Bioaccumulation and effects of selenium from surface coal mining in an aquatic songbird.

Open-pit mining operations are hailed for safe working conditions for miners as well as economically and logistically favourable outcomes for mining companies. However, ecological impacts of these operations may persist for decades. Expansions of open-pit coal mining in British Columbia of Western Canada are planned. Governmental regulation of background contaminants leached from these mines into nearby water systems were established to mitigate environmental impacts associated with these operations. We analyzed water, periphyton, invertebrate, and American dipper (Cinclus mexicanus) blood, egg, and feather samples for selenium and mercury exposure related to mining operations in the Elk Valley. We also quantified effects associated with exposure on clutch size and hatching success. Selenium concentrations in water, periphyton, and invertebrates were significantly higher downstream of mines compared to reference sites within and outside of the Elk River watershed. Selenium concentrations in water from exposed sites exceeded current regulatory levels established to protect wildlife by up to 20 times. Mercury concentrations were below toxic levels for birds in all media and did not factor into determination of selenium exposure or effects. Egg selenium concentrations were on average 0.9 times the regulatory threshold. Our stable isotope analyses showed that diets of nesting females included a negligible proportion of higher order consumers. We did not detect a significant effect of selenium on the rate of hatching success or clutch size of dippers nesting in exposed sites. We conclude that at the time of the investigation, selenium exposure was not impairing hatching of American dippers at the sampled locations.

Development of 3D MRI-Based Anatomically Realistic Models of Breast Tissues and Tumours for Microwave Imaging Diagnosis.

Breast cancer diagnosis using radar-based medical MicroWave Imaging (MWI) has been studied in recent years. Realistic numerical and physical models of the breast are needed for simulation and experimental testing of MWI prototypes. We aim to provide the scientific community with an online repository of multiple accurate realistic breast tissue models derived from Magnetic Resonance Imaging (MRI), including benign and malignant tumours. Such models are suitable for 3D printing, leveraging experimental MWI testing. We propose a pre-processing pipeline, which includes image registration, bias field correction, data normalisation, background subtraction, and median filtering. We segmented the fat tissue with the region growing algorithm in fat-weighted Dixon images. Skin, fibroglandular tissue, and the chest wall boundary were segmented from water-weighted Dixon images. Then, we applied a 3D region growing and Hoshen-Kopelman algorithms for tumour segmentation. The developed semi-automatic segmentation procedure is suitable to segment tissues with a varying level of heterogeneity regarding voxel intensity. Two accurate breast models with benign and malignant tumours, with dielectric properties at 3, 6, and 9 GHz frequencies have been made available to the research community. These are suitable for microwave diagnosis, i.e., imaging and classification, and can be easily adapted to other imaging modalities.

Induction of influenza-specific local CD8 T-cells in the respiratory tract after aerosol delivery of vaccine antigen or virus in the Babraham inbred pig.

There is increasing evidence that induction of local immune responses is a key component of effective vaccines. For respiratory pathogens, for example tuberculosis and influenza, aerosol delivery is being actively explored as a method to administer vaccine antigens. Current animal models used to study respiratory pathogens suffer from anatomical disparity with humans. The pig is a natural and important host of influenza viruses and is physiologically more comparable to humans than other animal models in terms of size, respiratory tract biology and volume. It may also be an important vector in the birds to human infection cycle. A major drawback of the current pig model is the inability to analyze antigen-specific CD8+ T-cell responses, which are critical to respiratory immunity. Here we address this knowledge gap using an established in-bred pig model with a high degree of genetic identity between individuals, including the MHC (Swine Leukocyte Antigen (SLA)) locus. We developed a toolset that included long-term in vitro pig T-cell culture and cloning and identification of novel immunodominant influenza-derived T-cell epitopes. We also generated structures of the two SLA class I molecules found in these animals presenting the immunodominant epitopes. These structures allowed definition of the primary anchor points for epitopes in the SLA binding groove and established SLA binding motifs that were used to successfully predict other influenza-derived peptide sequences capable of stimulating T-cells. Peptide-SLA tetramers were constructed and used to track influenza-specific T-cells ex vivo in blood, the lungs and draining lymph nodes. Aerosol immunization with attenuated single cycle influenza viruses (S-FLU) induced large numbers of CD8+ T-cells specific for conserved NP peptides in the respiratory tract. Collectively, these data substantially increase the utility of pigs as an effective model for studying protective local cellular immunity against respiratory pathogens.

