Genetic diversity, growth and heart function of Auckland Island pigs, a potential source for organ xenotransplantation.

One of the prerequisites for successful organ xenotransplantation is a reasonable size match between the porcine organ and the recipient's organ to be replaced. Therefore, the selection of a suitable genetic background of source pigs is important. In this study, we investigated body and organ growth, cardiac function, and genetic diversity of a colony of Auckland Island pigs established at the Center for Innovative Medical Models (CiMM), LMU Munich. Male and female Auckland Island pig kidney cells (selected to be free of porcine endogenous retrovirus C) were imported from New Zealand, and founder animals were established by somatic cell nuclear transfer (SCNT). Morphologically, Auckland Island pigs have smaller body stature compared to many domestic pig breeds, rendering their organ dimensions well-suited for human transplantation. Furthermore, echocardiography assessments of Auckland Island pig hearts indicated normal structure and functioning across various age groups throughout the study. Single nucleotide polymorphism (SNP) analysis revealed higher runs of homozygosity (ROH) in Auckland Island pigs compared to other domestic pig breeds and demonstrated that the entire locus coding the swine leukocyte antigens (SLAs) was homozygous. Based on these findings, Auckland Island pigs represent a promising genetic background for organ xenotransplantation.

A pilot trial of doxorubicin containing phosphatidyldiglycerol based thermosensitive liposomes in spontaneous feline soft tissue sarcoma.

PURPOSE: Doxorubicin (DOX)-loaded phosphatidyldiglycerol-based thermosensitive liposomes (DPPG2-TSL-DOX) combined with local hyperthermia (HT) was evaluated in cats with locally advanced spontaneous fibrosarcomas (soft tissue sarcoma [STS]). The study was designed to evaluate the safety and pharmacokinetic profile of the drug. Results from four dose-levels are reported. METHODS: Eleven client-owned cats with advanced STS were enrolled. Five cats received escalating doses of 0.1-0.4 mg/kg DOX (group I), three received 0.4 mg/kg constantly (group II) and three 0.6 mg/kg (group III) IV over 15 min. HT with a target temperature of 41.5 °C was started 15 min before drug application and continued for a total of 60 min. Six HT treatments were applied every other week using a radiofrequency applicator. Tumour growth was monitored by magnetic resonance imaging (MRI) and for dose level III also with 18F-FDG PET. RESULTS: Treatment was generally well tolerated and reasons for premature study termination in four cats were not associated with drug-induced toxicity. No DPPG2-TSL-DOX based hypersensitivity reaction was observed. One cat showed simultaneous partial response (PR) in MRI and positron emission tomography (PET) whereas one cat showed stable disease in MRI and PR in PET (both cats in dose level III). Pharmacokinetic measurements demonstrated DOX release triggered by HT. CONCLUSION: DPPG2-TSL-DOX + HT is a promising treatment option for advanced feline STS by means of targeted drug delivery. As MTD was not reached further investigation is warranted to determine if higher doses would result in even better tumour responses.

Sensitivity and specificity of a real-time reverse transcriptase polymerase chain reaction detecting feline coronavirus mutations in effusion and serum/plasma of cats to diagnose feline infectious peritonitis.

BACKGROUND: Feline coronavirus (FCoV) exists as two pathotypes, and FCoV spike gene mutations are considered responsible for the pathotypic switch in feline infectious peritonitis (FIP) pathogenesis. The aim of this study was to evaluate sensitivity and specificity of a real-time reverse transcriptase polymerase chain reaction (RT-PCR) specifically designed to detect FCoV spike gene mutations at two nucleotide positions. It was hypothesized that this test would correctly discriminate feline infectious peritonitis virus (FIPV) and feline enteric coronavirus (FECV). METHODS: The study included 63 cats with signs consistent with FIP. FIP was confirmed in 38 cats. Twenty-five control cats were definitively diagnosed with a disease other than FIP. Effusion and/or serum/plasma samples were examined by real-time RT-PCR targeting the two FCoV spike gene fusion peptide mutations M1058 L and S1060A using an allelic discrimination approach. Sensitivity, specificity, negative and positive predictive values including 95% confidence intervals (95% CI) were calculated. RESULTS: FIPV was detected in the effusion of 25/59 cats, one of them being a control cat with chronic kidney disease. A mixed population of FIPV/FECV was detected in the effusion of 2/59 cats; all of them had FIP. RT-PCR was negative or the pathotype could not be determined in 34/59 effusion samples. In effusion, sensitivity was 68.6% (95% CI 50.7-83.2), specificity was 95.8% (95% CI 78.9-99.9). No serum/plasma samples were positive for FIPV. CONCLUSIONS: Although specificity of the test in effusions was high, one false positive result occurred. The use of serum/plasma cannot be recommended due to a low viral load in blood.

