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.

Outcomes Following Balloon Dilation of Benign Urethral Strictures in Dogs: Eight Cases (2005-2018).

Balloon dilation has been described infrequently as a treatment for benign urethral strictures in dogs but is often a first-line therapeutic option for humans. Additional evidence is needed to evaluate the potential role of this procedure in veterinary medicine. The aim of the study was to describe the techniques used and evaluate the response to balloon dilation of benign urethral strictures in dogs. Medical records were reviewed from eight client-owned dogs who underwent balloon dilation of a benign urethral stricture over a 13 yr period in this retrospective case series. Clinical signs improved for five of eight dogs after a single balloon dilation during a follow-up period of 1 wk to 3 yr. After a second procedure, an additional dog demonstrated improvement for 5.5 yr. Adverse outcomes included urinary incontinence in two dogs and recurrent bacteriuria in four dogs. Findings suggest that balloon dilation is an effective, minimally invasive procedure for the treatment of benign urethral strictures in dogs. Urinary incontinence, urinary tract infection, and stricture recurrence are potential outcomes for dogs undergoing this procedure either as a result of the nature of the underlying disease or as a result of the procedure.

Antimicrobial Activity of Ceftiofur and Penicillin With Gentamicin Against Escherichia coli and Streptococcus equi Subspecies zooepidemicus in an Ex Vivo Model of Equine Postpartum Uterine Disease.

The use of antimicrobials for the management of equine uterine disease is commonplace, with antibiotic selection generally based on empirical evidence or in vitro sensitivity results. However, the potential disconnect between these laboratory results and clinical efficacy in the mare raises concern for antibiotic failure and subsequent development of resistant organisms. In this work, we attempt to bridge this gap by using an ex vivo model of the equine postpartum uterus to quantitatively evaluate the antimicrobial activity of two commonly used antibiotic treatments in the mare (ceftiofur and penicillin with gentamicin). The activity of both of these treatments was evaluated in two different fluid environments (standard bacterial culture broth and equine postpartum uterine fluid) against clinical isolates of E. coli and S. zooepidemicus. Although treatment with ceftiofur was effective at reducing growth of S. zooepidemicus in equine postpartum uterine fluid, it did not reduce bacterial growth of E. coli. Treatment with procaine penicillin G with gentamicin achieved at least bacteriostatic activity against E. coli in both fluid types, and bactericidal activity against S. zooepidemicus in both fluid types. The intrauterine infusion of procaine penicillin G with gentamicin in cases of postpartum uterine disease caused by E. coli or S. zooepidemicus is supported by the results of this work.

Retrospective analysis of the effect of acid-suppressant therapy on clinicopathologic parameters of cats with chronic kidney disease.

Objectives The aim was to retrospectively evaluate the effects of acid-suppressant therapy in a population of cats with chronic kidney disease (CKD). The study objectives were to evaluate the effects of acid-suppressant therapy on clinicopathologic variables and progression of CKD over time. Methods The databases of two institutions were searched over an 11 year time span for cats fitting inclusion criteria for CKD. A total of 89 cats met the criteria for inclusion and were grouped according to either early (ie, stages 1-2) or advanced (ie, stages 3-4) CKD. Variables were statistically analyzed before and after treatment with either: (1) proton pump inhibitors (PPIs; n = 17), (2) histamine-2 receptor antagonists (H2RAs; n = 30), (3) combined acid-suppressant therapy (PPI + H2RA; n = 6) or (4) no acid-suppressant therapy (n = 36). Shapiro-Wilk testing and Q-Q plots were used to assess normality and variance, respectively. A complete randomized design with a mixed-effects repeated measures ANOVA was used to evaluate for differences in stage, treatment and time, as well as the interaction between these effects. Results A significant increase in blood creatinine concentration was found over time independent of severity of CKD and treatment group ( P = 0.0087). A significant increase in blood sodium concentration (change of 3.12 mmol/l) was found independent of stage in cats receiving PPI therapy ( P = 0.0109). A significant decrease in total blood magnesium (change of 0.15 mmol/l) was detected in two cats with early CKD receiving combined acid suppressants ( P = 0.0025). Conclusions and relevance Results of this retrospective study suggest that cats with CKD receiving PPI therapy may develop alterations in blood sodium concentrations but do not experience more rapid progression of CKD.

Gait scoring in dogs with thoracolumbar spinal cord injuries when walking on a treadmill.

