Training the equine respiratory muscles: Inspiratory muscle strength.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

Effect of prespecified therapy escalation on plasma NT-proBNP concentrations in dogs with stable congestive heart failure due to myxomatous mitral valve disease.

BACKGROUND: Treatment targeted to achieve reduction in N-terminal pro-B-type natriuretic peptide (NT-proBNP) improves outcomes in human congestive heart failure (CHF) patients. HYPOTHESIS: A pre-specified therapeutic algorithm that increased diuretic or pimobendan usage will reduce plasma NT-proBNP concentrations in dogs with CHF secondary to myxomatous mitral valve disease (MMVD). ANIMALS: Twenty-six dogs with clinically stable CHF secondary to MMVD. METHODS: Prospective, controlled before-and-after study. Dogs were examined up to 3 times over 21 days. Treatment was prescribed based on NT-proBNP as follows: <1500 pmol/L at baseline, no treatment adjustment at any point during the study (group 1); ≥1500 pmol/L and creatinine ≤3.0 mg/dL at baseline or SC visits, treatment escalated according to the algorithm (group 2); ≥1500 pmol/L at baseline, no treatment adjustment (group 3). RESULTS: N-terminal pro-B-type natriuretic peptide decreased significantly in group 2 (mean change = -1736 pmol/L (95% CI, -804 to -2668), P < .001) but not in groups 1 or 3 (623 pmol/L [-631 to 1877 pmol/L], P = .14 and 685 pmol/L [-304 to 1068 pmol/L], P = .46, respectively). Serum BUN and creatinine did not change significantly between visit 0 and visit 2 in group 1 (median = 23 mg/dL [range 13-32] versus 19 mg/dL [12-38], P = .72 and 1.15 mg/dL [0.70-1.40] versus 0.95 mg/dL [0.70-1.10], P = .10, respectively) or group 2 (28 mg/dL [18-87] versus 43.5 mg/dL [21-160], P = .092 and 1.10 mg/dL [0.90-2.50] versus 1.55 mg/dL [0.90-3.30], P = .062, respectively). CONCLUSIONS AND CLINICAL IMPORTANCE: Use of this treatment escalation algorithm allows effective targeting of treatment for CHF in dogs against an objective criterion.

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

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

The effects of storage conditions on measurements of canine N-terminal pro-B-type natriuretic peptide.

OBJECTIVES: The study aims were to assess the temporal stability following storage at room temperature, the effect of up to 4 freeze-thaw cycles and the effect of simulated freezer failure on measurements of canine N-terminal pro-B-type natriuretic peptide (NT-proBNP) in serum and protease-inhibited (PI) plasma. ANIMALS: Twenty-five blood samples were collected from 16 dogs with myxomatous mitral valve disease. METHODS: Aliquots of canine serum and PI plasma were stored at room temperature (17-26 °C) for 30 min, 6, 24, 48 and 72 h, respectively. Further aliquots were subjected to between 1 and 4 freeze-thaw cycles. A further aliquot was transferred to storage at 4 °C for 24 h while a paired aliquot remained at -80 °C. All samples were returned to storage at -80 °C until subsequent analysis. N-terminal pro-B-type natriuretic peptide was measured in serum and PI plasma samples using first- and second-generation versions of a commercially-available ELISA. Repeated measures models were used to assess change in NT-proBNP measurements. Wilcoxon signed ranks were used to compare paired measurements. RESULTS: N-terminal pro-B-type natriuretic peptide concentrations declined over time in all samples stored at room temperature. Of the four situations tested, the rate of decrease was lowest for PI plasma samples measured using the second-generation assay. N-terminal pro-B-type natriuretic peptide is stable in samples subjected to up to 4 freeze-thaw cycles and in previously-frozen samples stored at 4 °C for 24 h. CONCLUSIONS: Use of the second-generation assay, compared with the first-generation, resulted in significantly higher recovery of NT-proBNP measured in PI plasma stored at room temperature. Transport of serum at room temperature for NT-proBNP measurement is not recommended.

Associations between N-terminal procollagen type III, fibrosis and echocardiographic indices in dogs that died due to myxomatous mitral valve disease.

OBJECTIVES: To evaluate associations between N-terminal procollagen type III (PIIINP), a serum biomarker of collagen biosynthesis, and myocardial fibrosis in dogs with naturally-occurring myxomatous mitral valve disease (MMVD). ANIMALS: Twenty-two dogs with echocardiographically-confirmed MMVD were prospectively recruited from a hospital population. All died as a result of MMVD and their hearts were available for post mortem examination. METHODS: Echocardiographic measurements and serum PIIINP concentrations were obtained from all dogs prior to death or euthanasia. Serum PIIINP concentrations (µg/mL) were measured using a validated commercially available radioimmunoassay. Myocardial tissue samples were collected post mortem and myocardial fibrosis was scored. The average fibrosis score for all cardiac sites in the heart was designated the global fibrosis score (GFS). The average fibrosis score for all papillary muscle sites was designated the papillary fibrosis score (PFS). Univariate and multivariate linear regression analyses were used separately to evaluate associations between GFS and PFS, respectively, and PIIINP and echocardiographic variables. RESULTS: Left ventricular end-diastolic diameter normalized for body weight (LVEDDN) and PIIINP were weakly independently positively associated with both GFS and PFS. LVEDDN and PIIINP were weakly negatively correlated. CONCLUSIONS: Both LVEDDN and serum PIIINP increase with increasing fibrosis score, although these relationships were not strong enough to be clinically useful. Although LVEDDN and PIIINP were positively correlated with fibrosis, PIIINP decreased with increasing LVEDDN, suggesting a complex interplay between fibrosis and remodeling in MMVD.

