Optical projection tomography as a tool for 3D microscopy and gene expression studies

Current techniques for three-dimensional (3D) optical microscopy (deconvolution, confocal microscopy, and optical coherence tomography) generate 3D data by "optically sectioning" the specimen. This places severe constraints on the maximum thickness of a specimen that can be imaged. We have developed a microscopy technique that uses optical projection tomography (OPT) to produce high-resolution 3D images of both fluorescent and nonfluorescent biological specimens with a thickness of up to 15 millimeters. OPT microscopy allows the rapid mapping of the tissue distribution of RNA and protein expression in intact embryos or organ systems and can therefore be instrumental in studies of developmental biology or gene function.     

Evaluation of Urethral Stent Placement for Benign Urethral Obstructions in Dogs

Background

Benign urethral obstructions (BUO) in dogs result in substantial morbidity because of challenges with conventional therapies. Treatment of malignant urethral obstructions with intraluminal urethral stents is reported to successfully relieve obstructions.

Hypothesis/Objectives

To evaluate the efficacy and outcome of urethral stent placement for treatment of BUO in dogs.

Animals

Eleven client‐owned animals with urethral stents placed for treatment of BUO.

Methods

Retrospective study in which medical records were reviewed in dogs diagnosed with BUO and treated with a metallic urethral stent. Data collected included signalment, cause of benign obstruction, procedure time, size and type of stent, complications, and short‐ and long‐term outcome.

Results

Eleven dogs with 15 urethral stents were included. Intraluminal urethral stent(s) relieved the obstructions in all dogs. Four dogs had 2 stents placed in separate procedures because of incomplete patency after treatment (n = 1), inadvertent compression of the stent (n = 1), or tissue ingrowth through the stent (n = 2). The median continence score after stent placement was 10 of 10 (range 3–10) with 6 dogs being continent, 3 mildly incontinent, and 1 each moderately and severely incontinent. All owners considered their dog to have an excellent long‐term clinical outcome with long‐term urethral patency. The median follow‐up time was 24 months (range 4–48).

Conclusions and Clinical Importance

Urethral stents appear to be an effective treatment for benign urinary obstructions. Moderate to severe incontinence developed in a minority (12.5%) of dogs. Stents relieved obstructions in all dogs with an excellent long‐term outcome.     

Balloon dilatation of nasopharyngeal stenosis in a dog

CASE DESCRIPTION: A dog was examined because of a 6-month history of upper airway stridor that began after postoperative regurgitation of gastric contents. CLINICAL FINDINGS: Constant stridor was evident during inspiration and expiration, although it was worse during inspiration. The stridor was no longer evident when the dog's mouth was manually held open. Computed tomography, rhinoscopy, and fluoroscopy were used to confirm a diagnosis of nasopharyngeal stenosis. TREATMENT AND OUTCOME: The dog was anesthetized, and balloon dilatation of the stenosis was performed. Prednisone was prescribed for 4 weeks after the procedure to decrease fibrous tissue formation. Although the dog was initially improved, signs recurred 3.5 weeks later, and balloon dilatation was repeated. This time, however, triamcinolone was injected into the area of stenosis at the end of the dilatation procedure. Two months later, although the dog did not have clinical signs of stridor, a third dilatation procedure was performed because mild stenosis was seen on follow-up computed tomographic images; again, triamcinolone was injected into the area of stenosis at the end of the dilatation procedure. Three and 6 months after the third dilatation procedure, the dog reportedly was clinically normal. CLINICAL RELEVANCE: Findings suggest that balloon dilatation may be an effective treatment for nasopharyngeal stenosis in dogs.     

Use of cystoscopic-guided laser ablation for treatment of intramural ureteral ectopia in male dogs: four cases (2006-2007)

OBJECTIVE: To describe and evaluate the outcome of cystoscopic-guided laser ablation of intramural ureteral ectopia in male dogs. DESIGN: Retrospective case series. ANIMALS: 4 incontinent male dogs with intramural ureteral ectopia. PROCEDURES: Intramural ectopic ureters were diagnosed via preoperative computed tomography-IV urography and subsequent cystoscopy. Transurethral cystoscopic-guided laser ablation (diode laser [n = 3 dogs] and holmium:yttrium aluminum garnet laser [1]) was performed to proximally relocate the ectopic ureteral orifice to the urinary bladder. Fluoroscopy was used during the procedures to confirm that the ureteral tract was intramural and the ureteral orifice was intravesicular after the procedure. In 1 dog with bilateral ureteral ectopia, staged laser ablation was performed at 6-week intervals because of difficulty viewing the second ureter on the first attempt. All ureteral orifices were initially located in the middle to proximal portion of the prostatic portion of the urethra. Six weeks after surgery, imaging was repeated in 3 of 4 dogs. RESULTS: Postoperative dysuria or hematuria did not develop. All dogs were immediately continent after laser treatment and remained so at a median follow-up period of 18 months (range, 15 to 20 months) without medical management. Conclusions and CLINICAL RELEVANCE: Ureteral ectopia can cause urinary incontinence in male dogs and is usually associated with other urinary tract abnormalities. Cystoscopic-guided laser ablation provided an effective and minimally invasive alternative to surgical management of intramural ureteral ectopia.     

Effects of tramadol and o‐desmethyltramadol on canine innate immune system function

ObjectiveTramadol is a commonly used opioid analgesic in dogs, particularly in dogs with a compromised immune system. An opioid may be selected for its immunomodulatory effects. Consequently, the objective of this study was to investigate the effects of tramadol on immune system function by evaluating the effect of tramadol and o-desmethyltramadol (M1) on the function of canine leukocytes in vitro. The hypothesis was that tramadol and M1 would not alter polymorphonuclear leukocyte (PMN) phagocytosis, PMN oxidative burst, or stimulated leukocyte cytokine production capacity of tumor necrosis factor (TNF)-a, interleukin (IL)-6, and IL-10.Study designIn vitro pharmacodynamic study.AnimalsSix healthy dogs.MethodsBlood from six dogs was obtained and incubated with various concentrations of tramadol and M1. Phagocytosis and oxidative burst were assessed using flow cytometry, and lipopolysaccharide (LPS), lipoteichoic acid (LTA) and peptidoglycan (PG)-stimulated leukocyte production of TNF, IL-6, and IL-10 were measured using a canine specific multiplex assay.ResultsNo differences were detected in phagocytosis or oxidative burst with any drug concentration. Tramadol did not alter leukocyte cytokine production, however, M1 significantly blunted IL-10 production.ConclusionsTramadol and its metabolite M1 were sparing to PMN phagocytosis and oxidative burst in dogs in vitro. Tramadol did not alter leukocyte cytokine production, however, M1 blunted IL-10 production at clinically achievable concentrations suggesting that M1 may promote a proinflammatory shift.Clinical relevanceThese data suggest that tramadol has minimal effect on phagocytosis and oxidative burst, and may promote a proinflammatory shift. Therefore, tramadol may be an ideal opioid analgesic in dogs at high risk of infection. Further investigation in vivo is warranted.