BACKGROUND: An inexpensive method of generating continuous data on hind limb function in dogs with spinal cord injury is needed to facilitate multicentre clinical trials. This study aimed to define normal fore limb, hind limb coordination in dogs walking on a treadmill and then to determine whether reliable data could be generated on the frequency of hind limb stepping and the frequency of coordinated stepping in dogs with a wide range of severities of thoracolumbar spinal cord injury. RESULTS: Sixty-nine neurologically normal dogs of different body sizes including seven lame dogs were videotaped walking on the treadmill without prior training and all used the lateral gait of right fore, left hind, left fore, right hind (RF-LH-LF-RH). Severely paraparetic dogs were able to walk on the treadmill for a minimum of 75 seconds, scoring of which generated data representative of function in animals with extremely variable gaits. Fifty consecutive stepping cycles were scored by three observers in 18 dogs with a wide range of disability due to acute thoracolumbar spinal cord injury using a stepping score (hind limb steps/fore limb steps ×100), and a coordination score (coordinated hind limb steps/total hind limb steps ×100). Dogs were also scored using a previously validated ordinal open field score (OFS). Inter- and intraobserver agreement was high as assessed with Cronbach's alpha test for internal reliability. The stepping and coordination scores were significantly correlated to each other and to the OFS. CONCLUSIONS: Dogs with naturally occurring spinal cord injury can walk on a treadmill without prior training and their hind limb function can be scored reliably using a stepping score and coordination score. The only requirements for data acquisition are a treadmill and appropriately positioned video camera and so the system can be used in multicentre clinical trials to generate continuous data on neurologic recovery in dogs.

Canine hereditary ataxia in old english sheepdogs and gordon setters is associated with a defect in the autophagy gene encoding RAB24.

Old English Sheepdogs and Gordon Setters suffer from a juvenile onset, autosomal recessive form of canine hereditary ataxia primarily affecting the Purkinje neuron of the cerebellar cortex. The clinical and histological characteristics are analogous to hereditary ataxias in humans. Linkage and genome-wide association studies on a cohort of related Old English Sheepdogs identified a region on CFA4 strongly associated with the disease phenotype. Targeted sequence capture and next generation sequencing of the region identified an A to C single nucleotide polymorphism (SNP) located at position 113 in exon 1 of an autophagy gene, RAB24, that segregated with the phenotype. Genotyping of six additional breeds of dogs affected with hereditary ataxia identified the same polymorphism in affected Gordon Setters that segregated perfectly with phenotype. The other breeds tested did not have the polymorphism. Genome-wide SNP genotyping of Gordon Setters identified a 1.9 MB region with an identical haplotype to affected Old English Sheepdogs. Histopathology, immunohistochemistry and ultrastructural evaluation of the brains of affected dogs from both breeds identified dramatic Purkinje neuron loss with axonal spheroids, accumulation of autophagosomes, ubiquitin positive inclusions and a diffuse increase in cytoplasmic neuronal ubiquitin staining. These findings recapitulate the changes reported in mice with induced neuron-specific autophagy defects. Taken together, our results suggest that a defect in RAB24, a gene associated with autophagy, is highly associated with and may contribute to canine hereditary ataxia in Old English Sheepdogs and Gordon Setters. This finding suggests that detailed investigation of autophagy pathways should be undertaken in human hereditary ataxia.

Genome-wide linkage study of atopic dermatitis in West Highland White Terriers.

BACKGROUND: Canine atopic dermatitis (AD) is a common, heritable, chronic allergic skin condition prevalent in the West Highland White Terrier (WHWT). In canine AD, environmental allergens trigger an inflammatory response causing visible skin lesions and chronic pruritus that can lead to secondary bacterial and yeast infections. The disorder shares many of the clinical and histopathological characteristics of human AD and represents an animal model of this disorder that could be used to further elucidate genetic causes of human AD. Microsatellite markers genotyped in families of WHWTs affected with AD were used to perform a genome-wide linkage study in order to isolate chromosomal regions associated with the disorder. RESULTS: Blood samples and health questionnaires were collected from 108 WHWTs spanning three families. A linkage simulation using these 108 dogs showed high power to detect a highly penetrant mutation. Ninety WHWTs were genotyped using markers from the Minimal Screening Set 2 (MSS-2). Two hundred and fifty six markers were informative and were used for linkage analysis. Using a LOD score of 2.7 as a significance threshold, no chromosomal regions were identified with significant linkage to AD. LOD scores greater than 1.0 were located in a 56 cM region of chromosome 7. CONCLUSIONS: The study was unable to detect any chromosomal regions significantly linked to canine AD. This could be a result of factors such as environmental modification of phenotype, incorrect assignment of phenotype, a mutation of low penetrance, or incomplete genome coverage. A genome-wide SNP association study in a larger cohort of WHWTs may prove more successful by providing higher density coverage and higher statistical power.

