Cardiac ion channel expression in the equine model - In-silico prediction utilising RNA sequencing data from mixed tissue samples.

Understanding cardiomyocyte ion channel expression is crucial to understanding normal cardiac electrophysiology and underlying mechanisms of cardiac pathologies particularly arrhythmias. Hitherto, equine cardiac ion channel expression has rarely been investigated. Therefore, we aim to predict equine cardiac ion channel gene expression. Raw RNAseq data from normal horses from 9 datasets was retrieved from ArrayExpress and European Nucleotide Archive and reanalysed. The normalised (FPKM) read counts for a gene in a mix of tissue were hypothesised to be the average of the expected expression in each tissue weighted by the proportion of the tissue in the mix. The cardiac-specific expression was predicted by estimating the mean expression in each other tissues. To evaluate the performance of the model, predicted gene expression values were compared to the human cardiac gene expression. Cardiac-specific expression could be predicted for 91 ion channels including most expressed Na+ channels, K+ channels and Ca2+ -handling proteins. These revealed interesting differences from what would be expected based on human studies. These differences included predominance of NaV 1.4 rather than NaV 1.5 channel, and RYR1, SERCA1 and CASQ1 rather than RYR2, SERCA2, CASQ2 Ca2+ -handling proteins. Differences in channel expression not only implicate potentially different regulatory mechanisms but also pathological mechanisms of arrhythmogenesis.

Detecting paroxysmal atrial fibrillation from normal sinus rhythm in equine athletes using Symmetric Projection Attractor Reconstruction and machine learning.

Background: Atrial fibrillation (AF) is a common cardiac arrhythmia in both human and equine populations. It is associated with adverse outcomes in humans and decreased athletic performance in both populations. Paroxysmal atrial fibrillation (PAF) presents with intermittent, self-terminating AF episodes, and is difficult to diagnose once sinus rhythm resumes. Objective: We aimed to detect PAF subjects from normal sinus rhythm equine electrocardiograms (ECGs) using the Symmetric Projection Attractor Reconstruction (SPAR) method to encapsulate the waveform morphology and variability as the basis of a machine learning classification. Methods: We obtained ECG signals from 139 active equine athletes (120 control, 19 with a PAF diagnosis). The SPAR method was applied to 9 short (20-second) ECG strips for each subject. An optimal SPAR feature set was determined by forward feature selection for input to a machine learning model ensemble of 3 different classifiers (k-nearest neighbors, linear support vector machine, and radial basis function kernel support vector machine). Imbalanced data were handled by upsampling the minority (PAF) class. A final subject classification was made by taking a majority vote over results from the 9 ECG strips. Results: Our final cross-validated classification for a subject gave an accuracy of 89.0%, sensitivity of 94.8%, specificity of 87.1%, and receiver operating characteristic area under the curve of 0.98, taking PAF as the positive class. Conclusion: The SPAR method and machine learning generated a final model with high sensitivity, suggesting that PAF can be discriminated from short equine ECG strips. This preliminary study indicated that SPAR analysis of human ECG could support patient monitoring, risk stratification, and clinical decision-making.

Fundamentals of arrhythmogenic mechanisms and treatment strategies for equine atrial fibrillation.

Atrial fibrillation (AF) is the most common pathological arrhythmia in horses. Although it is not usually a life-threatening condition on its own, it can cause poor performance and make the horse unsafe to ride. It is a complex multifactorial disease influenced by both genetic and environmental factors including exercise training, comorbidities or ageing. The interactions between all these factors in horses are still not completely understood and the pathophysiology of AF remains poorly defined. Exciting progress has been recently made in equine cardiac electrophysiology in terms of diagnosis and documentation methods such as cardiac mapping, implantable electrocardiogram (ECG) recording devices or computer-based ECG analysis that will hopefully improve our understanding of this disease. The available pharmaceutical and electrophysiological treatments have good efficacy and lead to a good prognosis for AF, but recurrence is a frequent issue that veterinarians have to face. This review aims to summarise our current understanding of equine cardiac electrophysiology and pathophysiology of equine AF while providing an overview of the mechanism of action for currently available treatments for equine AF.

Nasal high flow oxygen therapy in hospitalised neonatal foals.

