Utility and safety of nafamostat mesilate for anticoagulation in dogs.

BACKGROUND: Surgical interventions are recommended for cases of advanced mitral regurgitation, however, limited facilities are available. The most prominent complication in such procedures is heparin-derived bleeding. An alternative anticoagulant to heparin, nafamostat mesilate (NM), can reduce the occurrence of complications associated with heparin such as bleeding or shock. OBJECTIVES: This study aimed to evaluate the utility and safety of using NM during anaesthesia in canines. METHODS: Six healthy adult Beagle dogs were anaesthetised, and NM was administered intravenously as a 10 mg/kg bolus dose over 5 min, followed by a continuous infusion of 10 mg/kg/h over 20 min. Blood tests and blood pressure measurements were performed at 0, 5, 25 and 55 min after NM administration. RESULTS: Activated thromboplastin times at 0, 25 and 55 min were 13.0 ± 0.7 s, 106.7 ± 13.3 s and 28.2 ± 2.9 s, respectively, with a significant difference between 0 and 25 min (p < 0.01) only. No significant differences were observed in prothrombin time, antithrombin, fibrinogen and fibrin degradation product concentrations between timepoints. Activated clotting times (ACTs) at 0, 5, 25 and 55 min were 119.5 ± 9.6 s, 826.7 ± 78.6 s, 924.8 ± 42.4 s and 165.2 ± 13.5 s, respectively. Significant differences were observed between 0 and 5 min (p < 0.05) and between 0 and 25 min (p < 0.05). Blood pressure changes occurred in four dogs (66.7%). No other serious adverse effects were observed. CONCLUSIONS: ACT results indicated that NM use in anaesthetised healthy dogs was sufficient to obtain procedural anticoagulation with minimal adverse effects. However, these preliminary data require validation in further studies on cardiopulmonary bypass surgery.

Reference Values of M-mode Echocardiographic Parameter in Adult Toy Breed Dogs.

Introduction: Myxomatous mitral valve disease (MMVD) is one of the most common heart diseases in dogs, and there is a dearth of reports that have investigated reference values for left ventricular end-diastolic internal diameter corrected for body weight (LVIDDN) exclusively in toy breeds. Animals: Eighty-six client-owned healthy dogs weighing <5 kg, including Toy Poodles, Chihuahuas, Yorkshire Terriers, Papillon, and other small breeds or small mixed breeds (mixed breed, Pomeranian, dachshund, Shih Tzu, and Maltese). In this retrospective single-center study, data were collected from dogs attending clinic for annual checkup between April 2014 and March 2021. Materials and Methods: Experienced echocardiographers performed transthoracic echocardiography, with reference ranges established using healthy dogs. Measurements of body weight (BW), heart rate, and several echocardiographic variables were obtained. The association between BW and echocardiographic parameters was assessed by linear regression analyses. M-mode measurements were obtained and normalized using equations developed from the regression analyses. Results: The LVIDDN value for 95% of dogs weighing <5 kg was achieved by dividing the M-mode measurement by BW raised to the power 0.332. The upper limit of the prediction interval for breeds weighing <5 kg was much lower than the value currently applied. Conclusions: We propose a reference LVIDDN value of ≥1.6 for the diagnosis of stage B2 MMVD in toy breed dogs. The results of our study will guide clinicians in deciding when to start treatment for MMVD in small breed dogs.

Evaluation method for cell-free in situ tissue-engineered vasculature monitoring: Proof of growth and development in a canine IVC model.

We previously developed a non-cell-dependent biodegradable scaffold to create in situ tissue-engineered vasculature (iTEV) and tested it in a canine inferior vena cava (IVC) model. As iTEV features change dramatically during tissue generation, practical, simple, and accurate methods to evaluate iTEV are needed. The present study examined the usefulness of a novel method to evaluate iTEV growth and remodeling according to a simple formula using angiography: hepatic vein (HV) index = (IVC-HV junction angle) ÷ (π × [minimal internal iTEV diameter ÷ 2]2). HV index strongly correlated with the pressure gradient across iTEV, which tended to improve during the tissue generation period up to 12 months post-implantation. Time-course changes in HV index reflected iTEV tissue development and in-vivo characteristics, such as hemodynamic congestion. In conclusion, HV index is useful to assess iTEV graft function because it represents both the morphometrics and hemodynamics of iTEV with only diagnostic imaging data.

Case Report: Tricuspid Annuloplasty for Right-Sided Congestive Heart Failure Secondary to Pulmonary Hypertension in a Dog.

