Myocardial Adiponectin Isoform Shift in Dogs with Congestive Heart Failure-A Comparison to Hibernating Brown Bears (Ursus arctos horribilis).

Adiponectin is the most abundant plasma adipokine, and is well known for its role in energy homeostasis and cardiac protection. In humans with dilated cardiomyopathy, myocardial adiponectin protein expression is reduced compared to normal hearts and has been implicated in the pathology of cardiomyopathy. Serum adiponectin levels are often conflicting, with higher levels associated with poor survival in humans with congestive heart failure (CHF). We evaluated adiponectin serum concentrations and myocardial protein expression in dogs with naturally occurring myxomatous mitral valve disease and CHF. We compared the findings to active and hibernating brown bears as bears are adapted to endure an extreme period of low cardiac output during their annual hibernation. Bears exhibited largely the active high-molecular weight (HMW) versus the low-molecular weight isoforms of myocardial adiponectin (HMW:LMW = 6.3) during both the active period and hibernation, while healthy dogs exhibited a more balanced mix of isoforms. Dogs with CHF expressed predominately HMW isoforms of adiponectin (HMW:LMW = 12.5), appearing more similar to bears. In contrast to humans, serum adiponectin was significantly lower in dogs with CHF and lowest levels in the severest CHF class. In both dogs and bears, myocardial adiponectin was expressed independent of circulating adiponectin concentrations, suggesting a local regulatory mechanism within the heart.

Reduced high intensity training distance had no effect on VLa4 but attenuated heart rate response in 2-3-year-old Standardbred horses.

BACKGROUND: Training of Standardbred race horses aims to improve cardiovascular and metabolic functions but studies on the effects of different training strategies from breaking till racing are lacking. Sixteen horses with the goal to race as 3-year-olds were studied from breaking (1-year-olds) to December as 3-year-olds. Horses were allocated to either a control (C) or reduced (R) training program from 2 years of age. The aim was to evaluate the effect of reducing the distance of high intensity exercise by 30% with respect to velocity at lactate concentration 4 mmol/l (VLa4), blood lactate and cardiovascular response. All training sessions were documented and heart rate (HR) was recorded. A standardized exercise test of 1,600 m was performed 10 times and a VLa4 test was performed five times. RESULTS: C horses initially exercised for a longer time with a HR >180 beats per minute compared to R horses (P < 0.05) but after 6-9 months, time with HR >180 bpm decreased in C and were similar in the two groups (P > 0.05). Over the 2-year period, recovery HR after the 1,600 m-test decreased in both groups but was within 2 months lower in C than in R (P < 0.05). C horses also had lower resting HR as 3-year-olds (P < 0.01) than R horses. In C, post exercise hematocrit was higher than in R (P < 0.05). There was a tendency (P < 0.1) towards a larger aortic diameter in C as 3-year-olds (C: 1.75 ± 0.05, R: 1.70 ± 0.05 cm/100 kg BW). Left ventricle diameter and blood volume (in December as 2-year-olds) did not differ between groups. There were no differences between groups in post exercise blood lactate concentration or in VLa4. Both groups were equally successful in reaching the goal of participation in races. CONCLUSIONS: Horses subjected to a reduced distance of high intensity training from the age of 2 showed an attenuated heart rate response, but were able to maintain the same VLa4 and race participation as horses subjected to longer training distances.

Increased NT-proANP predicts risk of congestive heart failure in Cavalier King Charles spaniels with mitral regurgitation caused by myxomatous valve disease.

OBJECTIVES: To evaluate the predictive value of plasma N-terminal pro-atrial natriuretic peptide (NT-proANP) and nitric oxide end-products (NOx) as markers for progression of mitral regurgitation caused by myxomatous mitral valve disease. ANIMALS: Seventy-eight privately owned Cavalier King Charles spaniels with naturally occurring myxomatous mitral valve disease. METHODS: Prospective longitudinal study comprising 312 measurements over a 4.5 year period. Clinical values were recorded, NT-proANP concentrations were measured by radioimmunoassay, and NOx were analyzed colorimetrically. To predict congestive heart failure (CHF), Cox proportional hazards models with time-varying covariates were constructed. RESULTS: The hazard ratio for NT-proANP (per 1000 pmol/l increase) to predict future CHF was 6.7 (95% confidence interval, 3.6-12.5; p < 0.001). The median time to CHF for dogs with NT-proANP levels >1000 pmol/l was 11 months (95% confidence interval, 5.6-12.6 months), compared to 54 months (46 - infinity) for dogs with concentrations ≤ 1000 pmol/l (p < 0.001). Due to intra- and inter-individual variability, most corresponding analyses for NOx were insignificant but dogs reaching CHF had a lower mean NOx concentration than dogs not reaching CHF (23 vs. 28 µmol/l, p = 0.016). Risk of CHF increased with increase in heart rate (>130 beats per minute) and grade of murmur (≥ 3/6). CONCLUSIONS: The risk of CHF due to mitral regurgitation is increased in dogs with blood NT-proANP concentrations above 1000 pmol/l. Measurement of NT-proANP can be a valuable tool to identify dogs that may develop CHF within months.

