Acquired dysfunction of CFTR underlies cystic fibrosis-like disease of the canine gallbladder.

Mucocele formation in dogs is a unique and enigmatic muco-obstructive disease of the gallbladder caused by the amassment of abnormal mucus that bears striking pathological similarity to cystic fibrosis. We investigated the role of cystic fibrosis transmembrane conductance regulatory protein (CFTR) in the pathogenesis of this disease. The location and frequency of disease-associated variants in the coding region of CFTR were compared using whole genome sequence data from 2,642 dogs representing breeds at low-risk, high-risk, or with confirmed disease. Expression, localization, and ion transport activity of CFTR were quantified in control and mucocele gallbladders by NanoString, Western blotting, immunofluorescence imaging, and studies in Ussing chambers. Our results establish a significant loss of CFTR-dependent anion secretion by mucocele gallbladder mucosa. A significantly lower quantity of CFTR protein was demonstrated relative to E-cadherin in mucocele compared with control gallbladder mucosa. Immunofluorescence identified CFTR along the apical membrane of epithelial cells in control gallbladders but not in mucocele gallbladder epithelium. Decreases in mRNA copy number for CFTR were accompanied by decreases in mRNA for the Cl-/[Formula: see text] exchanger SLC26A3, K+ channels (KCNQ1, KCNN4), and vasoactive intestinal polypeptide receptor (VIPR1), which suggest a driving force for change in secretory function of gallbladder epithelial cells in the pathogenesis of mucocele formation. There were no significant differences in CFTR gene variant frequency, type, or predicted impact comparing low-risk, high-risk, and definitively diagnosed groups of dogs. This study describes a unique, naturally occurring muco-obstructive disease of the canine gallbladder, with uncanny similarity to cystic fibrosis, and driven by the underlying failure of CFTR function.NEW & NOTEWORTHY Cystic fibrosis transmembrane conductance regulatory protein (CFTR) genomic variants and expression of mRNA, protein, and electrogenic anion secretory activity of CFTR were characterized in dog gallbladder. Acquired inhibition of CFTR expression by gallbladder epithelium was identified as underpinning a naturally occurring muco-obstructive disease of the dog gallbladder that bears striking pathological similarity to animal models of cystic fibrosis.

Increased lipogenesis and lipidosis of gallbladder epithelium in dogs with gallbladder mucocele formation.

BACKGROUND: Gallbladder disease in people is frequently associated with disorders of lipid metabolism and metabolic syndrome. A recently emergent gallbladder disease of dogs, referred to as mucocele formation, is characterized by secretion of abnormal mucus by the gallbladder epithelium and is similarly associated with hyperlipidemia, endocrinopathy, and metabolic dysfunction. The cause of gallbladder mucocele formation in dogs is unknown. METHODS: A prospective case-controlled study was conducted to gain insight into disease pathogenesis by characterization of plasma lipid abnormalities in 18 dogs with gallbladder mucocele formation and 18 age and breed matched control dogs using direct infusion mass spectrometry for complex plasma lipid analysis. This analysis was complemented by histochemical and ultrastructural examination of gallbladder mucosa from dogs with gallbladder mucocele formation and control dogs for evidence of altered lipid homeostasis of the gallbladder epithelium. RESULTS: Gallbladder mucocele formation in dogs carried a unique lipidomic signature of increased lipogenesis impacting 50% of lipid classes, 36% of esterified fatty acid species, and 11% of complex lipid species. Broad enrichment of complex lipids with palmitoleic acid (16:1) and decreased abundance within complex lipids of presumptive omega-3 fatty acids eicosapentaenoic (20:5) and docosahexaenoic (22:6) was significant. Severe lipidosis of gallbladder epithelium pinpoints the gallbladder as involved causally or consequently in abnormal lipid metabolism. CONCLUSION: Our study supports a primary increase in lipogenesis in dogs with mucocele formation and abnormal gallbladder lipid metabolism in disease pathogenesis.

