Numerical sensitivity modeling for the detection of skin tumors by using tetrapolar probe

The measurement of electrical impedance of skin using surface electrodes permits the assessment of changes in local properties of the skin and can be used in the detection of tumors. The sensitivity of this technique depends mainly on the geometry of the probe and the size of the tumor. In this article, the impedance method was used to estimate the sensitivity of a tetrapolar probe in detecting small regions of increased conductivity in a stratified model of human skin. The impedance method was used to model the potential distribution using fasorial analysis to solve the node equations of the equivalent circuit. Interpolation was applied to reduce discretization error. The skin was modeled as a three-layer structure with different conductivity and permittivity obtained from the literature. A tumor was modeled as a small volume with admittivity four times higher than the normal tissue. Sensitivity calculation was made as a function of electrode diameter and separation, tumor size, and excitation frequency. The simulations indicated that by inserting a one square millimeter tumor in the epidermis, the load impedance to the current source varies about 1% while the transfer impedance varied 8%. The sensitivity also increases nonlinearly with increasing tumor area and thickness. Additionally, it was found that the sensitivity of the transfer impedance has a maximum value when the electrodes are separated by 1.8 mm. The results show that transfer impedance measurements of the skin may detect small skin tumors with a reasonable sensitivity by using an appropriate tetrapolar probe.     

A non-invasive method for an in vivo assessment of corneal epithelium permeability through tetrapolar impedance measurements

The permeability of the cornea epithelial layer has an important role in optimal function of the cornea. To assess this property quantitatively, methods must be based on the passive electrical properties of living tissues, as they can take advantage of the fundamental role that ionic permeability plays in such properties. For such techniques, measurement of the translayer electrical resistance (TER) has been consistently used to examine the ion transport mechanisms in the corneal epithelial cells; however, this technique has been only possible in vitro. To enhance the applications of this method, in this work we present a novel sensor to perform non-invasive in vivo TER measurements. Herein, the epithelial permeability was assessed using non-invasive tetrapolar impedance measurements that were performed with four electrodes placed on the corneal surface. The geometry of these electrodes was previously optimized to maximize the sensitivity of the corneal epithelium. To evaluate the feasibility of this sensor, the permeability of a rabbit corneal epithelium was monitored by applying a solution of benzalkonium chloride (0.05% BAC). The results validate the capability of the sensor to evaluate the cornea epithelial permeability in vivo.     

Changes of electrodermal properties in the "acupuncture points" on men and rats

The measurements of skin resistance to electrical current performed in rats and men indicated the occurrence of small skin areas, in which the conductivity for DC and AC was sharply increased. In healthy men, the anatomical localization of these areas of increased skin conductivity (AISC) corresponded to the localization of the so-called "acupuncture points" (AP). In patients, the AISC were also found outside the ordinary AP, mainly in areas of referred pain. The measurements of the size of AISC by multiple electrodes indicated the approximate size of AISC cca 350 microM in the rat, ccs 450 microns in man. The recording of skin conductivity were taken from : a) AISC in man and the rat, b) skin in the close neighbourhood of AISC, c) from the sweating human skin.     

On-chip microdialysis system with flow-through sensing components

Microdialysis probes have been used for diabetes treatment as continuous monitoring system coupled to a glucose sensor. An on-chip microdialysis system with in-line sensing electrodes is demonstrated. As a first step towards greater biosensor integration with this miniaturized microdialysis system, a stacked system with in-line sensing electrodes was developed. Impedance electrodes sputtered within the microchannels were used to determine fluid electrical resistance from a dialyzed phosphate buffered saline (PBS) solution, which characterizes solution conductivity as a function of PBS concentration. The permeability of the membrane to the salt ions was obtained as 0.246+/-0.028 microm/s (15 nm pores). Subsequently, experiments measuring PBS dialysis in the time-domain at 64.4% recovery were conducted. The PBS concentration of the reservoir was changed in both a step response and sinusoidally with an 800 s period. The subsequently measured impedance indicates that the system is able to continuously track concentration changes in the reservoir with a 210 s system response delay. Most of this delay is due to the dead volume within the tubing between the syringe pumps and the microsystem. In addition, the predicted response was modeled using linear systems theory and matches the experimental measurements (r=0.98). This system is expected to have the proper sensitivity to track physiologically relevant concentration changes of biomolecules such as glucose (which has a physiological maximum change rate of approximately 4 mg/dl min with a periodicity of 1h or greater) with minimal lag time and amplitude reduction.     