An 11-year-old, 12.3-kg, female Miniature Dachshund was presented to our institution with ascites of unknown etiology. The dog had been administered moxidectin for 3 years to treat a heartworm infection. Thoracic radiographs showed enlargement of the right heart. Echocardiography revealed right atrial and ventricular dilatation as well as flattening of the interventricular septum. Heartworm was identified in the main pulmonary artery, which was dilated. Tricuspid regurgitation (TR) was observed using color Doppler ultrasonography, and 2.5 L of ascites were removed. The dog was diagnosed with pulmonary hypertension, severe TR, and right-sided congestive heart failure. Except at the initial site, heartworm was not detected using echocardiography, and the antigen test was negative. However, pharmacological treatment did not improve the right-sided congestive heart failure. Instead, De Vega tricuspid annuloplasty (TAP) was performed on the beating heart under cardiopulmonary bypass with the owner's consent. Sutures terminated between the two commissures in the middle of the annulus and were secured using another pledget. Annular reduction was performed by tying down the plication suture while the cylindrical sizer was inserted into the tricuspid valve orifice. The size of the cylindrical sizer was 16 mm, which was set based on the height and width of the septal leaflet. A 6-month follow-up showed a reduction of TR and right-sided volume overload with no evidence of ascites retention/recurrence or any other complication. Our findings indicate that TAP may be a valid treatment option for dogs with right-sided congestive heart failure caused by secondary TR.

Comparison of N-terminal pro-atrial natriuretic peptide and three cardiac biomarkers for discriminatory ability of clinical stage in dogs with myxomatous mitral valve disease.

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

Evaluation of atrial natriuretic peptide and cardiac troponin I concentrations for assessment of disease severity in dogs with naturally occurring mitral valve disease.

OBJECTIVE: To evaluate and compare the clinical usefulness of plasma atrial natriuretic peptide (ANP) and cardiac troponin-I (cTnI) concentrations for assessment of disease severity in dogs with naturally occurring mitral valve disease (MVD). ANIMALS: 316 dogs with MVD and 40 healthy control dogs. PROCEDURES: Each dog underwent a physical examination and echocardiographic and thoracic radiographic assessments. Blood samples were obtained and processed for measurement of plasma ANP and cTnI concentrations. Dogs with MVD were categorized into 3 groups (stages B1 [no clinical signs or evidence of cardiac enlargement], B2 [no clinical signs with evidence of cardiac enlargement], and C [history of congestive heart failure and pulmonary edema]) on the basis of American College of Veterinary Internal Medicine guidelines. Receiver operating characteristic curve analysis was used to evaluate the accuracy of plasma ANP and cTnI concentrations for assessment of MVD severity. RESULTS: Plasma ANP and cTnI concentrations increased as disease severity increased. Median plasma ANP concentrations for all 3 MVD groups and median plasma cTnI concentrations for the stage B2 and C groups were significantly greater than the corresponding concentrations for the control group. Plasma ANP concentration, but not cTnI concentration, appeared to be useful for detection of dogs with subclinical (stages B1 and B2) MVD, whereas both concentrations appeared useful for detection of dogs with stage C MVD. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that plasma ANP and cTnI concentrations should not be used independently to diagnose MVD but can be used to assess MVD severity and supplement echocardiographic findings.

Epidemiological study of indirect blood pressure measured using oscillometry in clinically healthy cats at initial evaluation.

We investigated the clinical characteristics of healthy cats in accordance with the target organ damage (TOD) risk category, on the basis of systolic blood pressure (SBP). This prospective multi-center study included 137 healthy cats. Indirect blood pressure was measured using an oscillometric technique. The median SBP in all cats was 147 mmHg (interquartile range: 134-158). On the basis of the TOD risk category, 57.7, 19.7, 21.9, and 0.7% of the cats were classified into categories I-IV, respectively. Age, sex, and body weight did not affect the SBP. This study provides basic information on the distribution of TOD risk categories in clinically healthy cats.

Effects of the angiotensin-converting enzyme inhibitor alacepril in dogs with mitral valve disease.

Alacepril is a relatively novel angiotensin-converting enzyme inhibitor; however, the safety, tolerance, and efficacy of alacepril in terms of cough suppression in dogs with mitral valve disease (MVD) remain unknown. The aim of this study was to investigate the safety, tolerance, and cough suppression efficacy of alacepril in dogs with MVD. This was a multi-center, prospective study. Forty-two dogs with echocardiographic or radiographic evidence of cardiac enlargement in addition to cough were enrolled. Dogs were treated with alacepril (1.0-3.0 mg/kg/day) for at least 4 weeks. One dog (2.4%) developed complications, including appetite loss, lethargy, and vomiting. Thirty-six dogs were re-evaluated after 4 weeks of treatment. Cough resolved or improved in 20 dogs (55.6%) after treatment. Based on the efficacy of alacepril, the dogs were divided into an effective group (n=20) and an ineffective group (n=16). After treatment, the left ventricular end-diastolic internal diameter corrected for body weight was significantly increased from baseline in the ineffective group but was significantly decreased in the effective group. Univariate binomial logistic regression analyses showed that high atrial natriuretic peptide level, N-terminal pro-B-type natriuretic peptide level, and E wave velocity at baseline were significantly correlated with alacepril inefficacy. Alacepril as treatment for MVD is well tolerated in most dogs, and different conditions of cardiac loading may influence the effect of the drug. Alacepril is expected to improve the quality of life of dogs with early stage MVD.