Evaluation of thoracic duct ligation and unilateral subphrenic pericardiectomy via a left fourth intercostal approach in normal canine cadavers.

OBJECTIVE: To investigate a left-sided fourth intercostal approach to thoracic duct (TD) ligation and unilateral subphrenic pericardiectomy in dogs. STUDY DESIGN: Retrospective computed tomography (CT) review and cadaveric study. ANIMALS: Thirteen dogs with idiopathic chylothorax and 10 canine cadavers. METHODS: A retrospective study of CT lymphangiograms in client-owned dogs with idiopathic chylothorax evaluated location and branching of the TD at the left fourth intercostal space. A cadaveric study evaluated the efficacy of TD ligation at this site. Following methylene blue mesenteric lymph node injection, TDs were identified through a left fourth intercostal thoracotomy, ligated, and sealed. Unilateral subphrenic pericardiectomy was performed through the same incision. Computed tomography scans were performed to determine the success of TD ligation. RESULTS: A review of lymphangiograms revealed a single TD in 10/13 clinical cases at the fourth intercostal space. Three cases had additional branches. Thoracic duct ligation via a left fourth intercostal thoracotomy was successful in nine out of 10 cadavers. A single branch was noted intraoperatively in six out of 10, and two branches were noted in four out of 10 cadavers. All branches were observed on the left side of the esophagus. CONCLUSION: TD ligation at the left fourth intercostal space was successfully performed in 9/10 canine cadavers and appeared feasible in a retrospective review of 10/13 clinical cases. Unilateral subphrenic pericardiectomy can also be performed via this approach. CLINICAL SIGNIFICANCE: Fewer thoracic duct branches at this location in comparison with the standard caudal location may simplify TD ligation. If elected, unilateral subphrenic pericardiectomy can be performed through the same incision. Further investigation in clinical patients is warranted.

A postnatal network of co-hepato/pancreatic stem/progenitors in the biliary trees of pigs and humans.

A network of co-hepato/pancreatic stem/progenitors exists in pigs and humans in Brunner's Glands in the submucosa of the duodenum, in peribiliary glands (PBGs) of intrahepatic and extrahepatic biliary trees, and in pancreatic duct glands (PDGs) of intrapancreatic biliary trees, collectively supporting hepatic and pancreatic regeneration postnatally. The network is found in humans postnatally throughout life and, so far, has been demonstrated in pigs postnatally at least through to young adulthood. These stem/progenitors in vivo in pigs are in highest numbers in Brunner's Glands and in PDGs nearest the duodenum, and in humans are in Brunner's Glands and in PBGs in the hepato/pancreatic common duct, a duct missing postnatally in pigs. Elsewhere in PDGs in pigs and in all PDGs in humans are only committed unipotent or bipotent progenitors. Stem/progenitors have genetic signatures in liver/pancreas-related RNA-seq data based on correlation, hierarchical clustering, differential gene expression and principal component analyses (PCA). Gene expression includes representative traits of pluripotency genes (SOX2, OCT4), endodermal transcription factors (e.g. SOX9, SOX17, PDX1), other stem cell traits (e.g. NCAM, CD44, sodium iodide symporter or NIS), and proliferation biomarkers (Ki67). Hepato/pancreatic multipotentiality was demonstrated by the stem/progenitors' responses under distinct ex vivo conditions or in vivo when patch grafted as organoids onto the liver versus the pancreas. Therefore, pigs are logical hosts for translational/preclinical studies for cell therapies with these stem/progenitors for hepatic and pancreatic dysfunctions.

Gallbladder microbiota in healthy dogs and dogs with mucocele formation.