Evaluation of plasma activity of matrix metalloproteinase-2 and -9 in dogs with myxomatous mitral valve disease.

OBJECTIVE: To investigate whether plasma activity of matrix metalloproteinase (MMP)-2 and -9 was associated with severity of myxomatous mitral valve disease (MMVD) in dogs and to assess potential associations between MMP activity and dog characteristics, echocardiographic variables, systolic arterial blood pressure (SAP), heart rate, cardiac troponin I (cTnI) concentration, and C-reactive protein concentration. ANIMALS: 75 client-owned dogs. PROCEDURES: Severity of MMVD was assessed by use of echocardiography. Plasma activity of latent (pro-MMP) and active MMP-2 and -9 was analyzed via zymography. Plasma concentration of cTnI was analyzed with a high-sensitivity cTnI assay, and C-reactive protein concentration was analyzed with a canine-specific ELISA. RESULTS: Pro-MMP-9, active MMP-9, and pro-MMP-2 were detected, but active MMP-2 was not. No significant differences were found in MMP concentrations among the 4 MMVD severity groups. Activity of pro-MMP-9 decreased with decreases in SAP and was higher in male dogs than in female dogs. Activity of MMP-9 decreased with increases in left ventricular end-systolic dimension and with decreases in SAP and cTnI concentration. Left ventricular end-systolic dimension was the variable most strongly associated with MMP-9 activity. No associations were found between the activity of pro-MMP-2 and investigated variables. CONCLUSIONS AND CLINICAL RELEVANCE: Plasma MMP-9 activity decreased with increases in the end-systolic left ventricular internal dimension and decreases in SAP. Hence, evaluation of MMP-9 activity has the potential to provide unique information about the myocardial remodeling process in dogs with MMVD.

Evaluation of the Swedish breeding program for cavalier King Charles spaniels.

A breeding program with the aim of reducing the prevalence of mitral regurgitation (MR) caused by myxomatous mitral valve disease (MMVD) in Cavalier King Charles Spaniels (CKCS) is currently ongoing in Sweden. In this investigation 353 CKCS were selected as a sample of the population and 150 were examined by auscultation for heart murmurs when they reached the age of six years in 2007 and 2009. The aim with this investigation was to study the prevalence of heart murmurs in six-year-old CKCS and to estimate if prevalence has decreased since the breeding program was introduced 2001. The effect of the breeding program was evaluated by comparing the prevalence of heart murmurs in the two groups. In 2007, the prevalence of heart murmurs was 52% (50% for females and 54% for males) and in 2009, the prevalence was 55% (44% for females and 67% for males). No significant difference was found in the prevalence of heart murmurs between 2007 and 2009 (P=0.8). For all six-year-old CKCS, the prevalence of heart murmur was 53% (females 46% and males 61%), which is higher than previous Swedish investigations.

Use of signal analysis of heart sounds and murmurs to assess severity of mitral valve regurgitation attributable to myxomatous mitral valve disease in dogs.

OBJECTIVE: To investigate use of signal analysis of heart sounds and murmurs in assessing severity of mitral valve regurgitation (mitral regurgitation [MR]) in dogs with myxomatous mitral valve disease (MMVD). ANIMALS: 77 client-owned dogs. PROCEDURES: Cardiac sounds were recorded from dogs evaluated by use of auscultatory and echocardiographic classification systems. Signal analysis techniques were developed to extract 7 sound variables (first frequency peak, murmur energy ratio, murmur duration > 200 Hz, sample entropy and first minimum of the auto mutual information function of the murmurs, and energy ratios of the first heart sound [S1] and second heart sound [S2]). RESULTS: Significant associations were detected between severity of MR and all sound variables, except the energy ratio of S1. An increase in severity of MR resulted in greater contribution of higher frequencies, increased signal irregularity, and decreased energy ratio of S2. The optimal combination of variables for distinguishing dogs with high-intensity murmurs from other dogs was energy ratio of S2 and murmur duration > 200 Hz (sensitivity, 79%; specificity, 71%) by use of the auscultatory classification. By use of the echocardiographic classification, corresponding variables were auto mutual information, first frequency peak, and energy ratio of S2 (sensitivity, 88%; specificity, 82%). CONCLUSIONS AND CLINICAL RELEVANCE: Most of the investigated sound variables were significantly associated with severity of MR, which indicated a powerful diagnostic potential for monitoring MMVD. Signal analysis techniques could be valuable for clinicians when performing risk assessment or determining whether special care and more extensive examinations are required.

