Investigation of red blood cell and platelet indices in adult dogs suffered from myxomatous mitral valve disease with and without pulmonary hypertension.

Background: Pulmonary hypertension (PH) is a common complication of cardiopulmonary disease. In dogs, PH commonly occurs secondary to myxomatous mitral valve disease (MMVD). Red blood cell and platelet indices including mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red cell distribution width (RDW), mean platelet volume (MPV) and platelet distribution width (PDW), have previously been found to be indicators for predicting and prognosing PH in humans. Therefore, this study aimed to investigate whether these indices are associated with MMVD and/or PH in dogs. Methods: Two hundred and forty-six dogs were retrospectively recruited for the study and classified into 4 groups: normal (n = 49), MMVD (n =102), PH (n =17), MMVD+PH (n =78). A sub-analysis was performed in dogs with MMVD without evidence of PH according to stage B1 (n =20), stage B2 (n =15), stage C (n =67). The data are expressed as median (interquartile range). Results and discussion: No significant differences (p < 0.05) were found in MCV, RDW and MPV among all groups (normal, MMVD, PH and MMVD+PH). However, decreases in MCH and MCHC were found in MMVD [22.40 (20.90-23.50) pg and 35.25 (33.08-36.90) g/dL], MMVD+PH [22.25 (20.85-23.98) pg and 35.65 (33.30-37.33) g/dL] and PH groups [21.20 (20.60-22.20) pg and 33.80 (32.75-35.70) g/dL] compared to the normal dogs [24.29 (23.55-24.90) pg and 38.20 (37.50-39.05) g/dL] (p < 0.001). Decreases in PDW were found in dogs in the MMVD+PH [15.10 (14.98-15.30) %] groups compared to dogs in the normal group [15.30 (15.10-15.50) %] (p = 0.004). Sub-analysis of MMVD dogs without PH showed a decrease in MCH in dogs with stage B2 MMVD [21.00 (20.50-22.90) pg] and stage C MMVD [22.40 (20.90-23.20) pg] compared to normal dogs [24.29 (23.55-24.90) pg] (p < 0.001). MCHC of dogs with stage B1 [36.55 (33.53-37.78) g/dL] (p = 0.004), B2 [32.90 (32.00-35.00) g/dL] (p < 0.001) and C MMVD [35.30 (33.30-36.80) g/dL] (p < 0.001) were lower than those of normal dogs [38.20 (37.50-39.05) g/dL]. PDW in the stage C MMVD group [15.10 (15.00-15.30) %] was reduced compared to the normal group [15.30 (15.10-15.50) %] (p = 0.042) and the stage B1 MMVD group [15.35 (15.23-15.68) %] (p = 0.002). MCH, MCHC and PDW were negatively correlated with the left atrial and left ventricular size. Conclusion: Decreases in MCH and MCHC are related to MMVD, precapillary PH and postcapillary PH while PDW are associated with MMVD severity but not with the presence of PH.

Assessment of microcirculation variables and endothelial glycocalyx using sidestream dark field videomicroscopy in anesthetized dogs undergoing cardiopulmonary bypass.

Introduction: To evaluate microcirculation and endothelial glycocalyx (eGC) variables using sidestream darkfield (SDF) videomicroscopy in canine cardiopulmonary bypass (CPB). Methods: Dogs undergoing CPB for surgical correction of naturally-occurring cardiac disease were prospectively included. Variables collected included patient demographics, underlying cardiac disease, red blood cell flow (Flow), 4-25 µm vessel density (Density), absolute capillary blood volume (CBVabs), relative capillary blood volume (CBVrel) and eGC width assessed by perfused boundary region (PBR). Anesthetized healthy dogs were used as control. Microcirculation and eGC variables were compared at baseline under anesthesia (T0), on CPB prior to cross clamping (T1), after cross clamp removal following surgical correction (T2) and at surgical closure (T3). Results: Twelve dogs were enrolled, including 10 with a complete dataset. Median Flow was 233.9, 79.9, 164.3, and 136.1 µm/s at T0, T1, T2, and T3, respectively, (p = 1.00). Median Density was 173.3, 118.4, 121.0 and 155.4 mm/mm2 at T0, T1, T2, and T3, respectively, (p = 1.00). Median CBVabs decreased over time: 7.4, 6.6, 4.8 and 4.7 103µm3 at T0, T1, T2, and T3, respectively, (p < 0.01). Median CBVrel increased over time: 1.1, 1.5,1.1, and 1.3 103µm3 at T0, T1, T2, and T3, respectively, (p < 0.001). Median PBR increased over time: 1.8, 2.1, 2.4, 2.1 µm at T0, T1, T2, and T3, respectively, (p < 0.001). Compared to control dogs (n = 8), CPB dogs had lower CBVabs at T0. Conclusion: Alterations in eGC thickness and microvascular occur in dogs undergoing CPB for naturally-occurring cardiac disease.