To date studies have not investigated the culture-independent microbiome of bile from dogs, a species where aseptic collection of bile under ultrasound guidance is somewhat routine. Despite frequent collection of bile for culture-based diagnosis of bacterial cholecystitis, it is unknown whether bile from healthy dogs harbors uncultivable bacteria or a core microbiota. The answer to this question is critical to understanding the pathogenesis of biliary infection and as a baseline to exploration of other biliary diseases in dogs where uncultivable bacteria could play a pathogenic role. A pressing example of such a disease would be gallbladder mucocele formation in dogs. This prevalent and deadly condition is characterized by excessive secretion of abnormal mucus by the gallbladder epithelium that can eventually lead to rupture of the gallbladder or obstruction of bile flow. The cause of mucocele formation is unknown as is whether uncultivable, and therefore unrecognized, bacteria play any systematic role in pathogenesis. In this study we applied next-generation 16S rRNA gene sequencing to identify the culture-negative bacterial community of gallbladder bile from healthy dogs and gallbladder mucus from dogs with mucocele formation. Integral to our study was the use of 2 separate DNA isolations on each sample using different extraction methods and sequencing of negative control samples enabling recognition and curation of contaminating sequences. Microbiota findings were validated by simultaneous culture-based identification, cytological examination of bile, and fluorescence in-situ hybridization (FISH) performed on gallbladder mucosa. Using culture-dependent, cytological, FISH, and 16S rRNA sequencing approaches, results of our study do not support existence of a core microbiome in the bile of healthy dogs or gallbladder mucus from dogs with mucocele formation. Our findings further document how contaminating sequences can significantly contribute to the results of sequencing analysis when performed on samples with low bacterial biomass.

The use of autologous skeletal muscle progenitor cells for adjunctive treatment of presumptive urethral sphincter mechanism incompetence in female dogs.

BACKGROUND: Urethral sphincter mechanism incompetence (USMI) is a common problem in female dogs, but some dogs fail to achieve continence with standard treatment. Urethral submucosal injection of autologous skeletal muscle progenitor cells (skMPCs) previously has been shown to restore urethral function in a canine model of USMI. HYPOTHESIS/OBJECTIVE: To determine if urethral submucosal injection of skMPC alters continence in dogs with USMI that had previously failed standard medical management. We hypothesized that the injections would lead to improved continence. ANIMALS: Fifteen client-owned dogs with USMI that had failed standard medical management. METHODS: Dogs were prospectively enrolled into a single-armed clinical trial. Once enrolled, a triceps muscle of each dog was biopsied; the tissue specimens were digested, cultured, and expanded to 100 million cells before injection into the urethral submucosa using a surgical approach. Continence was assessed at baseline and 3, 6, 12, and 24 months post-injection using continence scores and urethral pressure profilometry. RESULTS: Median continence scores increased significantly from baseline at 3, 6, 12, and 24 months. Increases were seen in 14 of 15 dogs with 7, 6 or 1 dog achieving scores of 5, 4 or 3, respectively. Additional medication was required to achieve continence in all but 2 dogs. CONCLUSIONS AND CLINICAL IMPORTANCE: Urethral submucosal injection of skMPC can be used adjunctively to improve continence in dogs with difficult to manage USMI. The procedure is labor intensive but well tolerated; most dogs will require continued medication to remain continent.

Patch grafting of organoids of stem/progenitors into solid organs can correct genetic-based disease states.

Patch grafting, a novel strategy for transplantation of stem/progenitor organoids into porcine livers, has been found successful also for organoid transplantation into other normal or diseased solid organs in pigs and mice. Each organoid contained ∼100 cells comprised of biliary tree stem cells (BTSCs), co-hepato/pancreatic stem/progenitors, and partnered with early lineage stage mesenchymal cells (ELSMCs), angioblasts and precursors to endothelia and stellate cells. Patch grafting enabled transplantation into livers or pancreases of ≥108th (pigs) or ≥106th-7th (mice) organoids/patch. Graft conditions fostered expression of multiple matrix-metalloproteinases (MMPs), especially secretory isoforms, resulting in transient loss of the organ's matrix-dictated histological features, including organ capsules, and correlated with rapid integration within a week of organoids throughout the organs and without emboli or ectopic cell distribution. Secondarily, within another week, there was clearance of graft biomaterials, followed by muted expression of MMPs, restoration of matrix-dictated histology, and maturation of donor cells to functional adult fates. The ability of patch grafts of organoids to rescue hosts from genetic-based disease states was demonstrated with grafts of BTSC/ELSMC organoids on livers, able to rescue NRG/FAH-KO mice from type I tyrosinemia, a disease caused by absence of fumaryl acetoacetate hydrolase. With the same grafts, if on pancreas, they were able to rescue NRG/Akita mice from type I diabetes, caused by a mutation in the insulin 2 gene. The potential of patch grafting for cell therapies for solid organs now requires translational studies to enable its adaptation and uses for clinical programs.

