Search

Post-operative new-onset atrial fibrillation (POAF) is a possible complication following cardiac surgery. Digoxin is a drug with positive inotropic and negative chronotropic effects and is listed among antiarrhythmic drugs that can be prescribed in dogs with atrial fibrillation. This report aims at describing the use of digoxin in two dogs with persistent POAF after mitral valve repair. Case 1 was a Toy Poodle, neutered male, 12 years old, weighing 5.0 kg. The patient was diagnosed with the American College of Veterinary Internal Medicine (ACVIM) Stage D myxomatous mitral valve disease without arrhythmia. The day of the surgery was defined as Day 1. Persistent POAF was diagnosed on Day 30 (1-min mean heart rate of 206 bpm by in-clinic electrocardiogram), and treatment with digoxin was started at a dose of 0.0040 mg/kg, p12h. The resolution of atrial fibrillation (AF) was confirmed at the follow-up visit on Day 58 (1-min mean heart rate of 162 bpm by in-clinic electrocardiogram). Case 2 was a mixed-breed dog, neutered male, 12 years old, weighing 7.6 kg. The patient was diagnosed with ACVIM Stage B2 myxomatous mitral valve disease without arrhythmia. After surgery, the patient had a normal sinus rhythm, but a paroxysmal ectopic atrial rhythm was observed on Day 32. Persistent POAF was diagnosed on Day 130 (1-min mean heart rate of 216 bpm by in-clinic electrocardiogram), and treatment with digoxin was started at a dose of 0.0037 mg/kg, p12h. The resolution of AF was confirmed on Day 151 (1-min mean heart rate of 107 bpm by in-clinic electrocardiogram). In this report, the authors suspected that digoxin therapy could have contributed, among other factors, to cardioversion. However, specifically designed studies are needed to confirm this preliminary hypothesis., (© 2025 The Author(s). Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

Background: Mitral valvuloplasty (MVP) is a surgical procedure for treating severe mitral regurgitation in dogs. Although MVP is considered highly invasive, the extent of myocardial injury, postoperative complications, and recovery has not been evaluated., Aim: This study examined the degree of MVP invasiveness, the extent of myocardial damage, postoperative complications, cardiomyocyte recovery, and timing of hospital discharge., Methods: Cardiac troponin I (cTnI) was used to investigate the myocardial damage caused by cardiac arrest associated with a surgical approach to the myocardium in 13 patients with MVP and five controls with patent ductus arteriosus (PDA) who underwent similar anesthesia and thoracotomy., Results: The level of cTnI peaked 1 day after surgery and was significantly higher in the MVP group (median, 19.90 ng/ml) than in the control group (median, 1.50 ng/ml p < 0.001). At day 7, the cTnI level was significantly higher in the MVP group (1.9 ng/ml) than in the control group (0.1 ng/ml) ( p < 0.001), and recovery to the preoperative level took 10 days in the MVP group but returned to the preoperative level at day 7 in the control group. Although the mean arterial pressure of cardiopulmonary bypass (CPB) at the time of use was 42.92 mmHg, the peak cTnI levels in the two patients who exhibited a temporary decrease of 20 mmHg or less (46.03 ng/ml) were significantly higher than in the other 11 patients (19.70 ng/ml) ( p < 0.05). Preoperative cTnI levels were correlated with the severity of postoperative complications ( P = 0.03, F = 0.71)., Conclusion: The results showed that MVP caused temporary greater myocardial tissue damage than thoracotomy, but postoperative recovery was smoother. A high preoperative cTnI level requires relatively more careful postoperative management, and measuring the level of cTnI over time after surgery can provide information about the extent of myocardial damage and recovery from surgery and help determine the time of discharge., Competing Interests: The authors declare that there is no conflict of interest.

Background: Myxomatous mitral valve disease (MMVD) is prevalent in dogs. Specialized diagnostics (radiography and echocardiography) may be unavailable in some veterinary settings. Cardiac biomarkers offer potential alternatives., Aim: This study evaluated the diagnostic value of N -terminal fragments of pro-brain natriuretic peptides (NT-proBNPs), atrial natriuretic peptides (ANPs), and cardiac troponin I (cTnI) levels in dogs with MMVD., Methods: 69 dogs with MMVD (asymptomatic and symptomatic) and 19 healthy controls were assessed. Biomarker levels were measured using commercial kit rapid tests., Results: Our results showed that the median NT-proBNP level in the symptomatic group was higher than those in the asymptomatic ( p < 0.001) and control ( p < 0.001) groups. Moreover, the median NT-proBNP level in the asymptomatic group was higher than that in the control group ( p < 0.001). The cTnI level in the control group was lower than those in the asymptomatic ( p = 0.039) and symptomatic ( p = 0.001) groups. No statistically significant difference in the cTnI level was noted between the asymptomatic and symptomatic groups. The best cutoff value of the NT-proBNP level to differentiate the normal controls from dogs with MMVD with or without congestive heart failure was > 505.65 pmol/l [sensitivity, 76.8%; specificity, 89.5%; and area under the curve (AUC), 0.862]. The suggested cutoff value of the NT-proBNP level to differentiate symptomatic MMVD from asymptomatic MMVD was >787.65 pmol/l (sensitivity, 78.38%; specificity, 72.55%; and AUC, 0.792)., Conclusion: NT-proBNP and cTnI may serve as point-of-care tests for dyspneic dogs, aiding MMVD assessment where specialized diagnostics are limited., Competing Interests: The authors declare no conflict of interest. While this study received funding from the Faculty of Veterinary Science Mahidol University (Thailand), Scarecrow Inc (Japan), T.J. Animal Health Co., Ltd. (Thailand), and Pharmalink International Ltd. (Hong Kong), these entities had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Background: Myxomatous mitral valve disease (MMVD) is the most common acquired cardiovascular disease in small breed dogs. In contrast to human patients with heart failure (HF), iron deficiency (ID) prevalence in dogs with MMVD is weakly known. The study aimed to assess the usability of ID markers in serum and reticulocyte parameters from whole blood of dogs with MMVD to evaluate early ID symptoms., Results: Sixty-eight dogs (43 male and 25 female) were included in the study. MMVD dogs were assigned according to the 2019 ACVIM guidelines for groups B1 (n = 9), B2 (n = 10), C (n = 27) and D (n = 10). Groups were also combined into B1 and B2 as non-symptomatic HF and C with D as symptomatic HF. Healthy controls were 12 dogs. Serum iron concentration below the reference range in dogs with MMVD was 12.5%. Other ID indices, such as %SAT, UIBC, and TIBC were similar in the MMVD groups and healthy controls (p > 0.05 for all parameters). Statistical comparison between control group and 4 groups of different stages of MMVD showed that significant differences occur only in serum transferrin. The assessment of ferritin and soluble transferrin receptors using Western Blotting did not show differences between control (n = 7) and MMVD (n = 33) dogs. Study has shown positive correlation between ID parameters and echocardiographic indices such as LA/Ao and LVIDdN, and some biochemical parameters. A significant increase in reticulocytes percentage, assessed manually, was observed in the HF group of animals (p = 0.027) compared to the control group., Conclusions: Studies have shown that ID parameters in serum are not significantly different in dogs with MMVD compared to healthy dogs. However, there is a clear correlation between atrial size and normalised left ventricular size to body size and some biochemical parameters, including ID parameters and therefore the severity of MMVD., (© 2024. The Author(s).)

Background: Differentiation of the subclinical phases of myxomatous mitral valve disease (MMVD) in dogs relies heavily on echocardiography. Focused cardiac ultrasonography (FCU) is a point-of-care technique that can assess heart size., Hypothesis/objectives: Veterinary students trained in FCU can differentiate dogs with subclinical MMVD based on left ventricular (LV) and left atrial (LA) dimensions., Animals: Forty-eight dogs with subclinical MMVD., Methods: Veterinary students were trained to measure LV dimension and LA-to-aortic root dimension ratio (LA : Ao) using FCU. Dogs were categorized into 2 cohorts based on whether or not the LV normalized internal diastolic dimension was ≥1.7 and LA : Ao was ≥1.6. Agreement between FCU and echocardiographic studies performed by cardiologists was evaluated., Results: One-hundred and forty-six FCU examinations were performed by 58 veterinary students on 48 dogs. Overall agreement between students and cardiologists was moderate (Fleiss' kappa, 0.54; 95% confidence interval [CI], 0.39-0.69; P < .001). Percentage accuracy in observations with heart dimensions less than the cutoffs (86/89, 97%) was significantly higher than in observations in with larger hearts (31/57, 54%; P < .001). Agreement increased from moderate to good as heart sizes became more extreme. Degree of confidence by students in performing FCU was significantly higher at the end vs start of the study., Conclusions and Clinical Importance: Categorization of dogs with subclinical MMVD by veterinary students using FCU was associated with moderate to good agreement with echocardiography. Focused cardiac ultrasonography is a point-of-care method that can help assess clinical stage in dogs with subclinical MMVD., (© 2024 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.)