Predictive value of natriuretic peptides in dogs with mitral valve disease.

Natriuretic peptides are useful in diagnosing heart failure in dogs. However, their usefulness in detecting early stages of myxomatous mitral valve disease (MMVD) has been debated. This study evaluated N-terminal (NT) fragment pro-atrial natriuretic peptide (NT-proANP) and NT-pro-brain natriuretic peptide (NT-proBNP) in 39 Cavalier King Charles Spaniels (CKCS) with pre-clinical mitral valve regurgitation (MR), sixteen dogs with clinical signs of heart failure (HF) and thirteen healthy control dogs. Twenty seven CKCS and ten control dogs were re-examined 4 years after the initial examination and the status of the dogs 5 years after the initial examination was determined by telephone calls to the owner. All dogs were evaluated by clinical examination and echocardiography. CKCS with severe MR had higher NT-proANP and NT-proBNP compared to controls and CKCS with less severe MR. Dogs with clinical signs of HF had markedly elevated NT-proANP and NT-proBNP. Plasma concentrations of the natriuretic peptides measured at re-examination could predict progression in regurgitant jet size.

Time-frequency and complexity analyses for differentiation of physiologic murmurs from heart murmurs caused by aortic stenosis in Boxers.

OBJECTIVE: To investigate whether time-frequency and complexity analyses of heart murmurs can be used to differentiate physiologic murmurs from murmurs caused by aortic stenosis (AS) in Boxers. ANIMALS: 27 Boxers with murmurs. PROCEDURES: Dogs were evaluated via auscultation and echocardiography. Analyses of time-frequency properties (TFPs; ie, maximal murmur frequency and duration of murmur frequency > 200 Hz) and correlation dimension (T(2)) of murmurs were performed on phonocardiographic sound data. Time-frequency property and T(2) analyses of low-intensity murmurs in 16 dogs without AS were performed at 7 weeks and 12 months of age. Additionally, TFP and T(2) analyses were performed on data obtained from 11 adult AS-affected dogs with murmurs. RESULTS: In dogs with low-intensity murmurs, TFP or T(2) values at 7 weeks and 12 months did not differ significantly. For differentiation of physiologic murmurs from murmurs caused by mild AS, duration of murmur frequency > 200 Hz was useful and the combination assessment of duration of frequency > 200 Hz and T(2) of the murmur had a sensitivity of 94% and a specificity of 82%. Maximal murmur frequency did not differentiate dogs with AS from those without AS. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that assessment of the duration of murmur frequency > 200 Hz can be used to distinguish physiologic heart murmurs from murmurs caused by mild AS in Boxers. Combination of this analysis with T(2) analysis may be a useful complementary method for diagnostic assessment of cardiovascular function in dogs.

Contrast echocardiography in Boxer dogs with and without aortic stenosis.

OBJECTIVES: The aim of this study was to investigate whether contrast echocardiography could enhance the subcostal Doppler signal for aortic flow measurements and achieve myocardial opacification, in Boxer dogs with and without AS. BACKGROUND: In evaluating dogs for aortic stenosis (AS) subcostal Doppler echocardiography was used for measurement of the aortic flow velocity, a measurement that can sometimes be difficult to perform in Boxer dogs. ANIMALS, MATERIALS, AND METHODS: Cardiac auscultation, phonocardiographic and echocardiographic examinations, including a contrast study with Optison, were performed on 29 Boxer dogs selected based on previous examinations. RESULTS: The initial subcostal Doppler signal was weak in 66% of the dogs and a marked improvement was seen in all dogs after contrast injection. The peak aortic flow velocity increased 5% from 2.58+/-1.42 m/s before contrast to 2.71+/-1.54 m/s after contrast (p=0.003). This corresponds to a 2.8 mmHg increase in the pressure gradient from 26.6 mmHg before to 29.4 mmHg after contrast. A dose of 0.05-0.1 mL of Optison administered intravenously resulted in approximately 4 min of Doppler signal enhancement. With the present technique contrast echocardiography did not achieve myocardial opacification. CONCLUSIONS: Single use of the contrast agent Optison can be recommended for enhancement of the subcostal Doppler signal in dogs, in which plain Doppler signals are difficult to obtain. Albeit statistically significant, the mild increase in peak aortic flow velocity after contrast was not considered biologically or clinically significant.