A lateral surgical approach to the larynx may impair cervical esophageal function in dogs.

OBJECTIVE: To determine whether muscle-sparing laryngoplasty results in fewer changes in swallowing function compared to standard surgical treatment for laryngeal paralysis. ANIMALS: 12 clinically normal sexually intact male Beagles. PROCEDURES: Group A dogs (n = 4) had a standard approach to the larynx, with left arytenoid cartilage lateralization. Group B dogs (n = 4) had a muscle-sparing laryngoplasty performed with the thyropharyngeus muscle fibers bluntly separated, and the cricoarytenoideus dorsalis muscle spared. Pre- and 24-hour postoperative fluoroscopic swallowing studies were performed and graded. Larynges were harvested after humane euthanasia, and glottic area was measured. Group C dogs (n = 4) acted as controls, with surgical dissection ending lateral to the thyropharyngeus muscle, arytenoid lateralization not performed, and the dogs not euthanized. The study was performed between October 15, 2011 and May 15, 2021. RESULTS: Changes in pharyngeal and upper esophageal sphincter function were not detected in any group. There was no difference in glottic area between treatment groups. Aspiration of liquid was not a consistent finding. Two dogs in each treatment group developed moderate to severe cervical esophageal paresis. This did not occur in control dogs. CLINICAL RELEVANCE: We found no evidence to support our hypothesis that muscle-sparing laryngoplasty results in less severe changes in swallowing function compared to a standard technique. The cervical esophageal paresis identified in both treatment groups could increase the risk of postoperative aspiration pneumonia in dogs treated for laryngeal paralysis via a lateral approach to the larynx. Further study to determine the frequency, cause, and duration of esophageal dysfunction is warranted.

Cranial internal hemipelvectomy (iliectomy) with limb sparing for a dog with ilial chondrosarcoma: A case report.

Cranial internal hemipelvectomy can be successful for excision of ilial CSA with minimal complications. Iliectomy with adjuvant radiation therapy was well tolerated in a dog with grade II ilial CSA. The dog survived 1,271 days postoperatively and supposedly succumbed to a disease process unrelated to the CSA.

Development of an ultrasound-guided transgluteal injection of the pudendal nerve in cats: a cadaveric study.

OBJECTIVE: To develop an ultrasound-guided interfascial plane technique for injection of the pudendal nerve near its sacral origin in cats. STUDY DESIGN: Prospective, randomized, anatomical study. ANIMALS: A group of 12 feline cadavers. METHODS: Gross and ultrasound anatomy of the ischiorectal fossa, the pudendal nerve relationship with parasacral structures, and the interfascial plane were described. Computed tomography was employed to describe a cranial transgluteal approach to the pudendal nerve. Bilateral ultrasound-guided injections were performed in eight cadavers using low [(LV) 0.1 mL kg-1] or high volume [(HV) 0.2 mL kg-1] of ropivacaine-dye solution. Dissections were performed to determine successful staining of the pudendal nerve (>1 cm) and inadvertent staining of the sciatic nerve, and any rectal, urethral, or intravascular puncture. Pudendal nerve staining in groups LV and HV were compared using Fisher's exact and Wilcoxon rank-sum test as appropriate (p = 0.05). RESULTS: The pudendal nerve and its rectal perineal and sensory branches coursed through the ischiorectal fossa, dorsomedial to the ischiatic spine. The pudendal nerve was not identified ultrasonographically, but the target plane was identified between the sacral transverse process, the ischiatic spine, the pelvic fascia and the rectum, and it was filled with dye solution. Both branches of the pudendal nerve were completely stained 75% and 87.5% in groups LV and HV, respectively (p = 1.00). The dorsal aspect of the sciatic nerve was partially stained in 37% of injections in group HV. Rectal or urethral puncture and intravascular injection were not observed. CONCLUSIONS AND CLINICAL RELEVANCE: In cats, ultrasound-guided cranial transgluteal injection successfully stained the pudendal nerve in at least 75% of attempts, regardless of injectate volume. Group HV had a greater probability of sciatic nerve staining.