Objective: Cardioplegic solutions are indispensable for open-heart surgeries, including mitral valve repair (MVR), a potentially curative treatment for myxomatous mitral valve disease in dogs. However, procedural methodologies are not fully established, and complications are yet to be comprehensively understood. Cardioplegic solutions contain various substances to protect the myocardium under temporal cardiac arrest. Nevertheless, ventricular fibrillation (VF) occurs as a common complication after releasing the crossclamp. Based on these backgrounds, the search for optimal cardioplegic solutions in dogs undergoing MVR is an urgent issue. This study aims to evaluate the occurrence of VF in dogs treated with blood cardioplegia (BCP) versus crystalloid cardioplegia (CCP) during MVR., Animals: A total of 251 client-owned dogs who underwent MVR from November 2015 to November 2017 were included., Methods: We retrospectively assessed the relationship between VF and type of cardioplegia (CCP or BCP) based on surgical records, including VF incidence, transfusion use, crossclamp time, and echocardiographic measurements., Results: Logistic regression analysis showed that the CCP group was associated with the occurrence of VF (OR, 2.378; CI, 1.133-4.992; P = .022). In addition, the CCP group was associated with transfusion use (OR, 2.586; CI, 1.232-5.428, P = .022). There was no difference between the groups for the pre- and postoperative echocardiographic measurements., Clinical Relevance: The BCP group had a lower incidence of VF and less transfusion use than the CCP group. This finding indicates that BCP may be a superior cardioplegic technique for MVR in dogs.

Objective: To determine quality-of-life changes in owners of dogs undergoing mitral valve repair for myxomatous mitral valve disease, up to 12 months postoperatively., Sample: Owners of 26 dogs undergoing mitral valve repair at a single UK veterinary referral hospital., Methods: Dogs underwent mitral valve repair under cardiopulmonary bypass as previously described. Owner quality of life was assessed by self-completion of a previously validated questionnaire preoperatively and at 1, 3, 6, and 12 months postoperatively., Results: There was a statistically significant improvement in quality-of-life scores from preoperatively up to 3 months postoperatively and a statistically significant improvement in individual question scores up to 6 months postoperatively., Clinical Relevance: Results suggested that owner quality of life is significantly improved following surgical repair of their pet's myxomatous mitral valve disease, and this improvement continues beyond the immediate postoperative period. These results may be useful when counseling owners of surgical candidates and is another useful outcome measure.

Objective: To determine the effects of cardiac cachexia on the metabolomic profile in dogs with myxomatous mitral valve disease (MMVD)., Animals: 3 groups of dogs with MMVD enrolled between November 30, 2018, and April 7, 2022: (1) Dogs with congestive heart failure (CHF) and cachexia (CHF-cachexia group; n = 10); (2) dogs with CHF that had no cachexia (CHF-no cachexia group; n = 10); and (3) dogs with asymptomatic disease (American College of Veterinary Internal Medicine [ACVIM] Stage B2) with no cachexia (B2 group; n = 10)., Methods: Metabolomic profiles were analyzed from serum samples using ultra-high-performance liquid chromatography-tandem mass spectroscopy. Dogs in the 3 groups were compared, with statistical significance defined as P < .05 with a low false discovery rate (q < .10) and nominal statistical significance defined as P < .05 but q > .10., Results: Numerous metabolites were significantly (n = 201) or nominally significantly (n = 345) different between groups. For example, when comparing the CHF-cachexia vs CHF-no cachexia groups, lipids were the predominant metabolite differences, including many medium- and long-chain dicarboxylates and dicarboxylate acylcarnitines. For comparisons of the CHF-cachexia vs B2 groups and the CHF-no cachexia vs B2 groups, amino acids, nucleotides, and cofactors/vitamins were the predominant metabolite differences., Clinical Relevance: Some significant metabolite differences were identified between dogs with and without cardiac cachexia.

Objective: To assess the feasibility of isometric myography in pet dogs with myxomatous mitral valve disease (MMVD) to determine its use in quantifying endothelial dysfunction., Animals: 9 dogs euthanized for medical reasons., Methods: Femoral, renal, and mesenteric arteries were collected postmortem and stored in physiological saline solution at 4 °C for myography. Mitral valves were scored for myxomatous degeneration (grades 1 to 4). Sections of arteries were mounted in wells, immersed in physiological saline solution perfused with 95% O2 and 5% CO2 at 37 °C, and stretched to an internal circumference (IC) that generated the maximal difference between active and passive wall tension (IC1). Normalization factors were calculated by dividing the IC1 by the IC at which the passive wall tension was 100 mm Hg (IC100). Vasoconstriction to phenylephrine and vasodilation to acetylcholine (endothelial dependent) and sodium nitroprusside (endothelial independent) were assessed by cumulative dose-response curves., Results: Median MMVD grade was 3. Mean values of normalization factors were 1.00 ± 0.14 (renal, n = 15), 1.00 ± 0.10 (femoral, 8), and 1.05 ± 0.12 (mesenteric, 6). Responses to phenylephrine were similar between dogs (P = .14). Reduced responses to acetylcholine compared with sodium nitroprusside were identified in 15 arteries, suggesting endothelial dysfunction., Clinical Relevance: Isometric myography of arteries from pet dogs is feasible and can identify loss of endothelial-dependent relaxation in dogs with MMVD postmortem. Its use in further research can lead to a better understanding of the pathophysiology mechanisms of this disease.

Pulmonary hypertension (PH), an increase in pulmonary arterial pressure (PAP), may occur in dogs affected with myxomatous mitral valve disease (MMVD). Recent studies suggest that an accumulation of perivascular inflammatory cells may be involved with medial thickening which is a sign of the pulmonary artery remodelling in PH. The aim of this study was to characterise perivascular inflammatory cells in the surrounding pulmonary arteries of dogs with PH due to MMVD compared to MMVD dogs and healthy control dogs. Nineteen lung samples were collected from cadavers of small-breed dogs (control n = 5; MMVD n = 7; MMVD + PH n = 7). Toluidine blue stain and multiple IHC targeting α-SMA, vWF, CD20, CD68 and CD3 was performed to examine intimal and medial thickening, assess muscularisation of the small pulmonary arteries and characterise perivascular leucocytes. Medial thickening without intimal thickening of pulmonary arteries and muscularisation of normally non-muscularised small pulmonary arteries was observed in the MMVD and MMVD + PH groups compared with the control group. The perivascular numbers of B lymphocytes, T lymphocytes and macrophages was significantly increased in the MMVD + PH group compared with the MMVD and control groups. In contrast, the perivascular number of mast cells was significantly higher in the MMVD group compared with the MMVD + PH and control groups. This study suggested that pulmonary artery remodelling as medial thickening and muscularisation of the normally non-muscular small pulmonary arteries is accompanied by the accumulation of perivascular inflammatory cells., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: Previous studies in dogs with degenerative mitral valve disease (DMVD) have identified altered myocardial energy metabolism and oxidation, which might contribute to cardiac hypertrophy. Diets rich in medium chain fatty acids and antioxidants are a potential means of treatment. A previous clinical study found significantly smaller left atrial diameter (LAD) and left atrium-to-aorta diameter ratio (LA : Ao) in dogs with subclinical DMVD fed a specially formulated diet vs control diet for 6 months., Hypothesis/objectives: A specially formulated diet will slow or arrest left heart enlargement in dogs with subclinical DMVD over 365 days., Animals: One hundred twenty-seven dogs with unmedicated subclinical DMVD; 101 dogs in the per protocol cohort., Methods: Randomized double-blinded controlled multicenter clinical trial., Results: The study's primary composite outcome measure was the sum of percentage change in LAD and left ventricular internal dimension at end-diastole (LVIDd) at day 365. In the per protocol cohort, the outcome measure increased by 8.0% (95% confidence interval [CI], 2.9%-13.1%) in dogs receiving the test diet vs 8.8% (95% CI, 5.1%-12.5%) in dogs receiving control diet (P = .79). Neither component of the primary outcome measure was significantly different between groups (LAD, P = .65; LVIDd, P = .92). No difference was found in mitral valve E wave velocity (P = .36) or the proportion of dogs withdrawn from the study because of worsening DMVD and heart enlargement (P = .41)., Conclusions and Clinical Importance: Feeding a specially formulated diet for 365 days was not associated with a significantly different rate of change of left heart size in dogs with subclinical DMVD as compared to control., (© 2023 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.)

Canine myxomatous mitral valve disease (MMVD) is similar to Barlow's form of MMVD in humans. These valvulopathies are complex, with varying speeds of progression. We hypothesized that the relative abundances of serum proteins would help identify the consecutive MMVD stages and discover new disease pathways on a systemic level. To identify distinction-contributing protein panels for disease onset and progression, we compared the proteomic profiles of serum from healthy dogs and dogs with different stages of naturally occurring MMVD. Dogs were divided into experimental groups on the basis of the left-atrium-to-aorta ratio and normalized left ventricular internal dimension in diastole values. Serum was collected from healthy (N = 12) dogs, dogs diagnosed with MMVD in stages B1 (N = 13) and B2 (N = 12) (asymptomatic), and dogs diagnosed with MMVD in chronic stage C (N = 13) (symptomatic). Serum biochemistry and selected ELISAs (galectin-3, suppression of tumorigenicity, and asymmetric dimethylarginine) were performed. Liquid chromatography-mass spectrometry (LC-MS), tandem mass tag (TMT) quantitative proteomics, and statistical and bioinformatics analysis were employed. Most of the 21 serum proteins with significantly different abundances between experimental groups ( p < 0.05, FDR ˂ 0.05) were classified as matrix metalloproteinases, protease inhibitors, scaffold/adaptor proteins, complement components, anticoagulants, cytokine, and chaperone. LC-MS TMT proteomics results obtained for haptoglobin, clusterin, and peptidase D were further validated analytically. Canine MMVD stages, including, for the first time, asymptomatic B1 and B2 stages, were successfully distinguished in dogs with the disease and healthy dogs on the basis of the relative abundances of a panel of specific serum proteins. Most proteins with significantly different abundances were involved in immune and inflammatory pathways. Their role in structural remodeling and progression of canine MMVD must be further investigated. Further research is needed to confirm the resemblance/difference with human MMVD. Proteomics data are available via ProteomeXchange with the unique dataset identifier PXD038475.