Direct electron transfer between cytochrome P450scc and gold nanoparticles on screen-printed rhodium-graphite electrodes

This paper is concerned with an investigation of electron transfer between cytochrome P450scc (CYP11A1) and gold nanoparticles immobilised on rhodium-graphite electrodes. Thin films of gold nanoparticles were deposited onto the rhodium-graphite electrodes by drop casting. Cytochrome P450scc was deposited onto both gold nanoparticle modified and bare rhodium-graphite electrodes. Cyclic voltammetry indicated enhanced activity of the enzyme at the gold nanoparticle modified surface. The role of the nanoparticles in mediating electron transfer to the cytochrome P450scc was verified using ac impedance spectroscopy. Equivalent circuit analysis of the impedance spectra was performed and the values of the individual components estimated. On addition of aliquots of cholesterol to the electrolyte bioelectrocatalytic reduction currents were obtained. The sensitivity of the nanoparticle modified biosensor to cholesterol was 0.13 microA microM-1 in a detection range between 10 and 70 microM of cholesterol. This confirms that gold nanoparticles enhance electron transfer to the P450scc when present on the rhodium-graphite electrodes.     

Metallic oxide CdIn2O4 films for the label free electrochemical detection of DNA hybridization

DNA functionalised semiconductor metallic oxide electrodes have been developed for the direct electrochemical detection of DNA hybridization, without labelling or the introduction of a redox couple. Conductive CdIn(2)O(4) thin films with controlled properties were deposited on glass substrates using an aerosol pyrolysis technique. The films exhibit a polycrystalline microstructure with a surface roughness of 1.5 nm (r.m.s.) and an electrical resistivity ranging between 1 and 3 x 10(-3) Omega cm. These electrodes were functionalised using hydroxylation and silanisation steps, to allow the binding of DNA probe sequences (20 bases). The electrical detection of DNA hybridization with complementary sequences has been performed using electrochemical impedance spectrometry (EIS) measuring the variation of impedance before and after hybridization. Two set-ups were used, a standard set-up including three electrodes and a set-up including two symmetrical electrodes. In both configurations, a significant increase of the impedance modulus, more particularly of the real part of the impedance (160-225% according to the electrochemical cell used) has been obtained over a frequency range of 10-10(5)Hz. DNA hybridization has also been systematically confirmed using the fluorescence spectrometry. This study emphasizes the high sensitivity of the CdIn(2)O(4) as a working electrode for the detection of biological events occurring at the electrode surface.     

Approaches to reduce the retroaction of long-term monitoring of bioelectric events in ergonomic field studies (author's transl)

When monitoring bioelectric signals the surface electrodes can cause a retroaction on the subject thereby introducing an error of measurement. There are two types of retroaction: physical and psycho-physiological. A physical retroaction due to the hydration process of the skin occurs if 'wet' electrodes are used for the recording of the skin conductance level (SCL) causing a continuous drift of the SCL and a decrease in sensitivity to SCL changes. Therefore a dry electrode was developed with improved performance: It exhibits less sensitivity to motion, is not subject to polarization, and features better SCL long-term stability. When recording the electrocardiogram or the electromyogram a psychophysiological retroaction occurs due to the annoyance caused by the skin-irritating abrading techniques in order to decrease the skin impedance and reduce the motion artifact. In an attempt to abandon the skin preparation whenever permissible without sacrificing the measurement accuracy a performance estimation procedure was developed. Basing on the information on the signal frequency content, the electrode contact area, the required accuracy of measurement and the amplifier input impedance a decision on the necessity of skin preparation is made. Moreover, the results of a study are reported investigating the reduction of motion artifacts by means of electrode design and appropriate electrode jelly formulation.     

