Arrhythmogenic late Ca2+ sparks in failing heart cells and their control by action potential configuration.

Sudden death in heart failure patients is a major clinical problem worldwide, but it is unclear how arrhythmogenic early afterdepolarizations (EADs) are triggered in failing heart cells. To examine EAD initiation, high-sensitivity intracellular Ca2+ measurements were combined with action potential voltage clamp techniques in a physiologically relevant heart failure model. In failing cells, the loss of Ca2+ release synchrony at the start of the action potential leads to an increase in number of microscopic intracellular Ca2+ release events ("late" Ca2+ sparks) during phase 2-3 of the action potential. These late Ca2+ sparks prolong the Ca2+ transient that activates contraction and can trigger propagating microscopic Ca2+ ripples, larger macroscopic Ca2+ waves, and EADs. Modification of the action potential to include steps to different potentials revealed the amount of current generated by these late Ca2+ sparks and their (subsequent) spatiotemporal summation into Ca2+ ripples/waves. Comparison of this current to the net current that causes action potential repolarization shows that late Ca2+ sparks provide a mechanism for EAD initiation. Computer simulations confirmed that this forms the basis of a strong oscillatory positive feedback system that can act in parallel with other purely voltage-dependent ionic mechanisms for EAD initiation. In failing heart cells, restoration of the action potential to a nonfailing phase 1 configuration improved the synchrony of excitation-contraction coupling, increased Ca2+ transient amplitude, and suppressed late Ca2+ sparks. Therapeutic control of late Ca2+ spark activity may provide an additional approach for treating heart failure and reduce the risk for sudden cardiac death.

Improved Ca2+ release synchrony following selective modification of Itof and phase 1 repolarization in normal and failing ventricular myocytes.

Loss of ventricular action potential (AP) early phase 1 repolarization may contribute to the impaired Ca2+ release and increased risk of sudden cardiac death in heart failure. Therefore, restoring AP phase 1 by augmenting the fast transient outward K+ current (Itof) might be beneficial, but direct experimental evidence to support this proposition in failing cardiomyocytes is limited. Dynamic clamp was used to selectively modulate the contribution of Itof to the AP and Ca2+ transient in both normal (guinea pig and rabbit) and in failing rabbit cardiac myocytes. Opposing native Itof in non-failing rabbit myocytes increased Ca2+ release heterogeneity, late Ca2+ sparks (LCS) frequency and AP duration. (APD). In contrast, increasing Itof in failing myocytes and guinea pig myocytes (the latter normally lacking Itof) increased Ca2+ transient amplitude, Ca2+ release synchrony, and shortened APD. Computer simulations also showed faster Ca2+ transient decay (mainly due to fewer LCS), decreased inward Na+/Ca2+ exchange current and APD. When the Itof conductance was increased to ~0.2 nS/pF in failing cells (a value slightly greater than seen in typical human epicardial myocytes), Ca2+ release synchrony improved and AP duration decreased slightly. Further increases in Itof can cause Ca2+ release to decrease as the peak of the bell-shaped ICa-voltage relationship is passed and premature AP repolarization develops. These results suggest that there is an optimal range for Itof enhancement that may support Ca2+ release synchrony and improve electrical stability in heart failure with the caveat that uncontrolled Itof enhancement should be avoided.

Endothelial glycocalyx is damaged in diabetic cardiomyopathy: angiopoietin 1 restores glycocalyx and improves diastolic function in mice.

