube clamping and liver dysfunction.

    PubMed Central

    Naoumov, N V; Tredger, J M; Steward, C M; O'Grady, J G; Grevel, J; Niven, A; Whiting, B; Williams, R

    1989-01-01

    Cyclosporin A pharmacokinetics were studied after oral (4-14 mg/kg body weight) and intravenous dosing (1.5-3.5 mg/kg) in 13 orthotopic liver transplant recipients before and after permanent clamping of the biliary T-tube. After T-tube clamping, cyclosporin A absorption was faster and more complete with the mean time of peak concentration, tmax, reduced to around three hours from around six hours and mean bioavailability rising from only 16.6% (n = 13) to 30% in the entire group (n = 11 after clamping) or to 35% after excluding two patients who developed severe cholestasis after the preclamping study. Bioavailability in these two patients fell below 8% and to around 1% in a further patient with severe graft dysfunction. Clamping reduced the metabolic clearance of cyclosporin A by only 25% from a mean before clamping of 2.9 ml/min/kg to 2.3 ml/min/kg (n = 11). Oral cyclosporin A becomes a reliable means of maintaining therapeutic drug concentrations only after bioavailability increases in association with T-tube clamping and in the absence of severe liver dysfunction or cholestasis. PMID:2651227

  3. Cyclosporin A pharmacokinetics in liver transplant recipients in relation to biliary T-tube clamping and liver dysfunction.

    PubMed

    Naoumov, N V; Tredger, J M; Steward, C M; O'Grady, J G; Grevel, J; Niven, A; Whiting, B; Williams, R

    1989-03-01

    Cyclosporin A pharmacokinetics were studied after oral (4-14 mg/kg body weight) and intravenous dosing (1.5-3.5 mg/kg) in 13 orthotopic liver transplant recipients before and after permanent clamping of the biliary T-tube. After T-tube clamping, cyclosporin A absorption was faster and more complete with the mean time of peak concentration, tmax, reduced to around three hours from around six hours and mean bioavailability rising from only 16.6% (n = 13) to 30% in the entire group (n = 11 after clamping) or to 35% after excluding two patients who developed severe cholestasis after the preclamping study. Bioavailability in these two patients fell below 8% and to around 1% in a further patient with severe graft dysfunction. Clamping reduced the metabolic clearance of cyclosporin A by only 25% from a mean before clamping of 2.9 ml/min/kg to 2.3 ml/min/kg (n = 11). Oral cyclosporin A becomes a reliable means of maintaining therapeutic drug concentrations only after bioavailability increases in association with T-tube clamping and in the absence of severe liver dysfunction or cholestasis.

  4. How to Properly Measure a Current-Voltage Relation?—Interpolation vs. Ramp Methods Applied to Studies of GABAA Receptors

    PubMed Central

    Yelhekar, Tushar D.; Druzin, Michael; Karlsson, Urban; Blomqvist, Erii; Johansson, Staffan

    2016-01-01

    The relation between current and voltage, I-V relation, is central to functional analysis of membrane ion channels. A commonly used method, since the introduction of the voltage-clamp technique, to establish the I-V relation depends on the interpolation of current amplitudes recorded at different steady voltages. By a theoretical computational approach as well as by experimental recordings from GABAA-receptor mediated currents in mammalian central neurons, we here show that this interpolation method may give reversal potentials and conductances that do not reflect the properties of the channels studied under conditions when ion flux may give rise to concentration changes. Therefore, changes in ion concentrations may remain undetected and conclusions on changes in conductance, such as during desensitization, may be mistaken. In contrast, an alternative experimental approach, using rapid voltage ramps, enable I-V relations that much better reflect the properties of the studied ion channels. PMID:26869882

  5. High-Voltage Droplet Dispenser

    NASA Technical Reports Server (NTRS)

    Eichenberg, Dennis J.

    2003-01-01

    An apparatus that is extremely effective in dispensing a wide range of droplets has been developed. This droplet dispenser is unique in that it utilizes a droplet bias voltage, as well as an ionization pulse, to release a droplet. Apparatuses that deploy individual droplets have been used in many applications, including, notably, study of combustion of liquid fuels. Experiments on isolated droplets are useful in that they enable the study of droplet phenomena under well-controlled and simplified conditions. In this apparatus, a syringe dispenses a known value of liquid, which emerges from, and hangs onto, the outer end of a flat-tipped, stainless steel needle. Somewhat below the needle tip and droplet is a ring electrode. A bias high voltage, followed by a high-voltage pulse, is applied so as to attract the droplet sufficiently to pull it off the needle. The voltages are such that the droplet and needle are negatively charged and the ring electrode is positively charged.

