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PubMed2002

Automated electrorotation to reveal dielectric variations related to HER-2/neu overexpression in MCF-7 sublines.

PURPOSE: Electrorotation (ROT) is a technique that allows for determination of the dielectric properties of living cells when exposed to a rotating electric field. We evaluated the ROT behavior of MCF/neo and p185(neu) transfectancts MCF/HER2-11 and MCF/HER2-18 to investigate whether differences in HER-2/neu expression were associated with differences in dielectric properties in these cells. EXPERIMENTAL DESIGN: P185(neu) was measured by Western blotting in MCF/neo cells and HER-2/neu transfectants MCF/HER2-11 and MCF/HER2-18. ROT spectra and cell membrane-specific capacitance were obtained for each cell line. RESULTS: The mean cell membrane-specific capacitance values for MCF/neo, MCF/HER2-11, and MCF/HER2-18 were 2.09, 1.70, and 2.56 microF/cm(2), respectively. The mean specific capacitance for MCF/neo was significantly different from that for MCF/HER2-11 (P = 0.006) and that for MCF/HER2-18 (P = 0.007). CONCLUSIONS: ROT is sufficiently sensitive to detect variations in dielectric properties in breast cancer cell lines overexpressing p185(neu). These differences may be related to the morphological alterations determined by HER-2/neu overexpression.

Automation↗
PubMed1987

Auditory-nerve single-neuron thresholds to electrical stimulation from scala tympani electrodes.

Single auditory-nerve neuron thresholds were studied in sensory-deafened squirrel monkeys to determine the effects of electrical stimulus shape and frequency on single-neuron thresholds. Frequency was separated into its components, pulse width and pulse rate, which were analyzed separately. Square and sinusoidal pulse shapes were compared. There were no or questionably significant threshold differences in charge per phase between sinusoidal and square pulses of the same pulse width. There was a small (less than 0.5 dB) but significant threshold advantage for 200 microseconds/phase pulses delivered at low pulse rates (156 pps) compared to higher pulse rates (625 pps and 2500 pps). Pulse width was demonstrated to be the prime determinant of single-neuron threshold, resulting in strength-duration curves similar to other mammalian myelinated neurons, but with longer chronaxies. The most efficient electrical stimulus pulse width to use for cochlear implant stimulation was determined to be 100 microseconds/phase. This pulse width delivers the lowest charge/phase at threshold. The single-neuron strength-duration curves were compared to strength-duration curves of a computer model based on the specific anatomy of auditory-nerve neurons. The membrane capacitance and resulting chronaxie of the model can be varied by altering the length of the unmyelinated termination of the neuron, representing the unmyelinated portion of the neuron between the habenula perforata and the hair cell. This unmyelinated segment of the auditory-nerve neuron may be subject to aminoglycoside damage. Simulating a 10 micron unmyelinated termination for this model neuron produces a strength-duration curve that closely fits the single-neuron data obtained from aminoglycoside deafened animals. Both the model and the single-neuron strength-duration curves differ significantly from behavioral threshold data obtained from monkeys and humans with cochlear implants. This discrepancy can best be explained by the involvement of higher level neurologic processes in the behavioral responses. These findings suggest that the basic principles of neural membrane function must be considered in developing or analyzing electrical stimulation strategies for cochlear prostheses if the appropriate stimulation of frequency specific populations of auditory-nerve neurons is the objective.

Animals↗
PubMed1969

Electrical properties and acetylcholine sensitivity of singly and multiply innervated avian muscle fibers.

Membrane constants and distribution of acetylcholine (ACh) receptors were determined for multiply innervated fibers of the anterior latissimus dorsi (ALD) and singly innervated fibers of the posterior latissimus dorsi (PLD) muscles of 3-6 month old chickens. The values of the various membrane constants were: length constant, 1.78 mm (mean) in ALD, 0.68 mm in PLD; time constant, 35 msec in ALD, 3.7 msec in PLD; transverse membrane resistance, 4388 ohm cm(2) in ALD, 561 ohm cm(2) in PLD; and membrane capacitance, 8.2 microF/cm(2) in ALD, 7.0 microF/cm(2) in PLD. Peaks of ACh sensitivity occurred at intervals of ca. 740 micro on ALD fibers with a low sensitivity remaining between peaks. Only one peak of ACh sensitivity was detected on PLD fibers. The maximum ACh sensitivity found was 5 +/- 4 mv/ncoul for fibers of the ALD and 77 +/- 60 mv/ncoul for fibers of the PLD. The distance over which this sensitivity fell to 0.1 was ca. 225 micro in the ALD and 140 micro in the PLD. The membranes of these two muscle fiber types differ widely regarding some electrical properties and the disposition of ACh-sensitive receptor sites.