AIMS/HYPOTHESIS: Diabetic cardiomyopathy (DCM) is a serious and under-recognised complication of diabetes. The first sign is diastolic dysfunction, which progresses to heart failure. The pathophysiology of DCM is incompletely understood but microcirculatory changes are important. Endothelial glycocalyx (eGlx) plays multiple vital roles in the microcirculation, including in the regulation of vascular permeability, and is compromised in diabetes but has not previously been studied in the coronary microcirculation in diabetes. We hypothesised that eGlx damage in the coronary microcirculation contributes to increased microvascular permeability and hence to cardiac dysfunction. METHODS: We investigated eGlx damage and cardiomyopathy in mouse models of type 1 (streptozotocin-induced) and type 2 (db/db) diabetes. Cardiac dysfunction was determined by echocardiography. We obtained eGlx depth and coverage by transmission electron microscopy (TEM) on mouse hearts perfusion-fixed with glutaraldehyde and Alcian Blue. Perivascular oedema was assessed from TEM images by measuring the perivascular space area. Lectin-based fluorescence was developed to study eGlx in paraformaldehyde-fixed mouse and human tissues. The eGlx of human conditionally immortalised coronary microvascular endothelial cells (CMVECs) in culture was removed with eGlx-degrading enzymes before measurement of protein passage across the cell monolayer. The mechanism of eGlx damage in the diabetic heart was investigated by quantitative reverse transcription-PCR array and matrix metalloproteinase (MMP) activity assay. To directly demonstrate that eGlx damage disturbs cardiac function, isolated rat hearts were treated with enzymes in a Langendorff preparation. Angiopoietin 1 (Ang1) is known to restore eGlx and so was used to investigate whether eGlx restoration reverses diastolic dysfunction in mice with type 1 diabetes. RESULTS: In a mouse model of type 1 diabetes, diastolic dysfunction (confirmed by echocardiography) was associated with loss of eGlx from CMVECs and the development of perivascular oedema, suggesting increased microvascular permeability. We confirmed in vitro that eGlx removal increases CMVEC monolayer permeability. We identified increased MMP activity as a potential mechanism of eGlx damage and we observed loss of syndecan 4 consistent with MMP activity. In a mouse model of type 2 diabetes we found a similar loss of eGlx preceding the development of diastolic dysfunction. We used isolated rat hearts to demonstrate that eGlx damage (induced by enzymes) is sufficient to disturb cardiac function. Ang1 restored eGlx and this was associated with reduced perivascular oedema and amelioration of the diastolic dysfunction seen in mice with type 1 diabetes. CONCLUSIONS/INTERPRETATION: The association of CMVEC glycocalyx damage with diastolic dysfunction in two diabetes models suggests that it may play a pathophysiological role and the enzyme studies confirm that eGlx damage is sufficient to impair cardiac function. Ang1 rapidly restores the CMVEC glycocalyx and improves diastolic function. Our work identifies CMVEC glycocalyx damage as a potential contributor to the development of DCM and therefore as a therapeutic target.

Three-Dimensional Printed Models of the Heart Represent an Opportunity for Inclusive Learning.

Three-dimensional (3D) printed models of anatomic structures offer an alternative to studying manufactured, "idealized" models or cadaveric specimens. The utility of 3D printed models of the heart for clinical veterinary students learning echocardiographic anatomy is unreported. This study aimed to assess the feasibility and utility of 3D printed models of the canine heart as a supplementary teaching aid in final-year vet students. We hypothesized that using 3D printed cardiac models would improve test scores and feedback when compared with a control group. Students (n = 31) were randomized to use either a video guide to echocardiographic anatomy alongside 3D printed models (3DMs) or video only (VO). Prior to a self-directed learning session, students answered eight extended matching questions as a baseline knowledge assessment. They then undertook the learning session and provided feedback (Likert scores and free text). Students repeated the test within 1 to 3 days. Changes in test scores and feedback were compared between 3DM and VO groups, and between track and non-track rotation students. The 3DM group had increased test scores in the non-track subgroup. Track students' test scores in the VO group increased, but not in the 3DM group. Students in the 3DM group had a higher completion rate, and more left free-text feedback. Feedback from 3DM was almost universally positive, and students believed more strongly that these should be used for future veterinary anatomy teaching. In conclusion, these pilot data suggest that 3D printed canine cardiac models are feasible to produce and represent an inclusive learning opportunity, promoting student engagement.

DETERMINATION OF VERTEBRAL HEART SCALE AND ECHOCARDIOGRAPHIC MEASUREMENTS IN HEALTHY ANESTHETIZED AFRICAN WILD DOGS (LYCAON PICTUS) IN HUMAN CARE.

Cardiac disease is an important cause of mortality in African wild dogs in human care. Vertebral heart scale (VHS) is a well-documented objective measure of cardiac size and is commonly used in domestic dogs. The VHS of 63 clinically healthy African wild dogs housed in zoological institutions was retrospectively calculated. Using the robust method of reference interval (RI) calculation, the RI for VHS in captive African wild dogs was 9.3-10.8. Echocardiographic measurements from 16 clinically healthy and 2 African wild dogs with preclinical dilated cardiomyopathy are reported. The cardiac biomarkers N-terminal pro-B-type natriuretic peptide (NT-proBNP) and cardiac troponin I (cTnI) were measured in a subset of African wild dogs. The median plasma NT-proBNP measurement was 845 pM/L (range 366-1,388) and the median serum cTnI measurement was 0.02 ng/ml (0.01-0.04). These data can be used for the assessment and identification of cardiac disease in this endangered species.

