Decrease In Electrical Impedance Research Articles

Potential role of periodontal pathogens in compromising epithelial barrier function by inducing epithelial-mesenchymal transition.

Epithelial-mesenchymal transition (EMT) is a process by which epithelial cells acquire a mesenchymal-like phenotype and this may be induced by exposure to gram-negative bacteria. It has been proposed that EMT is responsible for compromising epithelial barrier function in the pathogenesis of several diseases. However, the possible role of EMT in the pathogenesis of periodontitis has not previously been investigated. The aim of this study therefore was to investigate whether gram-negative, anaerobic periodontal pathogens could trigger EMT in primary oral keratinocytes in vitro. Primary oral keratinocytes were harvested from labial mandibular mucosa of Wistar Han rats. Cells were exposed to heat-killed Fusobacterium nucleatum and Porphyromonas gingivalis (100 bacteria/epithelial cell) and to 20μg/mL of Escherichia coli lipopolysaccharide over an 8-day period. Exposure to bacteria did not significantly change epithelial cell number or vitality in comparison with unstimulated controls at the majority of time-points examined. Expression of EMT marker genes was determined by semiquantitative RT-PCR at 1, 5, and 8days following stimulation. The expression of EMT markers was also assessed by immunofluorescence (E-cadherin and vimentin) and using immunocytochemistry to determine Snail activation. The loss of epithelial monolayer coherence, in response to bacterial challenge, was determined by measuring trans-epithelial electrical resistance. The induction of a migratory phenotype was investigated using scratch-wound and transwell migration assays. Exposure of primary epithelial cell cultures to periodontal pathogens was associated with a significant decrease in transcription (~3-fold) of E-cadherin and the upregulation of N-cadherin, vimentin, Snail, matrix metalloproteinase-2 (~3-5 fold) and toll-like receptor 4. Bacterial stimulation (for 8 days) also resulted in an increased percentage of vimentin-positive cells (an increase of 20% after stimulation with P.gingivalis and an increase of 30% after stimulation with F.nucleatum, compared with controls). Furthermore, periodontal pathogens significantly increased the activation of Snail (60%) and cultures exhibited a decrease in electrical impedance (P<.001) in comparison with unexposed controls. The migratory ability of the cells increased significantly in response to bacterial stimulation, as shown by both the number of migrated cells and scratch-wound closure rates. Prolonged exposure of primary rat oral keratinocyte cultures to periodontal pathogens generated EMT-like features, which introduces the possibility that this process may be involved in loss of epithelial integrity during periodontitis.

Cellular impedance measurement as a new tool for poxvirus titration, antibody neutralization testing and evaluation of antiviral substances

Impedance-based biosensing known as real-time cell electronic sensing (RT-CES) belongs to an emerging technology for analyzing the status of cells in vitro. In the present study protocols were developed for an RT-CES-based system (xCELLigence™, Roche Applied Science, ACEA Biosciences Inc.) to supplement conventional techniques in pox virology. First, proliferation of cells susceptible to orthopoxviruses was monitored. For virus titration cells were infected with vaccinia virus and cell status, represented by the dimensionless impedance-based cell index (CI), was monitored. A virus-dose dependent decrease in electrical impedance could be shown. Calculation of calibration curves at a suitable CI covering a dynamic range of 4 log enabled the quantification of virus titers in unknown samples. Similarly, antiviral effects could be determined as shown for anti-poxviral agents ST-246 and Cidofovir. Published values for the in vitro concentration that inhibited virus replication by 50% (IC 50) could be confirmed while cytotoxicity in effective concentrations was excluded in long-term incubation experiments. Finally, an RT-CES-based virus neutralization test was established. Various poxvirus-specific antibodies were examined for their neutralizing activity and a calculation mode for the neutralizing antibody titer was introduced. In summary, the presented RT-CES-based methods outmatch end-point assays by observing the cell population throughout the entire experiment while workload and time to result are reduced.