Local serotonin mediates cyclic strain-induced phenotype transformation, matrix degradation, and glycosaminoglycan synthesis in cultured sheep mitral valves.

This study addressed the following questions: 1) Does cyclic tensile strain induce protein expression patterns consistent with myxomatous degeneration in mitral valves? 2) Does cyclic strain induce local serotonin synthesis in mitral valves? 3) Are cyclic strain-induced myxomatous protein expression patterns in mitral valves dependent on local serotonin? Cultured sheep mitral valve leaflets were subjected to 0, 10, 20, and 30% cyclic strain for 24 and 72 h. Protein levels of activated myofibroblast phenotype markers, α-smooth muscle actin (α-SMA) and nonmuscle embryonic myosin (SMemb); matrix catabolic enzymes, matrix metalloprotease (MMP) 1 and 13, and cathepsin K; and sulfated glycosaminoglycan (GAG) content in mitral valves increased with increased cyclic strain. Serotonin was present in the serum-free media of cultured mitral valves and concentrations increased with cyclic strain. Expression of the serotonin synthetic enzyme tryptophan hydroxylase 1 (TPH1) increased in strained mitral valves. Pharmacologic inhibition of the serotonin 2B/2C receptor or TPH1 diminished expression of phenotype markers (α-SMA and SMemb) and matrix catabolic enzyme (MMP1, MMP13, and cathepsin K) expression in 10- and 30%-strained mitral valves. These results provide first evidence that mitral valves synthesize serotonin locally. The results further demonstrate that tensile loading modulates local serotonin synthesis, expression of effector proteins associated with mitral valve degeneration, and GAG synthesis. Inhibition of serotonin diminishes strain-mediated protein expression patterns. These findings implicate serotonin and tensile loading in mitral degeneration, functionally link the pathogeneses of serotoninergic (carcinoid, drug-induced) and degenerative mitral valve disease, and have therapeutic implications.

Surgical repair for canine tricuspid valve dysplasia: Technique and case report.

Tricuspid valve dysplasia (TVD) is a congenital malformation of the right atrioventricular valve characterized by restricted leaflet motion, annular dilation, and tricuspid regurgitation (TR). Severe cases typically exhibit progressive right-sided congestive heart failure, affecting the quality of life and survival. This article describes a technique for surgical repair of TVD and a case report with long-term follow-up. A 1.5-year-old intact male Labrador retriever with severe TR underwent surgical repair for TVD. Valve repair was performed under cardiopulmonary bypass and consisted of neochord mobilization of the valve leaflets and partial band annuloplasty. Transthoracic echocardiogram performed 5 days after surgery showed mild TR, a 93% decrease in anatomic regurgitant orifice area, and decreased right chamber dimensions. Forty-eight months after repair, the patient was free of clinical signs, did not have a heart murmur, and was receiving no cardiac medications. Based on this case, surgical repair of TVD is feasible with long-term durability, and the outcome suggests that the described technique may be a viable treatment option for patients with severe TVD.

Tissue-gel electrophoresis enhances antigen removal from porcine aortic valve and bovine pericardium.

