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Interpretation and physiological significance of diastolic coronary artery pressure-flow relationships in the canine coronary bed.

We analyzed the relationship between diastolic coronary artery pressure and flow in the canine coronary bed, using an electrical analog model of the coronary circulation based on the theory of critical closure. The model contains a voltage-dependent nonlinear resistance and capacitance. The behavior of the resistive element was described using experimental diastolic pressure-flow curves obtained in the absence of compliance effects. Compliance free zero flow pressure intercepts (Pf0) exceeded coronary venous pressure (Pv) by 2- to 5-fold and were related to initial diastolic coronary artery pressure Pa(0) and flow F(0), and Pv by: Pf0 = 14.3 [(Pa(0) - Pv)/F(0)] + Pv + 4.0 (r = 0.93). When coronary artery pressure was suddenly lowered to values less than or equal to the compliance-free Pf0, diastolic flow abruptly decreased and, after a transient reversal, remained at zero for up to 8 seconds. In the model, zero flow pressure represents critical closing pressure and the resistance regulating flow is the difference between coronary artery and venous pressure divided by flow. Theoretically predicted pressure-flow curves were in good agreement with existing experimental data, including the effects of elevating coronary venous pressure on zero flow pressure. Differences between compliance-free pressure-flow curves and those obtained with pressure gradually decreasing were explained by a coronary arterial compliance whose magnitude varies inversely with pressure and is dependent on vasomotor tone. In conclusion, the results of this study demonstrate the existence of a diastolic pressure gradient across the canine coronary bed at zero flow which is dependent on coronary vasomotor tone. A theoretical model of the coronary circulation based on the concept of critical closure describes the observed relationship between diastolic coronary artery pressure and flow during various experimental conditions.

Animals↗

An evaluation of safety guidelines to restrict exposure to stray radiofrequency radiation from short-wave diathermy units.

Short-wave diathermy (SWD), a form of radiofrequency radiation used therapeutically by physiotherapists, may be applied in continuous (CSWD) or pulsed (PSWD) mode using either capacitive or inductive methods. Stray radiation emitted by these units may exceed exposure guidelines close to the equipment. Discrepant guidelines exist on a safe distance from an operating unit for operators and other personnel. Stray electric (E-field) and magnetic (H-field) field strengths from 10 SWD units in six departments were examined using a PMM 8053 meter and two isotropic probes (EP-330, HP-032). A 5 l saline phantom completed the patient circuit. Measurements were recorded in eight directions between 0.5 m and 2 m at hip and eye levels while the units operated at maximum output and data compared to current guidelines. Results found stray fields from capacitive CSWD fell below operator limits at 2 m (E-field 4.8-39.8 V/m; H-field 0.015-0.072 A/m) and at 1 m for inductive CSWD (E-field 0-36 V/m; H-field 0.01-0.065 A/m). Capacitive PSWD fields fell below the limits at 1.5 m (E-field 1.2-19.9 V/m; H-field 0.002-0.045 A/m) and at 1m for inductive PSWD (E-field 0.74.0 V/m; H-field 0.009-0.03 A/m). An extra 0.5 m was required before fields fell below the guidelines for other personnel. These results demonstrate, under a worst case scenario, emissions from SWD exceed the guidelines for operators at distances currently recommended as safe. Future guidelines should include recommendations for personnel other than physiotherapists.

Diathermy↗

Effect of adherence, cell morphology, and lipopolysaccharide on potassium conductance and passive membrane properties of murine macrophage J774.1 cells.

The effects of adherence, cell morphology, and lipopolysaccharide on electrical membrane properties and on the expression of the inwardly rectifying K conductance in J774.1 cells were investigated. Whole-cell inwardly rectifying K currents (Ki), membrane capacitance (Cm), and membrane potential (Vm) were measured using the patch-clamp technique. Specific Ki conductance (GKi, whole-cell Ki conductance corrected for leak and normalized to membrane capacitance) was measured as a function of time after adherence, and was found to increase almost twofold one day after plating. Membrane potential (Vm) also increased from -42 +/- 4 mV (n = 32) to -58 +/- 2 mV (n = 47) over the same time period. GKi and Vm were correlated with each other; GL (leak conductance normalized to membrane capacitance) and Vm were not. The magnitudes of GKi and Vm 15 min to 2 hr after adherence were unaffected by the presence of 100 microM cycloheximide, but the increase in GKi and Vm that normally occurred between 2 and 8 hr after adherence was abolished by cycloheximide treatment. Membrane properties were analyzed as a function of cell morphology, by dividing cells into three categories ranging from small round cells to large, extremely spread cells. The capacitance of spread cells increased more than twofold within one day after adherence, which indicates that spread cells inserted new membrane. Spread cells had more negative resting membrane potentials than round cells, but GKi and GL were not significantly different. Lipopolysaccharide-(LPS; 1 or 10 micrograms/ml) treated cells showed increased Cm compared to control cells plated for comparable times. In contrast to the effect of adherence, LPS-treated cells exhibited a significantly lower GKi than control cells, indicating that the additional membrane did not have as high a density of functional GKi channels. We conclude that both adherence and LPS treatment increase the total surface membrane area of J774 cells and change the density of Ki channels. In addition, this study demonstrates that membrane area and density of Ki channels can vary independently of one another.