Background: Impaired renal function is 1 of the poor prognostic factors in dogs with myxomatous mitral valve disease (MMVD). However, the value of cystatin C (Cys-C), a marker of renal function, as a prognostic marker for MMVD in dogs has not yet been explored., Objective: This study aims to investigate the prognostic value of Cys-C in dogs with MMVD., Animals: Fifty client-owned small-breed dogs with MMVD were included in this study., Methods: This is a retrospective, cross-sectional study. The prognostic value of serum Cys-C concentration was assessed using univariable and multivariable Cox hazard regression analyses. Kaplan-Meier survival curves for MMVD-specific survival in dogs stratified into high and low Cys-C groups were generated and analyzed using the log-rank test., Results: Serum Cys-C concentrations were significantly associated with MMVD-related death (P < .01) in both univariable (hazard ratio [HR], 5.086; 95% confidence interval [CI], 1.950-13.270) and multivariable Cox hazard regression analysis (HR, 4.657; 95% CI, 1.767-12.270). The high Cys-C group (n = 14) had a significantly shorter MMVD-specific survival time than the low Cys-C group (n = 36; P < .01). In dogs with normal blood creatinine concentrations, the high Cys-C group (n = 10) had a significantly shorter MMVD-specific survival time than the low Cys-C group (n = 36; P < .01)., Conclusions and Clinical Importance: High serum Cys-C concentrations were associated with a worse prognosis of MMVD. Furthermore, serum Cys-C could be a predictor of MMVD prognosis even in dogs with normal blood creatinine concentration., (© 2023 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.)

Pulmonary hypertension (PH) is an important predictor of poor outcomes in dogs with mitral regurgitation (MR). The feasibility of radiography to predict PH in dogs with MR is unknown. This retrospective, observational, and analytic study aimed to identify a radiographic parameter to predict PH in dogs with MR. A total of 302 dogs diagnosed with MR on echocardiography were enrolled. Medical record and radiographic findings such as the size of the main pulmonary artery, left atrium, left ventricle, and right chamber, and cranial and caudal pulmonary arteries and veins were evaluated according to the presence of PH. The diameters of the cranial and caudal pulmonary vessels were compared to the fourth rib and the ninth rib, respectively, and the ratio of the pulmonary artery to the corresponding vein (CdPA/CdPV) was calculated. Pulmonary hypertension was diagnosed in 77 dogs (25.5%) and the prevalence of PH increased with MR grade. The CdPA/CdPV was significantly higher (P < 0.001) in the presence of PH. Multivariate analysis showed that the CdPA/CdPV was the only independent radiographic parameter that had a significant association with PH in dogs with MR (P = 0.028). The cut-off value of the CdPA/CdPV = 1.10 showed 90.6% specificity and 31.1% sensitivity for detecting PH in dogs with MR. In dogs with MR, PH can be predicted with high specificity when the caudal pulmonary artery is 1.1 times larger than the corresponding vein on radiographs., (© 2022 American College of Veterinary Radiology.)

Background: Information regarding the frequency of L waves and their prognostic relevance in dogs with secondary atrial fibrillation (AF) is limited., Hypothesis/objectives: To determine whether L waves occur and ascertain their prognostic role, as well as the role of other clinical and echocardiographic variables in dogs with AF., Animals: Fifty-five dogs with AF associated with myxomatous mitral valve disease or dilated cardiomyopathy., Methods: Retrospective, multicenter observational study. In addition to L waves analysis, other clinical and echocardiographic variables, including type of antiarrhythmic treatment, were evaluated. A survival analysis was performed to test for predictors of cardiac death and all-cause mortality using Cox proportional hazards regression models., Results: L waves were evident in 33/55 dogs (60%, 95% confidence interval [CI] = 47%-72%) but their presence did not influence outcome. Increased left ventricular end-systolic diameter normalized for body weight (LVSDn) was a significant predictor of both cardiac death (hazard ratio [HR] = 4.41, 95% CI = 1.18-16.54; P = .03) and all-cause mortality (HR = 9.39, 95% CI = 2.49-35.32; P < .001). Heart rate assessed during echocardiography (Echo-HR) represented an additional significant predictor of cardiac death (HR = 1.01, 95% CI = 1.00-1.01; P = .04) and all-cause mortality (HR = 1.01, 95% CI = 1.00-1.01; P = .04)., Conclusions and Clinical Importance: L waves occurred frequently in dogs with AF, but held no prognostic relevance. Conversely, LVSDn and Echo-HR represented independent predictors of negative outcome in these animals., (© 2022 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.)

Background: Myxomatous mitral valve disease (MMVD) is the most common diagnosed cardiovascular disease in dogs. Atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP) tests are used to diagnose congestive heart failure, but there are conflicting reports about their value in diagnosing the subclinical stages of MMVD in dogs. Moreover, the diagnostic value of blood lipoproteins in dogs with MMVD is still unclear. The purpose of this study was to assess the serum concentrations of ANP, BNP and lipoproteins of dogs with MMVD and to evaluate the correlation of the levels of ANP and BNP with lipoproteins., Results: This study was performed on 24 dogs with MMVD and 10 healthy dogs. Dogs with MMVD were classified in to stages B1 (n = 11), B2 (n = 6), C (n = 4) and D (n = 3) groups according to the classification suggested by American College of Veterinary Internal Medicine guidelines. Our results showed that the mean serum BNP levels were significantly increased for all MMVD groups compared to control dogs. The mean serum ANP levels for the stage B2, C and D groups were significantly higher than the control group, while the mean serum ANP concentrations did not differ significantly between the stage B1 and control groups. An increase in BNP level was observed in 87.5% of patients. Although BNP concentrations were elevated in 100% of dogs with stages C, D and B2, high BNP was observed in 72.72% of dogs with stage B1. Regarding ANP, 58.33% of patients had an increase in ANP. However, elevated ANP levels were found in only 27.27% of patients in stage B1, while increased ANP levels were observed in 66.66 and 100% of patients in stage B2 and C/D groups respectively. Also, in all patients with MMVD, the mean serum concentrations of high-density lipoprotein cholesterol (HDL-C) were approximately 1.7 to 2 times significantly lower than the control group. Additionally, the mean serum low-density lipoprotein cholesterol (LDL-C) increased significantly (1.9-2.7 times) compared to the control group. There was a significant inverse correlation between HDL-C and BNP, and HDL-C and ANP. LDL-C showed a significant positive correlation with BNP, and ANP. Also, LDL-C, but not HDL-C, had a significant positive correlation with LA/AO ratio, LVIDd, LVIDdN and VHS. BNP and ANP showed a significant positive correlation with LA/AO, LVIDd, LVIDdN and VHS., Conclusions: Serum BNP has a greater diagnostic value than serum ANP in dogs with MMVD. In addition, serum BNP can be used to determine the subclinical stages of B1 and B2 MMVD. This study also suggests that dogs with subclinical MMVD, showed an increase in BNP along with a decrease in HDL-C and an increase in LDL-C, which are known to be risk factors for cardiovascular diseases in human. However, it seems that high LDL-C is more involved in the pathogenesis of MMVD than low HDL-C. Therefore, periodic testing of serum lipoproteins is recommended in high-risk patients, even if total cholesterol levels are normal., (© 2022. The Author(s).)

The most common cardiovascular disease in domestic dogs is myxomatous mitral valve disease (MMVD), accounting for 75% of all cardiac disease. An increase in age is generally associated with increased incidence of the disease, but Cavalier King Charles Spaniels (CKCS) exhibit an unusually high prevalence of early-onset MMVD, and thus, potentially greater cardiac morbidity and mortality compared to other breeds. Previous research has suggested that selected candidate risk alleles for MMVD are fixed in CKCSs, including six locations within the Nebulette ( NEBL ) gene on CFA2. The current study analysed genotypes of 180 Australian CKCSs at the identified risk loci. Of these, 178 were phenotyped for severity of disease by echocardiographic measurements of left atrium to aortic root ratio (LA:Ao) and weight normalised left ventricular end diastolic diameter (LVIDdN). Genotyping array markers correctly predicted the genotype at the risk-variant loci in the CKCS population, and the NEBL1 , NEBL2 and NEBL3 variants were observed to be in perfect linkage disequilibrium in this cohort. The CKCS cohort included 6/178 dogs being heterozygous for the protective/wild-type alleles at the NEBL locus. The mean LA:Ao and LVIDdN scores of these dogs heterozygous at NEBL1-3 variants were significantly smaller, and with significantly lower variance compared to age-matched CKCSs that were homozygous for risk alleles. The lower cardiac measurements in the heterozygous dogs indicate a significantly reduced risk of severe MMVD disease. Our analysis suggests that despite relative fixation of the NEBL risk alleles, healthy reference alleles at NEBL1-3 exist in low frequency in the CKCS breed and can be used to reduce MMVD severity and mortality.

