R H Smallwood

Relation between tissue structure and imposed electrical current flow in cervical neoplasia

BACKGROUNDnWhen an electrical potential is applied to human tissue, the pattern of the resulting current flow is determined by the shapes, arrangements, and internal structure of the tissue cells. By measurement of the electrical current patterns over a range of frequencies, and use of an inverse modelling procedure, electrical variables describing the tissue structure can be calculated. We used this method to develop a screening technique for the detection of cervical precancers.nnnMETHODSnWe used a pencil probe (diameter 5 mm) to measure electrical impedance spectra from eight points on the cervix in 124 women with abnormal cervical smears. Variables that should be sensitive to the expected tissue changes were calculated. These were compared with the colposcopic results.nnnFINDINGSnThe measured electrical impedance changes were those predicted on the basis of the expected tissue structures. Measurements made on normal squamous tissues were well separated from those made on precancerous tissues. We constructed receiver-operating-characteristic curves, comparing measurements made on normal tissue and that showing cervical intraepithelial neoplasia grade 2/3; the area under the curve was 0.951. These groups of women could be separated with a sensitivity of 0.92 and a specificity of 0.92.nnnINTERPRETATIONnCharacteristics of the electrical impedance spectra of tissues can be explained by changes in cell arrangements (layering) and in the size of the nuclei. This relation opens the way to deriving tissue structure from electrical impedance spectral measurements. We show that this approach can be used to give good separation of normal and precancerous cervical tissues.

Agent-based computational modeling of wounded epithelial cell monolayers

Computational modeling of biological systems, or in silico biology, is an emerging tool for understanding structure and order in biological tissues. Computational models of the behavior of epithelial cells in monolayer cell culture have been developed and used to predict the healing characteristics of scratch wounds made to urothelial cell cultures maintained in low- and physiological [Ca/sup 2+/] environments. Both computational models and in vitro experiments demonstrated that in low exogenous [Ca/sup 2+/], the closure of 500-/spl mu/m scratch wounds was achieved primarily by cell migration into the denuded area. The wound healing rate in low (0.09 mM) [Ca/sup 2+/] was approximately twice as rapid as in physiological (2 mM) [Ca/sup 2+/]. Computational modeling predicted that in cell cultures that are actively proliferating, no increase in the fraction of cells in the S-phase would be expected, and this conclusion was supported experimentally in vitro by bromodeoxyuridine incorporation assay. We have demonstrated that a simple rule-based model of cell behavior, incorporating rules relating to contact inhibition of proliferation and migration, is sufficient to qualitatively predict the calcium-dependent pattern of wound closure observed in vitro. Differences between the in vitro and in silico models suggest a role for wound-induced signaling events in urothelial cell cultures.

Mk3.5: a modular, multi-frequency successor to the Mk3a EIS/EIT system

This paper describes the Sheffield Mk3.5 EIT/EIS system which measures both the real and imaginary part of impedance at 30 frequencies between 2 kHz and 1.6 MHz. The system uses eight electrodes with an adjacent drive/receive electrode data acquisition protocol. The system is modular, containing eight identical data acquisition boards, which contain DSPs to generate the drive frequencies and to perform the FFT used for demodulation. The current drive is in three sequentially applied packets, where each packet contains ten summed sine waves. The data acquisition system is interfaced to a host PC through an optically isolated high speed serial link (RS485) running at 2 Mbaud (2 Mbits s(-1)). Measurements on a saline filled tank show that the average signal to noise performance of the system is 40 dB measured across all frequencies and that this figure is independent of frequency of measurement. These results suggest that the current system is 10 dB better in absolute terms than the previous Sheffield (Mk3a) system.

A study of the morphological parameters of cervical squamous epithelium

Electrical impedance spectroscopy is a technique that has been investigated as a potential method for the diagnosis of epithelial carcinomas. Finite element modelling can provide an insight into the patterns of current flow in normal and pathological epithelium and hence aid in the process of probe design optimization. In order to develop a finite element model of the structure of normal and precancerous cervical squamous epithelium, it was first necessary to obtain the mean values and ranges of a number of morphological tissue parameters. The most important parameters in discriminating normal from neoplastic tissue were identified as being cell size and shape distribution, nuclear-to-cytoplasmic volume ratio and volume of extracellular space. A survey of the literature revealed an absence of reliable quantitative data for these parameters. We therefore present the results of our own basic image analysis on normal and pathological tissue sections, which we hope will be of use to other workers wishing to model cervical squamous epithelium, or other similar tissue structures.