Acetylcholine↗
PubMed2005

Cholinergic action on cortical glial cells in vivo.

This study aims at understanding complex interactions between cortical neurons, glia and blood supply developing during the transition from slow-wave sleep to wakefulness. In spite of essential advances from in vitro and culture preparations, the basic mechanisms of glial interactions with their cellular and ionic environment had remained uninvestigated in vivo. Here we approach this issue by performing simultaneous intracellular recordings of cortical neurons and glia, together with measurements of cerebral blood flow (CBF), extracellular K+ concentrations and local field potentials in both anesthetized (ketamine-xylazine) and naturally behaving cats. Under anesthesia, cortical activation was elicited with electric stimulation of cholinergic nuclei (pedunculopontine tegmental in the brainstem and/or nucleus basalis in the basal forebrain). Iontophoretic application of acetylcholine on the recorded cells was also used. In the vast majority of cases (> 80%) glial cells were hyperpolarized during electric stimulation or spontaneous activation. This result was also obtained in all cases where iontophoresis was used or when glutamatergic kainate/quisqualate receptors were blocked with 6-cyano-7-nitroquinoxaline-2,3-dione. The glial hyperpolarization was associated with steady neuronal depolarization, increased CBF, lower extracellular K+ concentration, increased membrane resistance, decreased membrane capacitance and persistent positive DC field potentials. In some cases of cortical activation (< 20%), glial cells displayed sustained depolarizing potentials, in parallel with neuronal depolarization, decreased CBF and more negative DC field potentials. The above-mentioned effects of cholinergic activation were blocked by the muscarinic antagonist scopolamine. We propose that the glial response to cholinergic activation results from the balance between the direct hyperpolarizing action of acetylcholine and the depolarizing modulation of glutamate from the neighboring neurons, in addition to the modulation of the interglial communication pathway and/or the ionic traffic across blood vessels.

Acetylcholine↗
PubMed1984

Electric current flow in a two-cell preparation from Chironomus salivary glands.

Conventional micro-electrode techniques were used to study the passive electrical properties of salivary glands from Chironomus nuditarsis insect larvae of the fourth instar stage. Linear cable analysis performed on intact glands revealed the following constants: axial intracellular resistance, Ri = 2730 omega cm; membrane resistance per unit apparent cylindrical area, Rm = 1350 omega cm2; membrane capacitance per unit apparent cylindrical area, Cm = 17.6 microF cm-2. The multicellular glands were reduced to intact two-cell preparations by destroying neighbouring cells mechanically. Each cell of a coupled cell pair was impaled with two micro-electrodes, one to pass rectangular current pulses and the other to monitor the resulting voltage deflexions. Internal consistency tests revealed that the experimental data under steady-state conditions may be described accurately by an equivalent circuit consisting of a delta configuration of three resistive elements: the resistances of the non-junctional membrane of cell 1 and cell 2 (r1 and r2), and the resistance of the gap junctional membrane connecting the two cells (rg). The current-voltage relation of the non-junctional membrane was found ohmic over a membrane potential ranging from -40 mV to + 10 mV. The mean value of Rm was 2020 omega cm2. The resistance function of the gap junctional membrane was also ohmic. There was no dependence of gap junctional resistance on voltage or direction of current flow, at least over the relatively narrow range of potentials tested (approximately +/- 10 mV). Individual values of rg varied from 20 to 3800 k omega, with an over-all mean of 1100 k omega. The lower values are thought to represent the physiological state of cellular coupling, whereas the higher ones may reflect partial uncoupling caused by local damage. The proposed cell pair is a suitable preparation for studying problems related to intercellular coupling.

Animals↗
PubMed2000

In vivo electrical characteristics of human skin, including at biological active points.

The aim is to compare the mean values of the in vivo electrical characteristics of biological active points (BAPs) with those of the surrounding human skin. The impedance measurements at BAPs and on the surrounding skin are carried out in vivo on ten young, healthy people. The results of the measurements show that the BAP resistance RP is smaller, and the capacitance CP is higher, than the corresponding values for skin, RS and CS, respectively, these differences are larger at low frequencies (at f = 3 Hz, RS/RP = 3.19 and CP/CS = 3.2). The mean values of the impedance measurements at the BAPs are different from those measured on the skin. The dependence of RP and CP on the pressing force, in the range of about 1-5 N, for the BAPs, has a smaller slope than that observed for the surrounding skin. An equivalent circuit for the BAPs is proposed that describes sufficiently well the experimental results obtained. These results show that the large dispersion in the observed impedance characteristics of the human body measurements in different body regions can be related to the influence of the BAPs present under the electrodes.