Short Treatment of 42 Days with Oral GS-441524 Results in Equal Efficacy as the Recommended 84-Day Treatment in Cats Suffering from Feline Infectious Peritonitis with Effusion-A Prospective Randomized Controlled Study.

In the past, feline infectious peritonitis (FIP) caused by feline coronavirus (FCoV) was considered fatal. Today, highly efficient drugs, such as GS-441524, can lead to complete remission. The currently recommended treatment duration in the veterinary literature is 84 days. This prospective randomized controlled treatment study aimed to evaluate whether a shorter treatment duration of 42 days with oral GS-441524 obtained from a licensed pharmacy is equally effective compared to the 84-day regimen. Forty cats with FIP with effusion were prospectively included and randomized to receive 15 mg/kg of GS-441524 orally every 24h (q24h), for either 42 or 84 days. Cats were followed for 168 days after treatment initiation. With the exception of two cats that died during the treatment, 38 cats (19 in short, 19 in long treatment group) recovered with rapid improvement of clinical and laboratory parameters as well as a remarkable reduction in viral loads in blood and effusion. Orally administered GS-441524 given as a short treatment was highly effective in curing FIP without causing serious adverse effects. All cats that completed the short treatment course successfully were still in complete remission on day 168. Therefore, a shorter treatment duration of 42 days GS-441524 15 mg/kg can be considered equally effective.

Detection and comparison of microRNA expression in the serum of Doberman Pinschers with dilated cardiomyopathy and healthy controls.

BACKGROUND: Dilated cardiomyopathy (DCM) is the most common heart disease in Doberman Pinschers. MicroRNAs (miRNAs) are short non-coding RNAs playing important roles in gene regulation. Different miRNA expression patterns have been described for DCM in humans and might represent potential diagnostic markers. There are no studies investigating miRNA expression profiles in canine DCM. The aims of this study were to screen the miRNA expression profile of canine serum using miRNA microarray and to compare expression patterns of a group of Doberman Pinschers with DCM and healthy controls. RESULTS: Eight Doberman Pinschers were examined by echocardiography and 24-hour-ECG and classified as healthy (n=4) or suffering from DCM (n=4). Total RNA was extracted from serum and hybridized on a custom-designed 8x60k miRNA microarray (Agilent) containing probes for 1368 individual miRNAs. Although total RNA concentrations were very low in serum samples, 404 different miRNAs were detectable with sufficient signal intensity on miRNA microarray. 22 miRNAs were differentially expressed in the two groups (p<0.05 and fold change (FC)>1.5), but did not reach statistical significance after multiple testing correction (false discovery rate adjusted p>0.05). Five miRNAs were selected for further analysis using quantitative Real-Time RT-PCR (qPCR) assays. No significant differences were found using specific miRNA qPCR assays (p>0.05). CONCLUSIONS: Numerous miRNAs can be detected in canine serum. Between healthy and DCM dogs, miRNA expression changes could be detected, but the results did not reach statistical significance most probably due to the small group size. miRNAs are potential new circulating biomarkers in veterinary medicine and should be investigated in larger patient groups and additional canine diseases.

Identification of novel genetic risk factors of dilated cardiomyopathy: from canine to human.