BACKGROUND: Respiratory disease is common in critically ill neonatal foals. Traditional oxygen therapy (TOT) with nasal insufflation of oxygen is often used to provide first-line respiratory support. Mechanical ventilation is used in foals which require a greater level of support but requires specialist expertise and can be associated with significant complications. Non-invasive ventilation (NIV) enables a greater level of respiratory support without the need for intubation. High flow oxygen therapy (HFOT) is a mode of NIV commonly used in human intensive care. OBJECTIVES: To describe the use of HFOT in hospitalised neonatal foals. STUDY DESIGN: Retrospective case series. METHODS: Hospital records of neonatal foals admitted between 2018 and 2019 that received treatment with HFOT were reviewed. Clinical data and complications were recorded. RESULTS: Fourteen foals were identified and the median duration of use was 43 hours (range 2-93 hours) with a median flow rate of 0.7L/kg/min (range 0.42-1.67). Ten foals survived to discharge. No significant complications associated with the technique were recorded. MAIN LIMITATIONS: A small study population which was retrospectively reviewed. CONCLUSIONS: This study provides preliminary information about the clinical use of HFOT in neonatal foals. The technique was well tolerated and no significant adverse effects were noted. However, further study is required to evaluate efficacy.

Atrial fibrillatory rate as predictor of recurrence of atrial fibrillation in horses treated medically or with electrical cardioversion.

BACKGROUND: The recurrence rate of atrial fibrillation (AF) in horses after cardioversion to sinus rhythm (SR) is relatively high. Atrial fibrillatory rate (AFR) derived from surface ECG is considered a biomarker for electrical remodelling and could potentially be used for the prediction of successful AF cardioversion and AF recurrence. OBJECTIVES: Evaluate if AFR was associated with successful treatment and could predict AF recurrence in horses. STUDY DESIGN: Retrospective multicentre study. METHODS: Electrocardiograms (ECG) from horses with persistent AF admitted for cardioversion with either medical treatment (quinidine) or transvenous electrical cardioversion (TVEC) were included. Bipolar surface ECG recordings were analysed by spatiotemporal cancellation of QRST complexes and calculation of AFR from the remaining atrial signal. Kaplan-Meier survival curve and Cox regression analyses were performed to assess the relationship between AFR and the risk of AF recurrence. RESULTS: Of the 195 horses included, 74 received quinidine treatment and 121 were treated with TVEC. Ten horses did not cardiovert to SR after quinidine treatment and AFR was higher in these, compared with the horses that successfully cardioverted to SR (median [interquartile range]), (383 [367-422] vs 351 [332-389] fibrillations per minute (fpm), P < .01). Within the first 180 days following AF cardioversion, 12% of the quinidine and 34% of TVEC horses had AF recurrence. For the horses successfully cardioverted with TVEC, AFR above 380 fpm was significantly associated with AF recurrence (hazard ratio = 2.4, 95% confidence interval 1.2-4.8, P = .01). MAIN LIMITATIONS: The treatment groups were different and not randomly allocated, therefore the two treatments cannot be compared. Medical records and the follow-up strategy varied between the centres. CONCLUSIONS: High AFR is associated with failure of quinidine cardioversion and AF recurrence after successful TVEC. As a noninvasive marker that can be retrieved from surface ECG, AFR can be clinically useful in predicting the probability of responding to quinidine treatment as well as maintaining SR after electrical cardioversion.

ECG Restitution Analysis and Machine Learning to Detect Paroxysmal Atrial Fibrillation: Insight from the Equine Athlete as a Model for Human Athletes.

Atrial fibrillation is the most frequent arrhythmia in both equine and human athletes. Currently, this condition is diagnosed via electrocardiogram (ECG) monitoring which lacks sensitivity in about half of cases when it presents in paroxysmal form. We investigated whether the arrhythmogenic substrate present between the episodes of paroxysmal atrial fibrillation (PAF) can be detected using restitution analysis of normal sinus-rhythm ECGs. In this work, ECG recordings were obtained during routine clinical work from control and horses with PAF. The extracted QT, TQ, and RR intervals were used for ECG restitution analysis. The restitution data were trained and tested using k-nearest neighbor (k-NN) algorithm with various values of neighbors k to derive a discrimination tool. A combination of QT, RR, and TQ intervals was used to analyze the relationship between these intervals and their effects on PAF. A simple majority vote on individual record (one beat) classifications was used to determine the final classification. The k-NN classifiers using two-interval measures were able to predict the diagnosis of PAF with area under the receiving operating characteristic curve close to 0.8 (RR, TQ with k ≥ 9) and 0.9 (RR, QT with k ≥ 21 or TQ, QT with k ≥ 25). By simultaneously using all three intervals for each beat and a majority vote, mean area under the curves of 0.9 were obtained for all tested k-values (3-41). We concluded that 3D ECG restitution analysis can potentially be used as a metric of an automated method for screening of PAF.