The prevalence of albuminuria in dogs and cats in an ICU or recovering from anesthesia.

OBJECTIVE: To evaluate the prevalence of albuminuria in dogs and cats admitted to the ICU or recovering from an anesthetic event. DESIGN: Prospective clinical study over a 10-week period in 2003. SETTING: Veterinary teaching hospital. ANIMALS: One hundred and five dogs and 22 cats. INTERVENTIONS: Urine was collected from dogs and cats admitted to the ICU or recovering from an anesthetic event. When possible, a second urine sample was collected approximately 48 hours later from those animals that had albuminuria during the initial screening. MEASUREMENTS AND MAIN RESULTS: All dog samples and most cat samples were screened for albumin using a commercial point-of-care immunoassay. Aliquots of samples that tested positive were stored at -20 °C until subsequent albumin quantification via antigen capture ELISA. Albuminuria was detected in 63 of 105 (60.0%) dogs and in 14 of 22 (63.6%) cats; the prevalence was higher in animals admitted to ICU than in those recovering from anesthesia. In subsequent samples from 26 dogs, urine albumin decreased in 20 (76.9%) when compared with the first sample; urine albumin was undetectable in 5 (19.2%). In subsequent samples from 6 cats, 4 (66.7%) had decreases in urine albumin when compared with the first sample; 1 (16.7%) was negative for urine albumin. Eleven of 12 dogs (91.7%) and 3 of 4 cats (75%) that died within 3 days of admission to the ICU had abnormal urine albumin; whereas 52 of 93 (55.9%) and 11 of 18 (61.1%) dogs and cats, respectively, who survived more than 3 days had abnormal urine albumin. Dogs with albuminuria were at increased risk of death. CONCLUSIONS: The prevalence of albuminuria in animals admitted to the ICU or recovering from anesthesia is higher than reported previously and transient in some patients. The presence of albuminuria may be a negative prognostic indicator in this population.

The use of pooled vs serial urine samples to measure urine protein:creatinine ratios.

BACKGROUND: Evaluation of serial urine protein:creatinine (UPC) ratios is important in prognosticating chronic kidney disease and monitoring response to therapeutic interventions. Owing to random biologic variation in dogs with stable glomerular proteinuria, multiple determinations of UPC ratios often are recommended to reliably assess urine protein loss. This can be cost-prohibitive. OBJECTIVE: The purpose of this study was to evaluate agreement between the mean of 3 UPC ratios obtained on 3 separate urine samples per dog and a single UPC ratio obtained when aliquots of the separate samples were pooled and analyzed as 1 sample. METHODS: Three separate urine samples were collected from each of 25 dogs, both client-owned and members of a research colony. Protein and creatinine concentrations were measured in the supernatant of each sample using a biochemical analyzer, and the mean of the 3 UPC ratios was calculated. A 1.0 mL aliquot of each of the 3 samples from each dog was pooled to create a fourth sample for that dog, and the UPC ratio of the pooled sample was similarly determined. Agreement and correlation between the mean and pooled UPC ratios were assessed using Bland-Altman difference plots and regression analysis, respectively. RESULTS: The UPC ratio in the pooled samples was highly correlated (r=.9998, P<.0001) with the mean UPC ratio of the 3 separate samples. Strong agreement between results was demonstrated; a UPC ratio from a pooled sample was at most +/-20% different than the mean UPC ratio obtained from 3 separate samples. CONCLUSIONS: Measuring the UPC ratio in a pooled sample containing equal volumes of several different urine specimens from the same dog provides a reliable and cost-effective alternative to assessing multiple UPC ratios on several specimens from the same dog.

Evaluation of perinuclear anti-neutrophilic cytoplasmic autoantibodies as an early marker of protein-losing enteropathy and protein-losing nephropathy in Soft Coated Wheaten Terriers.