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/kg 0.294 (1.26-2.21) vs 1.73 cm/kg 0.294 (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., (© 2022 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.)

Background: Endothelial cell-specific molecule-1 (ESM-1) has emerged as a potential biomarker for cardiovascular disease in humans. Myxomatous mitral valve disease (MMVD) is the most common heart disease in dogs, and we hypothesized that MMVD causes chronic inflammation that increases susceptibility to endothelial glycocalyx (eGCX) damage. In this study, we measured the concentration of ESM-1 in a group of dogs with MMVD and evaluated factors affecting eGCX damage., Results: Sixty-four dogs (control, n = 6; MMVD, n = 58) were enrolled in this study. There was no significant difference in serum ESM-1 concentrations among the MMVD stages. The serum ESM-1 concentration was significantly higher in the death group than in the alive group in MMVD dogs. (p = 0.006). In five dogs with MMVD, serum ESM-1 concentrations tended to decrease when the cardiac drug (pimobendan, furosemide, and digoxin) dose was increased., Conclusions: In cases where MMVD progressed to decompensated heart failure with clinical symptoms and resulted in death, the concentration of serum ESM-1 increased significantly. Therefore, ESM-1 could be utilized as a new potential negative prognostic factor in patients with MMVD., (© 2022. The Author(s).)

The aim of this work was to retrospectively evaluate the influence of pimobendan on the survival time (ST) of dogs with myxomatous mitral valve disease at different stages using an Inverse Probability Weighting (IPW) analysis. An IPW method was used to minimize confounding and IPW weighted time-repeated logistic model was used to approximate survival curves (SCs) and calculate survival differences. Subjects were allocated into exposed (E) and unexposed (U). Dogs in the American College of Veterinary Internal Medicine (ACVIM) B2 class treated with pimobendan (± ACE-inhibitors) were selected for the E group, as well as symptomatic patients (ACVIM class C) treated with triple (furosemide, ACE-inhibitor, pimobendan) or quadruple (furosemide, ACE-inhibitor, pimobendan and spironolactone) therapy. The U group included ACVIM class B2 dogs not treated with any medication and ACVIM C dogs treated with a combination of furosemide and ACE-inhibitor/spironolactone without pimobendan. The survival curve (SC) of the E group crossed the U group at 1634 days. The difference between the two SCs at the time of maximum survival difference in favor of the U group was 11.3% (CI 1.7%-20.9%) (significant), in favor of the E group was 3.9% (CI -8.6%-16.4%) (not significant) and at the mean ST was 3.6% (CI -8.5%-15.7%) (not significant) in favor of the E group. For times greater than 1634 days the survival was in favor of the E group, but there were no statistically significant differences in survival in favor of the E group in this clinical population., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Application of artificial intelligence (AI) to improve clinical diagnosis is a burgeoning field in human and veterinary medicine. The objective of this prospective, diagnostic accuracy study was to determine the accuracy, sensitivity, and specificity of an AI-based software for diagnosing canine cardiogenic pulmonary edema from thoracic radiographs, using an American College of Veterinary Radiology-certified veterinary radiologist's interpretation as the reference standard. Five hundred consecutive canine thoracic radiographs made after-hours by a veterinary Emergency Department were retrieved. A total of 481 of 500 cases were technically analyzable. Based on the radiologist's assessment, 46 (10.4%) of these 481 dogs were diagnosed with cardiogenic pulmonary edema (CPE+). Of these cases, the AI software designated 42 of 46 as CPE+ and four of 46 as cardiogenic pulmonary edema negative (CPE-). Accuracy, sensitivity, and specificity of the AI-based software compared to radiologist diagnosis were 92.3%, 91.3%, and 92.4%, respectively (positive predictive value, 56%; negative predictive value, 99%). Findings supported using AI software screening for thoracic radiographs of dogs with suspected cardiogenic pulmonary edema to assist with short-term decision-making when a radiologist is unavailable., (© 2022 American College of Veterinary Radiology.)

Introduction: Large breed (LB) dogs develop dilated cardiomyopathy (DCM) and myxomatous mitral valve disease (MMVD). Echocardiography is required for a definitive diagnosis but is not always available. Our objective was to assess the clinical utility of thoracic radiographs alone and in combination with physical examination and electrocardiography findings for the prediction of clinically important DCM or MMVD in LB dogs., Animals: Four hundred fifty-five client-owned dogs ≥20 kg with concurrent thoracic radiographs and echocardiogram., Materials and Methods: Medical records were reviewed and stored thoracic radiographs and echocardiographic images were measured to classify dogs as normal heart size (NHS), preclinical DCM, clinical DCM, preclinical MMVD (with cardiomegaly), clinical MMVD, or equivocal. Dogs with preclinical MMVD, without cardiomegaly, were classified as NHS. Vertebral heart size (VHS) and vertebral left atrial size (VLAS) were measured. Receiver operating characteristic curves and prediction models were derived., Results: Prevalence of MMVD (39.3%) was higher than the prevalence of DCM (24.8%), though most MMVD dogs (67.0%) lacked cardiomegaly and were classified as NHS for analysis. The area under the curve for VHS to discriminate between NHS and clinical DCM/MMVD or preclinical DCM/MMVD was 0.861 and 0.712, respectively, while for VLAS, it was 0.891 and 0.722, respectively. Predictive models incorporating physical examination and electrocardiography findings in addition to VHS/VLAS increased area under the curve to 0.978 (NHS vs. clinical DCM/MMVD) and 0.829 (NHS vs. preclinical DCM/MMVD)., Conclusions: Thoracic radiographs were useful for predicting clinically important DCM or MMVD in LB dogs, with improved discriminatory ability when physical examination abnormalities and arrhythmias were accounted for., Competing Interests: Conflict of Interest Statement The authors do not have any conflicts of interest to disclose., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

Myxomatous mitral valve disease (MMVD) is the most common cardiac disease in dogs. The association of QRS notching (nQRS) or fragmentation (fQRS) with disease severity is currently unknown. The study objective was to assess the prevalence of nQRS and fQRS in dogs with MMVD and its severity according to ACVIM classification and to compare the results with a group of healthy dogs. This retrospective cross-sectional study included 34 healthy control dogs and 155 dogs with spontaneous MMVD (42% of dogs in class B1, 23% in class B2 and 35% in class C). fQRS was defined as nQRS complexes in two contiguous leads in the frontal plane (leads I and aVL) and (II, III or aVF). A one-way ANOVA with Bonferroni post-hoc test was used to assess the differences in continuous data between control and MMVD groups. Of the MMVD group, 58% showed nQRS in at least one lead and 27% presented fQRS. There was no difference between the number of leads with a nQRS and disease severity ( p  = 0.75) nor did the number of leads with a nQRS correlate with left atrial size ( r  = 0.48; p  = 0.5). The number of dogs with fQRS did not differ among classes of MMVD ( p  = 0.21). nQRS and fQRS were more prevalent in dogs with MMVD compared to control dogs ( p  < 0.01). This study did not identify any relationship between the number of leads with a nQRS and disease severity. However, dogs with MMVD had a higher prevalence of nQRS and fQRS compared to control group.

Background: Angiotensin-converting enzyme inhibitors (ACEIs) are commonly prescribed in dogs, but the ideal dosage is unknown., Hypothesis/objectives: In dogs with cardiac disease, a dose-response relationship exists for ACEIs with respect to long-term outcome., Animals: One hundred forty-four dogs with cardiac disease, 63 with current or prior congestive heart failure., Methods: Retrospective medical record review. Cox proportional hazards models were used to determine variables associated with 2-year survival or survival from first-onset congestive heart failure (CHF)., Results: Median initial ACEI dosage was 0.84 (interquartile range [IQR], 0.56-0.98) mg/kg/day, and 108/144 (75%) of dogs received q12h dosing. No clinically relevant changes in renal function test results, serum electrolyte concentrations, or blood pressure occurred between initial prescription of ACEI and first reevaluation (median, 14 days later). In univariable analysis, higher ACEI dose was associated with increased survival from first-onset CHF (P = .005), and within the subgroup of dogs in CHF at the time of ACEI prescription, higher ACEI dose was associated with improved survival at 2 years (P = .04). In multivariable analysis, q12h dose frequency of ACEI (hazard ratio [HR], 0.30; 95% CI, 0.10-0.88; P = .03) and higher serum potassium concentration at visit 1 (HR, 0.39; 95% CI, 0.16-0.97; P = .04) were predictive of 2-year survival. The ACEIs were well-tolerated, with only 8/144 (5.6%) dogs having ACEI dose decreased or discontinued because of adverse effects., Conclusions and Clinical Importance: Twice daily dose frequency might optimize the cardioprotective benefit of ACEIs., (© 2021 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.)