The equine diaphragm: A novel technique for repeatable ultrasound measurement.

The diaphragm is an important respiratory muscle, playing a key role during exercise. In humans, diaphragm thickness increases in response to training and is correlated with inspiratory strength. In order to assess respiratory strength in the horse, new evaluation techniques are required and measurement of diaphragm thickness, in a non-invasive and repeatable manner, is a possible approach. The purposes of this four-part, prospective, pilot study were to develop and describe a repeatable method to measure the diaphragm thickness, using ultrasonography, in Thoroughbred racehorses. A standardized technique was developed whereby the ultrasound transducer was positioned 1 cm below a line between the cranioventral aspect of the tuber coxae and olecranon. The diaphragm thickness was measured on three occasions 1 week apart, by a single observer to determine the intraobserver repeatability, and by a second observer on one occasion to assess interobserver reproducibility. The diaphragm was observed in all intercostal spaces (ICS) from 7 to 17 on the left side, and 6 to 17 on the right side in a single horse. The thickest measurement (1.42 cm), obtained from 11 horses, was at ICS 11 on the left-side during inspiration. The narrowest measurement (0.56 cm) was obtained at ICS 16 on the right-side during expiration. There was no significant difference between the measurements obtained by a single observer on three occasions (P < .05). This is the first study to provide a detailed description of ultrasonographic imaging and measurement of the equine diaphragm. The novel technique developed to position the ultrasound transducer in a standardized location allowed examination and measurement of the diaphragm with good repeatability.

Training the equine respiratory muscles: Inspiratory muscle strength.

BACKGROUND: Little is known about the response of the equine respiratory muscles to training. OBJECTIVES: To measure an index of inspiratory muscle strength (IMSi) before and after a period of conventional exercise training (phase 1) and inspiratory muscle training (IMT), comparing high-load (treatment) and low-load (control) groups (phase 2). STUDY DESIGN: Prospective randomised controlled trial. METHODS: Phase 1: Twenty National Hunt Thoroughbred racehorses performed an inspiratory muscle strength test (IMST) twice on two occasions; when unfit at timepoint A (July), and when race fit at timepoint B (October). Phase 2: Thirty-five Thoroughbred racehorses at race fitness were randomly assigned into a high-load (treatment, n = 20) or low-load (control, n = 15) IMT group. The high-load group followed an IMT protocol that gradually increased the inspiratory pressure applied every 4 days. The low-load group underwent sham IMT with a low training load. The IMT was performed 5 days/week for 10 weeks. The IMST was performed twice on two occasions, timepoint B (October) and timepoint C (January). Conventional exercise training and racing continued during the study period. The peak IMSi values obtained from the different groups at timepoints A, B and C were compared using a Wilcoxon Signed Rank Test. RESULTS: Phase 1: There was a significant increase in IMSi from timepoint A: 22.5 cmH2 O (21-25) to timepoint B: 26 cmH2 O (24-30) (p = 0.015). Phase 2: From timepoint B to C there was a significant increase in IMSi for the high-load group 34 cmH2 O (28-36) (p = 0.001) but not the low-load group 26 cmH2 O (24-30) (p = 0.929). The peak IMSi at timepoint C was significantly higher for the high-load than low-load group (p = 0.019). MAIN LIMITATIONS: Single centre study with only National Hunt horses undergoing race-training included. CONCLUSIONS: In horses undergoing race training there is a significant increase in IMSi in response to conventional exercise training and high-load IMT.

Training the equine respiratory muscles: Ultrasonographic measurement of muscle size.

BACKGROUND: Limited information exists regarding changes in the size of respiratory and locomotor muscles in response to exercise training in the Thoroughbred racehorse. OBJECTIVES: To describe and compare the responses of the respiratory and locomotor muscles to conventional exercise training and inspiratory muscle training (IMT). STUDY DESIGN: Prospective randomised controlled trial. METHODS: Thoroughbred racehorses, in training for competition in National Hunt races, were recruited from two training establishments. Ultrasonographic images were obtained for selected muscles of the upper airway, diaphragm, accessory respiratory, and locomotor systems and their sizes measured. Examinations were performed at three timepoints: (A) when unfit, (B) following 12 weeks of conventional exercise training and (C) following 10-12 weeks continued training at race fitness. In addition, horses at yard 1 performed IMT, between timepoint B and C, and were randomly assigned into high-load (treatment) or low-load (control) group. Repeated measures models were constructed to compare the change in muscle measurements over time, and to investigate the effects of yard, previous airway surgery and IMT on the change in ultrasonographic size measurements obtained. RESULTS: Upper airway muscle size increased in response to conventional race training between timepoints A-C, and B-C. Diaphragm size increased in response to conventional exercise training between timepoints A and B. The diaphragm size of horses that undertook high-load IMT was either maintained or increased, whereas diaphragm size decreased in horses that undertook low-load IMT or no IMT between timepoints B and C. A significant interaction between gluteal muscle size and airway surgery status was observed, with greater gluteal muscle thicknesses measured in horses that had not previously undergone airway surgery (left gluteal 3.9%, p < 0.001; right 4.5%, p = 0.04). MAIN LIMITATIONS: Low number of horses underwent IMT. CONCLUSIONS: Respiratory and locomotor muscles increase in size in response to conventional exercise training, with a further change in diaphragm size in response to inspiratory muscle training.