Circulating concentrations of insulin-like growth factor-1 in dogs with naturally occurring mitral regurgitation.

Insulin-like growth factor-1 (IGF-1), which mediates most effects of growth hormone, has effects on cardiac mass and function, and plays an important role in the regulation of vascular tone. In humans, an inverse relationship between degree of heart failure (HF) and circulating IGF-1 concentrations has been found in several studies. In dogs with HF, few studies have focused on IGF-1. We examined circulating IGF-1 concentrations in dogs with mitral regurgitation (MR) caused by myxomatous mitral valve disease. Study 1 included 88 Cavalier King Charles Spaniels (CKCSs) with a broad range of asymptomatic MR (median serum IGF-1: 76.7 microg/L; 25-75 percentile, 59.8-104.9 microg/L). As expected, standard body weight and percentage under- or overweight correlated directly with IGF-1. MR (assessed in 4 different ways) did not correlate with IGF-1. In study 2, 28 dogs with severe MR and stable, treated congestive HF had similar serum IGF-1 concentrations (median, 100.8 g/L; 25-75 percentile, 74.9-156.5 microg/L) as 11 control dogs (79.6 microg/L; 25-75 percentile, 64.1-187.4 microg/L; P = .84). In study 3, the plasma IGF-1 concentration of 15 untreated CKCSs with severe MR was 16.4 +/- 24.2 microg/L lower (P = .02) at the examination when decompensated HF had developed (80.8 +/- 30.9 microg/L) than at a visit 1-12 months earlier (97.2 +/- 39.8 microg/L), possibly in part due to an altered state of nutrition. The studies document that circulating IGF-1 concentrations are not altered before development of congestive HF in dogs with naturally occurring MR, but decrease by approximately 20% with the development of untreated HE In treated HF, circulating IGF-1 concentrations apparently return to within the reference range.

Interobserver variability of vertebral heart size measurements in dogs with normal and enlarged hearts.

The vertebral heart size (VHS) method by Buchanan is based on anatomic landmarks. A potential source of variation among observers is differences in the selection of measurement points. The objective was to test variability between observers with different levels of training in thoracic radiology and small animal clinical practice. Fifty sets of thoracic radiographs of cavalier King Charles spaniels, were divided into five groups; (Normal) normal cardiopulmonary structures, (I) slight cardiomegaly, (II) moderate cardiomegaly, (II +) moderate cardiomegaly with congestive heart failure, and (III +) severe cardiomegaly with congestive heart failure. Cardiomegaly was confirmed by echocardiography to be caused by mitral regurgitation because of myxomatous mitral valve disease. Sixteen observers representing four levels of experience (four observers/level) evaluated the radiographs; (1) European Diplomates in Veterinary Diagnostic Imaging, (2) Experienced small animal clinicians, (3) Trainees in small animal clinical practice (4) Veterinary students. Almost identical mean VHS values were found among the four experience levels for each of the five groups of radiographs with a low coefficient of variation, range 1.5-3.2%. Mean difference among the 16 observers was 1.05 +/- 0.32 vertebrae (v). Mean difference among individuals in each observer group was approximately 0.5 v for all but the groups of trainees were the difference was 0.6-0.9 v. The conclusion is that VHS method for heart size is independent of observer experience but dependent of individual observers selection of reference points and transformation of long and short axis dimensions into VHS units.

New insights into degenerative mitral valve disease in dogs.

Degenerative mitral valve disease (DMVD) is the most common cardiac disease in dogs. Although the disease is frequently described in the veterinary literature, many aspects are still unknown or controversial. Based on recent research findings, this article addresses the etiology, pathogenesis, inheritance, diagnosis of early DMVD, diagnosis of mild decompensated heart failure, and efficacy of early medical intervention in clinically compensated dogs.

Increased pulmonary transit times in asymptomatic dogs with mitral regurgitation.