Comparison of Heterosubtypic Protection in Ferrets and Pigs Induced by a Single-Cycle Influenza Vaccine.

Influenza is a major health threat, and a broadly protective influenza vaccine would be a significant advance. Signal Minus FLU (S-FLU) is a candidate broadly protective influenza vaccine that is limited to a single cycle of replication, which induces a strong cross-reactive T cell response but a minimal Ab response to hemagglutinin after intranasal or aerosol administration. We tested whether an H3N2 S-FLU can protect pigs and ferrets from heterosubtypic H1N1 influenza challenge. Aerosol administration of S-FLU to pigs induced lung tissue-resident memory T cells and reduced lung pathology but not the viral load. In contrast, in ferrets, S-FLU reduced viral replication and aerosol transmission. Our data show that S-FLU has different protective efficacy in pigs and ferrets, and that in the absence of Ab, lung T cell immunity can reduce disease severity without reducing challenge viral replication.

Challenges of Post-measurement Histology for the Dielectric Characterisation of Heterogeneous Biological Tissues.

The dielectric properties of biological tissues are typically measured using the open-ended coaxial probe technique, which is based on the assumption that the tissue sample is homogeneous. Therefore, for heterogeneous tissue samples, additional post-measurement sample processing is conducted. Specifically, post-measurement histological analysis may be performed in order to associate the measured dielectric properties with the tissue types present in a heterogeneous sample. Accurate post-measurement histological analysis enables identification of the constituent tissue types that contributed to the measured dielectric properties, and their relative distributions. There is no standard protocol for conducting post-measurement histological analysis, which leads to high numbers of excluded tissue samples and inconsistencies in the resulting reported data for heterogeneous tissues. To this extent, this study examines the post-measurement histological process and the challenges in associating the acquired dielectric properties with the different tissue types present in heterogeneous samples. The results demonstrate that the histological process inevitably alters the morphology of samples, thus introducing errors in the interpretation of the dielectric properties acquired from heterogeneous biological samples. Notably, sample size was seen to shrink by up to 90% through the histological process, meaning that sensing volume determined from fresh tissues is not directly applicable to histology images.

Development of an Anthropomorphic Phantom of the Axillary Region for Microwave Imaging Assessment.

We produced an anatomically and dielectrically realistic phantom of the axillary region to enable the experimental assessment of Axillary Lymph Node (ALN) imaging using microwave imaging technology. We segmented a thoracic Computed Tomography (CT) scan and created a computer-aided designed file containing the anatomical configuration of the axillary region. The phantom comprises five 3D-printed parts representing the main tissues of interest of the axillary region for the purpose of microwave imaging: fat, muscle, bone, ALNs, and lung. The phantom allows the experimental assessment of multiple anatomical configurations, by including ALNs of different size, shape, and number in several locations. Except for the bone mimicking organ, which is made of solid conductive polymer, we 3D-printed cavities to represent the fat, muscle, ALN, and lung and filled them with appropriate tissue-mimicking liquids. Existing studies about complex permittivity of ALNs have reported limitations. To address these, we measured the complex permittivity of both human and animal lymph nodes using the standard open-ended coaxial-probe technique, over the 0.5 GHz-8.5 GHz frequency band, thus extending current knowledge on dielectric properties of ALNs. Lastly, we numerically evaluated the effect of the polymer which constitutes the cavities of the phantom and compared it to the realistic axillary region. The results showed a maximum difference of 7 dB at 4 GHz in the electric field magnitude coupled to the tissues and a maximum of 10 dB difference in the ALN response. Our results showed that the phantom is a good representation of the axillary region and a viable tool for pre-clinical assessment of microwave imaging technology.

Aerosol Delivery of a Candidate Universal Influenza Vaccine Reduces Viral Load in Pigs Challenged with Pandemic H1N1 Virus.

Influenza A viruses are a major health threat to livestock and humans, causing considerable mortality, morbidity, and economic loss. Current inactivated influenza vaccines are strain specific and new vaccines need to be produced at frequent intervals to combat newly arising influenza virus strains, so that a universal vaccine is highly desirable. We show that pandemic H1N1 influenza virus in which the hemagglutinin signal sequence has been suppressed (S-FLU), when administered to pigs by aerosol can induce CD4 and CD8 T cell immune responses in blood, bronchoalveolar lavage (BAL), and tracheobronchial lymph nodes. Neutralizing Ab was not produced. Detection of a BAL response correlated with a reduction in viral titer in nasal swabs and lungs, following challenge with H1N1 pandemic virus. Intratracheal immunization with a higher dose of a heterologous H5N1 S-FLU vaccine induced weaker BAL and stronger tracheobronchial lymph node responses and a lesser reduction in viral titer. We conclude that local cellular immune responses are important for protection against influenza A virus infection, that these can be most efficiently induced by aerosol immunization targeting the lower respiratory tract, and that S-FLU is a promising universal influenza vaccine candidate.