BACKGROUND AND AIM OF THE STUDY: A tissue-engineered heart valve could provide a living prosthesis with characteristics of an ideal valve replacement. One approach to scaffolding a tissue-engineered heart valve is through the 'decellularization' of xenogeneic tissues. Concerns regarding the completeness of antigen removal associated with current detergent-based decellularization treatments have been raised. The study aim was to evaluate antigen removal from candidate xenogeneic bioscaffolds using a novel tissue-gel electrophoresis (TGE) method. METHODS: Porcine aortic valve (PAV) conduit and bovine pericardium (BP) were treated sequentially with hypotonic lysis, sodium dodecyl sulfate (SDS), and TGE. The completeness of antigen removal was evaluated by immunoblot analysis of extractable soluble proteins using rabbit anti-PAV or anti-BP serum. Tissues were also evaluated by hematoxylin and eosin histology. RESULTS: TGE enhanced antigen removal from both the PAV and BP. The effects of TGE were shown to depend on the SDS concentration and voltage (60 versus 120 V), but to be independent of time after 4 h. The effects of TGE were detectable both before and after 96 h aqueous washout. Treatment with 1.0% SDS with TGE (120 V for 4 h) resulted in complete acellularity and no detectable soluble protein antigens from the PAV conduit. CONCLUSION: TGE is a promising adjunctive decellularization method for generating non-immunoreactive bioscaffolds from xenogeneic tissues.

Tryptophan hydroxylase 1 expression is increased in phenotype-altered canine and human degenerative myxomatous mitral valves.

BACKGROUND AND AIM OF THE STUDY: Serotonin is a known mediator of myxomatous pathology in heart valves. Tryptophan hydroxylase 1 (TPH1) is the limiting enzyme for peripheral serotonin synthesis, and its expression by valve interstitial cells (IC) could implicate an autocrine serotonin signaling mechanism in primary degenerative myxomatous mitral valve disease. Thus, the expression of TPH1 in canine and human myxomatous mitral valves was determined, and IC phenotypes expressing TPH1 identified. METHODS: TPH1 expression was determined in canine and human myxomatous and normal mitral valves by immunoblot (IB) and immunofluorescence microscopy (IFM). Co-localization of TPH1 expression with markers of IC phenotype transformation, alpha-smooth muscle actin (a-SMA) and non-muscle embryonic myosin (SMemb) was determined using double-IFM. RESULTS: TPH1 expression by IB was increased (p < 0.05) by three- to five-fold in canine early-stage and late-stage myxomatous valves, and in human surgically excised myxomatous valves compared to canine and human normal control valves, respectively. The number of TPH1 immunopositive cells per x400 field was increased (p < 0.005) in canine (14.9 +/- 1.2) and human (14.9 +/- 2.9) myxomatous valves compared to canine (5.0 +/- 2.4) and human (2.9 +/- 0.6) normal control valves, respectively. Patterns for alpha-SMA and SMemb IC phenotype transformation were distinctly different in myxomatous valves. TPH1 expression was more closely associated with the SMemb IC phenotype in canine and human myxomatous valves. CONCLUSION: An increased expression of TPH1 in canine and human myxomatous mitral valves implicates an autocrine serotonin signaling mechanism in primary degenerative myxomatous mitral valves. TPH1 expression is associated with the SMemb-positive IC phenotype.

Autocrine serotonin and transforming growth factor beta 1 signaling mediates spontaneous myxomatous mitral valve disease.

BACKGROUND AND AIM OF THE STUDY: Although serotonin and serotoninergic drugs are known to cause myxomatous-like valvulopathy, the role of serotonin in spontaneous myxomatous valve disease (MVD) remains unclear. Tryptophan hydroxylase 1 (TPH1) is the limiting enzyme for peripheral serotonin synthesis, and its expression in myxomatous valves could implicate an autocrine serotonin signaling mechanism. Studies in cultured cells demonstrate a close coupling between serotonin and transforming growth factor beta1 (TGFbeta1) signaling. The study aim was to investigate serotonin and TGFbeta1 signaling in spontaneous MVD. METHODS: In canine normal and myxomatous mitral valves, target signaling proteins including TPH1, serotonin 2B receptor (5HT(2B)R), serotonin transmembrane transporter (SERT), total and phosphorylated extracellular signaling-regulated kinase (ERK) 1/2, latent TGFbeta1 and TGFbeta1 receptors I and II, were studied using immunohistochemistry and immunoblot analysis. In human myxomatous valves, TPH1 was determined using immunofluorescence and immunoblot analysis. RESULTS: In canine mitral valves, both 5HT(2B)R and TPH1 were increased in myxomatous valves, whereas SERT, a key protein in serotonin metabolism, was decreased in myxomatous valves. Phosphorylated, but not total, ERK 1/2 was increased in myxomatous valves, consistent with an enhanced active serotonin signaling. The expression of TGFbeta1 receptors I and II, and of latent TGFbeta1, was increased in myxomatous valves. Human myxomatous mitral valves expressed TPH1. CONCLUSION: The expression of TPH1 by canine and human myxomatous valves demonstrates a capacity for local serotonin production. Key signaling protein expression patterns support active serotonin and TGFbeta1 signaling in canine myxomatous valves. These findings implicate an autocrine serotonin and TGFbeta1 mechanism in the pathogenesis of spontaneous MVD.