  6. High voltage MOSFET switching circuit

    DOEpatents

    McEwan, T.E.

    1994-07-26

    The problem of source lead inductance in a MOSFET switching circuit is compensated for by adding an inductor to the gate circuit. The gate circuit inductor produces an inductive spike which counters the source lead inductive drop to produce a rectangular drive voltage waveform at the internal gate-source terminals of the MOSFET. 2 figs.

  7. High voltage MOSFET switching circuit

    DOEpatents

    McEwan, Thomas E.

    1994-01-01

    The problem of source lead inductance in a MOSFET switching circuit is compensated for by adding an inductor to the gate circuit. The gate circuit inductor produces an inductive spike which counters the source lead inductive drop to produce a rectangular drive voltage waveform at the internal gate-source terminals of the MOSFET.

  8. High Voltage Space Solar Arrays

    NASA Technical Reports Server (NTRS)

    Ferguson, D. C.; Hillard, G. B.; Vayner, B. V.; Galofaro, J. T.; Lyons, Valerie (Technical Monitor)

    2002-01-01

    Recent tests performed at the NASA Glenn Research Center and elsewhere have shown promise in the design and construction of high voltage (300-1000 V) solar arrays for space applications. Preliminary results and implications for solar array design will be discussed, with application to direct-drive electric propulsion and space solar power.

  9. Voltage control of ferromagnetic resonance

    NASA Astrophysics Data System (ADS)

    Zhou, Ziyao; Peng, Bin; Zhu, Mingmin; Liu, Ming

    2016-05-01

    Voltage control of magnetism in multiferroics, where the ferromagnetism and ferroelectricity are simultaneously exhibiting, is of great importance to achieve compact, fast and energy efficient voltage controllable magnetic/microwave devices. Particularly, these devices are widely used in radar, aircraft, cell phones and satellites, where volume, response time and energy consumption is critical. Researchers realized electric field tuning of magnetic properties like magnetization, magnetic anisotropy and permeability in varied multiferroic heterostructures such as bulk, thin films and nanostructure by different magnetoelectric (ME) coupling mechanism: strain/stress, interfacial charge, spin-electromagnetic (EM) coupling and exchange coupling, etc. In this review, we focus on voltage control of ferromagnetic resonance (FMR) in multiferroics. ME coupling-induced FMR change is critical in microwave devices, where the electric field tuning of magnetic effective anisotropic field determines the tunability of the performance of microwave devices. Experimentally, FMR measurement technique is also an important method to determine the small effective magnetic field change in small amount of magnetic material precisely due to its high sensitivity and to reveal the deep science of multiferroics, especially, voltage control of magnetism in novel mechanisms like interfacial charge, spin-EM coupling and exchange coupling.

  10. LHCb calorimeters high voltage system

    NASA Astrophysics Data System (ADS)

    Gilitsky, Yu.; Golutvin, A.; Konoplyannikov, A.; Lefrancois, J.; Perret, P.; Schopper, A.; Soldatov, M.; Yakimchuk, V.

    2007-02-01

    The calorimeter system in LHCb aims to identify electrons, photons and hadrons. All calorimeters are equipped with Hamamatsu photo tubes as devices for light to signal conversion. Eight thousand R7899-20 tubes are used for electromagnetic and hadronic calorimeters and two hundred 64 channels multi-anode R7600-00-M64 for Scintillator-Pad/Preshower detectors. The calorimeter high voltage (HV) system is based on a Cockroft Walton (CW) voltage converter and a control board connected to the Experiment Control System (ECS) by serial bus. The base of each photomultiplier tube (PMT) is built with a high voltage converter and constructed on an individual printed circuit board, using compact surface mount components. The base is attached directly to the PMT. There are no HV cables in the system. A Field Programmable Gate Array (FPGA) is used on the control board as an interface between the ECS and the 200 control channels. The FPGA includes also additional functionalities allowing automated monitoring and ramp up of the high voltage values. This paper describes the HV system architecture, some technical details of the electronics implementation and summarizes the system performance. This safe and low power consumption HV electronic system for the photomultiplier tubes can be used for various biomedical apparatus too.

  11. Frequency-controlled voltage regulator

    NASA Technical Reports Server (NTRS)

    Mclyman, W. T.

    1980-01-01

    Converting input ac to higher frequency reduce size and weight and makes possible unique kind of regulation. Since conversion frequency is above range of human hearing, supply generated on audible noise. It also exploits highfrequency conversion features to regulate its output voltage in novel way. Circuit is inherently short-circuit proof.