Animals↗

Non-invasive determination of bacterial single cell properties by electrorotation.

So far, electrorotation and its application to the determination of single cell properties have been limited to eukaryotes. Here an experimental system is described that allows the recording of electrorotation spectra of single bacterial cells. The small physical dimensions of the developed measuring chamber combined with a single frame video analysis made it possible to monitor the rotation of objects as small as bacteria by microscopical observation despite Brownian rotation and cellular movement. Thus physical properties of distinct organelles of E. coli could be simultaneously determined in vivo at frequencies between 1 kHz and 1 GHz. Experimental data were evaluated following a three-shell model of the cell. Electrical conductivities of cytoplasm and outer membrane were determined to 4.4 mS/cm and 25 microS/cm, respectively, that of the periplasmic space was found to increase with the square root of the medium ionic strength. Specific capacitances of inner and outer membrane amounted to 1.4 microF/cm2 and 0.26 microF/cm2, respectively, the thickness of the periplasm to about 50 nm. Heat treatment of the cells lead to a reduction of cytoplasmic conductivity to 0.9 mS/cm, probably caused by an efflux of ions through the permeabilized inner membrane.

Cytoplasm↗

System identification of electrically coupled smooth muscle cells: the passive electrical properties.

A system model approach based on a network model is used to investigate the passive electrical properties of coupled smooth muscle cells. This approach makes use of a gradient method of optimization to estimate the passive electrical parameters directly from the magnitude of the input impedance or voltage transfer function of the network model. The need for subjective measurements of parameters and many intermediate steps involved in the analysis using the conventional signal model approach are eliminated. The coupling resistance and capacitance are estimated with sound theoretical and mathematical grounds directly from experimental data. From the simulated results using SPICE, it is evident that the system model approach is accurate, flexible, and reproducible. These properties grant the new approach excellent potential for future studies of drug actions on smooth muscle cells and their associated electrical coupling. Also, sensitivities of the network model with respect to its parameters can readily be obtained. This may provide new insight into the coupling mechanisms of smooth muscle cells.

Electric Conductivity↗

PGE(2) activation of apical membrane Cl(-) channels in A6 epithelia: impedance analysis.

Measurements of transepithelial electrical impedance of continuously short-circuited A6 epithelia were made at audio frequencies (0.244 Hz to 10.45 kHz) to investigate the time course and extent to which prostaglandin E(2) (PGE(2)) modulates Cl(-) transport and apical membrane capacitance in this cell-cultured model epithelium. Apical and basolateral membrane resistances were determined by nonlinear curve-fitting of the impedance vectors at relatively low frequencies (<50 Hz) to equations (Păunescu, T. G., and S. I. Helman. 2001. Biophys. J. 81:838--851) where depressed Nyquist impedance semicircles were characteristic of the membrane impedances under control Na(+)-transporting and amiloride-inhibited conditions. In all tissues (control, amiloride-blocked, and amiloride-blocked and furosemide-pretreated), PGE(2) caused relatively small (< approximately 3 microA/cm(2)) and rapid (<60 s) maximal increase of chloride current due to activation of a rather large increase of apical membrane conductance that preceded significant activation of Na(+) transport through amiloride-sensitive epithelial Na(+) channels (ENaCs). Apical membrane capacitance was frequency-dependent with a Cole-Cole dielectric dispersion whose relaxation frequency was near 150 Hz. Analysis of the time-dependent changes of the complex frequency-dependent equivalent capacitance of the cells at frequencies >1.5 kHz revealed that the mean 9.8% increase of capacitance caused by PGE(2) was not correlated in time with activation of chloride conductance, but rather correlated with activation of apical membrane Na(+) transport.

Amiloride↗

Isolated supporting cells from the organ of Corti: some whole cell electrical characteristics and estimates of gap junctional conductance.