BACKGROUND: Dilated cardiomyopathy (DCM) is a life-threatening heart disease and a common cause of heart failure due to systolic dysfunction and subsequent left or biventricular dilatation. A significant number of cases have a genetic etiology; however, as a complex disease, the exact genetic risk factors are largely unknown, and many patients remain without a molecular diagnosis. METHODS: We performed GWAS followed by whole-genome, transcriptome, and immunohistochemical analyses in a spontaneously occurring canine model of DCM. Canine gene discovery was followed up in three human DCM cohorts. RESULTS: Our results revealed two independent additive loci associated with the typical DCM phenotype comprising left ventricular systolic dysfunction and dilatation. We highlight two novel candidate genes, RNF207 and PRKAA2, known for their involvement in cardiac action potentials, energy homeostasis, and morphology. We further illustrate the distinct genetic etiologies underlying the typical DCM phenotype and ventricular premature contractions. Finally, we followed up on the canine discoveries in human DCM patients and discovered candidate variants in our two novel genes. CONCLUSIONS: Collectively, our study yields insight into the molecular pathophysiology of DCM and provides a large animal model for preclinical studies.

Right ventricular size and function evaluated by various echocardiographic indices in dogs with pulmonary hypertension.

BACKGROUND: Three-dimensional (3D) echocardiography and 2-dimensional (2D) strain measurements of the right ventricle (RV) are important indices in humans with pulmonary hypertension (PH) and need further evaluation in dogs with PH. OBJECTIVES: To evaluate various RV size and function indices in dogs with PH and to examine differences between pre- and postcapillary PH. ANIMALS: A total of 311 client-owned dogs: 100 dogs with PH, 31 with postcapillary and 69 with precapillary PH, and 211 healthy control dogs. METHODS: Retro- and prospective, multicenter study. Size and function of the RV was determined using several indices, derived using dedicated RV software, including 3D RV end-diastolic volume (EDVn), end-systolic volume (ESVn), ejection fraction, 2D global and free wall RV longitudinal strain (RVLS), end-diastolic area, end-systolic area, fractional area change, tricuspid annular plane systolic excursion, and tissue Doppler imaging-derived systolic myocardial velocity of the lateral tricuspid annulus (S'n). RESULTS: The EDVn (1.8 vs 2.5 mL/kg0.942 , P < .01) and ESVn (0.8 vs 1.2 mL/kg0.962 , P < .001) were significantly larger in the PH group compared to healthy controls. Free wall RVLS was decreased in dogs with severe PH compared to controls (-24% vs -29.6%, P < .001). Dogs with precapillary PH had worse RV systolic function than dogs with postcapillary PH. CONCLUSION: Three-dimensional echocardiography of the RV is a promising tool to detect RV changes in dogs with PH. Also, 2D strain measurements are able to detect decreased RV function and offer several advantages compared to conventional indices.

Echocardiographic reference intervals for right ventricular indices, including 3-dimensional volume and 2-dimensional strain measurements in healthy dogs.

BACKGROUND: There is currently a lack of reference intervals (RIs) for the novel measures like 3-dimensional (3D) echocardiography or speckle-tracking strain for assessment of right ventricular (RV) structure and function. OBJECTIVES: To generate RIs and to determine the influence of age, heart rate, and body weight (BW) on various RV function indices using a dedicated RV software for 3D RV end-diastolic volume (EDV), end-systolic volume (ESV), ejection fraction (EF), global and free wall RV longitudinal strain (RVLS), end-diastolic area (RVEDA), end-systolic area (RVESA), fractional area change (FAC), tricuspid annular plane systolic excursion (TAPSE), and tissue Doppler imaging (TVI)-derived systolic myocardial velocity of the lateral tricuspid annulus (S'). ANIMALS: Healthy adult client-owned dogs (n = 211) of various breeds and ages. METHODS: Prospective study. Reference intervals were estimated as statistical prediction intervals using allometric scaling for BW-dependent variables. Right-sided (upper limit) or left-sided (lower limit) 95% RIs were calculated for every variable. Inter- and intraobserver variability was determined. RESULTS: Most variables showed clinically acceptable repeatability with coefficient of variation less than 10. Upper or respectively lower RI after allometric scaling to normalize for different BWs were: EDVn ≤ 2.5 mL/kg0.942 , ESVn ≤ 1.2 mL/kg0.962 , TAPSEn ≥ 4.5 mm0.285 , RVEDAn ≤ 1.4 cm2 /kg0.665 , RVESAn ≤ 0.8 cm2 /kg0.695 , and TVI S'n ≥ 5.6 cm/s/kg0.186 . The calculated limits for indices without allometric normalization were: EF > 42.1%, FAC > 30.0%, free wall RVLS < -20.8%, and global RVLS < -18.3%. CONCLUSIONS: Echocardiographic RIs for RV structure and function are provided.