Temperature dependent alterations of the surface-EMG and ECG: an investigation of the electrical transfer characteristics of the human skin (author's transl)

The influence of local heating of the skin on the integrated EMG (registered with surface and subcutaneous electrodes), the amplitude of the ECG, the skin blood flow, the electrode impedance, the electrode-to-skin impedance, and the tissue impedance is investigated. Except for the increasing skin blood flow each of the variables exhibits a significant reduction with an increase in skin temperature. From these results the existence of two mechanisms is deduced mediating thermal influence on bioelectric signals picked up by surface electrodes: 1. An alteration of the signal source. 2. An alteration of the electric transfer characteristics of the tissue between signal source and electrode. Especially in quantitative surface electromyography the temperature dependence of the signal can be a source of error.     

Estimation of tumor characteristics in a breast tissue with known skin surface temperature

Presence of a tumor and its characteristics like location, size and properties are estimated. Estimation is based on the measurement of the skin surface temperature of the breast. Consideration is given to a 2-D breast tissue infected with a tumor. Heat transfer in the breast tissue modeled using the Pennes bioheat equation is solved by the finite volume method. Skin surface temperature profile of the breast is characteristic of the tumor location, its size and grade. In the inverse analysis, the objective function is minimized using the genetic algorithm. Exact estimation is obtained if one parameter is estimated at a time. However, the accuracies are acceptable even when blood perfusion rate, location and size are estimated simultaneously.     

基于电阻抗扫描的乳腺组织阻抗频谱特性测量实验

为了研究乳腺组织电导率特性及肿瘤生长方式对电阻抗扫描(electrical impedance scanning,EIS)肿瘤成像特征的影响,作者对照分析了69例乳腺肿瘤患者的EIS成像、X线钼靶摄影及离体乳腺组织的阻抗频谱特性。实验得到了40例恶性肿瘤、34例良性肿瘤、49例腺体组织和41例脂肪组织的离体电阻率特性,其中腺体组织和良性肿瘤的电阻率数值最小,其次是癌组织,脂肪组织的电阻率数值最大。恶性肿瘤中EIS检查24例高亮表现,11例暗区表现,5例无表现;良性肿瘤EIS检查多数无特征表现。实验表明,癌组织与其周围正常组织的电导率有显著性差异。因恶性肿瘤浸润组织的不同,组织间的电导率差异会出现正向或负向变化,EIS检查表现出亮、暗不同的特征成像。良性肿瘤的电导率较好,但其与周围组织电导率差异无统计学意义,EIS检查无显著成像。     

A method for continuous in vivo measurement of hemolymph conductivity in crabs

A miniaturized conductivity measuring device has been developed to monitor in vivo hemolymph conductivity in crabs. The device measures the conductivity by means of implanted platinum electrodes, and transmits the data via radiotelemetry to a receiving station through 6 ft of water. Crabs used in these investigations survived the insertion of the conductivity probe and appeared to behave and feed normally for up to one week with the probe in position. Blue crabs, Callinectes sapidus Rathbun, which were acclimated to either 5 or 35 ‰ were abruptly transferred to 35 and 5 ‰, respectively and the rate of change of hemolymph conductivity sucessfully monitored. The time required for the conductivity (ionic composition) change in the hemolymph of the crabs after transfer varied from 12 to 16 h for crabs transferred from 35 to 5 ‰ to less than an hour when the change was from 5 to 35 ‰.     