Heart rate variability during high-speed treadmill exercise and recovery in Thoroughbred racehorses presented for poor performance.

BACKGROUND: Heart rate variability (HRV) analysis measures the inter-beat interval variation of successive cardiac cycles. Measurement of these indices has been used to assess cardiac autonomic modulation and for arrhythmia identification in exercising horses. OBJECTIVES: To report HRV indices during submaximal exercise, strenuous exercise and recovery, and explore relationships with clinical conditions (arrhythmias, lameness, equine gastric ulcer syndrome [EGUS], lower airway inflammation and upper respiratory tract obstructions [URTOs]) in Thoroughbred racehorses. STUDY DESIGN: Retrospective, observational cross-sectional study. METHODS: One hundred and eighty Thoroughbred horses underwent a treadmill exercise test with simultaneous electrocardiographic recording. Time-domain HRV indices (standard deviation of the R-R interval [SDRR]; root mean square of successive differences [RMSSD]) were derived for submaximal and strenuous exercise and recovery segments. Clinical conditions (arrhythmia [during each phase of exercise], lameness, EGUS, lower airway inflammation and URTO) were assigned to binary categories for statistical analysis. Relationships between selected HRV indices and the clinical conditions were explored using linear regression models. RESULTS: During submaximal exercise, lameness was associated with decreased logRMSSD (B = -0.19 95% confidence interval [CI] -0.31 to -0.06, p = 0.006) and arrhythmia was associated with increased logRMSSD (B = 0.31 95% CI 0.01-0.608, p = 0.04). During strenuous exercise, arrhythmia was associated with increased HRV indices (logSDRR B = 0.51 95% CI 0.40-0.62, p < 0.001; RMSSD B = 0.60 95% CI 0.49-0.72, p < 0.001). During recovery, arrhythmia was associated with increased HRV indices (logSDRR B = 0.51 95% CI 0.40-0.62, p < 0.001, logRMSSD B = 0.60 95% CI 0.49-0.72, p < 0.001). MAIN LIMITATIONS: The main limitations of this retrospective study were that not every horse had the full range of clinical testing, therefore some horses may have had undetected abnormalities. CONCLUSIONS: The presence of arrhythmia increased HRV in both phases of exercise and recovery. Lameness decreased HRV during submaximal exercise.

Prevalence of ultrasonographic gastrointestinal wall changes in dogs with acute pancreatitis: A retrospective study (2012-2020).

BACKGROUND: Ultrasonographic gastrointestinal wall changes in dogs with acute pancreatitis (AP) are not well characterized in the literature. No detailed studies have described their prevalence, characteristics, distribution, or clinical relevance. HYPOTHESIS/OBJECTIVES: Describe the prevalence of ultrasonographic gastrointestinal wall changes in a population of dogs with AP and evaluate for associations between the presence of gastrointestinal wall changes and clinical or clinicopathological variables. ANIMALS: Referral population of 66 client-owned dogs with AP. METHODS: Retrospective search of clinical records to identify dogs with AP. Clinical variables, clinicopathological variables and ultrasonographic findings were reported using descriptive statistics. A binary logistic regression model was used to evaluate for associations between the presence of gastrointestinal wall changes and clinical or clinicopathological variables. RESULTS: Sixty-six dogs were included. Forty-seven percent of dogs (95% confidence interval [CI], 35.0%-59.0%; n = 31) with AP had ultrasonographic gastrointestinal wall changes. Gastrointestinal wall changes were most common in the duodenum and identified in 71% (n = 22) of affected dogs. Of dogs with gastrointestinal wall changes, 74.2% (n = 23) had wall thickening, 61.3% (n = 19) had abnormal wall layering, and 35.5% (n = 11) had wall corrugation. In the multivariable model, only heart rate remained an independent predictor of ultrasonographic gastrointestinal wall changes (P = .02). CONCLUSIONS AND CLINICAL IMPORTANCE: Ultrasonographic gastrointestinal wall changes in this population of dogs with AP were common. Increased heart rate was the only independent predictor of gastrointestinal wall changes, which might imply more severe disease. Additional studies are required to elucidate whether ultrasonographic gastrointestinal wall changes reflect disease severity in AP.