Exercise-associated rhythm disturbances in poorly performing Thoroughbreds: risk factors and association with racing performance.

BACKGROUND: Exercise-associated cardiac rhythm disturbances are common, but there is a lack of evidence-based criteria on which to distinguish clinically relevant rhythm disturbances from those that are not. OBJECTIVES: To describe and characterise rhythm disturbances during clinical exercise testing; to explore potential risk factors for these rhythm disturbances and to determine whether they influenced future racing. STUDY DESIGN: Retrospective cohort using a convenience sample. METHODS: Medical records were reviewed from two clinical services to identify horses with poor performance and/or respiratory noise with both exercise endoscopy and electrocardiography results. Respiratory and ECG findings recorded by the attending clinicians were described, and for polymorphic ventricular rhythms (n = 12), a consensus team agreed the final rhythm characterisation. Several statistical models analysing risk factors were built and racing records were reviewed to compare horses with and without rhythm disturbance. RESULTS: Of 245 racehorses, 87 (35.5%) had no ectopic/re-entrant rhythms, 110 (44.9%) had isolated premature depolarisations during sinus rhythm and 48 (19.6%) horses had complex tachydysrrythmias. Rhythm disturbances were detected during warm-up in 20 horses (8.2%); during gallop in 61 horses (24.9%) and during recovery in 124 horses (50.6%). Most complex rhythm events occurred during recovery, but there was one horse with a single couplet during gallop and another with a triplet during gallop. Fifteen horses (one with frequent isolated premature depolarisations and 14 complex rhythms) were considered by clinicians to be potentially contributing to poor performance. Treadmill exercise tests, the presence of exercise-associated upper respiratory tract obstructions and National Hunt racehorses were associated with rhythm disturbances. The proportion of horses racing again after diagnosis (82%) was similar in all groups and univariable analysis revealed no significant associations between subsequent racing and the presence of any ectopic/re-entrant rhythm, or the various sub-groups based on phase of exercise in which this was detected. MAIN LIMITATIONS: Reliance on retrospective data collection from medical records with no control group. Exercise ECGs were collected using only 1 or 2 leads. Variables examined as risk factors could be considered to be inter-related and our sub-groups were small. CONCLUSIONS: This study confirms a high prevalence of cardiac rhythm disturbances, including complex ectopic/re-entrant rhythms, in poorly performing racehorses. Detection of rhythm disturbances may vary with exercise test conditions and exercise-associated upper respiratory tract obstructions increase the risk of rhythm disturbances.

Real-time three-dimensional (3D) echocardiographic characterisation of an atrial septal defect in a horse.

Atrial septal defects have been well reported in humans and dogs and the principles of intervention have been well established. In contrast, there is very little information published about these congenital anomalies in horses. True ASDs are regarded as rare and little is known about the clinical significance of these defects in horses. An 11-year-old Thoroughbred gelding with a history of poor performance was diagnosed with an atrial septal defect, measuring approximately 2 cm in diameter, on 2D transthoracic echocardiography. Real-time three-dimensional (3D) echocardiography was used to map the structure of the defect and was useful in fully characterising the ASD in this case.

The complexity of clinically-normal sinus-rhythm ECGs is decreased in equine athletes with a diagnosis of paroxysmal atrial fibrillation.

Equine athletes have a pattern of exercise which is analogous to human athletes and the cardiovascular risks in both species are similar. Both species have a propensity for atrial fibrillation (AF), which is challenging to detect by ECG analysis when in paroxysmal form. We hypothesised that the proarrhythmic background present between fibrillation episodes in paroxysmal AF (PAF) might be detectable by complexity analysis of apparently normal sinus-rhythm ECGs. In this retrospective study ECG recordings were obtained during routine clinical work from 82 healthy horses and from 10 horses with a diagnosis of PAF. Artefact-free 60-second strips of normal sinus-rhythm ECGs were converted to binary strings using threshold crossing, beat detection and a novel feature detection parsing algorithm. Complexity of the resulting binary strings was calculated using Lempel-Ziv ('76 & '78) and Titchener complexity estimators. Dependence of Lempel-Ziv '76 and Titchener T-complexity on the heart rate in ECG strips obtained at low heart rates (25-60 bpm) and processed by the feature detection method was found to be significantly different in control animals and those diagnosed with PAF. This allows identification of horses with PAF from sinus-rhythm ECGs with high accuracy.

Prevalence of acute kidney injury in a population of hospitalized horses.