Detection and analysis of tumor fluorescence using a two-photon optical fiber probe

The utility of a two-photon optical fiber fluorescence probe (TPOFF) for sensing and quantifying tumor fluorescent signals was tested in vivo. Xenograft tumors were developed in athymic mice using MCA207 cells expressing green fluorescent protein (GFP). The TPOFF probe was able to detect ex vivo fluorescence from excised tumors containing as little as 0.3% GFP-expressing cells. TPOFF results were similar to both flow-cytometric analysis of tumor cells after isolation and suspension, and fluorescence determined by microscope images of cryosectioned tumors. TPOFF was then used to measure GFP fluorescence from tumors in live mice. The fiber probe detected fluorescently-labeled Herceptin antibody targeted to HER2-expressing tumors in severe combined immunodeficient mice. Dendrimer nanoparticles targeted by folic acid and having 6-TAMRA as a fluorescent probe were also used to label KB cell tumors in vivo. The fiber probe documented a fourfold increase in tumor fluorescence in animals that received the targeted dendrimer. These results suggest TPOFF can be used as a minimally invasive system for identifying tumor markers and monitoring drug therapy.     

Evolution in impedance at the electrode-skin interface of two types of surface EMG electrodes during long-term recordings.

The evolution in impedance at the electrode-skin interface of Beckman and Red Dot electrodes was assessed during long-term recordings. Impedance was measured between each pair of electrodes, arranged in a bipolar configuration on tibialis anterior (n=13). A waveform constructed of sinusoids of known frequencies, evenly distributed on a log scale from 1-16,384 Hz, was applied through the electrodes, and the result recorded by a DAQ system. SEMG signals were recorded at 1000 Hz during isometric dorsiflexion contractions of 30 s, performed every 15 min for 2 h. Impedance data were acquired at 65,536 Hz immediately before and after SEMG recordings. Large individual differences in impedance levels were observed at low frequencies. At high frequencies, impedance values depended only on the electrode type. Impedance decreased steadily with time for Beckman electrodes (p < 0.05), but did not decrease significantly for Red Dot electrodes. The magnitude of the reduction over time varied widely between individuals, and was related to the initial impedance values. The impedance-bandwidth product remained constant for each electrode type (95% confidence intervals 146.2-148.2 and 126.1-127.8 for Beckman and Red Dot electrodes respectively). When skin impedance is electrically modelled with a simple network containing a resistor and a capacitor, the capacitance varies with the properties of the electrode used, whereas resistance is dependent on the subject. Furthermore, the EMG spectrum is unaffected by impedance provided skin preparation is sufficient to reduce the impedance below 55 komega.     

A highly sensitive biocompatible spin probe for imaging of oxygen concentration in tissues

The development of an injectable probe formulation, consisting of perchlorotriphenylmethyl triester radical dissolved in hexafluorobenzene, for in vivo oximetry and imaging of oxygen concentration in tissues using electron paramagnetic resonance (EPR) imaging is reported. The probe was evaluated for its oxygen sensitivity, biostability, and distribution in a radiation-induced fibrosarcoma tumor transplanted into C3H mice. Some of the favorable features of the probe are: a single narrow EPR peak (anoxic linewidth, 41 microT), high solubility in hexafluorobenzene (>12 mM), large linewidth sensitivity to molecular oxygen ( approximately 1.8 microT/mmHg), good stability in tumor tissue (half-life: 3.3 h), absence of spin-spin broadening (up to 12 mM), and lack of power saturation effects (up to 200 mW). Three-dimensional spatial and spectral-spatial (spectroscopic) EPR imaging measurements were used to visualize the distribution of the probe, as well as to obtain spatially resolved pO(2) information in the mice tumor subjected to normoxic and hyperoxic treatments. The new probe should enable unique opportunities for measurement of the oxygen concentration in tumors using EPR methods.     