Adolescent↗
PubMed1998

Depletion of either ryanodine- or IP3-sensitive calcium stores activates capacitative calcium entry in mouse anococcygeus smooth muscle cells.

The properties of the calcium stores coupled to a depletion-operated cation current (IDOC) proposed to underlie capacitative calcium entry were studied in single smooth muscle cells isolated from the mouse anococcygeus using the whole-cell patch-clamp technique. Both caffeine (10 mM) and carbachol (50 microM) activated an initial, large ( approximately 200 pA), transient, calcium-dependent chloride current (IClCa) followed by a smaller ( approximately 10 pA) sustained, non-selective cation current (IDOC). Intracellular application of heparin (5 mg ml-1) abolished the response to carbachol but potentiated that to caffeine. Ryanodine (3 microM) activated IDOC but not IClCa; ryanodine (30 microM) failed to produce any response. Both concentrations of ryanodine abolished the response to caffeine and prevented activation of IClCa by carbachol. In the presence of 30 microM, but not 3 microM, ryanodine, carbachol was able to activate IDOC. Cyclopiazonic acid (CPA; 10 microM) abolished the response to carbachol; however, caffeine was still able to activate IClCa. In whole-muscle tension recordings, ryanodine at both 3 and 30 microM produced contractions of the tissue but only that in response to the lower concentration was maintained. Thus, depletion of either inositol 1,4, 5-trisphosphate-(IP3-) sensitive or ryanodine-sensitive calcium stores is able to activate IDOC, and, by extension, capacitative calcium entry in this tissue.

Animals↗
PubMed1998

An examination of the role of intracellular ATP in the activation of store-operated Ca2+ influx and Ca2+-dependent capacitance increases in rat basophilic leukaemia cells.

The role of ATP in both the activation of store-operated Ca2+ current ICRAC and in Ca2+-dependent vesicular fusion was examined in a study of rat basophilic leukaemia (RBL) cells using the whole-cell patch-clamp technique. Fusion was monitored via changes in plasma membrane capacitance. Following a decrease in the levels of intracellular ATP, achieved using the mitochondrial poison antimycin and the ATP synthase inhibitor oligomycin, as well as a reduction of glycolysis by removal of external glucose, ICRAC activated in a manner similar to control cells when stores are depleted by dialysis with a pipette solution containing either inositol 1,4, 5-trisphosphate (InsP3) or ionomycin together with a high concentration of EGTA. Dialysis of cells for 150 s with the non-hydrolysable ATP analogue 5'-adenylylimidodiphosphate (AMP-PNP) (2 mM) in addition to the mitochondrial inhibitors also failed to prevent activation of ICRAC following external application of ionomycin and thapsigargin, when compared with control recordings obtained with 2 mM ATP instead. Ca2+-dependent vesicular fusion was triggered by dialysing cells with 10 microM Ca2+ and guanosine-5'-O-(3-thiotriphosphate (GTP[gamma-S]). The capacitance increase was unaffected by inhibition of glycolysis, mitochondrial inhibitors or dialysis with either AMP-PNP or adenosine 5'-O-(3-thiotriphosphate) (ATP[gamma-S]) instead of ATP. We conclude that ATP hydrolysis does not seem to be necessary for the activation of ICRAC or for the capacitance increases elicited by high concentrations of intracellular Ca2+.

Adenine Nucleotides↗
PubMed1997

Genital tract burns during rollerball endometrial coagulation.

Thermal injuries occurred in three women after hysteroscopic rollerball endometrial ablation. Possible mechanisms of injury included insulation failure of the rollerball to the sheath of the resectoscope, current division with the sheath acting as an alternate return electrode, and capacitative leakage of current from the active electrode to the sheath.

Administration, Topical↗
PubMed1991

Blockade of reflex venous capacitance responses in liver and spleen by hexamethonium, atropine, and surgical section.