Myxomatous mitral valve disease (MMVD) is the most common heart disease and cause of cardiac death in domestic dogs. MMVD is characterised by slow progressive myxomatous degeneration from the tips of the mitral valves onwards with subsequent mitral valve regurgitation, and left atrial and ventricular dilatation. Although the disease usually has a long asymptomatic period, in dogs with severe disease, mortality is typically secondary to left-sided congestive heart failure. Although it is not uncommon for dogs to survive long enough in the asymptomatic period to die from unrelated causes; a proportion of dogs rapidly advance into congestive heart failure. Heightened prevalence in certain breeds, such as the Cavalier King Charles Spaniel, has indicated that MMVD is under a genetic influence. The genetic characterisation of the factors that underlie the difference in progression of disease is of strong interest to those concerned with dog longevity and welfare. Advanced genomic technologies have the potential to provide information that may impact treatment, prevalence, or severity of MMVD through the elucidation of pathogenic mechanisms and the detection of predisposing genetic loci of major effect. Here we describe briefly the clinical nature of the disorder and consider the physiological mechanisms that might impact its occurrence in the domestic dog. Using results from comparative genomics we suggest possible genetic approaches for identifying genetic risk factors within breeds. The Cavalier King Charles Spaniel breed represents a robust resource for uncovering the genetic basis of MMVD., (© 2021 Stichting International Foundation for Animal Genetics.)

Plasma N-terminal pro-atrial natriuretic peptide (NT-proANP) concentration increases with progression of myxomatous mitral valve disease (MMVD) in dogs. This multicentre, prospective study compared plasma NT-proANP, N-terminal pro-brain natriuretic peptide (NT-proBNP), ANP, and cardiac troponin I (cTnI) concentrations in dogs with MMVD for their characteristics and discriminatory ability to detect cardiac dilatation and congestive heart failure (CHF). Thirty-six healthy dogs and 69 dogs with MMVD were included. Clinical variables were obtained via physical examination, thoracic radiography, and echocardiography. The discriminatory ability of each cardiac biomarker (CB) to determine the presence or absence of cardiac dilatation (event 1) and CHF (event 2) was evaluated using the receiver operating characteristic curves. Plasma NT-proANP, NT-proBNP, and ANP concentrations showed a significant association with the left atrium/aorta ratio (P<0.01). The area under the curve of plasma NT-proANP and NT-proBNP concentrations were 0.72 and 0.75, respectively in event1 and 0.72 and 0.76, respectively in event2. Plasma NT-proANP and NT-proBNP concentrations showed sensitivity 80.0 and 80.0%; specificity 67.6 and 64.7% in event1 (cutoff value; 8,497.81 pg/ml and 1,453.00 pmol/l, respectively) and sensitivity 85.7 and 81.0%; specificity 60.4 and 64.6% in event2 (cutoff value; 8,684.33 pg/ml and 1,772.00 pmol/l, respectively). In dogs with MMVD, plasma NT-proANP, NT-proBNP, and ANP concentrations increase with left atrial enlargement. Particularly, plasma NT-proANP and NT-proBNP concentrations appeared to be equally useful in the discriminatory ability to detect cardiac dilatation and CHF.

Introduction: Information about real-time three-dimensional (3D) transesophageal echocardiography (TEE) for the evaluation of canine mitral valve morphology is lacking in veterinary medicine., Objectives: To evaluate the feasibility of 3D TEE for the evaluation of canine mitral valves and whether there was a difference in mitral valve morphology between American College of Veterinary Internal Medicine (ACVIM) stages., Animals: Thirty-one dogs were evaluated, including nine dogs classified as ACVIM stage B2, 15 as stage C, and seven as stage D., Materials and Methods: Three-dimensional TEE was performed after anesthetic induction for mitral valve surgery, and the 3D geometry of the mitral valve apparatus was measured., Results: The intraclass correlation coefficient was good in both inter- and intraobserver analyses of the 3D measurements of mitral valve annulus geometry and excellent in both inter- and intraobserver analyses in the 3D measurements of mitral valve annular and leaflet sizes. Annulus height to commissural width ratio of stage D dogs showed significantly lower values than B2 dogs (B2: 14.2% [9.1-20.5%]; C: 10.6% [6.5-24.1%]; D: 9.5% [4.7-13.8%]). The aortic-mitral angle of stages C and D were significantly flatter than stage B2 (B2: 122.32 ± 9.39; C: 133.66 ± 8.43; D: 140.70 ± 10.70)., Conclusions: Real-time 3D echocardiography using TEE is a feasible method to evaluate the morphology of the mitral valve in dogs. The saddle shape of the mitral annulus and aortic-mitral angle were flatter in stage D. Further studies are required to understand the pathology of mitral valve disease in dogs., Competing Interests: Conflict of Interest Statement The authors do not have any conflicts of interest to disclose., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Platelets play a central role in the development of cardiovascular diseases and changes in their proteins are involved in the pathophysiology of heart diseases in humans. There is lack of knowledge about the possible role of platelets in congestive heart failure (CHF) in dogs. Thus, this study aimed to investigate the changes in global platelet proteomes in dogs with CHF, to clarify the possible role of platelets in the physiopathology of this disease. Healthy-dogs (n = 10) and dogs with acute CHF due to myxomatous mitral valve disease (MMVD, n = 10) were used. Acute CHF was defined based on the clinical (increased respiratory rate or difficulty breathing) and radiographic findings of pulmonary edema. Dogs Blood samples were collected into tubes with acid-citrate-dextrose, and platelet-pellets were obtained by centrifuge and washing steps. Platelet-proteomes were identified using LC-MS based label-free differential proteome expression analysis method and matched according to protein database for Canis lupus familiaris., Results: Totally 104 different proteins were identified in the platelets of the dogs being 4 out of them were significantly up-regulated and 6 down-regulated in acute CHF dogs. Guanine-nucleotide-binding protein, apolipoproteins (A-II and C-III) and clusterin levels increased, but CXC-motif-chemokine-10, cytochrome-C-oxidase-subunit-2, cathepsin-D, serine/threonine-protein-phosphatase-PP1-gamma-catalytic-subunit, creatine-kinase-B-type and myotrophin levels decreased in acute CHF dogs. These proteins are associated with several molecular functions, biological processes, signaling systems and immune-inflammatory responses., Conclusion: This study describes by first time the changes in the protein composition in platelets of dogs with acute CHF due to MMVD. Our findings provide a resource for increase the knowledge about the proteome of canine platelets and their roles in CHF caused by MMVD and could be a tool for further investigations about the prevention and treatment of this disease.

Although deep learning has been explored extensively for computer-aided medical imaging diagnosis in human medicine, very little has been done in veterinary medicine. The goal of this retrospective, pilot project was to apply the deep learning artificial intelligence technique using thoracic radiographs for detection of canine left atrial enlargement and compare results with those of veterinary radiologist interpretations. Seven hundred ninety-two right lateral radiographs from canine patients with thoracic radiographs and contemporaneous echocardiograms were used to train, validate, and test a convolutional neural network algorithm. The accuracy, sensitivity, and specificity for determination of left atrial enlargement were then compared with those of board-certified veterinary radiologists as recorded on radiology reports. The accuracy, sensitivity, and specificity were 82.71%, 68.42%, and 87.09%, respectively, using an accuracy driven variant of the convolutional neural network algorithm and 79.01%, 73.68%, and 80.64%, respectively, using a sensitivity driven variant. By comparison, accuracy, sensitivity, and specificity achieved by board-certified veterinary radiologists was 82.71%, 68.42%, and 87.09%, respectively. Although overall accuracy of the accuracy driven convolutional neural network algorithm and veterinary radiologists was identical, concordance between the two approaches was 85.19%. This study documents proof-of-concept for application of deep learning techniques for computer-aided diagnosis in veterinary medicine., (© 2020 The Authors. Veterinary Radiology & Ultrasound published by Wiley Periodicals LLC on behalf of American College of Veterinary Radiology.)

Background: Almost all elderly dogs develop myxomatous mitral valve disease by the end of their life, but the cavalier King Charles spaniel (CKCS) has a heightened susceptibility, frequently resulting in death at a young age and suggesting that there is a genetic component to the condition in this breed. Transcriptional profiling can reveal the impact of genetic variation through differences in gene expression levels. The aim of this study was to determine whether expression patterns were different in mitral valves showing myxomatous degeneration from CKCS dogs compared to valves from non-CKCS dogs., Results: Gene expression patterns in three groups of canine valves resulted in distinct separation of normal valves, diseased valves from CKCS and diseased valves from other breeds; the latter were more similar to the normal valves than were the valves from CKCS. Gene expression patterns in diseased valves from CKCS dogs were quite different from those in the valves from other dogs, both affected and normal. Patterns in all diseased valves (from CKCS and other breeds) were also somewhat different from normal non-diseased samples. Analysis of differentially expressed genes showed enrichment in GO terms relating to cardiac development and function and to calcium signalling canonical pathway in the genes down-regulated in the diseased valves from CKCS, compared to normal valves and to diseased valves from other breeds. F2 (prothrombin) (CKCS diseased valves compared to normal) and MEF2C pathway activation (CKCS diseased valves compared to non-CKCS diseased valves) had the strongest association with the gene changes. A large number of genes that were differentially expressed in the CKCS diseased valves compared with normal valves and diseased valves from other breeds were associated with cardiomyocytes including CASQ2, TNNI3 and RYR2., Conclusion: Transcriptomic profiling identified gene expression changes in CKCS diseased valves that were not present in age and disease severity-matched non-CKCS valves. These genes are associated with cardiomyocytes, coagulation and extra-cellular matrix remodelling. Identification of genes that vary in the CKCS will allow exploration of genetic variation to understand the aetiology of the disease in this breed, and ultimately development of breeding strategies to eliminate this disease from the breed.