Identification of 2 loci associated with development of myxomatous mitral valve disease in Cavalier King Charles Spaniels.

Myxomatous mitral valve disease (MMVD) is the most common heart disease in dogs. It is characterized by chronic progressive degenerative lesions of the mitral valve. The valve leaflets become thickened and prolapse into the left atrium resulting in mitral regurgitation (MR). MMVD is most prevalent in small to medium sized dog breeds, Cavalier King Charles Spaniels (CKCS) in particular. The onset of MMVD is highly age dependent, and at the age of 10 years, nearly all CKCS are affected. The incidence of a similar disease in humans-mitral valve prolapse-is 1-5%. By defining CKCSs with an early onset of MMVD as cases and old dogs with no or mild signs of MMVD as controls, we conducted a genome-wide association study (GWAS) to identify loci associated with development of MMVD. We have identified a 1.58 Mb region on CFA13 (P(genome) = 4.0 × 10(-5)) and a 1.68 Mb region on CFA14 (P(genome) = 7.9 × 10(-4)) associated with development of MMVD. This confirms the power of using the dog as a model to uncover potential candidate regions involved in the molecular mechanisms behind complex traits.

Echocardiographic reference intervals for volumetric measurements of the left ventricle using the Simpson's method of discs in 1331 dogs.

BACKGROUND: Echocardiographic measurements play an important role in detecting cardiac enlargement and assessing cardiac function. In human cardiology, M-mode measurements have been widely replaced by volumetric measurements of the left ventricle (LV) using Simpson's method of disc (SMOD). In veterinary cardiology, more large-scale studies are necessary to generate reference intervals (RIs) for SMOD LV volume measurements. OBJECTIVE: To generate body size independent RIs for LV volume measurements in dogs. ANIMALS: Healthy adult dogs (n = 1331) of variable size and somatotype. METHODS: Prospective study. The SMOD was measured from the right parasternal long axis and the left apical 4-chamber view in clinically healthy dogs. The SMOD measurements were normalized to various allometric scales (kg, kg2/3 , or kg1/3 ). RIs for LV end-diastolic volume (LVEDV) and LV end-systolic volume (LVESV) using SMOD were estimated as prediction intervals of both a linear and an additive regression model. Additionally, after normalization to body weight, 95% RIs were determined using nonparametric methods with 2.5 and 97.5 percentiles serving as the lower and upper limits. Separate analyses were performed for 120 sighthound breeds and 1211 other breeds. RESULTS: Echocardiographic LV volumes correlated best with weight in kilograms. The additive model proved to be more flexible and accurate than the other 2 methods to generate RIs. Separate RIs for sighthound and all other breeds are provided. CONCLUSIONS AND CLINICAL IMPORTANCE: Body size and breed-independent RIs for LV volume measurements using SMOD were generated prospectively from a large and diverse population of dogs and are available for clinical use.

Effects of pimobendan in cats with hypertrophic cardiomyopathy and recent congestive heart failure: Results of a prospective, double-blind, randomized, nonpivotal, exploratory field study.