Whole cell voltage clamp studies were performed upon isolated and small groups of supporting cells from the guinea pig organ of Corti in order to evaluate junctional and non-junctional membrane characteristics. Single Hensen cells have an average input resistance and capacitance of 1.03 G omega and 24.9 pF, respectively. I-V functions indicate an outward K+ rectification, which is blocked by external TEA, intracellular Cs, or photo-irradiation of intracellularly injected fluorescent dye. Voltage clamping of pairs or small groups of cells indicates that supporting cells 'share' K+ channels within the syncitium. The input impedance of coupled cells was studied during uncoupling with CO2 or octanol media. As expected, coupled cells showed an increase in input capacitance and a decrease in input resistance over single cell values. Input capacitance is a more sensitive indicator of cell coupling than dc input resistance. During uncoupling, input capacitance values drop to single cell levels prior to an increase of dc input resistance to single cell levels. Modeling the results indicates that Hensen cells are well-coupled under normal conditions and may have junctional resistances with values less than 0.1% of the non-junctional resistance. The sensitivity of the supporting cell syncitium's input impedance to small changes in junctional resistance markedly influences the syncitium's RC filter characteristics, and thus may control the frequency response of sound evoked electrical activity measurable in supporting cells in vivo.

Animals↗

Electroreceptor model of the weakly electric fish Gnathonemus petersii. I. The model and the origin of differences between A- and B-receptors.

We present an electroreceptor model of the A- and B-receptors of the weakly electric fish Gnathonemus petersii. The model consists of a sensory cell, whose membrane is separated into an apical and basal portions by support cells, and an afferent fiber. The apical membrane of the cell contains only leak channels, while the basal membrane contains voltage-sensitive Ca2+ channels, voltage-sensitive and Ca2+-activated K+ channels, and leak channels. The afferent fiber is described with the modified Hodgkin-Huxley equation, in which the voltage-sensitive gate of the K+ channels is a dynamic variable. In our model we suggest that the electroreceptors detect and process the information provided by an electric organ discharge (EOD) as follows: the current caused by an EOD stimulus depolarizes the basal membrane to a greatly depolarized state. Then the release of transmitter excites the afferent fiber to oscillate after a certain time interval. Due to the resistance-capacitance structure of the cells, they not only perceive the EOD intensity, but also sense the variation of the EOD waveform, which can be strongly distorted by the capacitive component of an object. Because of the different morphologies of A- and B-cells, as well as the different conductance of leak ion channels in the apical membrane and the different capacitance of A- and B-cells, A-receptors mainly respond to the EOD intensity, while B-receptors are sensitive to the variation of EOD waveform.

Animals↗

Dielectric properties of synaptosomes isolated from rat brain cortex.

Dielectric measurements were performed on the suspensions of synaptosomes isolated from rat brain cortex. The synaptosomes in buffered salt media showed typical dielectric dispersions caused by the presence of a thin limiting membrane of sufficiently low conductivity. An analysis of the dielectric data revealed that the electric conductivity of the synaptosome interior was about 37% of the external medium conductivity under isotonic conditions and that the dielectric constant for the interior phase was about 35. The membrane capacitance (0.7 muF cm-2) remained constant irrespective of nature and concentration of the univalents salts examined. Significant reduction in both the conductivity and the dielectric constant of the internal phase can be explained theoretically proveded that some intra-synaptosomal structural (synaptic vesicles and/or small mitochondria) of non-conducting nature occupies about 50% of the particulate volume, the remainder being in equilibrium with the external salt medium.

Animals↗

Solutions for transients in arbitrarily branching cables: IV. Nonuniform electrical parameters.

Solutions for transients in arbitrarily branching passive cable neurone models with a soma are extended to models with nonuniform electrical parameters and multiple dendritic shunts. The response to an injected current can again be represented as an infinite series of exponentially decaying components with system time constants obtained from the roots of a recursive transcendental equation. The reciprocity relations and global parameter dependencies are the same as for uniform models. Infinitely many "raw" electro-morphological models map onto a given "core" electrotonic model; local as well as global raw parameter trade-offs are now possible. The solutions are illustrated by means of biologically relevant examples: (i) the effects of nonuniform electrical parameters in a two-cylinder + soma cortical pyramidal cell model, (ii) the errors that can occur when uniformity is incorrectly assumed in a single cylinder model, (iii) nonsumming interactions between cells and electrodes that can dramatically increase the duration of the effective capacitative electrode artefact, and (iv) shunting inhibition and double impalements in a hippocampal CA1 pyramidal cell "cartoon" representation. These solutions should complement compartmental modelling techniques.

Animals↗

Rapid determination of intraepithelial resistance barriers by alternating current spectroscopy. II. Test of model circuits and quantification of results.