Immunoblot detection of soluble protein antigens from sodium dodecyl sulfate- and sodium deoxycholate-treated candidate bioscaffold tissues.

BACKGROUND AND AIMS OF THE STUDY: The detergent-based 'decellularization' of xenogeneic tissues is one approach to scaffolding a tissue-engineered heart valve construct; however, concern persists regarding the immunogenicity of decellularized xenogeneic bioscaffolds. The study aims were to: (i) develop a sensitive and robust immunoblot-based assay for the detection of soluble protein antigens in xenogeneic bioscaffolds; and (ii) evaluate the completeness of protein antigen removal from sodium dodecyl sulfate (SDS)- or sodium deoxycholate (SD)-treated bovine pericardium (BP) or porcine aortic valve (PAV) conduit. METHODS: Homogenized BP or PAV were injected into rabbits to generate immune serum towards these tissues. Soluble proteins were extracted from untreated BP and PAV. Immunoblot analyses of the extracts were performed using pre-immune and 14-, 28-, 42-, 56- and 70-day post-immune serum. BP and PAV were treated sequentially with 4 h hypotonic lysis; with 0, 0.01, 0.025, 0.05, 0.1, 0.25 or 0.5% SDS or SD for 24 h; and with 96 h of aqueous wash-out. Immunoblot analyses of protein extracts from treated tissues were performed using 70-day post-immune rabbit serum. RESULTS: Immunoblot analysis of untreated BP or PAV with pre-immune serum showed no immune banding. The immune banding density increased progressively when immunoblots were performed with 14-day through 70-day post-immune serum. The immunoblot analysis of treated BP and PAV showed that soluble protein antigen removal from SDS- or SD-treated tissues was incomplete. CONCLUSION: Immunoblot analysis is a sensitive and robust assay for detecting soluble protein xenogeneic antigens after the decellularization of xenogeneic bioscaffolds. Under the study conditions, hypotonic lysis, SDS or SD detergent treatment, and aqueous wash-out-based decellularization of bovine pericardium and porcine aortic valve conduit did not completely remove detectable protein antigens.

Does sodium dodecyl sulfate wash out of detergent-treated bovine pericardium at cytotoxic concentrations?

BACKGROUND AND AIM OF THE STUDY: The ionic detergent sodium dodecyl sulfate (SDS) is a proposed treatment for the removal of antigenic proteins from unfixed biological scaffolds used in tissue engineering. However, questions remain about possible cytotoxic effects of SDS-treated tissues. The study aims were to: (i) develop a sensitive SDS assay for physiological solutions; (ii) measure SDS concentrations in the washing media of SDS-treated tissue; and (iii) determine cytotoxic SDS concentrations in cultured ovine vascular cells. METHODS: An assay was developed to monitor SDS concentrations at microM levels, based on attenuated total reflectance infrared spectroscopy. Bovine pericardium was treated with SDS (1.0 to 0.01%) and washed for 96 h. The SDS concentration in the washing media was measured at 24-h intervals; data were expressed as microM/g tissue. Ovine vascular cells were cultured in DME media at 37 degrees C for 48 h in various SDS concentrations (10 to 1000 microM). The cells were then counted, and the percentage live cells expressed, based on trypan blue exclusion (n=5). RESULTS: SDS concentrations > or =10 microM significantly reduced (p < 0.05) the total cell number, while concentrations > or =100 microM reduced (p < 0.05) the percentage live cells of ovine vascular cell cultures. SDS was present in the washing media of SDS-treated bovine pericardium. SDS leaching from bovine pericardium was found to depend on the SDS concentration used for the treatment, and diminished with time. CONCLUSION: SDS leaches from SDS-treated bovine pericardium at concentrations that are potentially cytotoxic. An understanding of the dynamics of SDS washout, based on a sensitive SDS assay, may lead to the creation of protocols for the preparation of biological scaffolds that are free from cytotoxic leaching.