BACKGROUND: The benefits of pimobendan in the treatment of congestive heart failure (CHF) in cats with hypertrophic cardiomyopathy (HCM) have not been evaluated prospectively. HYPOTHESIS/OBJECTIVES: To investigate the effects of pimobendan in cats with HCM and recent CHF and to identify possible endpoints for a pivotal study. We hypothesized that pimobendan would be well-tolerated and associated with improved outcome. ANIMALS: Eighty-three cats with HCM and recently controlled CHF: 30 with and 53 without left ventricular outflow tract obstruction. METHODS: Prospective randomized placebo-controlled double-blind multicenter nonpivotal field study. Cats received either pimobendan (0.30 mg/kg q12h, n = 43), placebo (n = 39), or no medication (n = 1) together with furosemide (<10 mg/kg/d) with or without clopidogrel. The primary endpoint was a successful outcome (ie, completing the 180-day study period without a dose escalation of furosemide). RESULTS: The proportion of cats in the full analysis set population with a successful outcome was not different between treatment groups (P = .75). For nonobstructive cats, the success rate was 32% in pimobendan-treated cats versus 18.2% in the placebo group (odds ratio [OR], 2.12; 95% confidence interval [CI], 0.54-8.34). For obstructive cats, the success rate was 28.6% and 60% in the pimobendan and placebo groups, respectively (OR, 0.27; 95% CI, 0.06-1.26). No difference was found between treatments for the secondary endpoints of time to furosemide dose escalation or death (P = .89). Results were similar in the per-protocol sets. Adverse events in both treatment groups were similar. CONCLUSIONS AND CLINICAL IMPORTANCE: In this study of cats with HCM and recent CHF, no benefit of pimobendan on 180-day outcome was identified.

Accuracy of history, physical examination, cardiac biomarkers, and biochemical variables in identifying dogs with stage B2 degenerative mitral valve disease.

BACKGROUND: Treatment is indicated in dogs with preclinical degenerative mitral valve disease (DMVD) and cardiomegaly (stage B2). This is best diagnosed using echocardiography; however, relying upon this limits access to accurate diagnosis. OBJECTIVES: To evaluate whether cardiac biomarker concentrations can be used alongside other clinical data to identify stage B2 dogs. ANIMALS: Client-owned dogs (n = 1887) with preclinical DMVD prospectively sampled in Germany, the United Kingdom, and the United States. METHODS: Dogs that met inclusion criteria and were not receiving pimobendan (n = 1245) were used for model development. Explanatory (multivariable logistic regression) and predictive models were developed using clinical observations, biochemistry, and cardiac biomarker concentrations, with echocardiographically confirmed stage B2 disease as the outcome. Receiver operating characteristic curves assessed the ability to identify stage B2 dogs. RESULTS: Age, appetite, serum alanine aminotransferase activity, body condition, serum creatinine concentration, murmur intensity, and plasma N-terminal propeptide of B-type natriuretic peptide (NT-proBNP) concentration were independently associated with the likelihood of being stage B2. The discriminatory ability of this explanatory model (area under curve [AUC], 0.84; 95% confidence interval [CI], 0.82-0.87) was superior to NT-proBNP (AUC, 0.77; 95% CI, 0.74-0.80) or the vertebral heart score alone (AUC, 0.76; 95% CI, 0.69-0.83). A predictive logistic regression model could identify the probability of being stage B2 (AUC test set, 0.86; 95% CI, 0.81-0.91). CONCLUSION AND CLINICAL IMPORTANCE: Our findings indicate accessible measurements could be used to screen dogs with preclinical DMVD. Encouraging at-risk dogs to seek further evaluation could result in a greater proportion of cases being appropriately managed.

A scalable, clinically severe pig model for Duchenne muscular dystrophy.

Large-animal models for Duchenne muscular dystrophy (DMD) are crucial for the evaluation of diagnostic procedures and treatment strategies. Pigs cloned from male cells lacking DMD exon 52 (DMDΔ52) exhibit molecular, clinical and pathological hallmarks of DMD, but die before sexual maturity and cannot be propagated by breeding. Therefore, we generated female DMD+/- carriers. A single founder animal had 11 litters with 29 DMDY/-, 34 DMD+/- as well as 36 male and 29 female wild-type offspring. Breeding with F1 and F2 DMD+/- carriers resulted in an additional 114 DMDY/- piglets. With intensive neonatal management, the majority survived for 3-4 months, providing statistically relevant cohorts for experimental studies. Pathological investigations and proteome studies of skeletal muscles and myocardium confirmed the resemblance to human disease mechanisms. Importantly, DMDY/- pigs displayed progressive myocardial fibrosis and increased expression of connexin-43, associated with significantly reduced left ventricular ejection fraction, at 3 months. Furthermore, behavioral tests provided evidence for impaired cognitive ability. Our breeding cohort of DMDΔ52 pigs and standardized tissue repositories provide important resources for studying DMD disease mechanisms and for testing novel treatment strategies.