Background: Torasemide is a potent loop diuretic with potential to treat congestive heart failure (CHF) in dogs., Objective: Evaluate the efficacy and safety of torasemide compared to furosemide in dogs with first occurrence of CHF caused by degenerative mitral valve disease (DMVD)., Animals: Three hundred and nineteen dogs with new onset CHF attributable to DMVD., Methods: Double-blinded randomized noninferiority study of PO torasemide vs furosemide in addition to standard CHF treatment. The primary efficacy criterion was decreased pulmonary edema and cough and no worsening of dyspnea or exercise tolerance at day 14. Secondary endpoints included clinical response at day 84 and time to death, euthanasia, or premature study withdrawal for cardiac reasons., Results: Torasemide q24h (n = 161) was noninferior to furosemide q12h (n = 158); percentage of dogs meeting primary efficacy criterion at day 14 was similar between groups (torasemide, 74.4% [95% confidence interval (CI), 66.8%-81.0%] vs. furosemide, 73.5% [95% CI, 65.7%-80.4%]; risk ratio [RR], 1.01; 95% CI, 0.89-1.15; P = .87). Efficacy at day 84 showed similar results (RR, 1.05; 95% CI, 0.88-1.25; P = .6). Dogs receiving torasemide had a longer time to endpoint and were less than half as likely to experience death, euthanasia, or premature study withdrawal (hazard ratio, 0.36; 95% CI, 0.19-0.65; P = .001) than dogs receiving furosemide at any time during the study., Conclusion and Clinical Importance: Torasemide was noninferior to furosemide as first line PO treatment for new onset CHF caused by DMVD. Torasemide significantly decreased risk of cardiac-related death or premature study withdrawal for cardiac reasons compared to furosemide., (© 2020 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC. on behalf of the American College of Veterinary Internal Medicine.)

Background: Evaluation of left heart size helps determine disease severity in dogs with myxomatous mitral valve disease (MMVD)., Hypothesis/objectives: Determine the ability of radiographic vertebral heart size (VHS) and vertebral left atrial size (VLAS) to predict LHE ECHO in dogs with preclinical MMVD., Animals: Seventy client-owned dogs with MMVD and no historical or present clinical or radiographic evidence of congestive heart failure (CHF)., Methods: Retrospective cross-sectional study of dogs with same-day echocardiography and thoracic radiography. Receiver-operating characteristic (ROC) curves were used to assess the ability of VHS, VLAS, and VHS + VLAS to discern dogs with and without LHE ECHO , and clinically relevant cutpoints for these radiographic measurements were selected., Results: The ability of VHS and VHS + VLAS to predict LHE ECHO was moderate (area under the curve [AUC] VHS = 0.851; 95% CI, 0.762-0.941; AUC VHS + VLAS = 0.865; 0.783-0.947), and performance of VLAS and VHS + VLAS was not different from that of VHS alone. A VHS cutpoint of >10.8 had sensitivity = 91.1% (76.3%-98.1%) and specificity = 69.4% (51.9%-83.7%) for predicting LHE ECHO . A cutpoint of >11.7 had sensitivity = 32.4% (17.4%-50.5%) and specificity = 97.2% (85.5%-99.9%) for predicting LHE ECHO . Thirty (43%) of the 70 dogs had a VHS value of 10.9 to 11.7., Conclusions and Clinical Importance: Vertebral heart size >11.7 identified dogs with LHE ECHO and VHS ≤ 10.8 excluded dogs with LHE ECHO . A large percentage of dogs had VHS values intermediate to these cutpoints., (© 2020 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC. on behalf of the American College of Veterinary Internal Medicine.)

Background: Cardiac disease in dogs impacts the quality of life (QoL) of their owners, but owners' QoL has not been comprehensively assessed in this population., Objectives: To develop, validate, and provide preliminary data from a health-related QoL (hrQoL) questionnaire for owners of dogs with cardiac disease., Subjects: A total of 141 owners of dogs with cardiac disease were studied., Methods: An owner hrQoL (O-hrQoL) questionnaire containing 20 items related to areas of a person's life that could be impacted by caring for a dog with cardiac disease was developed and administered to owners of dogs with cardiac disease. The highest possible total score was 100, with higher scores indicating a worse hrQoL. Readability, internal consistency, face and construct validity, and item-total correlations were assessed., Results: Median O-hrQoL score was 35 (range, 0-87). The questionnaire had good internal consistency (Cronbach's α = 0.933), construct validity (Spearman's r = 0.38-0.53; Kendall's τ = 0.30-0.43; P < .001), and item-total correlation (Spearman's r = 0.44-0.79; Kendall's τ = 0.34-0.66; all P < .001). Fifty percent of owners indicated a negative effect of dogs' cardiac disease on their own QoL, but all owners responded that caring for their dogs either had strengthened (n = 76; 53.9%) or had no effect on their relationship with their dog (n = 65; 46.1%)., Conclusions and Clinical Importance: The O-hrQoL questionnaire had good validity, and results suggest that owners' QoL is significantly impacted by caring for dogs with cardiac disease. Additional research on effective approaches to minimizing the negative effects of a dog's cardiac disease on the owner is warranted., (© 2020 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.)

In young Cavalier King Charles spaniels (CKCS), intermittent mitral regurgitation (iMR; defined as moderate to severe mitral regurgitation [MR] in a small proportion of heartbeats), has been associated with an increased risk of cardiac death due to myxomatous mitral valve disease (MMVD). It is associated with increased R-R interval variability. Little is known about response to physiological factors and whether iMR is a precursor for developing significant MR. The aim of this study was to determine the effect of stress testing on the presence of iMR and heart rate, and short-term (1-2 year) progression of MR in CKCS with and without iMR. In total, 52 CKCS were included. Substudy 1 enrolled six dogs with iMR and 11 dogs without iMR. Substudy 2 enrolled 14 dogs with iMR and 28 dogs without iMR. Substudy 1 prospectively assessed the influence of stress testing on the presence of iMR and heart rate. Substudy 2 retrospectively evaluated short-term progression of iMR. During stress testing, iMR disappeared in 50% of CKCS and no iMR was recorded at mean heart rates >150 beats/min. Heart rate response did not differ between CKCS with or without iMR. CKCS with iMR did not have a higher odds (odds ratio = 5.2; 95% confidence interval, 0.7-38.2) of MR progression compared to controls (P = 0.1). In conclusion, physical stress influenced the occurrence of iMR in CKCS, but heart rate response was not different from CKCS without iMR. Intermittent mitral regurgitation did not significantly predict short-term MR progression. In stressed CKCS with early disease, iMR may be overlooked., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Myxomatous mitral valve disease (MMVD) is the single most important acquired cardiovascular disease of the dog. Much is known about the cellular changes and the contribution of activated myofibroblasts (valve interstitial cells (aVICs) to the valve extra-cellular matrix remodelling characteristic of the disease. However, little is known on how aVIC survival might contribute to disease pathogenesis. This study examined the temporal (disease severity-dependent) and spatial distribution of aVICs in MMVD valves, the expression of a range of apoptosis-related genes in cultured VICs from both normal (quiescent VIC (qVIC) and diseased (aVIC) valves, and the differential effects of doxorubicin treatment, as a trigger of apoptosis, on expression of the same genes. Activated myofibroblasts were identified in normal valves at the valve base only (the area closest to the annulus), and then became more numerous and apparent along the valve length as the disease progressed, with evidence of cell survival at the valve base. There were no significant differences in basal gene expression comparing qVICs and aVICs for CASP3, FAS, BID, BAX, BCL2, CASP8, DDIAS, XIAP and BIRC5. After doxorubicin treatment (2 mM) for 8 h there was significant difference (P < .05) in the expression of BID, BCL2, DDIAS, and CASP8, but when assessed for interactions using a mixed model ANOVA only CASP8 was significantly different because of treatment (P < .05). These data suggest aVIC survival in MMVD valves may be a consequence of heightened resistance of aVICs to apoptosis, but would require confirmation examining expression of the relevant proteins., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The early asymptomatic stage of glomerular injury is a diagnostic challenge in the course of renal and extra-renal disease, e.g., heart insufficiency. It was found that podocin, a podocyte-specific protein present in the urine, may serve as a biomarker in the diagnosis of glomerular disease in humans and animals including glomerulonephritis, glomerulosclerosis, amyloidosis, or nephropathy. Therefore, there is a need of development of the sensitive and straightforward method of urinary podocin identification. In this work, we report our extended research under the glomerular injury investigation in dogs by application of clinical examination and LC-MS-MRM method in the identification of canine podocin in urine samples. The LC-MS-MRM method is based on the identification of podocin tryptic peptide with the 218 H-AAEILAATPAAVQLR-OH 232 sequence. The model peptide was characterized by the highest ionization efficiency of all the proposed model podocin tryptic peptides in a canine urine sediment according to the LC-MS/MS analysis. The obtained results revealed the presence of the model peptide in 40.9% of dogs with MMVD (active glomerular injury secondary to heart disease = cardiorenal syndrome-CRS) and 33.3% dogs with chronic kidney disease. The potential applicability of the developed methodology in the analysis of podocin in canine urine sediments was confirmed.