The impedance of Necturus gallbladder epithelium was measured with transepithelial and intracellular microelectrodes in different transport states. The data are analysed with five electrical equivalent circuits, which differ with respect to the configuration of the paracellular shunt path (lumped vs distributed model of the lateral space), and of the apical cell membrane (non-ideal capacitance or surface amplification by micro-tubular infoldings). Least square fits indicate: 1. that the lumped model cannot represent the epithelium properly, even under control conditions; 2. that the distributed model, which considers the lateral intracellular space separately as a cable-like structure, describes the data well, both under control conditions and during collapse of the lateral spaces; and 3. that the above indicated variations of the apical membrane configuration improve the fits, but have little effect on the magnitude of the calculated circuit parameters. Quantitatively the analysis of 214 measurements on 25 gallbladders under control conditions yields the following results: The resistances of the tight junctions, of the lateral intercellular space, and of the apical and basal cell membrane are Rj = 123, Rlis = 35.5, Ra approximately 3,500, and Rb = 225 (all in omega cm2), and the capacitances of the cell membranes are Ca = 4.95 and Cbl = 26.5 (mu F/cm2). In oxygen deficiency and after cessation of chamber perfusion transport decreased, the spaces collapsed, and Rlis increased to approximately 130 omega cm2. Although the accuracy of the estimated Ra values is still limited, the analysis shows that it is possible to determine Rj and Rlis separately and to follow their changes in response to experimental maneuvers.

Animals↗

Impedance characterization and modeling of electrodes for biomedical applications.

A low electrode-electrolyte impedance interface is critical in the design of electrodes for biomedical applications. To design low-impedance interfaces a complete understanding of the physical processes contributing to the impedance is required. In this work a model describing these physical processes is validated and extended to quantify the effect of organic coatings and incubation time. Electrochemical impedance spectroscopy has been used to electrically characterize the interface for various electrode materials: platinum, platinum black, and titanium nitride; and varying electrode sizes: 1 cm2, and 900 microm2. An equivalent circuit model comprising an interface capacitance, shunted by a charge transfer resistance, in series with the solution resistance has been fitted to the experimental results. Theoretical equations have been used to calculate the interface capacitance impedance and the solution resistance, yielding results that correspond well with the fitted parameter values, thereby confirming the validity of the equations. The effect of incubation time, and two organic cell-adhesion promoting coatings, poly-L-lysine and laminin, on the interface impedance has been quantified using the model. This demonstrates the benefits of using this model in developing better understanding of the physical processes occurring at the interface in more complex, biomedically relevant situations.

Biomedical Engineering↗

Influence of electrolytes in the QCM response: discrimination and quantification of the interference to correct microgravimetric data.

In this work we demonstrate that the presence of electrolytes in solution generates desorption-like transients when the resonance frequency is measured. Using impedance spectroscopy analysis and Butterworth-Van Dyke (BVD) equivalent electrical circuit modeling we demonstrate that non-Kanazawa responses are obtained in the presence of electrolytes mainly due to the formation of a diffuse electric double layer (DDL) at the sensor surface, which also causes a capacitor like signal. We extend the BVD equivalent circuit by including additional parallel capacitances in order to account for such capacitor like signal. Interfering signals from electrolytes and DDL perturbations were this way discriminated. We further quantified as 8.0+/-0.5 Hz pF-1 the influence of electrolytes to the sensor resonance frequency and we used this factor to correct the data obtained by frequency counting measurements. The applicability of this approach is demonstrated by the detection of oligonucleotide sequences. After applying the corrective factor to the frequency counting data, the mass contribution to the sensor signal yields identical values when estimated by impedance analysis and frequency counting.

Biosensing Techniques↗

Noninvasive neuroelectronic interfacing with synaptically connected snail neurons immobilized on a semiconductor chip.

A hybrid circuit of a semiconductor chip and synaptically connected neurons was implemented and characterized. Individual nerve cells from the snail Lymnaea stagnalis were immobilized on a silicon chip by microscopic picket fences of polyimide. The cells formed a network with electrical synapses after outgrowth in brain conditioned medium. Pairs of neurons were electronically interfaced for noninvasive stimulation and recording. Voltage pulses were applied to a capacitive stimulator on the chip to excite the attached neuron. Signals were transmitted in the neuronal net and elicited an action potential in a second neuron. The postsynaptic excitation modulated the current of a transistor on the chip. The implementation of the silicon-neuron-neuron-silicon circuit constitutes a proof-of-principle experiment for the development of neuroelectronic systems to be used in studies on neuronal signal processing, neurocomputation, and neuroprosthetics.

Animals↗

Charge-qubit operation of an isolated double quantum dot.

We have investigated coherent time evolution of pseudomolecular states of an isolated (leadless) silicon double quantum dot, where operations are carried out via capacitively coupled elements. Manipulation is performed by short pulses applied to a nearby gate, and measurement is performed by a single-electron transistor. The electrical isolation of this qubit results in a significantly longer coherence time than previous reports for semiconductor charge qubits realized in artificial molecules.

Journal Article↗