Left ventricular M-mode prediction intervals in 7651 dogs: Population-wide and selected breed-specific values.

BACKGROUND: Echocardiography is a common method to measure heart size in dogs. The heart dimensions are influenced by body weight (BW) and potentially by breed. OBJECTIVES: To establish BW-dependent prediction intervals (PIs) of the left ventricular (LV) linear dimensions in a population of dogs of many breeds in multicenter environment, and to identify breeds deviating from these intervals. DOGS: Seven thousand six hundred and fifty-one dogs. METHODS: Retrospectively, data from heart screens conducted between 2009 and 2016 were included. Cardiac dimensional PIs were generated using allometric scaling including all nonsighthound dogs and values were compared to previously published PIs. The values measured in dogs of respective breeds, including sighthounds, were then compared to the overall nonsighthound PIs to identify deviant breeds. The interobserver-variability of the measurements was determined using the explained residual variance. RESULTS: Prediction intervals for the nonsighthound dogs were in agreement with previously published cardiac PIs, although the upper limits of the generated PIs of our study were slightly below those currently applied (except the interventricular septum in systole and the left ventricular free wall in diastole below 10.0 kg and 15.0 kg, respectively). Values measured in the nonsighthound breed Newfoundland deviated for most dimensions. Most of the sighthound breeds analyzed had greater cardiac dimensions, with the exception of the Irish Wolfhound. CONCLUSION AND IMPORTANCE: Findings of our study reinforces the value of BW-dependent PIs for cardiac dimensions in dogs and suggest that these PIs are valid for most nonsighthound breeds, but not the sighthound breeds.

The aptamer BC 007 for treatment of dilated cardiomyopathy: evaluation in Doberman Pinschers of efficacy and outcomes.

AIMS: Aptamer BC 007, a 15-mer single-strand DNA oligonucleotide (5'-GGTTGGTGTGGTTGG-3'), was developed to neutralize functional autoantibodies that bind to the extracellular domains of G protein-coupled receptors (GPCR-AAB), leading to the modulation of receptor-mediated signalling cascades that induce pathophysiological states. Among the GPCR-AAB, there are those directed against the ß1-adrenergic receptor (ß1-AAB) that are highly present in patients with dilated cardiomyopathy (DCM) and are increasingly accepted as disease drivers. Using Doberman Pinschers (DP) with DCM, which possess similarities with human DCM among these ß1-AAB positivity for that the disease-driving role in DP DCM was demonstrated, the safety of BC 007, efficacy for neutralizing ß1-AAB, and the DP's outcome were investigated. METHODS AND RESULTS: Fourteen client-owned ß1-AAB-positive DP with electrocardiographically and echocardiographically indicated DCM were treated with BC 007. For controlling, two groups were created: 14 ß1-AAB-positive DP with DCM not treated with BC 007 (Control 1) and 14 DP with DCM closely matched to the BC 007-treated DP (Control 2), retrospectively selected from the institutional database of DP. After treatment, DP were monitored both echocardiographically, and for ß1-AAB, and survival curves were calculated. Based on clinical and laboratory examination, no adverse effects associated with BC 007 treatment were observed during the study. Forty-eight hours after treatment, the DP's blood was free of ß1-AAB, which led to a reduction or stabilization of left ventricular end-systolic volume (ESVI) during ß1-AAB free time in 10 of the treated DP. In one DP, where ß1-AAB returned after 3 months and ESVI worsened again, a second BC 007 treatment after 9 months again cleared the blood from ß1-AAB and improved the ESVI. Compared with the controls, DP treated with BC 007 showed a significantly longer survival time [572 days, interquartile range (IQR) 442-840 days] vs. Control group 1 (266 days, IQR 97-438 days; logrank: P = 0.009) and Control group 2 (229 days, IQR 174-319 days; logrank: P = 0.012). CONCLUSIONS: Treatment with BC 007 for ß1-AAB neutralization was safe, resulted in a long-lasting reduction of ß1-AAB combined with improved cardiac function and prolonged the survival of DP with DCM. Using a natural large animal model of DCM considered superior to small animal models of immunization-induced cardiomyopathy, combined with a study design comparable with clinical trials, we believe that our results provide the basis for optimism that treatment with BC 007 might also be effective in human patients with DCM.