Creation of a Continent Urinary Bladder Reservoir Vascularized by Omentum as a Possible Surgical Option for Canine Trigonal/Urethral Urothelial Carcinoma.

Surgical procedures that maintain continence with minimal complication following resection of trigono-urethral urothelial carcinoma (UC) are limited in canines; therefore, palliative options are often pursued. A feasible tumor resection option may improve disease control and survival. The study's objective was to evaluate a continent urine reservoir created from the urinary bladder body and vascularized solely by omentum. We hypothesized that a viable urine reservoir could be created, and staged omentalization would provide improved vascularity. Nine normal female Beagles were randomized to one of three groups. Group A urinary bladders were transected cranial to the ureteral papillae to create a closed bladder vesicle which was concomitantly omentalized. Group B underwent omentalization two weeks prior to vesicle creation. Based on Group A and B results, Group C underwent neoureterocystostomy and omentalization followed by neoreservoir formation and tube cystostomy 2 weeks later. Serial ultrasounds and histopathology confirmed adequate omental neovascularization in Groups B and C with continent Group C neoreservoirs maintained for 2 months. Some pylectasia and ureteral dilation was documented in all Group C dogs at variable timepoints. Progressive hydroureteronephrosis developed in 2/6 kidneys. Transient azotemia was noted in only 1 Group C dog, although all developed treatable urinary tract infections. The sample size is limited, and the efficacy of this technique in providing disease control for UC is unknown. However, this novel option could allow for primary UC resection while providing continence and limiting complications. Postoperative local or systemic adjuvant therapy, ultrasonographic neoreservoir monitoring, and BRAF analysis would be indicated.

Patch grafting, strategies for transplantation of organoids into solid organs such as liver.

Epithelial cell therapies have been at an impasse because of inefficient methods of transplantation to solid organs. Patch grafting strategies were established enabling transplantation of ≥107th organoids/patch of porcine GFP+ biliary tree stem/progenitors into livers of wild type hosts. Grafts consisted of organoids embedded in soft (~100 Pa) hyaluronan hydrogels, both prepared in serum-free Kubota's Medium; placed against target sites; covered with a silk backing impregnated with more rigid hyaluronan hydrogels (~700 Pa); and use of the backing to tether grafts with sutures or glue to target sites. Hyaluronan coatings (~200-300 Pa) onto the serosal surface of the graft served to minimize adhesions with neighboring organs. The organ's clearance of hyaluronans enabled restoration of tissue-specific paracrine and systemic signaling, resulting in return of normal hepatic histology, with donor parenchymal cells uniformly integrated amidst host cells and that had differentiated to mature hepatocytes and cholangiocytes. Grafts containing donor mature hepatocytes, partnered with endothelia, and in the same graft biomaterials as for stem/progenitor organoids, did not engraft. Engraftment occurred if porcine liver-derived mesenchymal stem cells (MSCs) were co-transplanted with donor mature cells. RNA-seq analyses revealed that engraftment correlated with expression of matrix-metalloproteinases (MMPs), especially secreted isoforms that were found expressed strongly by organoids, less so by MSCs, and minimally, if at all, by adult cells. Engraftment with patch grafting strategies occurred without evidence of emboli or ectopic cell distribution. It was successful with stem/progenitor organoids or with cells with a source(s) of secreted MMP isoforms and offers significant potential for enabling cell therapies for solid organs.