Monitoring of cell growth and assessment of cytotoxicity using electrochemical impedance spectroscopy

Electrochemical impedance spectroscopy (EIS) was used to monitor the growth of mammalian cancer cells and evaluate the cytotoxicity of chemicals using Fe(CN)6(3-/4-) as a redox probe. Cancer cells, the human hepatocarcinoma cell line (BEL7404), were grown on optically transparent indium tin oxide (ITO) semiconductor slides, which were used as the working electrodes in electrochemical experiments. Attachment and proliferation of cancer cells on ITO surfaces resulted in increase of electron-transfer resistance (R(et)) between the redox probe of Fe(CN)6(3-/4-) in electrolyte solution and ITO electrode surface. For cytotoxicity assessment, cells grown on ITO substrates were further cultured in the presence of different cytotoxicants and electrochemical impedance measurements were carried out at different time intervals. Gemcitabine, a promising antineoplastic drug showing activity against a wide spectrum of human solid tumors, was selected as a model for long-term cytotoxicity effect study, whereas mercury chloride represented a model for acute toxicants. The inhibitions of gemcitabine and mercury chloride on the viability and proliferation of BEL7404 cells were observed from the electrochemical impedance experiments, and the different action modes were discriminated. Additionally, microscope images were also used to observe the effects of these two chemicals on the morphology of the cells. General consistency has been found between the electrochemical impedance response and the morphological observation. Such an impedance method provides a simple and inexpensive way for in vitro assessment of chemical cytotoxicity.     

Which Disposable Chest Electrode?

Chest electrodes are preferred to limb electrodes for cardiac monitoring, as limb movements are not restricted and produce less interference of the E.C.G. trace. Eight types of disposable chest electrodes were investigated to compare their performance, skin reactions, cost, ease of application, size, and skin–electrode impedance.Elema-Schonander electrodes were found to be the most efficient and the most expensive. In their application care was required to avoid severe skin reactions.Dracard electrodes were simple to attach, worked well without severe skin reactions, and were cheap. They are recommended for routine use. Smith and Nephew electrodes, a type of “multipoint electrodes” which do not require electrode jelly, frequently produced severe skin reactions, making them unsuitable for monitoring for periods exceeding 12 hours.     

Validation of tetrapolar bioelectrical impedance method to assess human body composition

This study was conducted to validate the relationship between bioelectrical conductance (ht2/R) and densitometrically determined fat-free mass, and to compare the prediction errors of body fatness derived from the tetrapolar impedance method and skinfold thicknesses, relative to hydrodensitometry. One-hundred and fourteen male and female subjects, aged 18-50 yr, with a wide range of fat-free mass (34-96 kg) and percent body fat (4-41%), participated. For males, densitometrically determined fat-free mass was correlated highly (r = 0.979), with fat-free mass predicted from tetrapolar conductance measures using an equation developed for males in a previous study. For females, the correlation between measured fat-free mass and values predicted from the combined (previous and present male data) equation for men also was strong (r = 0.954). The regression coefficients in the male and female regression equations were not significantly different. Relative to hydrodensitometry, the impedance method had a lower predictive error or standard error of the estimates of estimating body fatness than did a standard anthropometric technique (2.7 vs. 3.9%). Therefore this study establishes the validity and reliability of the tetrapolar impedance method for use in assessment of body composition in healthy humans.     

CTL induction of tumoricidal nitric oxide production by intratumoral macrophages is critical for tumor elimination