Efficacy of adding ramipril (VAsotop) to the combination of furosemide (Lasix) and pimobendan (VEtmedin) in dogs with mitral valve degeneration: The VALVE trial.

BACKGROUND: Triple therapy (TT) consisting of furosemide, pimobendan, and an angiotensin-converting enzyme inhibitor (ACEI) frequently is recommended for the treatment of congestive heart failure (CHF) attributable to myxomatous mitral valve disease (MMVD). However, the effect of adding an ACEI to the combination of pimobendan and furosemide (dual therapy [DT]) so far has not been evaluated prospectively. HYPOTHESIS: Triple therapy will extend survival time compared to DT in dogs with CHF secondary to MMVD. ANIMALS: Client-owned dogs presented with the first episode of CHF caused by MMVD. METHODS: Prospective, single-blinded, randomized multicenter study. One-hundred and fifty-eight dogs were recruited and prospectively randomized to receive either DT (furosemide and pimobendan) or TT (furosemide, pimobendan, and ramipril). The primary endpoint was a composite of cardiac death, euthanasia for heart failure, or treatment failure. RESULTS: Seventy-seven dogs were randomized to receive DT and 79 to receive TT. Two dogs were excluded from analysis. The primary endpoint was reached by 136 dogs (87%; 66 dogs, DT; 70 dogs, TT). Median time to reach the primary endpoint for all dogs in the study was 214 days (95% confidence interval [CI], 168-259 days). Median time to reach the primary endpoint was not significantly different between the DT group (227 days; interquartile range [IQR], 103-636 days) compared with TT group (186 days; IQR, 72-453 days; P = .42). CONCLUSIONS AND CLINICAL IMPORTANCE: Addition of the ACEI ramipril to pimobendan and furosemide did not have any beneficial effect on survival time in dogs with CHF secondary to MMVD.

Temporal changes in clinical and radiographic variables in dogs with preclinical myxomatous mitral valve disease: The EPIC study.

BACKGROUND: The Evaluation of pimobendan in dogs with cardiomegaly caused by preclinical myxomatous mitral valve disease (EPIC) study monitored dogs with myxomatous mitral valve disease (MMVD) as they developed congestive heart failure (CHF). OBJECTIVES: To describe the changes in clinical and radiographic variables occurring as dogs with MMVD and cardiomegaly develop CHF, compared to similar dogs that do not develop CHF. ANIMALS: One hundred and thirty-five, and 73 dogs that did or did not develop CHF, respectively. MATERIALS AND METHODS: The following variables were evaluated in 2 groups of dogs (dogs that did or did not develop CHF): Heart rate (HR), clinic respiratory rate (RR), home-measured resting respiratory rate (RRR), rectal temperature (RT), body weight (BW), and vertebral heart sum (VHS). Absolute value and rate of change of each variable were calculated for each day a dog was in study. Daily means were calculated and plotted against time. The onset of CHF or last visit before leaving the study were set as reference time points. RESULTS: The most extreme values and rate of change occurred in variables immediately before onset of CHF. Vertebral heart sum increased earliest. Heart rate, RR, and RRR also increased. Rectal temperature and BW decreased. Increases in RR and RRR were most extreme and occurred immediately before CHF. CONCLUSIONS AND CLINICAL IMPORTANCE: Dogs with MMVD and cardiomegaly experience increases in HR, RR, RRR, and VHS, and decreases in BW and RT as they develop CHF. The variables with highest absolute change and rate of change were RR and RRR. These findings reinforce the value of RR and RRR as indicators of impending or incipient CHF.