A randomized, prospective, comparative trial of a variable-angle versus fixed-angle endoscope for exploratory thoracoscopy in canine cadavers.

OBJECTIVE: To compare the efficacy of a variable-angle endoscope (VAE) for canine thoracoscopic exploration to a traditional fixed-angle endoscope (FAE). STUDY DESIGN: Randomized, prospective, comparative study. SAMPLE POPULATION: Five fresh canine cadavers. METHODS: Twelve predetermined anatomical locations were labeled after median sternotomy in each cadaveric thorax. Two board-certified veterinary surgeons performed thoracoscopic evaluation of each thorax using a fixed-angle (30°) and a variable-angle (0°-120°) endoscope with and without lungs mechanically ventilated. The order of surgeon, lung ventilation, and endoscope were determined using a randomized block design. Time to visualize each anatomical location was compared for surgeon, endoscope, and lung ventilation status. Primary outcome measures were time to individual anatomical location, total simulated thoracoscopic exploration time, and ability to identify anatomical location within the designated time period. RESULTS: Lung ventilation (difference = 184 seconds, P = .015, 95% CI = 45-342 seconds) and endoscope type (difference = 112 seconds, P = .029; 95% CI = 10-213 seconds) had an effect on the cumulative time for complete thoracoscopic exploration. The VAE shortened the time to identify three of the 12 anatomical locations when controlling for the effects of lung ventilation. Use of the VAE did not improve time to identification for any locations compared to the FAE when lungs were not ventilated. The VAE facilitated significantly shorter cumulative thoracoscopic exploration time compared with the FAE. Failure to identify predetermined locations was more common with the FAE than with the VAE. CONCLUSION: Use of a rigid VAE decreased cumulative thoracoscopic exploration time and provided an alternative to one-lung ventilation for circumventing the visual impediments of lung ventilation. CLINICAL SIGNIFICANCE: This cadaveric study provides evidence that one-lung ventilation and use of a VAE may improve surgeon efficiency during exploratory thoracoscopy.

Ligation of the Maxillary Artery Prior to Caudal Maxillectomy in the Dog-A Description of the Technique, Retrospective Evaluation of Blood Loss, and Cadaveric Evaluation of Maxillary Artery Anatomy.

Two different surgical techniques have been described for performing caudal maxillectomies in dogs including the intraoral (IO) and combined dorsolateral and intraoral (DL-IO) approach. Hemorrhage is the most common intraoperative complication reported during these procedures as maxillary arterial ligation is not performed until after all osteotomies and mobilization of tumor-bearing bone. The objectives of this study were to describe a modified approach for caudal maxillectomy in the dog involving preligation of the maxillary artery, to retrospectively evaluate the ability of this modified approach to limit hemorrhage in a cohort of 22 dogs, and to clarify the vascular anatomy of the maxillary artery and its branches in relation to associated nerves. Medical records were retrospectively reviewed for cases that had caudal maxillectomy via a combined approach (with or without preligation of the maxillary artery) from January 1, 2004 to December 31, 2019. Twenty-two cases were identified, six without, and 16 with arterial preligation, respectively. Osteotomies were completed with a high-speed handpiece and rotary bur (n = 18), or oscillating bone saw (n = 4). All six (100%) dogs in the traditional DL-IO group developed hypotension under general anesthesia. Four (67%) of these required intraoperative blood transfusions, one of which required an additional postoperative blood transfusion. In contrast, only one of 16 (6%) dogs in the modified DL-IO group required an intraoperative blood transfusion, and only three (19%) developed hypotension. Moreover, a significant association was detected between postoperative PCV and the two different surgical approaches (P = 0.021). These results demonstrate the effectiveness of preligation of the maxillary artery in preventing hemorrhage in caudal maxillectomies in dogs and this represents an improvement in outcome over previously reported studies. Decreased intraoperative hemorrhage may improve surgical exposure and decrease overall patient morbidity.

