The future of canine glaucoma therapy.

Canine glaucoma is a group of disorders that are generally associated with increased intraocular pressure (IOP) resulting in a characteristic optic neuropathy. Glaucoma is a leading cause of irreversible vision loss in dogs and may be either primary or secondary. Despite the growing spectrum of medical and surgical therapies, there is no cure, and many affected dogs go blind. Often eyes are enucleated because of painfully high, uncontrollable IOP. While progressive vision loss due to primary glaucoma is considered preventable in some humans, this is mostly not true for dogs. There is an urgent need for more effective, affordable treatment options. Because newly developed glaucoma medications are emerging at a very slow rate and may not be effective in dogs, work toward improving surgical options may be the most rewarding approach in the near term. This Viewpoint Article summarizes the discussions and recommended research strategies of both a Think Tank and a Consortium focused on the development of more effective therapies for canine glaucoma; both were organized and funded by the American College of Veterinary Ophthalmologists Vision for Animals Foundation (ACVO-VAF). The recommendations consist of (a) better understanding of disease mechanisms, (b) early glaucoma diagnosis and disease staging, (c) optimization of IOP-lowering medical treatment, (d) new surgical therapies to control IOP, and (e) novel treatment strategies, such as gene and stem cell therapies, neuroprotection, and neuroregeneration. In order to address these needs, increases in research funding specifically focused on canine glaucoma are necessary.

APHAKIC PHACOEMULSIFICATION AND AUTOMATED ANTERIOR VITRECTOMY, AND POSTRETURN MONITORING OF A REHABILITATED HARBOR SEAL (PHOCA VITULINA RICHARDSI) PUP.

A female harbor seal pup rescued along the coast of San Diego on 13 June 2012 was diagnosed with bilateral mature cataracts, apparently congenital, in association with vitreal herniation in the anterior chamber of each eye. The cataracts were surgically removed on 1 August 2012 with single-port aphakic phacoemulsification and automated anterior vitrectomy. Postoperative monitoring during the next several weeks indicated that vision had been functionally repaired and that she could visually orient to and capture live fish in three different environments and in the presence of other animals. Consequently, we equipped the seal with a satellite-linked radio transmitter and returned her to the Pacific Ocean on 21 November 2012, and then monitored her movements until radio contact ended on 2 March 2013. She remained along the San Diego coast from 21 November until 5 December 2012 when she relocated to the Coronado Islands and remained there until 26 December. She then traveled directly to San Clemente Island and remained foraging in the near-shore kelp beds there through 2 March 2013, when radio contact ended. To our knowledge, this is the first published report of cataract treatment in a marine mammal using high-frequency ultrasound to emulsify the lenses followed by suction removal of the emulsified microfragments (i.e., phacoemulsification). Moreover, the rapid postoperative recovery of the seal and its quick acclimation, orientation, navigation, and foraging in marine habitats after return to the Pacific Ocean indicates that these surgical procedures can be safe and effective treatments for cataracts in seals, with substantially reduced postsurgical complications relative to other types of lens fragmentation and removal procedures.

Efficacy of prophylactic antiglaucoma and anti-inflammatory medications in canine primary angle-closure glaucoma: a multicenter retrospective study (2004-2012).

PURPOSE: To evaluate long-term efficacy of antiglaucoma medications with or without combined topical anti-inflammatory treatment in preventing increased intraocular pressure and clinical signs of glaucoma in eyes considered at risk of the development of the disease. METHODS: Retrospective analysis identified 88 canine patients presenting with unilateral acute congestive primary angle-closure glaucoma (IOP > 25 mm Hg) and gonioscopic findings of pectinate ligament dysplasia and/or narrow or closed iridocorneal angle in the contralateral nonglaucomatous eye. Patients with histopathologic confirmation of pectinate ligament dysplasia or angle closure in the initial glaucomatous eye receiving prophylactic medical therapy in the contralateral eye were included. Time to medical failure for each antiglaucoma medication and efficacy of the combination therapy were evaluated. RESULTS: The most commonly affected pure-breds were the American Cocker Spaniel (20.4%) and Basset Hound (11.36%). The patients receiving demecarium bromide 0.125% had the longest estimated median time to medical failure at 330.0 days, followed by latanoprost 0.005%, dorzolamide hydrochloride 2.0%, and demecarium bromide 0.25% at 284.0 days, 272.5 days, and 143.0 days, respectively. The estimated median time to medical failure for patients receiving topical antiglaucoma and anti-inflammatory medication was 324.0 days versus 195.0 days in patients receiving antiglaucoma medication alone. Survival analysis showed no statistical significance. CONCLUSIONS: None of the four antiglaucoma medications evaluated statistically delayed medical failure when compared to each other. Although significance was not achieved, our data suggest that adjunctive use of topical anti-inflammatory medications may be beneficial in these cases.

Evaluation of intraorbital prosthetic pigmentation using 0.3 and 1.5 Tesla magnetic resonance imaging and computed tomography.