Associations among serum N-terminal procollagen type III concentration, urinary aldosterone-to-creatinine ratio, and ventricular remodeling in dogs with myxomatous mitral valve disease.

OBJECTIVE: To assess relationships among serum N-terminal procollagen type III concentration, urinary aldosterone-to-creatinine concentration ratio (UAC), and clinical variables in dogs with myxomatous mitral valve disease (MMVD) and healthy dogs. ANIMALS: 162 dogs with MMVD and 24 healthy control dogs of comparable age and body weight. PROCEDURES: Blood and urine samples were collected from each dog. Dogs with MMVD underwent echocardiography and ECG. Ventricular diameter measurements were normalized for body weight. Serum N-terminal procollagen type III and urinary aldosterone concentrations were measured via radioimmunoassay. Each dog was examined on 1 to 3 occasions. Examinations were repeated at approximately 6-month intervals. RESULTS: Serum N-terminal procollagen type III concentration decreased with increasing severity of MMVD and was negatively associated with age and left ventricular end-diastolic and end-systolic diameters. The UAC increased with prior percentage change in left ventricular end-diastolic diameter per month, subsequent percentage change in left ventricular end-systolic diameter per month, and treatment with diuretics and was negatively associated with age. Both UAC and serum N-terminal procollagen type III concentration were higher in Cavalier King Charles Spaniels than in other breeds when other measured variables were controlled for. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs with MMVD, echocardiographic indicators of left ventricular remodeling appeared to be associated with a decrease in serum concentration of a marker of collagen type III turnover and an increase in urinary aldosterone concentration.

Selected echocardiographic variables change more rapidly in dogs that die from myxomatous mitral valve disease.

OBJECTIVES: To determine if echocardiographic measurements change at a greater rate in dogs with myxomatous mitral valve disease (MMVD) that die of cardiac mortality. ANIMALS: Client-owned dogs (n = 242) with MMVD of varying severity were recruited from first opinion private practice. Only dogs which died during the study period (n = 102) were included in statistical analyses. METHODS: Prospective cohort study comparing the rate of change of echocardiographic variables between dogs that experienced cardiac mortality and those that experienced non-cardiac mortality. Measurements were repeated approximately every 6 months and repeated measures linear models were constructed to estimate the rate of change of each variable over time. RESULTS: Left ventricular (LV) end-diastolic diameter, normalized for body weight (LVEDDN) increased over time in both mortality groups. LV end-systolic diameter, normalized for body weight (LVESDN), LV end-diastolic diameter to LV free wall thickness in diastole (LVEDD/LVFWd) ratio, E wave velocity, E- to A-wave velocity ratio and left atrial to aortic root diameter ratio all increased over time in the cardiac mortality group, but did not change in the non-cardiac mortality group. MR velocity decreased over time in the cardiac mortality group but did not change in the non-cardiac mortality group. Tricuspid regurgitation jet velocity increased over time in both mortality groups. A wave velocity and fractional shortening did not change over time in either mortality group. CONCLUSIONS: Serial echocardiographic examination every 6-12 months is useful to identify dogs with progressive MMVD that are at increased risk of cardiac mortality.

Ultrasonographic measurement of flow-mediated vasodilation in dogs with chronic valvular disease.

OBJECTIVES: To measure flow-mediated vasodilation (FMD) in healthy dogs and in client-owned dogs with chronic valvular disease (CVD) and to investigate possible correlations between markers of CVD severity and FMD. ANIMALS: Twelve dogs with CVD and 11 healthy weight-matched dogs. METHODS: Brachial artery FMD following 5 min inflation of a cuff around the antebrachium was measured in 12 dogs with CVD and 11 healthy weight-matched dogs. Measurements were also obtained in the healthy dogs 5 min after cuff placement but without inflation ('sham cuff placement'). Dogs with CVD underwent echocardiography to confirm and characterize their disease. RESULTS: In healthy dogs (median age 4 [2-6] years), median FMD was 7.7% versus 3.4% with sham cuff placement (P = 0.003). In dogs with CVD (median age 8 [4-16] years) median FMD was 5.5% versus 7.7% in healthy dogs (P = 0.131). FMD showed an inverse correlation with left ventricular end-diastolic diameter normalized for body weight (r = -0.76, P = 0.0043). CONCLUSIONS: Brachial FMD in dogs with early CVD inversely correlates with severity of left ventricular remodelling.