Limitations of using feline coronavirus spike protein gene mutations to diagnose feline infectious peritonitis.

Feline infectious peritonitis (FIP) is a fatal disease of cats, and a sequela of systemic feline coronavirus (FCoV) infection. Mutations in the viral spike (S) gene have been associated with FCoVs found in tissues from cats with FIP, but not FCoVs found in faeces from healthy cats, and are implicated in monocyte/macrophage tropism and systemic spread. This study was designed to determine whether S gene mutation analysis can reliably diagnose FIP. Cats were categorised as with FIP (n = 57) or without FIP (n = 45) based on gross post-mortem and histopathological examination including immunohistochemistry for FCoV antigen. RNA was purified from available tissue, fluid and faeces. Reverse-transcriptase quantitative-PCR (RT-qPCR) was performed on all samples using FCoV-specific primers, followed by sequencing of a section of the S gene on RT-qPCR positive samples. Samples were available from a total of 102 cats. Tissue, fluid, and faecal samples from cats with FIP were more likely to be FCoV RT-qPCR-positive (90.4, 78.4 and 64.6% respectively) than those from cats without FIP (7.8, 2.1 and 20% respectively). Identification of S gene mutated FCoVs as an additional step to the detection of FCoV alone, only moderately increased specificity for tissue samples (from 92.6 to 94.6%) but specificity was unchanged for fluid samples (97.9%) for FIP diagnosis; however, sensitivity was markedly decreased for tissue (from 89.8 to 80.9%) and fluid samples (from 78.4 to 60%) for FIP diagnosis. These findings demonstrate that S gene mutation analysis in FCoVs does not substantially improve the ability to diagnose FIP as compared to detection of FCoV alone.

Distinct immune responses and virus shedding in pigs following aerosol, intra-nasal and contact infection with pandemic swine influenza A virus, A(H1N1)09.

Influenza virus infection in pigs is a major farming problem, causing considerable economic loss and posing a zoonotic threat. In addition the pig is an excellent model for understanding immunity to influenza viruses as this is a natural host pathogen system. Experimentally, influenza virus is delivered to pigs intra-nasally, by intra-tracheal instillation or by aerosol, but there is little data comparing the outcome of different methods. We evaluated the shedding pattern, cytokine responses in nasal swabs and immune responses following delivery of low or high dose swine influenza pdmH1N1 virus to the respiratory tract of pigs intra-nasally or by aerosol and compared them to those induced in naturally infected contact pigs. Our data shows that natural infection by contact induces remarkably high innate and adaptive immune response, although the animals were exposed to a very low virus dose. In contacts, the kinetics of virus shedding were slow and prolonged and more similar to the low dose directly infected animals. In contrast the cytokine profile in nasal swabs, antibody and cellular immune responses of contacts more closely resemble immune responses in high dose directly inoculated animals. Consideration of these differences is important for studies of disease pathogenesis and assessment of vaccine protective efficacy.

Genotyping coronaviruses associated with feline infectious peritonitis.

Feline coronavirus (FCoV) infections are endemic among cats worldwide. The majority of infections are asymptomatic or result in only mild enteric disease. However, approximately 5 % of cases develop feline infectious peritonitis (FIP), a systemic disease that is a frequent cause of death in young cats. In this study, we report the complete coding genome sequences of six FCoVs: three from faecal samples from healthy cats and three from tissue lesion samples from cats with confirmed FIP. The six samples were obtained over a period of 8 weeks at a single-site cat rescue and rehoming centre in the UK. We found amino acid differences located at 44 positions across an alignment of the six virus translatomes and, at 21 of these positions, the differences fully or partially discriminated between the genomes derived from the faecal samples and the genomes derived from the tissue lesion samples. In this study, two amino acid differences fully discriminated the two classes of genomes: these were both located in the S2 domain of the virus surface glycoprotein gene. We also identified deletions in the 3c protein ORF of genomes from two of the FIP samples. Our results support previous studies that implicate S protein mutations in the pathogenesis of FIP.

Tris(2-butoxyethyl)phosphate and triethyl phosphate alter embryonic development, hepatic mRNA expression, thyroid hormone levels, and circulating bile acid concentrations in chicken embryos.