Use of fat-fed rats to study the metabolic and vascular sequelae of obesity and beta-adrenergic antagonism.

Obesity-associated cardiovascular disease exerts profound human and monetary costs, creating a mounting need for cost-effective and relevant in vivo models of the complex metabolic and vascular interrelationships of obesity. Obesity is associated with endothelial dysfunction and inflammation. Free fatty acids (FFA), generated partly through beta-adrenergic receptor-mediated lipolysis, may impair endothelium-dependent vasodilation (EDV) by proinflammatory mechanisms. beta-Adrenergic antagonists protect against cardiovascular events by mechanisms not fully defined. We hypothesized that beta antagonists may exert beneficial effects, in part, by inhibiting lipolysis and reducing FFA. Further, we sought to evaluate the fat-fed rat as an in vivo model of obesity-induced inflammation and EDV. Control and fat-fed rats were given vehicle or beta antagonist for 28 d. Serum FFA were measured to determine the association to serum IL6, TNFalpha, and C-reactive protein and to femoral artery EDV. Compared with controls, fat-fed rats weighed more and had higher FFA, triglyceride, leptin, and insulin levels. Unexpectedly, in control and fat-fed rats, beta antagonism increased FFA, yet inflammatory cytokines were reduced and EDV was preserved. Therefore, reduction of FFA is unlikely to be the mechanism by which beta antagonists protect the endothelium. These results reflect the need for validation of ex vivo models of obesity-induced inflammation and endothelial dysfunction, concurrent with careful control of dietary fat composition and treatment duration.

Epicardial pacemaker implantation in small animals.

Epicardial pacemaker implantation can be performed as a lone procedure or in combination with another thoracic or abdominal surgery. This article reviews the current literature and describes a minimally invasive approach for epicardial pacemaker implantation in small animals. The principal advantage of epicardial pacing is that it avoids contact with blood and intracardiac structures and thereby avoids uncommon but potentially devastating complications associated with endocardial pacemaker implantation. Epicardial pacing as a lone procedure can be accomplished via an abdominal transdiaphragmatic or minimal incision thoracotomy approach (minithoracotomy). A minithoracotomy offers the advantages of being less invasive and providing more direct access to the cardiac surface for suturing of epicardial electrodes. Epicardial pacing is a viable option for smaller animals, animals with pre-existing infections, animals at risk for thrombotic complications, or animals undergoing another thoracic or abdominal surgery.

Protein extraction and 2-DE of water- and lipid-soluble proteins from bovine pericardium, a low-cellularity tissue.

Bovine pericardium (BP) is an important biomaterial used in the production of glutaraldehyde-fixed heart valves and tissue-engineering applications. The ability to perform proteomic analysis on BP is useful for a range of studies, including investigation of immune rejection after implantation. However, proteomic analysis of fibrous tissues such as BP is challenging due to their relative low-cellularity and abundance of extracellular matrix. A variety of methods for tissue treatment, protein extraction, and fractionation were investigated with the aim of producing high-quality 2-DE gels for both water- and lipid-soluble BP proteins. Extraction of water-soluble proteins with 3-(benzyldimethylammonio)-propanesulfonate followed by n-dodecyl beta-D-maltoside extraction and ethanol precipitation for lipid-soluble proteins provided the best combination of yield, spot number, and resolution on 2-DE gels (Protocol E2). ESI-quadrupole/ion trap or MALDI-TOF/TOF MS protein identifications were performed to confirm bovine origin and appropriate subcellular prefractionation of resolved proteins. Twenty-five unique, predominantly cytoplasmic bovine proteins were identified from the water-soluble fraction. Thirty-two unique, predominantly membrane bovine proteins were identified from the lipid-soluble fraction. These results demonstrated that the final protocol produced high-quality proteomic data from this important tissue for both cytoplasmic and membrane proteins.