  12. Eugenol dilates rat cerebral arteries by inhibiting smooth muscle cell voltage-dependent calcium channels.

    PubMed

    Peixoto-Neves, Dieniffer; Leal-Cardoso, Jose Henrique; Jaggar, Jonathan H

    2014-11-01

    Plants high in eugenol, a phenylpropanoid compound, are used as folk medicines to alleviate diseases including hypertension. Eugenol has been demonstrated to relax conduit and ear arteries and reduce systemic blood pressure, but mechanisms involved are unclear. Here, we studied eugenol regulation of resistance-size cerebral arteries that control regional brain blood pressure and flow and investigated mechanisms involved. We demonstrate that eugenol dilates arteries constricted by either pressure or membrane depolarization (60 mM K) in a concentration-dependent manner. Experiments performed using patch-clamp electrophysiology demonstrated that eugenol inhibited voltage-dependent calcium (Ca) currents, when using Ba as a charge carrier, in isolated cerebral artery smooth muscle cells. Eugenol inhibition of voltage-dependent Ca currents involved pore block, a hyperpolarizing shift (∼-10 mV) in voltage-dependent inactivation, an increase in the proportion of steady-state inactivating current, and acceleration of inactivation rate. In summary, our data indicate that eugenol dilates cerebral arteries by means of multimodal inhibition of voltage-dependent Ca channels.

  13. Regulation of Na(+) channel inactivation by the DIII and DIV voltage-sensing domains.

    PubMed

    Hsu, Eric J; Zhu, Wandi; Schubert, Angela R; Voelker, Taylor; Varga, Zoltan; Silva, Jonathan R

    2017-03-06

    Functional eukaryotic voltage-gated Na(+) (NaV) channels comprise four domains (DI-DIV), each containing six membrane-spanning segments (S1-S6). Voltage sensing is accomplished by the first four membrane-spanning segments (S1-S4), which together form a voltage-sensing domain (VSD). A critical NaV channel gating process, inactivation, has previously been linked to activation of the VSDs in DIII and DIV. Here, we probe this interaction by using voltage-clamp fluorometry to observe VSD kinetics in the presence of mutations at locations that have been shown to impair NaV channel inactivation. These locations include the DIII-DIV linker, the DIII S4-S5 linker, and the DIV S4-S5 linker. Our results show that, within the 10-ms timeframe of fast inactivation, the DIV-VSD is the primary regulator of inactivation. However, after longer 100-ms pulses, the DIII-DIV linker slows DIII-VSD deactivation, and the rate of DIII deactivation correlates strongly with the rate of recovery from inactivation. Our results imply that, over the course of an action potential, DIV-VSDs regulate the onset of fast inactivation while DIII-VSDs determine its recovery.

  14. Characterization of a voltage-dependent conductance in the basolateral membrane of leech skin epithelium.

    PubMed

    Schnizler, M; Clauss, W

    1998-05-01

    Voltage clamp studies were performed on the dorsal integument of Hirudo medicinalis. Under apical calcium-free conditions an inward-directed component of transepithelial current was activated by changes of transepithelial voltage. Depolarization caused up to 50% increase of the transepithelial sodium current. Hyperpolarization had no comparable effects. With calcium (1.8 mM) or amiloride (100 microM) in the apical solution and in sodium-free solutions the inward-directed current failed to increase after depolarization. Activation also occurred under chloride-free conditions. Permeabilization of the apical membrane by nystatin (5 microM) increased the current activation significantly. After nystatin, calcium as well as amiloride lost their inhibitory effects. This indicates a basolateral localization of the voltage-dependent conductance. Vesicle insertion or cytoskeletal structures are probably not involved in regulation, as seen by the lack of effects of brefeldin A and the cytochalasins B and D. However, serosal hyposmolar solutions (170 mosmol.1(-1)) caused a reinforced activation of the current. Our results indicate a voltage-dependent conductance in a tight sodium-absorbing epithelium.

  15. Design, Control, and Modeling of a New Voltage Source Converter for HVDC System

    NASA Astrophysics Data System (ADS)

    Mohan, Madhan; Singh, Bhim; Ketan Panigrahi, Bijaya

    2013-05-01

    Abstract: A New Voltage Source Converter (VSC) based on neutral clamped three-level circuit is proposed for High Voltage DC (HVDC) system. The proposed VSC is designed in a multipulse configuration. The converter is operated by Fundamental Frequency Switching (FFS). A new control method is developed for achieving all the necessary control aspects of HVDC system such as independent real and reactive power control, bidirectional real and reactive power control. The basic of the control method is varying the pulse width and by keeping the dc link voltage constant. The steady state and dynamic performances of HVDC system interconnecting two different frequencies network are demonstrated for active and reactive power control. Total number of transformers used in this system are reduced to half in comparison with the two-level VSCs for both active and reactive power control. The performance of the HVDC system is improved in terms of reduced harmonics level even at fundamental frequency switching. The harmonic performance of the designed converter is also studied for different value of the dead angle (β), and the optimized range of the dead angle is achieved for varying reactive power requirement. Simulation results are presented for the designed three level multipulse voltage source converters with the proposed control algorithm.