BACKGROUND: Hospital-acquired acute kidney injury (AKI) in humans and dogs increases morbidity and nonsurvival. Azotemia at presentation has been associated with a poor outcome in horses; however, prevalence and consequences of hospital-acquired AKI are unreported. HYPOTHESIS/OBJECTIVES: To evaluate the prevalence of AKI in hospitalized horses, risk factors associated with AKI, and the effect of AKI on short-term survival. We hypothesized that the prevalence of AKI in horses is similar to that reported in other domestic mammalian species and would be associated with nonsurvival. ANIMALS: Adult horses hospitalized for >2 days from which a minimum of 2 measurements of serum creatinine concentration were available. METHODS: Retrospective cohort study. Clinical records were reviewed and horses grouped according to their baseline serum creatinine concentration and change in serum creatinine concentration from baseline. The associations between signalment, diagnosis, and treatment variables, and the presence of azotemia or AKI were assessed using multinomial logistic regression. The relationship between these conditions and survival to discharge was evaluated. RESULTS: Three hundred twenty-five horses were included; 4.3% (14/325) had azotemia at baseline and 14.8% (48/325) developed AKI. There were no significant associations between investigated risk factors and development of AKI. The presence of azotemia and AKI did not significantly affect survival to discharge (P = .08 and .81, respectively). CONCLUSIONS AND CLINICAL IMPORTANCE: The prevalence of AKI in this population of hospitalized horses is similar to that reported in dogs and humans; however, in this study population, there was less impact on morbidity and short-term survival.

Equine Acquired Valvular Disease.

Degenerative myxomatous disease is common and is associated with aging. Poor prognostic indicators for equine aortic regurgitation specifically include ventricular ectopy, increased pulse pressure, and hyperkinetic pulses. Valvular prolapse is a functional abnormality diagnosed echocardiographically, about which knowledge is limited. A better understanding of its role in valvular regurgitation is needed. Infective endocarditis presents with fever and other systemic signs accompanying valvular regurgitation. The prognosis is poor, warranting aggressive therapy. Other forms of valvular disease occur rarely, but often presenting with severe regurgitation. Management of horses with valvular disease is focused on assessment of severity and regular clinical, echocardiographic, and electrocardiographic monitoring.

The application of Lempel-Ziv and Titchener complexity analysis for equine telemetric electrocardiographic recordings.

The analysis of equine electrocardiographic (ECG) recordings is complicated by the absence of agreed abnormality classification criteria. We explore the applicability of several complexity analysis methods for characterization of non-linear aspects of electrocardiographic recordings. We here show that complexity estimates provided by Lempel-Ziv '76, Titchener's T-complexity and Lempel-Ziv '78 analysis of ECG recordings of healthy Thoroughbred horses are highly dependent on the duration of analysed ECG fragments and the heart rate. The results provide a methodological basis and a feasible reference point for the complexity analysis of equine telemetric ECG recordings that might be applied to automate detection of equine arrhythmias in equine clinical practice.

Cardiac electrophysiological adaptations in the equine athlete-Restitution analysis of electrocardiographic features.

Exercising horses uniquely accommodate 7-8-fold increases in heart rate (HR). The present experiments for the first time analysed the related adaptations in action potential (AP) restitution properties recorded by in vivo telemetric electrocardiography from Thoroughbred horses. The horses were subjected to a period of acceleration from walk to canter. The QRS durations, and QT and TQ intervals yielded AP conduction velocities, AP durations (APDs) and diastolic intervals respectively. From these, indices of active, λ = QT/(QRS duration), and resting, λ0 = TQ/(QRS duration), AP wavelengths were calculated. Critical values of QT and TQ intervals, and of λ and λ0 at which plots of these respective pairs of functions showed unity slope, were obtained. These were reduced by 38.9±2.7% and 86.2±1.8%, and 34.1±3.3% and 85.9±1.2%, relative to their resting values respectively. The changes in λ were attributable to falls in QT interval rather than QRS duration. These findings both suggested large differences between the corresponding critical (129.1±10.8 or 117.4±5.6 bpm respectively) and baseline HRs (32.9±2.1 (n = 7) bpm). These restitution analyses thus separately identified concordant parameters whose adaptations ensure the wide range of HRs over which electrophysiological activation takes place in an absence of heart block or arrhythmias in equine hearts. Since the horse is amenable to this in vivo electrophysiological analysis and displays a unique wide range of heart rates, it could be a novel cardiac electrophysiology animal model for the study of sudden cardiac death in human athletes.