Interstitial cells from dogs with naturally occurring myxomatous mitral valve disease undergo phenotype transformation.

BACKGROUND AND AIM OF THE STUDY: Myxomatous mitral valve disease is a common naturally occurring heart disease of dogs that is pathologically similar to myxomatous mitral valve disease in humans. It was hypothesized that interstitial cell phenotype transformation recently described in human myxomatous valves might also occur in dogs with myxomatous mitral valves, and correlate with disease severity. METHODS: Normal and early-, intermediate- and late-stage myxomatous canine mitral valves were examined histologically and immunohistochemically for cytoskeletal (vimentin, desmin, smooth muscle alpha-actin, smooth muscle myosin, and non-muscle myosin), collagenolytic (MMP-1, MMP-13), cell surface (CD-31, CD-45, CD-68) and proliferation (Ki-67) proteins. RESULTS: Normal canine mitral valve interstitial cells were positive for vimentin, but negative for alpha-actin, desmin and non-muscle myosin (i.e., fibroblast phenotype). Interstitial cells from myxomatous valves showed progressive positive staining for alpha-actin and desmin, but were negative for smooth muscle myosin (i.e., myofibroblast phenotype). Positive-staining cells first appeared as cellular clusters in the subendocardial region of the lamina atrialis and extended into deeper layers with increasing severity. Interstitial cells from myxomatous valves showed positive staining for non-muscle myosin (i.e., activated mesenchymal cell phenotype). Positive-staining cells increased with disease severity and were dispersed throughout the valve layers. The expression of MMP-1 and MMP-13 increased in myxomatous mitral valves and correlated with disease severity. Interstitial cellularity increased dramatically in degenerative mitral valves, though Ki-67 staining was only mildly increased. CONCLUSION: Two patterns of interstitial cell phenotype transformation were identified in dogs with myxomatous mitral valve disease, and both correlated with disease severity. Myofibroblast transformation characterized by positive staining for alpha-actin and desmin occurred in cellular clusters primarily in the lamina atrialis. Mesenchymal cell activation characterized by positive staining to non-muscle myosin occurred throughout the valve. The dog may be a natural model for studying the cell biology of progressive myxomatous valve disease.

Heterotopic implantation of a porcine bioprosthetic heart valve in a dog with aortic valve endocarditis.

CASE DESCRIPTION: A 5-year-old male German Shepherd Dog was evaluated because of a 5-month history of progressive lethargy, weight loss, and heart failure. CLINICAL FINDINGS: On physical examination, bounding femoral pulses and systolic and diastolic murmurs were detected. Echocardiography revealed severe aortic valve insufficiency (AVI) and a large vegetative lesion on the aortic valve consistent with aortic valve endocarditis. The AVI velocity profile half-time was 130 milliseconds; the calculated peak systolic pressure gradient across the aortic valve was 64 mm Hg. Left ventricular diameter during diastole was 63.6 mm (predicted range, 40.2 to 42 mm) and during systole was 42.9 mm (predicted range, 25.4 to 27 mm). Systolic, diastolic, and mean arterial blood pressures were 120, 43, and 65 mm Hg, respectively. TREATMENT AND OUTCOME: To palliate severe AVI, the descending aorta was occluded (duration, 16.75 minutes) and heterotopic implantation of a porcine bioprosthetic heart valve in that vessel was performed. After surgery, systolic, diastolic, and mean arterial blood pressures were 115, 30, and 61 mm Hg, respectively, in the forelimb and 110, 62, and 77 mm Hg, respectively, in the hind limb. Within 6 months, the AVI velocity profile half-time had increased to 210 milliseconds, indicating diminished severity of AVI. After 24 months, the dog was able to engage in vigorous exercise; no pulmonary edema had developed since surgery. CLINICAL RELEVANCE: Heterotopic bioprosthetic heart valve implantation into the descending aorta during brief aortic occlusion appears feasible in dogs and may provide substantial palliation for dogs with severe AVI.

Inflow Venous Occlusion for Intracardiac Resection of an Occluding Right Ventricular Tumor.