Diagnostic utility of cardiac troponin I in cats with hypertrophic cardiomyopathy.

BACKGROUND: Cardiac troponin I (cTnI) is useful for assessing hypertrophic cardiomyopathy (HCM) in cats. OBJECTIVE: To measure plasma cTnI concentrations in healthy cats and evaluate the clinical utility of cTnI in determining the severity of HCM. ANIMALS: Clinically healthy cats (n = 88) and cats with HCM (n = 93). METHODS: Multicenter prospective study. Cats with HCM, including hypertrophic obstructive cardiomyopathy at various stages, were diagnosed using echocardiography. Plasma cTnI concentrations were analyzed by a commercial laboratory. Receiver-operating characteristic curve analysis was used to evaluate the accuracy of plasma cTnI concentrations to detect HCM. RESULTS: The median cTnI concentration was 0.027 ng/mL (interquartile range, 0.012-0.048 ng/mL) in healthy cats. Concentrations were significantly higher in diseased cats than in healthy controls, and concentrations were significantly higher in cats with heart failure than in asymptomatic cats. A plasma cTnI concentration of 0.163 ng/mL had a sensitivity of 62.0% and specificity of 100% when used to distinguish normal cats from asymptomatic HCM cats without left atrial dilatation. A cutoff of 0.234 ng/mL had high sensitivity (95.0%) and specificity (77.8%) for assessing heart failure. The areas under the receiver-operating characteristic curves were 0.85 and 0.93, respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: Increased cTnI concentrations reflect the severity of HCM. If other causes of cardiac injury are ruled out, plasma cTnI concentration may be useful for predicting the severity of HCM in cats.

Relationship between indirect blood pressure and various stages of chronic kidney disease in cats.

Chronic kidney disease (CKD) is a common cause of secondary systemic hypertension in cats. We investigated the relationship between indirect blood pressure and the prevalence of systemic hypertension in various CKD stages in cats. Client-owned cats (24 control cats and 77 cats with CKD) were included. Biochemical examinations of plasma were conducted by a commercial laboratory. Diseased cats were divided into two groups based on the International Renal Interest Society (IRIS) guidelines (II and III-IV). Indirect blood pressure was measured using an oscillometric technique. Severe hypertension was diagnosed if systolic blood pressure (SBP) was ≥180 mmHg. Indirect blood pressures were significantly higher in IRIS stage III-IV than in the control cats. Of 77 cats with CKD, 25 (32.5%) had severe hypertension. The frequency of severe hypertension increased with an increase in IRIS stage; 0% in the controls, 27.6% in the IRIS stage II, and 47.4% in the IRIS stage III-IV, respectively. The indirect SBP was weakly correlated with urea nitrogen (r=0.27) and creatinine (r=0.23) concentrations in plasma. Binary logistic regression analysis showed that if plasma creatinine concentration is >3.7 mg/dl, cats with CKD had an increased risk for developing severe hypertension (P<0.001). Our results suggest that indirect blood pressure was correlated with the severity of CKD, and the prevalence of severe hypertension increased in cats with severe CKD. The risk of severe hypertension may be high in cats with severe CKD.

Histological maturation of vascular smooth muscle cells in in situ tissue-engineered vasculature.

The goal of regenerative medicine is to achieve histological and functional recovery to the level of the original tissue. For this purpose, we have developed a biodegradable scaffold to create cell-free in-situ tissue-engineered vasculature (iTEV) with good long-term results. However, the regeneration process of vascular smooth muscle cells (VSMCs) over time has yet to be examined. To evaluate the regeneration ability of VSMCs, the inferior vena cava of experimental animals was replaced with iTEV, and tested at 1, 3, 6, 12, and 24 months (n = 6 each) after implantation. Six animals were enrolled to compare 24-month iTEV and native vasculature in single individual samples. There were no complications throughout the study. Immunohistology, protein expression analysis, and biochemical findings indicate that iTEV can gradually regenerate and develop into a mature vessel within 24 months using our biodegradable scaffold. These results provide a time course for the regeneration of VSMCs within the tissue-engineered vascular autograft constructed using a biodegradable scaffold.

Long-term results of cell-free biodegradable scaffolds for in situ tissue engineering of pulmonary artery in a canine model.