The organophosphate flame retardants tris(2-butoxyethyl) phosphate (TBOEP) and triethyl phosphate (TEP) are used in a wide range of applications to suppress or delay the ignition and spread of fire. Both compounds have been detected in the environment and TBOEP was recently measured in free-living avian species. In this study, TBOEP and TEP were injected into the air cell of chicken embryos at concentrations ranging from 0 to 45,400 ng/g and 0 to 241,500 ng/g egg, respectively. Pipping success, development, hepatic mRNA expression of 9 target genes, thyroid hormone levels, and circulating bile acid concentrations were determined. Exposure to the highest doses of TBOEP and TEP resulted in negligible detection of the parent compounds in embryonic contents at pipping indicating their complete metabolic degradation. TBOEP exposure had limited effects on chicken embryos, with the exception of hepatic CYP3A37 mRNA induction. TEP exposure decreased pipping success to 68%, altered growth, increased liver somatic index (LSI) and plasma bile acids, and modulated genes associated with xenobiotic and lipid metabolism and the thyroid hormone pathway. Plasma thyroxine levels were decreased at all TEP doses, including an environmentally-relevant concentration (8 ng/g), and gallbladder hypotrophy was evident at ≥ 43,200 ng/g. Tarsus length and circulating thyroxine concentration emerged as potential phenotypic anchors for the modulation of transthyretin mRNA. The increase in plasma bile acids and LSI, gallbladder hypotrophy, and discoloration of liver tissue represented potential phenotypic outcomes associated with modulation of hepatic genes involved with xenobiotic and lipid metabolism.

1,2-Dibromo-4-(1,2-dibromoethyl)-cyclohexane and tris(methylphenyl) phosphate cause significant effects on development, mRNA expression, and circulating bile acid concentrations in chicken embryos.

1,2-Dibromo-4-(1,2-dibromoethyl)-cyclohexane (DBE-DBCH; formerly abbreviated as TBECH) and tris(methylphenyl) phosphate (TMPP; formerly abbreviated as TCP) are additive flame retardants that are detected in the environment and biota. A recent avian in vitro screening study of 16 flame retardants identified DBE-DBCH and TMPP as important chemicals for follow-up in ovo evaluation based on their effects on cytotoxicity and mRNA expression in avian hepatocytes. In this study, technical mixtures of DBE-DBCH and TMPP were injected into the air cell of chicken embryos at concentrations ranging from 0 to 54,900ng/g and from 0 to 261,400ng/g, respectively, to determine effects on pipping success, development, hepatic mRNA expression, thyroid hormone levels, and circulating bile acid concentrations. Both compounds were detectable in embryos at pipping and the ß-DBE-DBCH isomer was depleted more rapidly than the α-isomer in tissue samples. DBE-DBCH had limited effects on the endpoints measured, with the exception of the up-regulation of two phase I metabolizing enzymes, CYP3A37 and CYP2H1. TMPP exposure caused embryonic deformities, altered growth, increased liver somatic index (LSI) and plasma bile acid concentrations, and altered mRNA expression levels of genes associated with xenobiotic and lipid metabolism and the thyroid hormone pathway. Overall, TMPP elicited more adverse molecular and phenotypic effects than DBE-DBCH albeit at concentrations several orders of magnitude greater than those detected in the environment. The increase in plasma bile acid concentrations was a useful phenotypic anchor as it was associated with a concomitant increase in LSI, discoloration of the liver tissue, and modulation of hepatic genes involved with xenobiotic and lipid metabolism.

Amino acid changes in the spike protein of feline coronavirus correlate with systemic spread of virus from the intestine and not with feline infectious peritonitis.

Recent evidence suggests that a mutation in the spike protein gene of feline coronavirus (FCoV), which results in an amino acid change from methionine to leucine at position 1058, may be associated with feline infectious peritonitis (FIP). Tissue and faecal samples collected post mortem from cats diagnosed with or without FIP were subjected to RNA extraction and quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) to detect FCoV RNA. In cats with FIP, 95% of tissue, and 81% of faecal samples were PCR-positive, as opposed to 22% of tissue, and 60% of faecal samples in cats without FIP. Relative FCoV copy numbers were significantly higher in the cats with FIP, both in tissues (P < 0.001) and faeces (P = 0.02). PCR-positive samples underwent pyrosequencing encompassing position 1058 of the FCoV spike protein. This identified a methionine codon at position 1058, consistent with the shedding of an enteric form of FCoV, in 77% of the faecal samples from cats with FIP, and in 100% of the samples from cats without FIP. In contrast, 91% of the tissue samples from cats with FIP and 89% from cats without FIP had a leucine codon at position 1058, consistent with a systemic form of FCoV. These results suggest that the methionine to leucine substitution at position 1058 in the FCoV spike protein is indicative of systemic spread of FCoV from the intestine, rather than a virus with the potential to cause FIP.