Use of normothermic venous inflow occlusion enabled removal of an intracardiac tumor in a 4 yr old, 27 kg, spayed female Airedale terrier with a history of appendicular osteosarcoma and recent exertional syncope. Inflow venous occlusion via a median sternotomy thoracotomy without hypothermia was used to access the mineralized mass within the right ventricular outflow tract. Duration of circulatory arrest was 70 s for this beating heart surgery. A circumscribed intracardiac chondrosarcoma tumor was marginally resected in this dog, successfully alleviating exertional syncope and restoring a normal echogenic appearance of the right heart. Asymptomatic intracardiac chondrosarcoma recurrence and pulmonary metastasis was detected at 309 days and cardiopulmonary arrest occurred 372 days following intracardiac surgery. Use of inflow occlusion is a viable technique for select intracardiac tumors in dogs with preoperative planning.

Decellularization of bovine pericardium for tissue-engineering by targeted removal of xenoantigens.

BACKGROUND AND AIM OF STUDY: Assessment of decellularization of xenogeneic biological scaffolds for tissue engineering has relied primarily on histological cellularity, though this may not ensure the removal of known xenogeneic antigens such as galactose-alpha1,3-galactose (alpha-gal) and MHC I. METHODS: Bovine pericardium (BP) underwent standard (Std) decellularization consisting of hypotonic lysis and treatment with DNAse/RNAse. In addition to Std decellularization, tissues were treated for 24 h with either 0.5% Triton X-100, 0.5% sodium deoxycholate (SD), 0.1% sodium dodecyl sulfate (SDS), alpha-galactosidase (5 U/ml) or phospholipase (PL) A2 (150 U/ml). Tissues underwent a 96-h washout under gentle agitation at 27 degrees C, and then evaluated by light microscopy for % cellularity, and by immunohistochemistry and Western blot for alpha-gal, bovine MHC I and smooth muscle alpha-actin. RESULTS: Standard treatment of BP resulted in only partial removal histological cellularity and persistence of alpha-gal, MHC I and alpha-actin. Adding SD treatment resulted in apparent acellularity, but persistence of xenogeneic antigens. Only the addition of SDS resulted in complete histological acellularity and removal of xenogeneic antigens. Treatment with alpha-galactosidase selectively removed alpha-gal from BP. CONCLUSION: Histological cellularity is not an adequate end-point for assuring removal of antigenicity from xenogeneic biological scaffolds. However, known xenogeneic antigens can be targeted for removal by novel decellularization treatments such as alpha-galactosidase.

Evaluation of survival rate and prognostic indicators for surgical treatment of left-to-right patent ductus arteriosus in dogs: 52 cases (1995-2003).

OBJECTIVE: To determine factors associated with long-term survival in dogs treated surgically for patent ductus arteriosus (PDA). DESIGN: Retrospective case series. Animals-52 dogs treated surgically for left-to-right shunting PDA. PROCEDURE: Data pertaining to age, breed, sex, body weight, clinical examination findings, type and duration of medical treatment, results of thoracic radiography and echocardiography, and surgical and postoperative complications were collected from records. Follow-up information was obtained from medical records or telephone interviews with owners or referring veterinarians. RESULTS: 22 dogs had mitral valve regurgitation. Mean weight and age were not significantly different between dogs with or without mitral valve regurgitation. Twenty-four (46.2%) dogs had clinical signs related to cardiac insufficiency. Left atrial dilatation was observed in 56.3% of dogs that were radiographed. Sonographic imaging was used to diagnose left atrial dilatation in 23 dogs and left ventricular dilatation in 25 dogs. The 1- and 2-year survival rates were 92% and 87%, respectively. Diagnosis of mitral valve regurgitation before surgery was not associated with the probability of survival. Age, weight, lethargy, preoperative treatment with angiotensin-converting enzyme inhibitors, and right atrial dilatation on radiographs at the time of surgery were negatively associated with probability of survival. CONCLUSIONS AND CLINICAL RELEVANCE: Surgical treatment of PDA was curative in young dogs without clinical signs of heart failure. Surgical correction of PDA should be recommended as early as possible after diagnosis, and mitral valve regurgitation is not a contraindication for surgery.

Technique and outcome of mitral valve replacement in dogs.