  16. Regulation of Na+ channel inactivation by the DIII and DIV voltage-sensing domains

    PubMed Central

    Hsu, Eric J.; Zhu, Wandi; Voelker, Taylor; Varga, Zoltan

    2017-01-01

    Functional eukaryotic voltage-gated Na+ (NaV) channels comprise four domains (DI–DIV), each containing six membrane-spanning segments (S1–S6). Voltage sensing is accomplished by the first four membrane-spanning segments (S1–S4), which together form a voltage-sensing domain (VSD). A critical NaV channel gating process, inactivation, has previously been linked to activation of the VSDs in DIII and DIV. Here, we probe this interaction by using voltage-clamp fluorometry to observe VSD kinetics in the presence of mutations at locations that have been shown to impair NaV channel inactivation. These locations include the DIII–DIV linker, the DIII S4–S5 linker, and the DIV S4-S5 linker. Our results show that, within the 10-ms timeframe of fast inactivation, the DIV-VSD is the primary regulator of inactivation. However, after longer 100-ms pulses, the DIII–DIV linker slows DIII-VSD deactivation, and the rate of DIII deactivation correlates strongly with the rate of recovery from inactivation. Our results imply that, over the course of an action potential, DIV-VSDs regulate the onset of fast inactivation while DIII-VSDs determine its recovery. PMID:28232510

  17. Inhibition of voltage-gated potassium channels mediates uncarboxylated osteocalcin-regulated insulin secretion in rat pancreatic β cells.

    PubMed

    Gao, Jingying; Zhong, Xiangqin; Ding, Yaqin; Bai, Tao; Wang, Hui; Wu, Hongbin; Liu, Yunfeng; Yang, Jing; Zhang, Yi

    2016-04-15

    Insulin secretion from pancreatic β cells is important to maintain glucose homeostasis and is regulated by electrical activities. Uncarboxylated osteocalcin, a bone-derived protein, has been reported to regulate glucose metabolism by increasing insulin secretion, stimulating β cell proliferation and improving insulin sensitivity. But the underlying mechanisms of uncarboxylated osteocalcin-modulated insulin secretion remain unclear. In the present study, we investigated the relationship of uncarboxylated osteocalcin-regulated insulin secretion and voltage-gated potassium (KV) channels, voltage-gated calcium channels in rat β cells. Insulin secretion was measured by radioimmunoassay. Channel currents and membrane action potentials were recorded using the conventional whole-cell patch-clamp technique. Calcium imaging system was used to analyze intracellular Ca(2+) concentration ([Ca(2+)]i). The data show that under 16.7mmol/l glucose conditions uncarboxylated osteocalcin alone increased insulin secretion and [Ca(2+)]i, but with no such effects on insulin secretion and [Ca(2+)]i in the presence of a KV channel blocker, tetraethylammonium chloride. In the patch-clamp experiments, uncarboxylated osteocalcin lengthened action potential duration and significantly inhibited KV currents, but had no influence on the characteristics of voltage-gated calcium channels. These results indicate that KV channels are involved in uncarboxylated osteocalcin-regulated insulin secretion in rat pancreatic β cells. By inhibiting KV channels, uncarboxylated osteocalcin prolongs action potential duration, increases intracellular Ca(2+) concentration and finally promotes insulin secretion. This finding provides new insight into the mechanisms of osteocalcin-modulated insulin secretion.