To characterize mechanisms of CTL inhibition within an ocular tumor microenvironment, tumor-specific CTLs were transferred into mice with tumors developing within the anterior chamber of the eye or skin. Ocular tumors were resistant to CTL transfer therapy whereas skin tumors were sensitive. CTLs infiltrated ocular tumors at higher CTL/tumor ratios than in skin tumors and demonstrated comparable ex vivo effector function to CTLs within skin tumors indicating that ocular tumor progression was not due to decreased CTL accumulation or inhibited CTL function within the eye. CD11b(+)Gr-1(+)F4/80(-) cells predominated within ocular tumors, whereas skin tumors were primarily infiltrated by CD11b(+)Gr-1(-)F4/80(+) macrophages (Ms), suggesting that myeloid derived suppressor cells may contribute to ocular tumor growth. However, CD11b(+) myeloid cells isolated from either tumor site suppressed CTL activity in vitro via NO production. Paradoxically, the regression of skin tumors by CTL transfer therapy required NO production by intratumoral Ms indicating that NO-producing intratumoral myeloid cells did not suppress the effector phase of CTL. Upon CTL transfer, tumoricidal concentrations of NO were only produced by skin tumor-associated Ms though ocular tumor-associated Ms demonstrated comparable expression of inducible NO synthase protein suggesting that NO synthase enzymatic activity was compromised within the eye. Correspondingly, in vitro-activated Ms limited tumor growth when co-injected with tumor cells in the skin but not in the eye. In conclusion, the decreased capacity of Ms to produce NO within the ocular microenvironment limits CTL tumoricidal activity allowing ocular tumors to progress.     

Near-infrared pH-activatable fluorescent probes for imaging primary and metastatic breast tumors

Highly tumor selective near-infrared (NIR) pH-activatable probe was developed by conjugating pH-sensitive cyanine dye to a cyclic arginine-glycine-aspartic acid (cRGD) peptide targeting α(v)β(3) integrin (ABIR), a protein that is highly overexpressed in endothelial cells during tumor angiogenesis. The NIR pH-sensitive dye used to construct the probe exhibits high spectral sensitivity with pH changes. It has negligible fluorescence above pH 6 but becomes highly fluorescent below pH 5, with a pK(a) of 4.7. This probe is ideal for imaging acidic cell organelles such as tumor lysosomes or late endosomes. Cell microscopy data demonstrate that binding of the cRGD probe to ABIR facilitated the endocytosis-mediated lysosomal accumulation and subsequent fluorescence enhancement of the NIR pH-activatable dye in tumor cells (MDA-MB-435 and 4T1/luc). A similar fluorescence enhancement mechanism was observed in vivo, where the tumors were evident within 4 h post injection. Moreover, lung metastases were also visualized in an orthotopic tumor mouse model using this probe, which was further confirmed by histologic analysis. These results demonstrate the potential of using the new integrin-targeted pH-sensitive probe for the detection of primary and metastatic cancer.     

Label-free impedimetric immunosensor for ultrasensitive detection of cancer marker Murine double minute 2 in brain tissue

The detection of cancer biomarkers is as important tool for the diagnosis and prognosis of cancer such as brain cancer. Murine double minute 2 (MDM2) has been widely studied as prognostic marker for brain tumor. Here we describe development of a new sensitive label free impedimetric immunosensor for the detection of MDM2 based on cysteamine self assembled monolayers on a clean polycrystalline Au electrode surface. The amine-modified electrodes were further functionalized with antibody using homobifunctional 1,4-phenylene diisothiocyanate (PDITC) linker. The assembly processes of the immunosensor had been monitored with cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) techniques using Fe(CN)(6)(3-/4-) solution as redox probe. The impedance changes upon binding of MDM2 protein to the sensor surface was utilized for the detection of MDM2. The increase in relative electron-transfer resistance (ΔR/R(0)%) values was linearly proportional to the concentration of tumor marker MDM2 in the wide dynamic range of 1pg/ml-1μg/ml. The limit of detection was 0.29pg/ml in phosphate buffer saline (PBS) and 1.3pg/ml in mouse brain tissue homogenate, respectively. The immunosensor showed a good performance in comparison with ELISA for the analysis of the MDM2 in the cancerous mouse brain tissue homogenates. Moreover, the immunosensor had a good selectivity against epidermal growth factor receptor (EGFR) protein, long-storage stability and reproducibility. It might be become a promising assay for clinical diagnosis and early detection of tumors.