Effect of preoperative administration of atenolol to dogs with pulmonic stenosis undergoing interventional procedures.

BACKGROUND: Beta-blockade is sometimes used in dogs with pulmonic stenosis with the intent of reducing frequency of ventricular arrhythmias during right heart catheterization. OBJECTIVES: To evaluate if pretreatment with atenolol reduces frequency of ventricular arrhythmias, anesthetist interventions, or shortens procedure time. ANIMALS: Thirty dogs with pulmonic stenosis scheduled for interventional procedures. METHODS: Single center, prospective, randomized, open-label study. Dogs were randomized to treatment with atenolol or no treatment preoperatively for a minimum of 10 days. Variables recorded included heart rate, arrhythmias and complexity, total procedure time and administration of antiarrhythmic treatment, vasopressors, positive chronotropes, or fluid boluses. RESULTS: Fifteen dogs were enrolled in each group. Dogs receiving atenolol had lower mean heart rates during the procedure (atenolol 100 ± 11 bpm vs untreated 115 ± 19 bpm, P = .01). There were no significant differences between the atenolol and untreated groups in the frequency of ventricular ectopic complexes (535 [6-5296] vs 553 [79-2863], P = .9), ventricular couplets (46 [0-481] vs 29 [3-121], P = .59), ventricular triplets (20 [0-265] vs 16 [1-82], P = .67), ventricular tachycardia (8 [0-224] vs 8 [1-118], P = .99), proportion exhibiting R-on-T phenomenon (11/15 vs 14/15, P = .33), proportion receiving intraoperative lidocaine (1/15 vs 3/15, P = .6), vasopressors/positive chronotropes (11/15 vs 5/15, P = .06), or fluid boluses (12/15 vs 7/15, P = .13). The procedure time was similar (atenolol 41 [23-68] min vs untreated 35 [18-98] min, P = .91). CONCLUSIONS AND CLINICAL IMPORTANCE: No benefit of preoperative atenolol treatment was identified in this small group of dogs.

Validation of a focused echocardiographic training program in first opinion practice.

BACKGROUND: The EPIC study defines criteria, including echocardiographic assessments of left atrial to aortic ratio (LA: Ao) and left ventricular internal diameter in diastole normalized for body weight (LVIDdN), for dogs with preclinical myxomatous mitral valve disease (MMVD) likely to benefit from pimobendan therapy. Access to echocardiography by a cardiologist is not universally available. HYPOTHESIS/OBJECTIVES: Completion of a focused echocardiographic training program would result in accurate identification of dogs fulfilling the EPIC criteria by primary care veterinarians (PC). PARTICIPANTS: Six PCs with no previous echocardiographic experience. METHODS: Prospective diagnostic test accuracy study. After training, each PC evaluated ≤10 dogs that they believed to have preclinical MMVD. The evaluation was repeated by 1 of 3 cardiology diplomates, blinded to the PC's findings. Agreement between clinical assessments and echocardiographic measurements was assessed. RESULTS: Fifty-seven dogs were evaluated by PCs; 1 dog was withdrawn because of congestive heart failure. The median time between PC and cardiologist evaluation was 0 days (range, 0-8). One incorrect diagnosis of MMVD was made by a PC (this dog had dilated cardiomyopathy [DCM]); preclinical MMVD was confirmed by the cardiologist in 55 dogs. No difference in LA: Ao (P = .96; CV = 6.9%) was detected between PCs and cardiologists. LVIDdN (1.69 cm/kg0.294 (1.26-2.21) vs 1.73 cm/kg0.294 (1.32-2.73); P = .001; CV = 6.5%), was significantly lower when measured by PCs vs cardiologists. PCs and cardiologists agreed regarding assessment of EPIC criteria in 49/56 dogs (Alpha = .761, 95% confidence interval 0.697-0.922). CONCLUSIONS AND CLINICAL IMPORTANCE: The program effectively trained PCs to accurately assess EPIC criteria in dogs with preclinical MMVD.