Computed tomography in two recumbencies aides in the identification of pulmonary bullae in dogs with spontaneous pneumothorax.

Spontaneous pneumothorax presents a unique diagnostic and therapeutic challenge in veterinary medicine, specifically with regard to accurate identification of bullous lesions. Positioning of dogs with spontaneous pneumothorax during CT has not previously been evaluated. This retrospective, diagnostic accuracy study was performed to evaluate the sensitivity, positive predictive value (PPV), and interobserver variability for detection of pulmonary bullae with dogs positioned in multiple recumbencies. Dogs underwent CT in sternal and dorsal recumbency followed by thoracic exploration via median sternotomy. Three American College of Veterinary Radiology-certified veterinary radiologists blinded to surgical findings reviewed dorsal and sternal images simultaneously. Severity of pneumothorax, degree of atelectasis, lesion location and size, and view in which lesions were most confidently identified were compared to surgical and histologic findings. Sensitivities and PPVs for bulla detection ranged from 57.7% to 69.2% and 62.1% to 78.9%, respectively. For two of the 3 radiologists, the location of bullae in the thorax was significantly associated with the recumbency in which the lesion was best identified. Degree of atelectasis was found to be associated with the ability to identify lesions (P ≤ .02). The interobserver variability for identification was good (κ = 0.670). The sensitivity of CT when performed in both sternal and dorsal recumbency is similar to that previously reported. Because the distribution of bullae is unknown prior to advanced imaging and bulla location affects which recumbency is most useful for identification, acquisition of CT images in both sternal and dorsal recumbency may improve detection of bullous lesions and aid surgical planning.

An off-the-shelf artificial cardiac patch improves cardiac repair after myocardial infarction in rats and pigs.

Cell therapy has been a promising strategy for cardiac repair after injury or infarction; however, low retention and engraftment of transplanted cells limit potential therapeutic efficacy. Seeding scaffold material with cells to create cardiac patches that are transplanted onto the surface of the heart can overcome these limitations. However, because patches need to be freshly prepared to maintain cell viability, long-term storage is not feasible and limits clinical applicability. Here, we developed an off-the-shelf therapeutic cardiac patch composed of a decellularized porcine myocardial extracellular matrix scaffold and synthetic cardiac stromal cells (synCSCs) generated by encapsulating secreted factors from isolated human cardiac stromal cells. This fully acellular artificial cardiac patch (artCP) maintained its potency after long-term cryopreservation. In a rat model of acute myocardial infarction, transplantation of the artCP supported cardiac recovery by reducing scarring, promoting angiomyogenesis, and boosting cardiac function. The safety and efficacy of the artCP were further confirmed in a porcine model of myocardial infarction. The artCP is a clinically feasible, easy-to-store, and cell-free alternative to myocardial repair using cell-based cardiac patches.

Lower urinary tract transitional cell carcinoma in cats: Clinical findings, treatments, and outcomes in 118 cases.