  18. The effect of hyperinsulinaemic-euglycaemic clamp and exercise on bone remodeling markers in obese men

    PubMed Central

    Levinger, Itamar; Brennan-Speranza, Tara C; Jerums, George; Stepto, Nigel K; Serpiello, Fabio R; McConell, Glenn K; Anderson, Mitchell; Hare, David L; Byrnes, Elizabeth; Ebeling, Peter R; Seeman, Ego

    2015-01-01

    Bone remodelling markers (BRMs) are suppressed following a glucose load and during glucose infusion. As exercise increases indices of bone health and improves glucose handling, we hypothesised that, at rest, hyperinsulinaemic-euglycaemic clamp will suppress BRMs in obese men and that exercise prior to the clamp will prevent this suppression. Eleven obese nondiabetic men (age 58.1±2.2 years, body mass index=33.1±1.4 kg m−2 mean±s.e.m.) had a hyperinsulinaemic-euglycaemic clamp (HEC) at rest (Control) and 60 min post exercise (four bouts × 4 min cycling at 95% of hazard ratiopeak). Blood samples were analysed for serum insulin, glucose, bone formation markers, total osteocalcin (tOC) and procollagen type 1 N-terminal propeptide (P1NP), and the bone resorption marker, β-isomerised C-terminal telopeptides (β-CTx). In the control trial (no exercise), tOC, P1NP and β-CTx decreased with HEC by >10% compared with baseline (P<0.05). Fasting serum glucose, but not insulin, tended to correlate negatively with the BRMs (β range −0.57 to −0.66, p range 0.051–0.087). β-CTx, but not OC or P1NP, increased within 60 min post exercise (∼16%, P<0.01). During the post-exercise HEC, the glucose infusion rate was ∼30% higher compared with the no exercise trial. Despite this, BRMs were only suppressed to a similar extent as in the control session (10%). HEC suppressed BRMs in obese men. Exercise did not prevent this suppression of BRMs by HEC but improved glucose handling during the trial. It remains to be tested whether an exercise intervention of longer duration may be able to prevent the effect of HEC on bone remodelling. PMID:26331010

  19. Root hydraulic conductivity measured by pressure clamp is substantially affected by internal unstirred layers.

    PubMed

    Knipfer, Thorsten; Steudle, Ernst

    2008-01-01

    Using the root pressure probe in the pressure clamping (PC) mode, the impact of internal unstirred layers (USLs) was quantified for young corn roots, both in experiments and in computer simulations applying the convection/diffusion model of Knipfer et al. In the experiments, water flows (J(Vr)s) during PC were analysed in great detail, showing that J(Vr)s (and the apparent root hydraulic conductivity) were high during early stages of PC and declined rapidly during the first 80 s of clamping to a steady-state value of 40-30% of the original. The comparison of experimental results with simulations showed that, during PC, internal USLs at the inner surface of the endodermis substantially modify the overall force driving the water. As a consequence, J(Vr) and Lp(r) were inhibited. Effects of internal USLs were minimized when using the pressure relaxation mode, when internal USLs had not yet developed. Additional stop-clamp experiments and experiments where the endodermis was punctured to reduce the effect of internal USLs verified the existence of internal USLs during PC. Data indicated that the role of pressure propagation along the root xylem for both PC and pressure relaxation modes should be small, as should the effects of filling of the capacities during root pressure probe experiments, which are discussed as an alternative model. The results supported the idea that concentration polarization effects at the endodermis (internal USLs) cause a serious problem whenever relatively large amounts of water (xylem sap) are radially moved across the root, such as during PC or when using the high-pressure flow meter technique.

  20. Automated measurement of intestinal mucosa electrical parameters using a new digital clamp.

    PubMed

    Mathieu, Julien; Mammar, Saïd; Eto, Bruno

    2008-10-01

    Electrophysiological studies that include measurements of the electrical parameters of the epithelium offer insight into the cell's ability to react to different biological effectors and their functional viability. These parameters are commonly measured using a Ussing permeation chamber; however, most Ussing permeation chambers currently available must follow a strict operational protocol, and the type of electrodes used has to be taken into special consideration. The purpose of this study was to develop a new Ussing permeation chamber device with an automatic digital clamp which uses a microcontroller. Conventional electrodes, such as platinum or Ag/AgCl electrodes, are replaced by stainless steel 316L working electrodes. The electrode-electrolyte interface (inox-Ringer's) study was performed by impedance spectroscopy in the range of 1-10 kHz. The determination of Warburg's model electrical parameters was inferred from the Nyquist diagram. The model validation of the new digital clamp was performed experimentally on isolated segments of mouse jejunum. Two main study results should be mentioned. One is that impedance spectroscopy on stainless steel electrodes has provided Warburg's parameters, allowing the development of a transfer function model. The other is that the new digital clamp can simultaneously measure or calculate conductance, potential difference and short-circuit current. These results have also confirmed the great importance of Warburg's model for determining the electrical parameters of the electrode-electrolyte interface, and have shown that the measurement of intestinal mucosa electrical parameters can be achieved with a digital correction. Finally, the results suggest that stainless steel electrodes can be used successfully in a Ussing permeation chamber as working electrodes.