Clinical findings associated with N-terminal pro-B-type natriuretic peptide measurement in dogs and cats attending first opinion veterinary practices.

BACKGROUND: Clinical findings associated with N-terminal pro-B-type natriuretic peptide (NT-proBNP) measurement in dogs and cats in primary practice, and their relevance to published measurement indications, have not been described. METHODS: Using electronic health record data collected by the Small Animal Veterinary Surveillance Network, appointments in which NT-proBNP was measured were identified using keyword-based text filtering. For these appointments, clinical findings were manually identified from each patient's clinical narrative (CN) and their frequencies described. RESULTS: CNs of 3510 appointments (357 dogs and 257 cats) from 99practices were evaluated. The most frequently recorded clinical findings in dogs were: heart murmur (n = 147, 41.2% (95% confidence interval (CI) = 36.1%-46.3%), coughing (n = 83, 23.2% (95% CI = 18.8%-27.6%)) and panting (n = 58, 16.2% (95% CI = 12.4%-20.0%)) and in cats: heart murmur (n = 143, 55.6% (95% CI = 49.5%-61.7%)), suspected thromboembolism (n = 88, 34.2% (95% CI = 28.4%-40.0%)) and weight loss (n = 53, 20.6% (95% CI = 15.7%-25.5%)). Dyspnoea and tachypnoea were infrequently reported in dogs (n = 29, 8.1% (95% CI = 5.3%-10.9%) and n = 21, 5.9% (95% CI = 3.5%-8.3%), respectively) and cats (n = 26, 10.1% (95% CI = 6.4%-13.8%) and n = 36, 14.0% (95% CI = 9.8%-18.2%), respectively). CONCLUSION: Clinical findings referable to cardiac disease were recorded contemporaneously with NT-proBNP measurement and suggested both published and other indications (coughing (in dogs and cats), and serial measurements and thromboembolism (in cats)) for testing.

The microvascular endothelial glycocalyx: An additional piece of the puzzle in veterinary medicine.

The endothelial glycocalyx (eGlx) is a critically important structure lining the luminal surface of endothelial cells. There is increasing evidence, in human patients and animal models, for its crucial role in the maintenance of health. Moreover, its damage is associated with the pathogenesis of multiple disease states. This review provides readers with an overview of the eGlx; summarising its structure, essential functions, and evidence for its role in disease. We highlight the lack of studies regarding the eGlx in cats and dogs, particularly in naturally occurring diseases. Importantly, we discuss techniques to aid its study, which can be applied to veterinary species. Finally, we present targeted therapies aimed at preserving, and in some cases, restoring damaged eGlx.

Circulating hyaluronan as a marker of endothelial glycocalyx damage in dogs with myxomatous mitral valve disease and dogs in a hypercoagulable state.

The endothelial glycocalyx (eGlx) lines the luminal surface of endothelial cells, maintaining vascular health. Glycocalyx damage is pathophysiologically important in many diseases across species however few studies have investigated its breakdown in naturally occurring disease in dogs. The aims of the study were to investigate eGlx damage in dogs with myxomatous mitral valve disease (MMVD) diagnosed on echocardiography, and dogs in a hypercoagulable state diagnosed using thromboelastography (TEG), by measuring serum hyaluronan concentrations. Serum hyaluronan was quantified in dogs with MMVD (n = 27), hypercoagulability (n = 21), and in healthy controls dogs (n = 18). Serum hyaluronan concentrations were measured using a commercially-available ELISA validated for use in dogs. Hyaluronan concentrations were compared among groups using Kruskal-Wallis tests, and post-hoc with Dunn's tests. Serum hyaluronan concentrations (median [range]) were significantly increased in dogs with MMVD (62.4 [22.8-201] ng/mL; P = 0.031) and hypercoagulability (92.40 [16.9-247.6] ng/mL; P < 0.001) compared to controls (45.7 [8.7-80.2] ng/mL). Measurement of serum hyaluronan concentration offers a clinically applicable marker of eGlx health and suggests the presence of eGlx damage in dogs with MMVD and dogs in a hypercoagulable state.

Evaluation of serum cardiac troponin-I concentrations for diagnosis of infective endocarditis in dogs.