BACKGROUND: Lower urinary tract transitional cell carcinoma (TCC) is an important but rarely described disease of cats. OBJECTIVES: To report the clinical characteristics, treatments, and outcomes in a cohort of cats with lower urinary tract TCC and to test identified variables for prognostic relevance. ANIMALS: One-hundred eighteen client-owned cats with lower urinary tract carcinoma. METHODS: Medical records were retrospectively reviewed to obtain information regarding clinical characteristics, treatments, and outcomes. Recorded variables were analyzed statistically. RESULTS: Median age of affected cats was 15 years (range, 5.0-20.8 years) and median duration of clinical signs was 30 days (range, 0-730 days). The trigone was the most common tumor location (32/118; 27.1%) as assessed by ultrasound examination, cystoscopy, or both. Treatment was carried out in 73 of 118 (61.9%) cats. Metastatic disease was documented in 25 of 118 (21.2%) cats. Median progression-free survival and survival time for all cats were 113 days (95% confidence interval [CI], 69-153) and 155 days (95% CI, 110-222), respectively. Survival increased significantly (P < .001) when comparing cats across the ordered treatment groups: no treatment, treatment without partial cystectomy, and treatment with partial cystectomy. Partial cystectomy (hazard ratio [HR], 0.31; 95% CI, 0.17-0.87) and treatment with nonsteroidal anti-inflammatory drugs (HR, 0.55; 95% CI, 0.33-0.93) were significantly associated with longer survival times. CONCLUSIONS AND CLINICAL IMPORTANCE: The results support treatment using partial cystectomy and NSAIDs in cats with TCC.

Cardiac Stromal Cell Patch Integrated with Engineered Microvessels Improves Recovery from Myocardial Infarction in Rats and Pigs.

The vascularized cardiac patch strategy is promising for ischemic heart repair after myocardial infarction (MI), but current fabrication processes are quite complicated. Vascularized cardiac patches that can promote concurrent restoration of both the myocardium and vasculature at the injured site in a large animal model remain elusive. The safety and therapeutic benefits of a cardiac stromal cell patch integrated with engineered biomimetic microvessels (BMVs) were determined for treating MI. By leveraging a microfluidic method employing hydrodynamic focusing, we constructed the endothelialized microvessels and then encapsulated them together with therapeutic cardiosphere-derived stromal cells (CSCs) in a fibrin gel to generate a prevascularized cardiac stromal cell patch (BMV-CSC patch). We showed that BMV-CSC patch transplantation significantly promoted cardiac function, reduced scar size, increased viable myocardial tissue, promoted neovascularization, and suppressed inflammation in rat and porcine MI models, demonstrating enhanced therapeutic efficacy compared to conventional cardiac stromal cell patches. BMV-CSC patches did not increase renal and hepatic toxicity or exhibit immunogenicity. We noted a significant increase in endogenous progenitor cell recruitment to the peri-infarct region of the porcine hearts treated with BMV-CSC patch as compared to those that received control treatments. These findings establish the BMV-CSC patch as a novel engineered-tissue therapeutic for ischemic tissue repair.

Ability of positive and negative contrast computed tomographic peritoneography to delineate canine liver lobe fissures.

OBJECTIVE: To determine the efficacy of contrast peritoneography in delineating liver lobe fissures by using computed tomography (CT). STUDY DESIGN: Prospective cross-sectional study. ANIMALS: Canine cadavers (n = 10). METHODS: An abdominal CT image of each cadaver was obtained under three conditions: without peritoneal contrast (precontrast), after insufflation of the peritoneal cavity with CO2 (negative contrast), and after intraperitoneal administration of iohexol (positive contrast). Images were reconstructed in three planes. Three reviewers independently scored each plane/contrast combination. Observers measured the length of each fissure in each plane. The measurable length of fissures was compared between techniques but was not compared with the actual length on cadavers. RESULTS: When assessment of all fissures was combined, measurements were obtained in 96 to 108 of 120 (80%-90%) positive contrast studies, 56 to 96 of 120 (47%-80%) negative contrast studies, and 12 to 32 of 120 (10%-27%) precontrast CT images. All observers measured fissures in all planes that were approximately two times longer on positive compared with negative contrast studies and two times longer on negative contrast studies compared with CT images without contrast. Measurable fissure lengths were greater on positive contrast CT images (P < .0001) than on other CT images. CONCLUSION: Positive contrast CT peritoneography improved the delineation of hepatic interlobar fissures in normal canine cadavers. CLINICAL SIGNIFICANCE: These results justify further studies to determine the ability to locate large hepatic masses in dogs with positive CT. Positive contrast CT peritoneography may assist treatment planning in dogs with large hepatic tumors.