We previously developed a cell-free, biodegradable scaffold for in-situ tissue-engineering vasculature (iTEV) in a canine inferior vena cava (IVC) model. In this study, we investigated application of this scaffold for iTEV of the pulmonary artery (iTEV-PA) in a canine model. In vivo experiments were conducted to determine scaffold characteristics and long-term efficacy. Biodegradable scaffolds comprised polyglycolide knitted fibers and an l-lactide and ε-caprolactone copolymer sponge, with an outer glycolide and ε-caprolactone copolymer monofilament reinforcement. Tubular scaffolds (8 mm diameter) were implanted into the left pulmonary artery of experimental animals (n = 7) and evaluated up to 12 months postoperatively. Angiography of iTEV-PA after 12 months showed a well-formed vasculature without marked stenosis, aneurysmal change or thrombosis of iTEV-PA. Histological analysis revealed a vessel-like vasculature without calcification. However, vascular smooth muscle cells were not well-developed 12 months post-implantation. Biochemical analyses showed no significant difference in hydroxyproline and elastin content compared with native PA. Our long-term results of cell-free tissue-engineering of PAs have revealed the acceptable qualities and characteristics of iTEV-PAs. The strategy of using this cell-free biodegradable scaffold to create relatively small PAs could be applicable in pediatric cardiovascular surgery requiring materials.

Comparison of vascular smooth muscle cells in canine great vessels.

BACKGROUND: Elucidating the histological characteristics of normal vascular smooth muscle cells (VSMCs) is important for understanding mechanisms of development, disease etiology and the remodeling and/or regeneration process of the vessel. However, knowledge regarding VSMCs is focused primarily on the artery. Although the characteristics of each great vessel are documented, few studies have examined VSMCs in parallel within each great vessel. The present study focused on comparing characteristics of canine VSMCs within the aorta (Ao), branch pulmonary artery (bPA), main pulmonary artery (mPA) and inferior vena cava (IVC), simultaneously. RESULTS: Western blot and immunohistochemistry were used to determine VSMC protein content for alpha smooth muscle actin (ASMA), calponin, myosin heavy chain (MHC) and its isozyme SM2, and non-muscle myosin heavy chain B (SMemb). Thickness and ratio of the VSMC layer were also measured. Expression levels of ASMA, calponin and SM2 significantly differed between vessels, except between mPA and either bPA, Ao and IVC vessels. Expression levels of MHC were significantly different in all vessels, whilst expression of SMemb was significantly different in the Ao compared with either bPA and mPA vessels. All vessels were significantly different with respect to total wall and VSMC layer thickness. The ratio between VSMC layer and total wall thickness was significantly different for each vessel, except between bPA and mPA vessels. Histological analysis of the IVC revealed that the VSMC layer does not line evenly and continuously through the long axis or transverse sections. With respect to the pulmonary artery, calponin was expressed to a greater extent in the mPA compared with the bPA (P < 0.01*). In contrast, MHC and SM2 were expressed to a greater extent in the bPA compared with the mPA (P < 0.01*). Differences in VSMC distribution indicate structural differences in the proximal and distal pulmonary artery bifurcation. CONCLUSION: Our results show that the VSMC expression pattern in each great vessel is unique and suggestive of the developmental differences between great vessels. We believe this study provides basic data for the pathology, etiology and regenerative capability of the vessels.

Long-term results of cell-free biodegradable scaffolds for in situ tissue-engineering vasculature: in a canine inferior vena cava model.

We have developed a new biodegradable scaffold that does not require any cell seeding to create an in-situ tissue-engineering vasculature (iTEV). Animal experiments were conducted to test its characteristics and long-term efficacy. An 8-mm tubular biodegradable scaffold, consisting of polyglycolide knitted fibers and an L-lactide and ε-caprolactone copolymer sponge with outer glycolide and ε-caprolactone copolymer monofilament reinforcement, was implanted into the inferior vena cava (IVC) of 13 canines. All the animals remained alive without any major complications until euthanasia. The utility of the iTEV was evaluated from 1 to 24 months postoperatively. The elastic modulus of the iTEV determined by an intravascular ultrasound imaging system was about 90% of the native IVC after 1 month. Angiography of the iTEV after 2 years showed a well-formed vasculature without marked stenosis or thrombosis with a mean pressure gradient of 0.51 ± 0.19 mmHg. The length of the iTEV at 2 years had increased by 0.48 ± 0.15 cm compared with the length of the original scaffold (2-3 cm). Histological examinations revealed a well-formed vessel-like vasculature without calcification. Biochemical analyses showed no significant differences in the hydroxyproline, elastin, and calcium contents compared with the native IVC. We concluded that the findings shown above provide direct evidence that the new scaffold can be useful for cell-free tissue-engineering of vasculature. The long-term results revealed that the iTEV was of good quality and had adapted its shape to the needs of the living body. Therefore, this scaffold would be applicable for pediatric cardiovascular surgery involving biocompatible materials.