Lentiviral delivery of RNAi for in vivo lineage-specific modulation of gene expression in mouse lung macrophages.

Although RNA interference (RNAi) has become a ubiquitous laboratory tool since its discovery 12 years ago, in vivo delivery to selected cell types remains a major technical challenge. Here, we report the use of lentiviral vectors for long-term in vivo delivery of RNAi selectively to resident alveolar macrophages (AMs), key immune effector cells in the lung. We demonstrate the therapeutic potential of this approach by RNAi-based downregulation of p65 (RelA), a component of the pro-inflammatory transcriptional regulator, nuclear factor κB (NF-κB) and a key participant in lung disease pathogenesis. In vivo RNAi delivery results in decreased induction of NF-κB and downstream neutrophilic chemokines in transduced AMs as well as attenuated lung neutrophilia following stimulation with lipopolysaccharide (LPS). Through concurrent delivery of a novel lentiviral reporter vector (lenti-NF-κB-luc-GFP) we track in vivo expression of NF-κB target genes in real time, a critical step towards extending RNAi-based therapy to longstanding lung diseases. Application of this system reveals that resident AMs persist in the airspaces of mice following the resolution of LPS-induced inflammation, thus allowing these localized cells to be used as effective vehicles for prolonged RNAi delivery in disease settings.

Analysis of electrode arrangements for brain stroke diagnosis via electrical impedance tomography through numerical computational models.

Objective.Rapid stroke-type classification is crucial for improved prognosis. However, current methods for classification are time-consuming, require expensive equipment, and can only be used in the hospital. One method that has demonstrated promise in a rapid, low-cost, non-invasive approach to stroke diagnosis is electrical impedance tomography (EIT). While EIT for stroke diagnosis has been the topic of several studies in recent years, to date, the impact of electrode placements and arrangements has rarely been analyzed or tested and only in limited scenarios. Optimizing the location and choice of electrodes can have the potential to improve performance and reduce hardware cost and complexity and, most importantly, diagnosis time.Approach.In this study, we analyzed the impact of electrodes in realistic numerical models by (1) investigating the effect of individual electrodes on the resulting simulated EIT boundary measurements and (2) testing the performance of different electrode arrangements using a machine learning classification model.Main results.We found that, as expected, the electrodes deemed most significant in detecting stroke depend on the location of the electrode relative to the stroke lesion, as well as the role of the electrode. Despite this dependence, there are notable electrodes used in the models that are consistently considered to be the most significant across the various stroke lesion locations and various head models. Moreover, we demonstrate that a reduction in the number of electrodes used for the EIT measurements is possible, given that the electrodes are approximately evenly distributed.Significance.In this way, electrode arrangement and location are important variables to consider when improving stroke diagnosis methods using EIT.

Proof-of-Concept of Microwave-Based Bladder State Detection Using Realistic Pelvic Models.

Goal: Urinary incontinence (UI) affects a significant proportion of the population and is associated with negative physical and psychological side-effects. Microwave-based technologies may have the potential to monitor bladder volume, providing a proactive, low-cost and non-invasive tool to support individuals with UI. Methods: Studies to date on microwave bladder monitoring have been limited to highly simplified computational and experimental scenarios. In this work, we study the most realistic models to date (both male and female), which incorporate dielectrically and anatomically representative tissues of the pelvic region. Results: We examine the ability of detecting bladder fullness through both reflection and transmission-based parameters and, for the first time, study the effect of urine permittivity. As a proof-of-concept of bladder state detection, we further investigate reconstructing differential radar images of the bladder with two different volumes of urine. Conclusions: The results indicate that there is strong potential for monitoring and detecting the bladder state using microwave measurements.

A non-invasive approach to skin cancer diagnosis via graphene electrical tattoos and electrical impedance tomography.