BACKGROUND: Infective endocarditis (IE) in dogs is associated with severe disease and a high case fatality rate but often presents with nonspecific clinical signs. HYPOTHESIS/OBJECTIVES: Serum concentration of cardiac troponin-I (cTnI) is elevated in dogs with IE and can differentiate dogs with IE from dogs with other diseases with similar clinical features. Concentration of serum cTnI is negatively correlated with survival time in dogs with IE. ANIMALS: Seventy-two client-owned dogs; 29 with IE, 27 with stage-B myxomatous mitral valve disease (MMVD), and 16 with immune-mediated disease (IMD). METHODS: Retrospective clinical cohort study. Concentration of serum cTnI was measured in all dogs at time of diagnosis. Clinical findings and echocardiographic interpretation were also recorded. Statistical analyses included Kruskal-Wallis test, pairwise Mann-Whitney U tests, receiver operator characteristic, and Cox proportional hazards. RESULTS: Serum concentration of cTnI was significantly higher in the IE group (0.69 ng/mL [0.03-80.8]) than in the MMVD (0.05 ng/mL [0.02-0.11], P < .001) and IMD groups (0.05 ng/mL [0.03-0.57], P < .001). Increased cTnI was a moderately accurate predictor of IE (area under the curve 0.857 (95% confidence interval [CI] 0.745-0.968, P < .001). A cTnI cutoff of 0.625 ng/mL had 100% specificity (95% CI 90%-100%) and 52% sensitivity (95% CI 33%-70%) in this study sample. There was no association between cTnI concentration and survival time in dogs with IE (hazard ratio 1.013, 95% CI 0.993-1.034, P = .2). CONCLUSIONS AND CLINICAL IMPORTANCE: Cardiac troponin-I concentrations are higher in dogs with IE compared to dogs with preclinical MMVD or IMD. In dogs with a compatible clinical presentation, serum cTnI concentrations >0.625 ng/mL are supportive of IE.

Investigation of focused cardiac ultrasound in the emergency room for differentiation of respiratory and cardiac causes of respiratory distress in dogs.

OBJECTIVE: To determine whether focused cardiac ultrasound (FOCUS) performed by emergency and critical care (ECC) specialists or residents in training improves differentiation of cardiac (C) versus non-cardiac (NC) causes of respiratory distress in dogs compared to medical history and physical examination alone. DESIGN: Prospective cohort study (May 2014 to February 2016). SETTING: University hospital. ANIMALS: Thirty-eight dogs presenting with respiratory distress. INTERVENTIONS: FOCUS. MEASUREMENTS AND MAIN RESULTS: Medical history, physical examination, and FOCUS were obtained at presentation. Emergency and critical care clinicians, blinded to any radiographic or echocardiographic data, categorized each patient (C vs NC) before and after FOCUS. Thoracic radiography (within 3 h) and echocardiography (within 24 h) were performed. Percent agreement was calculated against a reference diagnosis that relied on agreement of a board-certified cardiologist and ECC specialist with access to all diagnostic test results. Reference diagnosis included 22 dogs with cardiac and 13 dogs with noncardiac causes of respiratory distress. In 3 dogs a reference diagnosis was not established. Prior to FOCUS, positive and negative percent agreement to detect cardiac causes was 90.9% (95% CI, 70.8-98.9) and 53.9% (25.1-80.8), respectively. Overall agreement occurred in 27 of 35 dogs (77.1%). Two C and 6 NC cases were incorrectly categorized. Following FOCUS, positive and negative percent agreement to detect cardiac causes was 95.5% (77.2-99.9) and 69.2% (38.6-90.9), respectively. Overall agreement occurred in 30 of 35 dogs (85.7%). Three dogs with discrepant pre-FOCUS diagnoses were correctly re-categorized post-FOCUS. One C and 4 NC cases remained incorrectly categorized. No correctly categorized dogs were incorrectly re-categorized following FOCUS. The proportions of dogs correctly classified pre- versus post-FOCUS were not significantly different (P = 0.25). CONCLUSIONS: FOCUS did not significantly improve differentiation of C vs NC causes of respiratory distress compared to medical history and physical examination alone.

Focused cardiac ultrasound and point-of-care NT-proBNP assay in the emergency room for differentiation of cardiac and noncardiac causes of respiratory distress in cats.