Impedance Characterization of Microarray Recording Electrodes in Vitro

The mechanisms underlying performance degradation of chronically implanted silicon microelectrode arrays in the central nervous system (CNS) remain unclear. Humoral and cellular components of the brain foreign body response were evaluated to determine whether their presence on the electrode surface results in increased electrical impedance. Iridium oxide microelectrode recording arrays were electrically characterized in saline, culture media with 10% fetal bovine serum, and coated with various CNS cell types isolated from rat brain. Electrochemical impedance spectroscopy and cyclic voltammetry were performed using a three-electrode system. Potential cycling caused an immediate decrease in electrical impedance, which increased with time toward precycling values, with the effect of cycling remaining significant for several days. The addition of serum caused a significant increase in impedance of up to 28% relative to the saline control. Microelectrodes coated with various cell types known to participate in the foreign body response caused a 20%-80% increase in impedance immediately after contact that remained constant or gradually increased for several weeks. Our findings suggest that the attachment of molecular and cellular species following microelectrode implantation into brain tissue likely contribute to increases in impedance, but do not appear sufficient to hinder recording performance.

Electrical impedance along connective tissue planes associated with acupuncture meridians

BackgroundAcupuncture points and meridians are commonly believed to possess unique electrical properties. The experimental support for this claim is limited given the technical and methodological shortcomings of prior studies. Recent studies indicate a correspondence between acupuncture meridians and connective tissue planes. We hypothesized that segments of acupuncture meridians that are associated with loose connective tissue planes (between muscles or between muscle and bone) visible by ultrasound have greater electrical conductance (less electrical impedance) than non-meridian, parallel control segments.MethodsWe used a four-electrode method to measure the electrical impedance along segments of the Pericardium and Spleen meridians and corresponding parallel control segments in 23 human subjects. Meridian segments were determined by palpation and proportional measurements. Connective tissue planes underlying those segments were imaged with an ultrasound scanner. Along each meridian segment, four gold-plated needles were inserted along a straight line and used as electrodes. A parallel series of four control needles were placed 0.8 cm medial to the meridian needles. For each set of four needles, a 3.3 kHz alternating (AC) constant amplitude current was introduced at three different amplitudes (20, 40, and 80 μAmps) to the outer two needles, while the voltage was measured between the inner two needles. Tissue impedance between the two inner needles was calculated based on Ohm's law (ratio of voltage to current intensity).ResultsAt the Pericardium location, mean tissue impedance was significantly lower at meridian segments (70.4 ± 5.7 Ω) compared with control segments (75.0 ± 5.9 Ω) (p = 0.0003). At the Spleen location, mean impedance for meridian (67.8 ± 6.8 Ω) and control segments (68.5 ± 7.5 Ω) were not significantly different (p = 0.70).ConclusionTissue impedance was on average lower along the Pericardium meridian, but not along the Spleen meridian, compared with their respective controls. Ultrasound imaging of meridian and control segments suggested that contact of the needle with connective tissue may explain the decrease in electrical impedance noted at the Pericardium meridian. Further studies are needed to determine whether tissue impedance is lower in (1) connective tissue in general compared with muscle and (2) meridian-associated vs. non meridian-associated connective tissue.

Tumor necrosis factor decreases thrombin receptor expression in endothelial cells.

We examined the effects of the proinflammatory cytokine, tumor necrosis factor-alpha (TNF alpha) on the expression of proteolytically activated thrombin receptor (PATR) in human umbilical vein endothelial cells (HUVEC). PATR mRNA and protein levels were measured in confluent HUVEC monolayers after challenge with TNF alpha. Northern analysis indicated that TNF alpha treatment resulted in 2- to 3-fold decrease in PATR mRNA in a time- and dose-dependent manner. PATR mRNA level returned to the control level within 6 hr. The nuclear run-on assay indicated that the decreased mRNA signal was due to reduction in the transcription rate. Immunoblotting experiments indicated that the decrease in expression of PATR protein followed in time the decrease in mRNA; the lowest level of protein expression was achieved at 22 hr after TNF alpha treatment. PATR protein returned to basal value within 40 hr after TNA alpha challenge. To assess alterations in endothelial cell function after TNF alpha treatment, we measured thrombin-induced increase in cytosolic Ca2+ ([Ca2+]i) and the cell shape change (measured by decrease in electrical impedance of endothelial cell monolayer). In HUVEC treated with TNF alpha (100 U/ml for 22 hr), the rise in [Ca2+]i after thrombin challenge was approximately 2-fold less than in control thrombin-treated cells. The decrease in electrical impedance of HUVEC monolayers in response to thrombin after TNF alpha treatment was also significantly reduced. However, the rise in [Ca2+]i in response to histamine was not altered by TNF alpha pretreatment. In conclusion, TNF alpha exposure of endothelial cells decreased both mRNA and protein expression of PATR, which explain the decreased activation of thrombin generated signals after the TNF alpha exposure.