Eight dogs with naturally occurring severe mitral regurgitation underwent mitral valve replacement with a mechanical valve prosthesis during cardiopulmonary bypass. Dogs received warfarin orally after surgery to maintain a prothrombin time-based international normalized ratio from 2.5 to 3.5. Seven dogs survived surgery. Left ventricular diastolic volume index decreased significantly from 206 +/- 91 mL/m2 before surgery to 121 +/- 47 mL/m2 after surgery. Left atrium-to-aorta ratio decreased significantly from 2.66 +/- 0.4 before surgery to 1.73 +/- 0.65 after surgery. Left ventricular systolic volume index was not significantly different after surgery (56 +/- 36 mL/m2), compared with before surgery (40 +/- 32 mL/m2). Median survival after surgery was 4.5 months (range, 0.75 months to 5.25 years). Six dogs died of confirmed or suspected thrombosis of the valve prosthesis. Dogs with severe mitral regurgitation tolerated mitral valve replacement well, but a high incidence of prosthetic valve thrombosis limited long-term outcome.

Linear, logarithmic, and polynomial models of M-mode echocardiographic measurements in dogs.

OBJECTIVE: To determine whether logarithmic and polynomial models are superior to simple linear models for predicting reference values for M-mode echocardiographic variables in dogs with a wide range of body weights. ANIMALS: 69 apparently healthy adult male and female dogs of various breeds, ages (range, 1 to 12 years; median, 3.5 years), and body weights (range, 3.9 to 977 kg; median, 25.4 kg). PROCEDURE: Echocardiographic M-mode measurements of the interventricular septum, left ventricular dimension (LVD), left ventricular wall, aorta, and left atrium were obtained. Simple linear, second-order polynomial, third-order polynomial, and logarithmic regression models were determined by use of the least-squares method to describe the relationship between M-mode measurements and body weight. Differences in adjusted R2 values of logarithmic and polynomial models were tested for significance of contribution, compared with the simple linear model. RESULTS: Significant differences in adjusted R2 were found when comparing simple linear with logarithmic or polynomial models for LVD-diastole, LVD-systole, aorta, and left atrium. Differences in adjusted R2 between second-order polynomial, third-order polynomial, and logarithmic models were not significant for any M-mode measurement. CONCLUSIONS AND CLINICAL RELEVANCE: In this study, logarithmic or second-order polynomial models predicted reference values of M-mode measurements for size of the cardiac chambers better than simple linear models for dogs with a wide range of body weights. Logarithmic and polynomial models were not superior to simple linear models for M-mode measurements of cardiac wall thickness.

Evaluation of techniques and outcomes of mitral valve repair in dogs.

OBJECTIVE: To describe surgical techniques for and assess outcome of treatment of mitral regurgitation in dogs. DESIGN: Uncontrolled prospective study. ANIMALS: 18 dogs with naturally occurring mitral regurgitation. PROCEDURE: All dogs weighed > 5 kg (11 lb) and had severe mitral regurgitation, congestive heart failure (CHF), and no serious noncardiac disease. Left ventricular volume indices, left atrial size, and degree of mitral regurgitation were determined echocardiographically before and after surgery. Repair techniques included circumferential annuloplasty, placement of artificial chordae, chordal fenestration and papillary muscle splitting, and edge-to-edge repair. Factors predictive for surgery survival and resolution of CHF were determined. RESULTS: 12 dogs survived surgery. Factors predictive for surgery survival included weight > 10 kg (22 lb) and CHF of less than 6 months' duration. In 9 dogs, CHF resolved for a median period of 1 year (range, 4 months to 3 years) after surgery. One dog had stable CHF at 12 months. One dog died as a result of progressive CHF; another was euthanatized for a noncardiac reason. Left ventricular diastolic volume index was 226.9 +/- 117.7 cm3/m2 before surgery and 134.9 +/- 70.4 cm3/m2 at 6 months after surgery (n = 10). Factors predictive for resolution of CHF included left ventricular diastolic volume index < 250 cm3/m2 and systolic volume index < 70 cm3/m2. CONCLUSION AND CLINICAL RELEVANCE: Mitral valve repair may resolve CHF in dogs with severe mitral regurgitation, particularly in dogs that weigh > 10 kg and are treated within 6 months of the onset of CHF.