Determination of the relationship between the pH and conductivity of gastric juice

Studies of gastric secretion were carried out on 14 subjects, some of whom had taken acid secretion inhibitors. In vitro studies were performed in an attempt to ascertain the effect of H+ and Na+ ions on conductivity. There is a strong correlation between intragastric pH and conductivity for pH < 2, but none of the gastric samples were isotonic. The measured conductivity of the samples was therefore considerably lower than predicted for isotonic gastric juice.

Modelling of epithelial tissue impedance measured using three different designs of probe

Impedance measurement is a promising technique for detecting pre-malignant changes in epithelial tissue. This paper considers how the design of the impedance probe affects the ability to discriminate between tissue types. To do this, finite element models of the electrical properties of squamous and glandular columnar epithelia have been used. The glandular tissue model is described here for the first time. Glandular mucosa is found in many regions of the gastrointestinal tract, such as the stomach and intestine, and has a large effective surface area. Firstly, the electrical properties of a small section of gland, with epithelial cells and supportive tissue, are determined. These properties are then used to build up a three-dimensional model of a whole section of mucosa containing many thousands of glands. Measurements using different types of impedance probe were simulated by applying different boundary conditions to the models. Transepithelial impedance, and tetrapolar measurement with a probe placed on the tissue surface have been modelled. In the latter case, the impedance can be affected by conductive fluid, such as mucus, on the tissue surface. This effect has been investigated, and a new design of probe, which uses a guard electrode to counteract this potential source of variability, is proposed.

Modelled current distribution in cervical squamous tissue.

The electrical properties of cervical squamous epithelium have been modelled in the frequency range 100 Hz to 10 MHz. The hierarchical modelling process comprises a cellular level stage, which includes detailed models of cells typical of different depths within the epithelium and a tissue model, which utilizes electrical properties obtained from the cellular models. The fit between the modelled and measured impedance spectra and the distribution of current with depth depends on the macroscopic model structure. Both the properties of the basement membrane and the presence of a surface mucus layer are shown to have a significant effect. The best fit with measured data is obtained when a 10 microm thick, high-conductivity surface layer is included in the tissue model.

A comparison of neonatal and adult lung impedances derived from EIT images

An objective method of extracting respiratory data from lung images is presented, together with a technique for automatically generating regions of interest delineating the anterior and posterior regions of the lungs. The method is used to extract data on the change in lung impedance with frequency, and on calculated Cole parameters, from 19 normal neonates (gestational age 32 to 42 weeks) and 8 normal adults (age 21 to 82 years). A comparison of the impedance properties of neonatal and adult lungs was made. The variation of lung impedance with frequency in neonates, as derived from EIT images, is significantly different from that found for adults. The implications for a model of the electrical impedance of lung tissue are discussed.

Multifrequency and parametric EIT images of neonatal lungs

The aims of the study were to investigate the problems involved in making multifrequency EIT measurements on neonates and to compare the images obtained with the results from a group of normal adults. The Sheffield electrical impedance tomographic spectroscopy (EITS) system acquires multifrequency data using a set of eight drive and eight receive electrodes. EITS measurements were made on an inhomogeneous group of 10 neonates admitted to the special care baby unit for observation and feeding. R/S, characteristic frequency, RC and SC parameters were generated using the Cole equation. Comparisons of the parameters were made with data collected from normal adults in another study. We have shown that it is possible to obtain EITS parametric images of neonatal lungs and that there are some differences in Cole parameters between the adult and neonatal groups.

Electrical bioimpedance readings increase with higher pressure applied to the measuring probe

Electrical bioimpedance spectroscopy (EBIS) is a technique that uses a probe to calculate the transfer impedance from tissues. This transfer impedance can give information about the normal or pathological condition of the tissue. To take readings, pressure has to be applied to the probe in order to get a good contact between the electrodes and the tissue. We have been using EBIS to investigate the early diagnosis of dysplasia and cancer in the human cervix, oesophagus and bladder. We have found that, with increasing pressure (range used here was approximately 1 kPa to approximately 50 kPa), the resistivity readings increase in a consistent way up to 80%. In this paper, we show how this is a case in three different tissue types (oesophageal, gastric and vesical samples). These increases can be higher than those associated with the pathological changes that we are investigating (non-inflamed columnar tissue, for instance, shows values 50% higher than dysplastic columnar tissue). Finite-element modelling was also used to investigate the effect of volume reduction in the connective tissue or stroma. This simulation suggests no strong correlation between reduction of this structure and increase in resistivity. We hypothesize therefore that these changes may be mainly associated with the squeezing of water from the extracellular space. Finally, as pressure is difficult to control by hand, we raise the issue of the necessity of considering this variable when making EIS measurements.