OBJECTIVE: To assess the accuracy of focused cardiac ultrasound (FOCUS) and point-of-care N-terminal proBNP assay in the emergency setting for differentiation of cardiac from noncardiac causes of respiratory distress in cats. DESIGN: Prospective diagnostic accuracy study between 2014 and 2016. SETTING: Emergency room at an urban university teaching hospital. ANIMALS: Forty-one client-owned cats presenting for evaluation of respiratory distress. INTERVENTIONS: Emergency clinicians made an initial diagnosis of noncardiac or cardiac cause of respiratory distress based on physical examination (PE) findings and history. The diagnoses were updated after performing FOCUS and point-of-care N-terminal B-type natriuretic peptide (POC-BNP). Reference standard diagnosis was determined by agreement of a board-certified cardiologist and critical care specialist with access to subsequent radiographs and echocardiograms. MEASUREMENTS AND MAIN RESULTS: Forty-one cats were enrolled. Three cats with incomplete data and 1 cat with an uncertain reference standard diagnosis were excluded. The remaining 37 cats were used for analysis: 21 cardiac and 16 noncardiac cases. The ratio of left atrial to aortic root diameter (LA:Ao) measured by FOCUS was significantly correlated with LA:Ao measured by echocardiography (R = 0.646, P < 0.0001). Emergency clinicians correctly diagnosed 27 of 37 (73.0%), yielding a PE positive percent agreement = 76.2% (95% CI, 52.8-91.8%) and negative percent agreement = 68.8% (95% CI, 41.3-89.0%). Five noncardiac and 5 cardiac cats were misdiagnosed. Post FOCUS, overall percent agreement improved to 34 of 37 (91.9%), with positive percent agreement = 95.2% (95% CI, 76.2-99.9%) and negative percent agreement = 87.5% (95% CI, 61.7-98.5%). The POC-BNP yielded an overall percent agreement = 32/34 (94.1%), positive percent agreement = 100% (95% CI, 82.4-100.0%), and negative percent agreement = 86.7% (95% CI, 59.5-98.3%) in differentiating cardiac versus noncardiac cases. CONCLUSIONS: FOCUS evaluation of basic cardiac structure and LA:Ao by trained emergency clinicians improved accuracy of diagnosis compared to PE in cats with respiratory distress. FOCUS and POC-BNP are useful diagnostics in the emergent setting.

Exploring early life events including diet in cats presenting for gastrointestinal signs in later life.

Our study aimed to determine if certain early life events were more prevalent in cats presenting to veterinary practices specifically for gastrointestinal signs on at least two occasions between six months and 30 months of age. Data from an owner-completed questionnaire for 1212 cats before 16 weeks of age and subsequent questionnaires for the same cats between six months and 30 months of age were reviewed. Of the 1212 cats included, 30 visited a veterinary practice for gastrointestinal signs on two or more occasions. Of the early life events recorded, cats reported with vomiting, diarrhoea or both, and/or those not exclusively fed commercial diet(s) that meets the World Small Animal Veterinary Association (WSAVA) Global Nutrition Committee (GNC) guidelines before 16 weeks of age were more likely to visit veterinary practices specifically for gastrointestinal signs on at least two occasions between six months and 30 months of age (P<0.001, odds ratio (OR)=2.64, 95 per cent confidence interval (CI)=1.66-4.22 and P=0.030, OR=1.51, 95 per cent CI=1.04-2.22, respectively). Ensuring cats exclusively consume commercial diet(s) that meets the WSAVA GNC guidelines and further studies identifying specific aetiologies for vomiting and diarrhoea before 16 weeks of age to enable prevention may reduce the number of cats subsequently presenting to primary care veterinary practices for repeated gastrointestinal signs.

Sample Size Determination for Evaluation of Time Domain Heart Rate Variability Indices in Canine Lameness.

Heart rate variability (HRV) is a physiologic phenomenon that occurs due to changing autonomic tone resulting in variable RR intervals. A reduction in HRV is used as an index of pain in neonatal human patients. Objective measures of pain would be valuable in the evaluation of canine patients and assessment of response to pain management strategies. We hypothesized that dogs with diseases associated with discomfort (osteoarthritis and bone neoplasia) would have reduced HRV compared with normal, healthy dogs. The aim of the study was to calculate the sample size necessary to investigate this hypothesis. Seventeen dogs from the Ryan Veterinary Hospital of the University of Pennsylvania patient population or owned by Ryan Veterinary Hospital of the University of Pennsylvania staff were enrolled in this single-blind, prospective pilot study. A 30 min electrocardiogram (ECG) was obtained from each dog using an ambulatory electrocardiographic monitor. All ECGs were obtained between 10 a.m. and 2 p.m. ECGs were analyzed and time-domain HRV indices computed. Sample size calculations suggest that 207 dogs would be necessary to ascertain if HRV is reduced in dogs experiencing discomfort or pain (50 in the arthritis group, 79 in the bone cancer group, and 78 in the control group).