Objective.Making up one of the largest shares of diagnosed cancers worldwide, skin cancer is also one of the most treatable. However, this is contingent upon early diagnosis and correct skin cancer-type differentiation. Currently, methods for early detection that are accurate, rapid, and non-invasive are limited. However, literature demonstrating the impedance differences between benign and malignant skin cancers, as well as between different types of skin cancer, show that methods based on impedance differentiation may be promising.Approach.In this work, we propose a novel approach to rapid and non-invasive skin cancer diagnosis that leverages the technologies of difference-based electrical impedance tomography (EIT) and graphene electronic tattoos (GETs).Main results.We demonstrate the feasibility of this first-of-its-kind system using both computational numerical and experimental skin phantom models. We considered variations in skin cancer lesion impedance, size, shape, and position relative to the electrodes and evaluated the impact of using individual and multi-electrode GET (mGET) arrays. The results demonstrate that this approach has the potential to differentiate based on lesion impedance, size, and position, but additional techniques are needed to determine shape.Significance.In this way, the system proposed in this work, which combines both EIT and GET technology, exhibits potential as an entirely non-invasive and rapid approach to skin cancer diagnosis.

Repository of MRI-derived models of the breast with single and multiple benign and malignant tumors for microwave imaging research.

The diagnosis of breast cancer through MicroWave Imaging (MWI) technology has been extensively researched over the past few decades. However, continuous improvements to systems are needed to achieve clinical viability. To this end, the numerical models employed in simulation studies need to be diversified, anatomically accurate, and also representative of the cases in clinical settings. Hence, we have created the first open-access repository of 3D anatomically accurate numerical models of the breast, derived from 3.0T Magnetic Resonance Images (MRI) of benign breast disease and breast cancer patients. The models include normal breast tissues (fat, fibroglandular, skin, and muscle tissues), and benign and cancerous breast tumors. The repository contains easily reconfigurable models which can be tumor-free or contain single or multiple tumors, allowing complex and realistic test scenarios needed for feasibility and performance assessment of MWI devices prior to experimental and clinical testing. It also includes an executable file which enables researchers to generate models incorporating the dielectric properties of breast tissues at a chosen frequency ranging from 3 to 10 GHz, thereby ensuring compatibility with a wide spectrum of research requirements and stages of development for any breast MWI prototype system. Currently, our dataset comprises MRI scans of 55 patients, but new exams will be continuously added.

Common injuries and medical problems in singles figure skaters.

Figure skating continues to increase in popularity as both a recreational and competitive sport. Skaters continue to "push the envelope" in perfecting more technically difficult skills. With their strenuous training programs, skaters are at risk from chronic overuse injuries, which tend to plague singles skaters more often than acute injuries. The majority of injuries occur in the lower extremities or low back. Figure skaters also are at risk for particular medical problems, such as exercise-induced bronchospasm and disordered eating. Sports Medicine physicians are sure to encounter figure skaters and their "Sports-specific problems" in their practice and should be knowledgeable about the common maladies that can plague figure skaters. This article reviews the most common injuries and medical problems in singles figure skaters.

Applied machine learning for stroke differentiation by electrical impedance tomography with realistic numerical models.

Electrical impedance tomography (EIT) may have potential to overcome existing limitations in stroke differentiation, enabling low-cost, rapid, and mobile data collection. Combining bioimpedance measurement technologies such as EIT with machine learning classifiers to support decision-making can avoid commonly faced reconstruction challenges due to the nonlinear and ill-posed nature of EIT imaging. Therefore, in this work, we advance this field through a study integrating realistic head models with clinically relevant test scenarios, and a robust architecture consisting of nested cross-validation and principal component analysis. Specifically, realistic head models are designed which incorporate the highly conductive layers of cerebrospinal fluid in the subarachnoid space and ventricles. In total, 135 unique models are created to represent a large patient population, with normal, haemorrhagic, and ischemic brains. Simulated EIT voltage data generated from these models are used to assess the classification performance of support vector machines. Parameters explored include driving frequency, signal-to-noise ratio, kernel function, and composition of binary classes. Classifier accuracy at 60 dB signal-to-noise ratio, reported as mean and standard deviation, are (79.92% ± 10.82%) for lesion differentiation, (74.78% ± 3.79%) for lesion detection, (77.49% ± 15.90%) for bleed detection, and (60.31% ± 3.98%) for ischemia detection (after ruling out bleed). The results for each method were obtained with statistics from 3 independent runs with 17,280 observations, polynomial kernel functions, and feature reduction of 76% by PCA (from 208 to 50 features). While results of this study show promise for stroke differentiation using EIT data, our findings indicate that the achievable accuracy is highly dependent on the classification scenario and application-specific classifiers may be necessary to achieve acceptable accuracy.