Myxomatous mitral valve disease (MMVD) is the most common acquired cardiac disorder found in dogs. The disease process can lead to heart failure (HF) and has been found to be associated with oxidative stress and inflammation. Statins exert antioxidant and anti-inflammatory effects in human HF patients. However, the beneficial effects of statins in MMVD dogs are still unclear. Thirty MMVD dogs were enrolled in the study and were divided into two groups: MMVD without HF dogs (n = 15) and MMVD with HF dogs (n = 15). Atorvastatin (8 mg kg -1  day -1 ) was administered orally to all dogs for 4 weeks. All dogs underwent physical examination and cardiac examination at the beginning and end of the experiment, including baseline values for hematology, blood chemistry profile, lipid profile, N-terminal pro B-type natriuretic peptide, oxidative stress marker (8-isoprostane), and inflammatory marker (tumor necrosis factor alpha). The results showed that atorvastatin reduced plasma cholesterol levels in both groups. In addition, plasma concentrations of 8-isoprostane, tumor necrosis factor alpha, and N-terminal pro B-type natriuretic peptide were significantly lower after atorvastatin administration, but only in MMVD dogs in the HF group. Atorvastatin found to be associated with possible antioxidant and inflammatory effects in dogs with HF secondary to MMVD. The potential benefits of statins in dogs with HF merits further investigation in larger, placebo-controlled studies., (© 2019 John Wiley & Sons Ltd.)

Myxomatous mitral valve disease (MMVD) is a cardiac condition commonly found in older dogs. The disease process can lead to heart failure (HF). In HF, an increase of reactive oxygen species (ROS) and abnormal mitochondrial activity, as well as apoptosis, have been reported. Humanin (HN) is a polypeptide that has a cardioprotective effect against apoptosis and oxidative stress. The purposes of this study were (1) to investigate the potential role of plasma HN as a cardiac biomarker to predict disease progression of MMVD, and (2) to compare plasma HN concentrations with plasma NT-pro BNP concentrations. Thirty-one dogs were included in the study. The dogs were separated into four groups: Group 1 was healthy dogs (n = 8), Group 2 was MMVD class B (n = 8), Group 3 was MMVD class C (n = 8), and Group 4 was MMVD class D (n = 7). All dogs were given a physical examination, thoracic radiography, echocardiography, and samples of their blood were collected for hematology and blood chemistry analysis. Levels of plasma HN and plasma NT-proBNP were also investigated. The results showed that plasma HN levels were lower in the dogs with MMVD and that lower plasma HN levels were associated with greater severity of MMVD-induced HF. It was possible to observe changes in plasma HN levels at a less severe disease stage than plasma NT-proBNP in dogs with MMVD. These findings sug- gest that a decreased plasma HN level can be used as a biomarker to identify dogs with MMVD-induced HF., (Copyright© by the Polish Academy of Sciences.)

Objectives: To create reference intervals for right ventricular outflow tract fractional shortening (RVOT-FS) in healthy dogs and examine diagnostic performance of this index in dogs with pulmonary hypertension (PH). In addition, we examine the impact of myxomatous mitral valve disease (MMVD) without PH on RVOT-FS., Animals, Materials and Methods: The study population included 52 healthy adult dogs, 51 dogs with MMVD but without PH, and 51 dogs with PH. This is a prospective study. Complete echocardiographic evaluations were performed on all dogs. Right ventricular outflow tract fractional shortening was obtained by two-dimensional guided M-mode recordings from the right parasternal short axis view. Right ventricular outflow tract fractional shortening was evaluated in healthy dogs of various breeds, and reference intervals were generated. We examined the effect of PH on RVOT-FS with receiver operating characteristic analysis and evaluated the effect of MMVD on RVOT-FS in dogs without PH. Intraobserver and interobserver reproducibility was calculated., Results: Healthy dogs had RVOT-FS > 44%. Right ventricular outflow tract fractional shortening values of healthy dogs and MMVD dogs without PH did not differ (p=0.84). In dogs with PH, RVOT-FS decreased with increasing tricuspid regurgitation velocity (p<0.0001). Pimobendan use in dogs with PH increased RVOT-FS as PH worsened. Right ventricular outflow tract fractional shortening was acquired with clinically acceptable intraobserver and interobserver reproducibility., Conclusions: Right ventricular outflow tract fractional shortening is a novel, easy applicable, and repeatable index for evaluating RV systolic function. Studies comparing this index with common echocardiographic indices used to assess RV function in dogs are needed., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In humans, iron deficiency represents a relevant occurrence in heart failure (HF), with or without anaemia, and is associated with the worst outcome. Moreover, chronic kidney disease (CKD) is a well-known comorbidity of HF and is strongly associated with the risk of developing anaemia. The most common cause of HF in dogs is myxomatous mitral valve disease (MMVD). To the best of our knowledge, no studies have examined the iron status in dogs with HF, with and without CKD. The aim of this retrospective study was to evaluate the iron status in dogs affected by MMVD and how strong is the relation with HF. The retrospective study included 54 dogs with complete case records, echocardiography and laboratory analyses. Iron status was evaluated by measuring serum iron concentration (SIC), un- saturated iron binding capacity (UIBC), total iron binding capacity (TIBC), and percentage of saturation (%SAT). The prevalence of dogs showing low serum iron concentration (SIC) was 18% in the whole population, 33% in symptomatic patients, 100% in dogs with acute decompensated HF. No signif- icant differences in SIC, UIBC, TIBC and %SAT median values were found among dogs classi- fied in different ACVIM (American College of Veterinary Internal Medicine) classes, between symptomatic and non-symptomatic patients, and among IRIS (International Renal Interest Soci- ety) classes. Azotemic and non-azotemic patients presented a significant difference in SIC mean values (p=0.02). Generalised linear model (GLM) revealed that dogs with low SIC were at high- er risk of being included in a higher ACVIM class (OR=6.383, p-value=0.014). Log-rank analysis showed shorter survival in dogs with low SIC (p=0.020), multivariate Cox analysis revealed that only HF symptoms can affect survival., (Copyright© by the Polish Academy of Sciences.)

Myxomatous mitral valve disease is the most common cardiac disease of the dog, but examination of the associated cellular and molecular events has relied on the use of cadaveric valve tissue, in which functional studies cannot be undertaken. The aim of this study was to develop a three-dimensional (3D) cell co-culture model as an experimental platform to examine disease pathogenesis. Mitral valve interstitial (VIC) and endothelial (VEC) cells were cultured from normal and diseased canine (VIC only) valves. VICs were embedded in a fibrin-based hydrogel matrix and one surface was lined with VECs. The 3D static cultures (constructs) were examined qualitatively and semiquantitatively by light microscopy, immunofluorescence microscopy and protein immunoblotting. Some constructs were manipulated and the endothelium damaged, and the response examined. The construct gross morphology and histology demonstrated native tissue-like features and comparable expression patterns of cellular (α-smooth muscle actin [SMA] and embryonic smooth muscle myosin heavy chain [SMemb]) and extracellular matrix associated markers (matrix metalloproteinase [MMP]-1 and MMP-3), reminiscent of diseased valves. There were no differences between constructs containing normal valve VICs and VECs (type 1) and those containing diseased valve VICs and normal valve VECs (type 2). Mechanical manipulation and endothelial damage (type 3) tended to decrease α-SMA and SMemb expression, suggesting reversal of VIC activation, but with retention of SMemb + cells adjacent to the wounded endothelium consistent with response to injury. Fibrin-based 3D mitral valve constructs can be produced using primary cell cultures derived from canine mitral valves, and show a phenotype reminiscent of diseased valves. The constructs demonstrate a response to endothelial damage indicating their utility as experimental platforms., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