Since hexamethonium and surgical section have been used to prevent reflex splanchnic capacitance responses, we examined the effectiveness of these procedures in blocking responses to direct stimulation of preganglionic fibres in the splanchnic nerves. Liver blood volume was measured by plethysmography and splenic blood volume by weighing in cats anesthetized by pentobarbital. The cats were adrenalectomized to prevent adrenal catecholamine secretion in response to splanchnic nerve stimulation. Hexamethonium (10 and 20 mg/kg) alone or atropine (1 mg/kg) alone caused only a small variable block of the responses to preganglionic nerve stimulation. A combination of the two drugs essentially produced a complete block of the liver capacitance response, but a significant response still persisted in the spleen. Surgical section of the postganglionic nerve bundles around the hepatic and splenic arteries completely abolished the responses to preganglionic stimulation. It is concluded that a relatively complete block of reflex splanchnic capacitance responses requires either a combination of hexamethonium and atropine or surgical section of the postganglionic nerves.

Adrenal Glands↗
PubMed2002

Dendritic resonance in rat neocortical pyramidal cells.

Dendritic integration of synaptic signals is likely to be an important process by which nerve cells encode synaptic input into spike output. However, the response properties of dendrites to time-varying inputs are largely unknown. Here, I determine the transfer impedance of the apical dendrite in layer V pyramidal cells by dual whole cell patch-clamp recordings in slices of rat somatosensory cortex. Sinusoidal current waveforms of linearly changing frequencies (0.1-25 Hz) were alternately injected into the soma or apical dendrite and the resulting voltage oscillations recorded by the second electrode. Dendrosomatic and somatodendritic transfer impedances were calculated by Fourier analysis. At near physiological temperatures (T approximately 35 degrees C), the transfer impedance had a maximal magnitude at low frequencies (f(res) approximately 6 Hz). In addition, voltage led current up to approximately 3 Hz, followed by a current lead over voltage at higher frequencies. Thus the transfer impedance of the apical dendrite is characterized by a low-frequency resonance. The frequency of the resonance was voltage dependent, and its strength increased with dendritic distance. The resonance was completely abolished by the I(h) channel blocker ZD 7288. Dendrosomatic and somatodendritic transfer properties of the apical dendrite were independent of direction or amplitude of the input current, and the responses of individual versus distributed inputs were additive, thus implying linearity. For just threshold current injections, action potentials were generated preferentially at the resonating frequency. I conclude that due to the interplay of a sag current (I(h)) with the membrane capacitance, layer V pyramids can act as linear band-pass filters with a frequency preference in the theta frequency band.

Action Potentials↗
PubMed1978

Arterial and venous responses to hypothalamic stimulation in the dog.

Stimulation of the ventro-medial nucleus of the hypothalamus induces active constriction of both pre- and post-capillary vessels in the dog's hindlimb. Alpha-adrenolytic agents reduce these responses, indicating that they are mediated by the sympathetic nervous system. Stimulation of the paraventricular nucleus dilates both resistance and capacitance vessels. The present study demonstrates that hypothalamic neurones can control venomotor tone.

Animals↗
PubMed1998

The breakdown of cell membranes by electrical and mechanical stress.

We attempted to determine whether mechanical tension and electrical stress couple to cause membrane breakdown in cells. Using cell-attached patches from HEK293 cells, we estimated the mechanically produced tension from the applied pressure and geometry of the patch. Voltage pulses of increasing amplitude were applied until we observed a sudden increase in conductance and capacitance. For pulses of 50 micros duration, breakdown required >0.5 V and was dependent on the tension. For pulses of 50-100 ms duration, breakdown required 0.2-0.4 V and was independent of tension. Apparently two physically different processes can lead to membrane breakdown. We could explain the response to the short, high-voltage pulses if breakdown occurred in the lipid bilayer. The critical electromechanical energy per unit area for breakdown by short pulses was approximately 4 dyne/cm, in agreement with earlier results on bilayers. Our data suggest that, at least in a patch, the bilayer may hold a significant fraction (approximately 40%) of the mean tension. To be compatible with the large, nonlytic area changes of patches, the bilayer appears to be pulled toward the pipette tip, perhaps by hydrophobic forces wetting membrane proteins bound to the glass. Although breakdown voltages for long pulses were in agreement with earlier work on algae, the mechanism(s) for this breakdown remain unclear.

Biophysical Phenomena↗
PubMed2002

Diagnostic potential of targeted electrical impedance scanning in classifying suspicious breast lesions.