Tumor necrosis factor decreases thrombin receptor expression in endothelial cells

We examined the effects of the proinflammatory cytokine, tumor necrosis factor-α (TNFα), on the expression of proteolytically activated thrombin receptor (PATR) in human umbilical vein endothelial cells (HUVEC). PATR mRNA and protein levels were measured in confluent HUVEC monolayers after challenge with TNFα. Northern analysis indicated that TNFα treatment resulted in 2- to 3-fold decrease in PATR mRNA in a time- and dose-dependent manner. PATR mRNA level returned to the control level within 6 hr. The nuclear run-on assay indicated that the decreased mRNA signal was due to reduction in the transcription rate. Immunoblotting experiments indicated that the decrease in expression of PATR protein followed in time the decrease in mRNA; the lowest level of protein expression was achieved at 22 hr after TNFα treatment. PATR protein returned to basal value within 40 hr after TNFα challenge. To assess alterations in endothelial cell function after TNFα treatment, we measured thrombin-induced increase in cytosolic Ca2+ ([Ca2+]i) and the cell shape change (measured by decrease in electrical impedance of endothelial cell monolayer). In HUVEC treated with TNFα (100 U/ml for 22 hr), the rise in [Ca2+]i after thrombin challenge was ∼2-fold less than in control thrombin-treated cells. The decrease in electrical impedance of HUVEC monolayers in response to thrombin after TNFα treatment was also significantly reduced. However, the rise in [Ca2+]i in response to histamine was not altered by TNFα pretreatment. In conclusion, TNFα exposure of endothelial cells decreased both mRNA and protein expression of PATR, which explain the decreased activation of thrombin generated signals after the TNFα exposure. © 1996 Wiley-Liss, Inc.

Measurement of vertebral cortical integrity during pedicle exploration for intrapedicular fixation.

This study determined the predictive ability of electrical impedance measurement in detecting cortical perforation in a porcine model of pedicular exploration. This study tested the hypothesis that a large decrease in electrical impedance would occur as a result of perforation of the vertebral cortex by the pedicle probe. The resistivity of cortical bone has been reported to be 25 to 100 times greater than that of soft tissues. A total of 42 pedicles of the lumbar spines of six swine were explored using the instrumented pedicle probes. Using a 1 microAmp 30-Hz current source, measurement of electrical impedance predicted cortical rupture with a sensitivity, specificity, and accuracy of 95%. Maximum applied voltages of 2.8 mV did not result in myogenic stimulus. Electrical impedance measurement provides an accurate real-time measurement of cortical perforation. This technique is adapted readily for use with pedicular screws and screw tape. Further investigation to determine the clinical use of this technique is recommended.

Acetylcholinesterase activity in Nitella.

IT is generally accepted that the process of excitation starts with a localized depolarization of the conducting cell membrane. The adjacent region is then depolarized by the resultant current flow. This process repeats itself at successive points and thereby the impulse is propagated. These electrical phenomena have been widely investigated, but the underlying mechanism is still under discussion. Nachmansohn1 has suggested a theory according to which the acetylcholine system plays an essential part in bioelectrogenesis. Much evidence has been accumulated in favour of this theory during recent years. The evidence is based on experiments with animal cells, mostly nerve and muscle fibres of vertebrates and invertebrates. However, there are plants which resemble nerve and muscle in being able to propagate impulses by electric currents. There is an excitable membrane separating the cell from its environment, in which a sharp decrease in electrical impedance occurs during the passage of an impulse.