Validation of DE50-MD dogs as a model for the brain phenotype of Duchenne muscular dystrophy

Duchenne muscular dystrophy (DMD), a fatal musculoskeletal disorder, is associated with neurodevelopmental disorders and cognitive impairment caused by brain dystrophin deficiency. Dog models of DMD represent key translational tools to study dystrophin biology and to develop novel therapeutics. However, characterization of dystrophin expression and function in the canine brain is lacking. We studied the DE50-MD canine model of DMD that has a missense mutation in the donor splice site of exon 50. Using a battery of cognitive tests, we detected a neurocognitive phenotype in DE50-MD dogs including reduced attention, problem-solving and exploration of novel objects. Through a combination of capillary immunoelectrophoresis, immunolabelling, qPCR and RNAScope in situ hybridization we show that regional dystrophin expression in the adult canine brain reflects that of humans, and that the DE50-MD dog lacks full length dystrophin (Dp427) protein expression but retains expression of the two shorter brain-expressed isoforms, Dp140 and Dp71. Thus, the DE50-MD dog is a translationally-relevant pre-clinical model to study the consequences of Dp427 deficiency in the brain and to develop therapeutic strategies for the neurological sequelae of DMD.

Improved Ablation Efficiency in PVI Guided by Contact Force and Local Impedance: Chronic Canine Model

Background: The purpose of this study was to assess the effect local impedance (LI) has on an ablation workflow when combined with a contact force (CF) ablation catheter.Methods: Left pulmonary vein isolation was performed in an in vivo canine model (N = 8) using a nominal (30 W) or an elevated (50 W) power strategy with a CF catheter. The catheter was enabled to measure LI prior to and during ablation. LI was visible for only one of the vein isolations.Results: Chronic block was achieved in all animals when assessed 30 ± 5 days post-ablation procedure with a median LI drop during RF ranging from 23.0 to 34.0 Ω. In both power cohorts, the median radiofrequency (RF) duration decreased if LI was visible to the operator (30 W only CF: 17.0 s; 30 W CF + LI: 14.0 s, p = 0.009; 50 W only CF: 6.0 s; 50 W CF + LI: 4.0 s, p = 0.019). An inverse relationship between the LI prior to RF delivery and the RF duration required to achieve an effective lesion was observed. There was no correlation between the magnitude of the applied force and the drop in LI, once at least 5 g was achieved.Conclusions: An elevated power strategy with the context of CF and LI led to the most efficient titration of successful RF energy delivery. The combination of feedback allows for customization of the ablation strategy based on local tissue variation rather than a uniform approach that could potentially lead to overtreatment. Higher LI drops were more readily achievable when an elevated power strategy was utilized, especially in conditions where the catheter was coupled against tissue with low resistivity. Clinical study is warranted to determine if there is an additive safety benefit to visualizing the dynamics of the tissue response to RF energy with LI when an elevated power strategy is used.

Immuno-histopathologic evaluation of mineralized plasmatic matrix in the management of horizontal ridge defects in a canine model (a split-mouth comparative study)

Our research aimed to investigate the effect of combining biphasic calcium phosphate (BCP) alloplast with mineralized plasmatic matrix (MPM) as compared with platelet-rich fibrin (PRF) on the quality and quantity of bone formation and maturation at surgically created horizontal critical-sized ridge defects (HRDs) in a canine model. We used a split-mouth design using the third and fourth mandibular premolars of the mongrel dogs. Twelve defects on the left side (experimental group, I) were managed with MPM composite mixed with BCP alloplast, MPM compact layer. On the right side (control group, II), another 12 defects were managed with PRF mixed with BCP alloplast, followed by the application of PRF compact strips. Finally, both were covered by a collagen membrane. Dogs were euthanized at 4, 8, and 12 weeks, and the studied defects were processed to evaluate treatment outcome, including mean percentage of bone surface area, collagen percentage, and osteopontin (OPN) immunoreaction. Our results revealed that the mean percentage of bone surface area was significantly increased in the experimental group treated with MPM at all time intervals as compared with the PRF group. Decreased collagen percentage and increased OPN immunoreactivity showed significant results in the MPM group as compared with PRF at 4 and 8 weeks postoperatively, respectively. In conclusion, MPM accelerates the formation of superior new bone quality when used in the treatment of HRDs.

Urine and Fecal Microbiota in a Canine Model of Bladder Cancer

Introduction: Urothelial carcinoma (UC) is the tenth most diagnosed cancer in humans worldwide. Dogs are a robust model for invasive UC as tumor development and progression is similar in humans and dogs. Recent studies on urine microbiota in humans revealed alterations in microbial diversity and composition in individuals with UC; however, the potential role of microbiota in UC has yet to be elucidated. Dogs could be valuable models for this research, but microbial alterations in dogs with UC have not been evaluated. Objective: The objective of this this pilot study was to compare the urine and fecal microbiota of dogs with UC (n = 7) and age-, sex-, and breed-matched healthy controls (n = 7). Methods: DNA was extracted from mid-stream free-catch urine and fecal samples using Qiagen Bacteremia and PowerFecal kits, respectively. 16S rRNA gene sequencing was performed followed by sequence processing and analyses (QIIME 2 and R). Results: Canine urine and fecal samples were dominated by taxa similar to those found in humans. Significantly decreased microbial diversity (Kruskal-Wallis: Shannon, p = 0.048) and altered bacterial composition were observed in the urine but not feces of dogs with UC (PERMANOVA: Unweighted UniFrac, p = 0.011). The relative abundances of Fusobacterium was also increased, although not significantly, in the urine and feces of dogs with UC. Conclusion: This study characterizes urine and fecal microbiota in dogs with UC, and it provides a foundation for future work exploring host-microbe dynamics in UC carcinogenesis, prognosis, and treatment.

Subcutaneous nerve stimulation in canine model of persistent atrial fibrillation v1

This animal protocol was approved by the Institutional Animal Care and Use Committee of the Indiana University School of Medicine and the Methodist Research Institute, Indianapolis, IN, and conformed to the Guide for Care and Use of Laboratory Animals. The surgical procedure is performed under anesthesia and should incorporate all local requirements for standards of animal experimentation, including methods of anesthesia, surgical environment, and post-operative monitoring and care.