Evaluation of a high-sensitivity cardiac troponin I assay compared to a first-generation cardiac troponin I assay in Doberman Pinschers with and without dilated cardiomyopathy.

BACKGROUND: Echocardiography and 24-hour ECG are the gold standard tests to diagnose dilated cardiomyopathy (DCM) in Doberman Pinschers (DP), but myocardial damage might be detected earlier using a high-sensitivity cardiac troponin I (hs-cTnI) assay. OBJECTIVE: To evaluate and compare an hs-cTnI assay (Advia Centaur TnI-Ultra assay) with a conventional cTnI assay in DP with different stages of DCM and in healthy DP. ANIMALS: Three hundred forty-five examinations from 162 DP with and 179 DP without DCM. METHODS: Prospective longitudinal study. Dogs were allocated into 6 groups based on echocardiographic and 24-hour ECG criteria: (1) healthy group (179 dogs), (2) last-normal group (29 dogs), which included dogs that were considered to be healthy at the time of their examination but were assigned to the last-normal group retrospectively when DCM was diagnosed at their next examination within 1.5 years, (3) only arrhythmias (45 dogs, 119 examinations), (4) only echocardiographic changes (24 dogs, 61 examinations), (5) echocardiographic changes with ventricular premature complexes (41 dogs, 100 examinations), and (6) decompensated (23 dogs, 36 examinations). Hs-cTnI and conventional cTnI concentration measurements were performed and compared. RESULTS: A cutoff value of hs-cTnI concentration >0.113 ng/mL had a sensitivity of 81.2% and a specificity of 73.2% to identify the presence of DCM. The conventional cTnI assay showed a similar test performance, but the hs-cTnI assay identified more dogs (21/29 dogs, 72%) in the last-normal group compared to the conventional cTnI test (18/29 dogs, 62%). CONCLUSIONS AND CLINICAL IMPORTANCE: The hs-cTnI is an additional test with good potential to identify early DCM.

Retrospective analysis of pleural effusion in cats.

OBJECTIVES: Pleural effusion is a common presenting cause for feline patients in small animal practice. The objectives of this study were to identify possible correlations between the aetiology of effusion and clinical and laboratory findings. METHODS: In this retrospective study of 306 cats diagnosed with pleural effusion of established aetiology, cats were divided into six major groups: cardiac disease (CD), feline infectious peritonitis (FIP), neoplasia, pyothorax, chylothorax and miscellaneous. Clinical, laboratory and radiographic parameters were compared between groups. RESULTS: CD was the most common aetiology (35.3%), followed by neoplasia (30.7%), pyothorax (8.8%), FIP (8.5%), chylothorax (4.6%) and miscellaneous diseases (3.7%). In 26 (8.5%) cats, more than one underlying disease was diagnosed as a possible aetiology for pleural effusion. Cats with FIP were significantly younger than those with CD (P <0.001) and neoplasia (P <0.001). Cats with CD were presented with a significantly lower body temperature compared with cats with FIP (P = 0.022). Cats with CD had significantly higher serum alanine aminotransferase activity compared with all other cats (FIP and pyothorax, P <0.001; neoplasia and chylothorax, P = 0.02) and serum alkaline phosphatase activity compared with the pyothorax (P <0.001) and FIP groups (P = 0.04), and significantly lower protein concentrations (FIP, pyothorax and neoplasia, P <0.001; chylothorax, P = 0.04) and nucleated cell counts in the effusion than all other groups (pyothorax and neoplasia, P <0.001; chylothorax, P = 0.02; FIP, P = 0.04). The glucose level in the effusion of cats with pyothorax was significantly lower than glucose levels in patients with CD, neoplasia and chylothorax (P <0.001). Of 249 cats with a follow-up of at least 10 days, 55.8% died or were euthanased during that time. CONCLUSIONS AND RELEVANCE: CD and neoplasia were the most common causes for feline pleural effusion. Age, liver enzymes, as well as cell count, protein and glucose levels in the effusion can aid in the investigation of underlying aetiologies.