OBJECTIVE: To evaluate perinuclear anti-neutrophilic cytoplasmic autoantibody (pANCA) status in Soft Coated Wheaten Terriers (SCWTs) and SCWT-Beagle crossbred dogs and to correlate pANCA status of dogs with clinicopathologic variables of protein-losing enteropathy (PLE), protein-losing nephropathy (PLN), or both. ANIMALS: 13 SCWTs and 8 SCWT-Beagle crossbred dogs in a research colony and a control group comprising 7 dogs with X-linked hereditary nephropathy and 12 healthy SCWTs > 9 years old. PROCEDURES: Samples were obtained from dogs in the research colony every 6 months. At each sample-collection time point, serum concentrations of albumin, globulin, creatinine, and urea nitrogen; fecal concentration of alpha-proteinase inhibitor; and urinary protein-to-creatinine ratios were determined and correlated with pANCA status. RESULTS: 20 of 21 dogs in the research colony had positive results for pANCAs at a minimum of 2 time points, and 18 of 21 dogs had definitive evidence of disease. None of the control dogs had positive results for pANCAs. A positive result for pANCAs was significantly associated with hypoalbuminemia, and pANCAs preceded the onset of hypoalbuminemia on an average of 2.4 years. Sensitivity and specificity for use of pANCAs to predict development of PLE or PLN were 0.95 (95% confidence interval, 0.72 to 1.00) and 0.8 (95% confidence interval, 0.51 to 0.95), respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Most dogs in this study affected with PLE, PLN, or both had positive results for pANCAs before clinicopathologic evidence of disease was detected. Thus, pANCAs may be useful as an early noninvasive test of disease in SCWTs.

Recovery of pelvic limb function in dogs following acute intervertebral disc herniations.

Chondrodystrophoid breeds of dog are prone to explosive herniation of mineralized disc material into the thoracolumbar spinal canal. The resulting acute spinal cord injury may represent an excellent spontaneous model of acute traumatic spinal cord injury. The aims of this study were to quantify the recovery of dogs following acute disc herniations, to evaluate external factors that influence recovery, and to identify a group of dogs suitable for use in clinical trials on neuroprotective drugs. The gait of 88 dogs with thoracolumbar disc herniations was scored at the time of injury and 2, 4, and 12 weeks after surgical decompression. Dogs were placed into four groups dependent on the severity of presenting signs; dogs in group 1 had the most severe injury severity. Group 1 dogs showed a variable but incomplete recovery by 12 weeks. Dogs in groups 2 and 3 recovered uniformly but more completely, while dogs in group 4 made a rapid and excellent recovery and were deemed unsuitable for clinical trials. Combining dogs in groups 1, 2 and 3 produced a population of dogs with incomplete recovery by 12 weeks. Power analysis revealed that 87 such dogs would be needed per treatment group to detect a 20% change in function with a power of 95%. The number needed reduced drastically to 19 by eliminating dogs in group 1, but this produced less room for functional improvement. External factors did not appear to influence outcome. We conclude that dogs with spontaneous disc herniations provide a useful model of acute spinal cord injury for clinical trials.

Long-term functional outcome of dogs with severe injuries of the thoracolumbar spinal cord: 87 cases (1996-2001).

OBJECTIVE: To determine long-term (> 6 months) outcome of dogs with paraplegia and loss of hind limb deep pain perception (DPP) resulting from intervertebral disk herniation or trauma. DESIGN: Retrospective study. ANIMALS: 87 dogs. PROCEDURE: Outcome was determined as successful or unsuccessful. The association of neuroanatomic localization, breed, age, weight, sex, and (for dogs with intervertebral disk herniation) speed of onset of signs and duration of paraplegia prior to surgery with outcome was evaluated. Owners were contacted by telephone to identify long-term health problems. RESULTS: Nine of 17 dogs with traumatic injuries were treated, and 2 regained the ability to walk; none of the 17 dogs regained DPP. Sixty-four of 70 dogs with intervertebral disk herniation underwent surgery; 9 (14%) were euthanatized within 3 weeks after surgery (7 because of ascending myelomalacia), 37 (58%) regained DPP and the ability to walk, 7 (11%) regained the ability to walk without regaining DPP, and 11 (17%) remained paraplegic without DPP. Outcome was not associated with any of the factors evaluated, but speed of recovery of ambulation was significantly associated with body weight and age. Fifteen (41%) and 12 (32%) dogs that regained DPP had intermittent fecal and urinary incontinence, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that the prognosis for paraplegic dogs without DPP because of trauma was guarded, while dogs with disk herniation had a better chance of recovering motor function. A third of the dogs that recovered motor function had intermittent incontinence. Persistent loss of DPP did not preclude recovery of motor function, but such dogs remained incontinent.