RATIONALE AND OBJECTIVE: To evaluate the potential of targeted electrical impedance scanning (EIS) for classifying suspicious breast lesions. METHODS: EIS was performed in full knowledge of mammographic findings and findings of clinical breast examination. One hundred seventeen patients with a total of 129 breast lesions were examined with EIS before breast biopsy (surgical excision or vacuum core biopsy). Diagnostic indexes of targeted EIS were calculated depending on major lesion characteristics. Capacitance and conductivity of all positive spots (S) and the surrounding normal breast tissue (NBT) were quantified using ROI measurements. The ratio S/NBT was calculated to compare true positive (n = 44) and false positive (n = 18) spots. RESULTS: With respect to histology, of the 129 lesions 71 were malignant and 58 lesions were benign. Overall sensitivity of targeted EIS was 62%, specificity 69%, PPV 71%, and NPV 60%. Sensitivity of EIS varied depending on the tumor size, which was between 48% (> 20 mm) and 71% (11-20 mm). Highest specificity (86%) was observed for large lesions (> 20 mm); however, the NPV was only 35% for lesions of that size. NPV was higher for nonpalpable lesions (74%) and clusters of microcalcifications (85.7%) compared with palpable lesions (39%) and solid lesions (44%). There was no statistical difference of S/NBT ratio neither for conductivity nor capacitance of true and false positive spots. Compared with true positive spots a trend of a higher conductivity ratio at 100 Hz and 200 Hz was seen for false positive spots. CONCLUSION: EIS showed mediocre overall diagnostic accuracy for classifying suspicious breast lesions. Quantitative analysis of positive EIS findings did not help to differentiate between false and true positive spots.

Biopsy↗
PubMed2004

Use of bipolar parallel electrodes for well-controlled microstimulation in a mouse hippocampal brain slice.

In a hippocampal brain slice two types of stimulating electrodes [single (SE) or monopolar and parallel bipolar (PE)] were used to determine the optimal protocol for single pulse microstimulation. We show that even for a constant-current power source the amplitude of stimulating current (SC) is not constant, especially for short pulse widths (PW) (<200 micros). Recording the stimulating current and computing the amount of electric charge that is passed through the microelectrode gives the best estimate of the strength of electrical stimulation. For SE the evoked response is obstructed for a time interval larger than three times the PW. The stimulus artifact (SA) substantially decreases when a PE is used. The orientation of the stimulating current relative to the position of the targeted fibers (Schaffer collaterals) was controlled when using a PE. The use of PEs allowed the accurate recording of the physiological response that contains three clearly defined peaks. Stimulation can be elicited at PW as short as 30 micros when the main current is capacitive. The charge needed to elicit physiological responses was in the range of 1-40 nC (the lower values for the PE) suggesting that use of PEs is most advantageous for well-controlled microstimulation studies in brain slices.

Action Potentials↗
PubMed2005

Mechanosensitive channels in the lateral wall can enhance the cochlear outer hair cell frequency response.

We present the results of a modeling study on the impact of mechanosensitive channels in the lateral wall of the outer hair cell on the cell frequency response. The model includes the electrical properties of the cell membrane, piezoelectricity associated with a membrane motor mechanism, and mechanosensitive channels in the cell lateral wall. The outer hair cell is loaded by the vibrating basilar and tectorial membranes, and this loading generates strain in the lateral wall. Our analysis reveals a property, the strain rate sensitivity, that, in concert with the piezoelectric effect, can enhance the cell frequency response. We discuss possible viscoelastic-type mechanisms of the channel's strain rate sensitivity that is consistent with the organization of the composite cell lateral wall. The parameters of our model are chosen on the basis of the previously estimated electrical and piezoelectric properties as well as typical conductance and density of the mechanosensitive channels in cells. We found that the strain rate sensitivity of the channels can result in receptor potentials greater than those predicted by the RC (resistance and capacitance) analysis. The effect of the channels is especially significant in an intermediate range of sound frequencies, and the channel-related gain is up to 3-4 times between 3 and 15 kHz.

Acoustics↗
PubMed2001

Changes of structural and dynamic properties of model lipid membranes induced by alpha-tocopherol: implication to the membrane stabilization under external electric field.

The effects of alpha-tocopherol on electric properties of bilayer lipid membranes were investigated. Planar bilayer membranes formed by the Mueller-Rudin method were used. Voltammetric and chronopotentiometric measurements were performed using a four-electrode potentiostat-galvanostat. It was demonstrated that registration of membrane capacitance, resistance, and voltammetric characteristics provided information about the change in the structure and permeability of bilayer lipid membranes. The results suggested that incorporation of alpha-tocopherol into lipid membrane destabilized its structure and facilitated the electrogeneration of pores. The possible role of observed changes in physiological functions of alpha-tocopherol was discussed.

Electromagnetic Fields↗