A 14-year-old American Staffordshire terrier was presented for episodes of exercise-induced syncope. At admission, atrial flutter coupled to third-degree atrioventricular block was diagnosed electrocardiographically. On the second day of hospitalization, surface electrocardiogram revealed spontaneous conversion to sinus rhythm with persistence of atrioventricular block. Complete transthoracic echocardiograms were performed after each electrocardiographic examination. The combined use of conventional echocardiography with tissue Doppler imaging-based modalities allowed to investigate the atrial electromechanical correlation and function during typical atrial flutter and after its resolution., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Objectives: To assess the prevalence of functional ejection murmurs and murmurs of mitral regurgitation (MR) due to myxomatous mitral valve disease in healthy whippets; to assess the diagnostic value of auscultation to detect MR; and investigate the relationship between age and presence of echocardiographically documented MR (MR echo )., Animals: A total of 200 healthy client-owned Whippets, recruited at national shows between 2005 and 2009 were involved in this study., Methods: Cross-sectional study. Dogs were examined by auscultation by one examiner and Doppler echocardiography by another, and results were compared. Prevalence of types of murmurs and MR echo were calculated and correlated to age. Accuracy of auscultation to predict MR echo was calculated., Results: Left-sided systolic heart murmurs were detected in 185/200 (93%) of dogs. Left apical systolic murmurs (L apex ) were detected in 57/200 (29%) and left basilar systolic murmurs (L base ) in 128/200 of the dogs (64%). MR echo was present in 76/200 (38%) dogs. Prevalence MR echo was correlated with age (r = 0.96, p=0.0028). Mitral regurgitation detected by echocardiography was present in 12/78 (15%) of the dogs ≤ 2 years of age and in 59% of the dogs at 7-8 years old. Detection of L apex predicted MR echo with sensitivity 65%, specificity 94%, positive predictive value 86%, and negative predictive value 81%; and accuracy improved when only dogs with more intense L apex (grade ≥ 3/6) were considered., Conclusions: Systolic murmurs are common in North American Whippets and this breed exhibits a high prevalence of MR echo , which may be documented at a relatively early age. Whippets with non-clinical MR echo may not be identifiable by auscultation alone; echocardiographic examination may be required to exclude a diagnosis of MR. Louder heart murmurs allow more accurate localization in this population., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Development and progression of myxomatous mitral valve disease (MMVD) in dogs are difficult to predict. Identification at a young age of dogs at high risk of adverse outcome in the future is desirable., Hypothesis/objectives: To study the predictive value of selected clinical and echocardiographic characteristics associated with MMVD obtained at a young age for prediction of long-term cardiac and all-cause mortality in Cavalier King Charles Spaniels (CKCS)., Animals: 1125 privately owned CKCS., Methods: A retrospective study including CKCS examined at the age of 1-3 years. Long-term outcome was assessed by telephone interview with owners. The value of variables for predicting mortality was investigated by Cox proportional hazard and Kaplan-Meier analyses., Results: Presence of moderate to severe mitral regurgitation (MR) (hazard ratio (HR) = 3.03, 95% confidence interval (95% CI) = 1.48-6.23, P = 0.0025) even intermittent moderate to severe MR (HR = 2.23, 95% CI = 1.48-6.23, P = 0.039) on color flow Doppler echocardiography was significantly associated with increased hazard of cardiac death. An interaction between MR and sex was significant for all-cause mortality (P = 0.035), showing that males with moderate to severe MR had a higher all-cause mortality compared to males with no MR (HR = 2.38, 95% CI = 1.27-4.49, P = 0.0071), whereas no difference was found between female MR groups. The risk of cardiac (HR = 1.37, 95% CI = 1.14-1.63, P < 0.001) and all-cause (HR = 1.13, 95% CI = 1.02-1.24, P = 0.016) mortality increased with increasing left ventricular end-systolic internal dimension normalized for body weight (LVIDS N )., Conclusions and Clinical Importance: Moderate to severe MR, even if intermittent, and increased LVIDS N in dogs <3 years of age were associated with cardiac death later in life in CKCS., (Copyright © 2017 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.)

Myxomatous mitral valve disease (MMVD) is the single most common acquired heart disease of the dog, but is also of emerging importance in human medicine, with some features of the disease shared between both species. There has been increased understanding of this disease in recent years, with most research aiming to elucidate the cellular and molecular events of disease pathogenesis. For gross and histological changes, much of our understanding is based on historical studies and there has been no comprehensive reappraisal of the pathology of MMVD. This paper reviews the gross, histological, ultrastructural, cellular and molecular changes in canine MMVD., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Objectives: To determine the prevalence of mitral valve regurgitation (MR) in asymptomatic Swedish Norfolk terriers., Animals: Seventy-nine privately owned Norfolk terriers., Materials and Methods: A prospective observational study was conducted where dogs were recruited via the Swedish Norfolk terrier club. All dogs were examined using the same protocol including physical examination and Doppler echocardiography., Results: Fifteen dogs (19%) had a murmur at the time of the examination. A total of 35 dogs (44%) had MR, including 23 dogs (29%) with both MR and tricuspid valve regurgitation and 12 dogs (15%) with MR only, identified on Doppler echocardiography. In addition, 7 dogs (9%) had tricuspid valve regurgitation only. The prevalence of MR increased with increasing age (p < 0.0001)., Conclusions: Mitral valve regurgitation is common in asymptomatic Norfolk terriers with and without murmurs and the prevalence increases with age. The impact of MR in this breed on survival remains to be elucidated by a longitudinal study., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Introduction: Dilated cardiomyopathy (DCM) and myxomatous mitral valve disease (MMVD) are common heart conditions in dogs. They have different etiology and pathogenesis and although other studies focused on changes in the left ventricles of the affected hearts, the aim of our study was to assess the expressions of some intrinsic proteins in the enlarged left atria., Material and Methods: We performed an immunohistochemical analysis of left atrial specimens obtained from 15 dogs with DCM, 35 dogs with MMVD and six control dogs. We assessed the expression of following proteins: SERCA1, SERCA2, sarcomeric actinin, smooth muscle actin, and dystrophin., Results: We noted a higher percentage of SERCA1-positive cells in the MMVD group and lower percentage of dystrophin-positive cells in the DCM group as compared to control group. The expression of other proteins was similar in the hearts of control dogs and dogs with heart diseases., Conclusions: The observed changes in the expression patterns of some proteins in the atria of dogs with DCM and MMVD suggest that atrial enlargement relies not only on volume overload, but also alterations of the intrinsic proteins can contribute to the pathogenesis of dilated cardiomyopathy.

Objectives: To describe pulmonary transit time (nPTT) and myocardial perfusion (nMP) normalised to heart rate in dogs with stable ACVIM stage C myxomatous mitral valve disease (MMVD) and to assess short-term effects of pimobendan on these variables. We hypothesised that nPTT and nMP would increase in dogs with MMVD compared with normal dogs. Additionally, we hypothesised that treatment with pimobendan would decrease nMP and nPTT in dogs with MMVD., Design: Prospective, single-blind study involving 6 normal dogs and 12 dogs with MMVD., Methods: Dogs with MMVD were treated with enalapril and furosemide for at least 1 month prior to examination. All dogs underwent standard and contrast echocardiographic examinations at the beginning of the study (T0). At this time, MMVD dogs were randomly assigned to receive either pimobendan (0.4-0.6 mg/kg) or not. All dogs with MMVD were re-evaluated by standard and contrast echocardiography after 1 week (T1) and nPTT and nMP were measured., Results: nPTT was significantly increased in dogs with MMVD (P = 0.0063), compared with normal dogs. It was significantly decreased at T1 in dogs receiving pimobendan (P = 0.0250). The nMP was not significantly different in dogs with MMVD, compared with healthy dogs (P = 0.2552), and it was not significantly different at T1 in the treatment group (P = 0.8798)., Conclusions: Contrast echocardiography was a valid, complementary tool for echocardiographic analysis of dogs with MMVD. Pimobendan decreased nPTT in dogs affected by MMVD. Myocardial perfusion was not different in dogs with severe MMVD., (© 2016 Australian Veterinary Association.)

Epithelial to mesenchymal transition (EMT) and the cardiovascular equivalent, endothelial to mesenchymal transition (EndoMT), contribute to a range of chronic degenerative diseases and cancer metastasis. Chronic valvulopathies exhibit some features of EndoMT and activation of developmental signalling pathways, such as osteogenesis and chondrogenesis, expression of cell differentiation markers, basement membrane damage and endothelial transformation. The aim of the present study was to investigate the potential role of developmental mechanisms in canine myxomatous mitral valve disease (MMVD) using a combination of transcriptomic array technology, RT-PCR and immunohistochemistry. There was significant differential expression for genes typically associated with valvulogenesis and EndoMT, including markers of inflammation (IL6, IL18 and TLR4), basement membrane disarray (NID1, LAMA2 and CTSS), mesenchymal and endothelial cell differentiation (MYH11 and TAGLN) and EndoMT (ACTA2, SNAI1, CTNNB1, HAS2, CDH5, and NOTCH1), with fold changes from +15.35 (ACTA2) to -5.52 (LAMA2). These changes in gene expression were confirmed using RT-PCR, except for HAS2. In silico analysis identified important gene networks and canonical pathways in MMVD that have associations with development and organogenesis, including inflammation, valve morphogenesis and EMT, as well as components of the basement membrane and extra-cellular matrix. Immunohistochemistry identified changes in the expression of hyaluronic acid synthase (Has2), Snai1, α-smooth muscle actin (α-SMA) and VE-cadherin (CDH5), and co-expression of Has2 with α-SMA. These research findings strongly suggest involvement of developmental signalling pathways and mechanisms, including EndoMT, in the pathogenesis of canine MMVD., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

MicroRNAs (miRNAs) are small non-coding RNAs that have shown promise as noninvasive biomarkers in cardiac disease. This study was undertaken to investigate the miRNA expression profile in dogs with myxomatous mitral valve disease (MMVD). 277 miRNAs were quantified using RT-qPCR from six normal dogs (American College of Veterinary Internal Medicine Stage A), six dogs with MMVD mild to moderate cardiac enlargement (ACVIM Stage B1/B2) and six dogs with MMVD and congestive heart failure (ACVIM Stage C/D). Eleven miRNAs were differentially expressed (False Discovery Rate < 0.05). Dogs in Stage B1/B2 or C/D had four upregulated miRNAs, including three cfa-let-7/cfa-miR-98 family members, while seven others were downregulated, compared to Stage A. Expression of six of the 11 miRNAs also were significantly different between dogs in Stage C/D and those in Stage B1/B2. The expression changes were greater as disease severity increased. These miRNAs may be candidates for novel biomarkers and may provide insights into genetic regulatory pathways in canine MMVD.