Pulmonary transit time (PTT) normalized to heart rate (nPTT) is a measure of the pulmonary blood volume (PBV) to stroke volume ratio (PBV/SV). It is an index of cardiac performance. To determine the effect of compensated mitral regurgitation (CMR) and decompensated mitral regurgitation (DMR) caused by valvular endocardiosis on the index nPTT, we measured nPTT by first-pass radionuclide angiocardiography and ECG in 13 normal dogs, 18 dogs with CMR, and 13 dogs with DMR. PTT was measured as time between onset of appearance of activity at the pulmonary trunk and the left atrium. In the normal dogs, the relationship between PTT and mean R-R interval (mRR) was PTT = 4.08 x mRR + 0.15 (R2 = 0.71). Normal nPTT was 4.4 +/- 0.6 (SD) (range. 3.6-5.3). in CMR, 6.3 +/- 1.6 (SD) (range, 4.0-9.7). and in DMR, 11.9 +/- 3.4 (SD) (range, 8.0-18.8). The differences among all groups were significant. Heart rates were 110 +/- 22 bpm in normal dogs, 111 +/- 20 in dogs with CMR, and 144 +/- 18 in dogs with DMR (P < .001 for difference between DMR group and normal and CMR groups). Increased nPTT in CMR indicates preclinical heart pump dysfunction. Heart rate-normalized pulmonary transit times may be a useful index of heart function in mitral regurgitation.

Left atrial to aortic root indices using two-dimensional and M-mode echocardiography in cavalier King Charles spaniels with and without left atrial enlargement.

Two-dimensional (2-D) echocardiographic measurement of the left atrium (LA) has the potential to be more accurate than the standard M-mode method, because the LA body can be measured. We evaluated a 2-D method for measuring LA and aorta (AO) in a right parasternal short-axis view and compared it to the M-mode method. An index for LA size (LA/AO) was calculated in 166 cavalier King Charles spaniels, 56 normal and 110 dogs with mitral regurgitation (MR) of varying degrees secondary to chronic valvular disease. In normal dogs, the AO-2-D and LA/AO-2-D did not correlate to body weight (BW) or BW2; whereas, all M-mode values and the LA-2-D were significantly (p < .05) related to both BW parameters. In normal dogs, there was no difference between M-mode and 2-D indices. For all dogs (normal and dogs with MR) there was an 11% bias between the M-mode and 2-D index with the LA/AO-2-D being higher than the LA/AO-M. The association between the mean and the difference of the indices demonstrated a quadratic relationship. Dogs with a mean LA/AO of 2.0-2.5 showed the largest difference between the two indices. Small values for the 2-D coefficients of variation for respiration and stage of diastole were found; 3.4 and 3.1%, respectively. The 2-D index is more sensitive to LA enlargement than the M-mode index.

Efficacy of enalapril for prevention of congestive heart failure in dogs with myxomatous valve disease and asymptomatic mitral regurgitation.

We evaluated the long-term effect of early angiotensin-converting enzyme (ACE) inhibition (enalapril maleate) as monotherapy to postpone or prevent congestive heart failure (CHF) in asymptomatic dogs with mitral regurgitation (MR) attributable to myxomatous valvular disease (MVD) in a prospective, randomized, double-blinded, placebo-controlled multicenter trial involving 14 centers in Scandinavia. Two hundred twenty-nine Cavalier King Charles (CKC) Spaniels with MR attributable to MVD but no signs of CHF were randomly allocated to treatment with enalapril 0.25-0.5 mg daily (n = 116) or to placebo groups (n = 113). Each dog was evaluated by physical examination, electrocardiography, and thoracic radiography at entry and every 12 months (+/-30 days). The number of dogs developing heart failure was similar in the treatment and placebo groups (n = 50 [43%] and n = 48 [42%], respectively; P = .99). The estimated means, adjusted for censored observations, for the period from initiation of therapy to heart failure were 1,150 +/- 50 days for dogs in the treatment group and 1,130 +/- 50 days for dogs in the placebo group (P = .85). When absence or presence of cardiomegaly at the entrance of the trial was considered, there were still no differences between the treatment and placebo groups (P = .98 and .51, respectively). Multivariate analysis showed that enalapril had no significant effect on the time from initiation of therapy to heart failure (P = .86). Long-term treatment with enalapril in asymptomatic dogs with MVD and MR did not delay the onset of heart failure regardless of whether or not cardiomegaly was present at initiation of the study.