PURPOSE: To investigate the magnetic susceptibility artifact associated with pigmented intraorbital prosthetics when performing magnetic resonance imaging (MRI) and computed tomography (CT). Potential artifact reduction techniques were also investigated. STUDY DESIGN: Prospective study. METHODS: Five different-colored 20-millimeter small animal silicone intraorbital prosthetics and two equine prosthetics were evaluated using 0.3 and 1.5 Tesla (T) MRI and CT. MRI sequences included T1- (T1WI) and T2-weighted spin echo (T2WI), T2 gradient echo (T2*), short tau inversion recovery (STIR), and fluid-attenuated inversion recovery (FLAIR). When present, artifact size was measured using computerized software by three separate observers. Artifact reduction techniques included alterations in receiver bandwidth, field of view, slice thickness, and matrix size. RESULTS: The ferrous brown-pigmented prosthetic resulted in a magnetic susceptibility artifact with MRI. No artifact was observed on CT images. Interobserver variability was not statistically significant. For both the 0.3T and 1.5T MRI, the T2* sequence exhibited the largest artifact surface area followed by T2WI, T1WI, STIR, and FLAIR. Decreasing slice thickness showed a decrease in artifact size; however, this difference was not statistically significant. CONCLUSIONS: The ferrous substances in the brown intraorbital prosthetic resulted in a significant magnetic susceptibility artifact when performing MRI. Artifact reduction techniques did not significantly decrease artifact surface area. The use of ferrous brown-pigmented prosthetics and their potential to affect future MR imaging studies should be adequately discussed with pet owners.

Major breed distribution of canine patients enucleated or eviscerated due to glaucoma following routine cataract surgery as well as common histopathologic findings within enucleated globes.

PURPOSE: To identify the most significantly represented breeds that were enucleated or eviscerated due to intractable glaucoma following elective cataract surgery and to describe the major histopathologic features of enucleated globes. METHODS: A retrospective review of the Comparative Ocular Pathology Laboratory of Wisconsin database revealed 153 canine evisceration or enucleation submissions that met inclusion criteria. The most commonly represented breeds were identified, and enucleated globes of these breeds were further evaluated histologically (n = 69). Following standard hematoxylin & eosin staining, globes were evaluated for the presence of; corneal pathology, goniodysgenesis, pigment dispersion, pre-iridal fibrovascular membranes (PIFVMs), peripheral anterior, and/or posterior synechiation, inflammatory infiltrates, lens pathology, retinal pathology, and/or endophthalmitis. RESULTS: The most significantly represented breeds were the Cocker Spaniel (11.7%), Boston Terrier (10.5%), Labrador Retriever (6.5%), Bichon Frise (6.5%), Shih Tzu (5.8%), and Jack Russell Terrier (5.8%). Common histopathologic findings included inflammatory keratitis, the presence of PIFVMs, and lymphoplasmacytic uveal infiltrates. CONCLUSIONS: The Cocker Spaniel, Boston Terrier, Labrador Retriever, Bichon Frise, Shih Tzu, and Jack Russell Terrier may be at increased risk of the development of glaucoma following elective cataract surgery. These findings have potential clinical implications.

A review of orbital and intracranial magnetic resonance imaging in 79 canine and 13 feline patients (2004-2010).

OBJECTIVE: To review the distribution of orbital and intracranial disease in canine and feline patients undergoing magnetic resonance imaging (MRI) following referral to a veterinary ophthalmologist and to correlate results of MRI with pathologic conditions including neoplasia, suspected optic neuritis (ON) and orbital cellulitis. Recognized and emerging imaging techniques are reviewed. PROCEDURE: Medical records of 79 canine and 13 feline patients were reviewed. RESULTS: Neoplasia was diagnosed in 53/92 (57.6%) of patients. The most prevalent types of neoplasia were carcinoma (16/53, 30.1%), sarcoma (11/53, 20.8%), lymphoma (8/53, 15.1%) and presumptive meningioma (9/53, 17.0%). Carcinomas and sarcomas were characterized by bony lysis and intracranial/sinonasal extension. Lymphoma was generally unilateral, less invasive and originated from the ventromedial orbit. Intracranial masses representing presumptive meningiomas frequently exhibited a 'dural tail' sign. Diagnosis of suspected ON was made in 13 of 92 (14.1%) patients. Results of MRI in patients with suspected ON included unilateral optic nerve hyperintensity (3/13, 23.0%), bilateral optic nerve hyperintensity (1/13, 7.7%) and optic chiasmal hyperintensity (3/13, 23.0%). Seven suspected ON patients demonstrated intracranial multifocal patchy contrast enhancement (7/13, 53.8%). Diagnosis of orbital cellulitis was made in 12/92 (13.0%) patients. CONCLUSIONS: Orbital neoplasia was the most common pathologic condition detected. Essential Roentgen characteristics are helpful when diagnosing pathologic processes and providing prognoses in cases of orbital or intracranial disease. Magnetic resonance imaging comprises an important diagnostic component in cases of suspected ON. Emerging contrast and functional MRI techniques as well as SI data may increase our ability to characterize disease processes.

Development of a microarray chip for gene expression in rabbit ocular research.

PURPOSE: To develop a microarray for the rabbit that can be used for ocular gene expression research. METHODS: Messenger RNA was isolated from anterior segment tissues (cornea, conjunctiva, and iris) and posterior segment tissues (lens, retina, and sclera) of rabbit eyes and used to create two independent cDNA libraries through the NEIBank project. Clones from each of these libraries were sequenced from both the 5' and 3' ends. These sequences and those from the National Center for Biotechnology Information (NCBI) taxonomy database for rabbit were combined and electronically assembled into a set of unique nonoverlapping continuous sequences (contigs). For each contig, a homology search was performed using BLASTX and BLASTN against both the NCBI NR and NT databases to provide gene annotation. Unique contigs were sent to Agilent Technologies, where 60 base oligonucleotide probes were designed and synthesized, in situ, on two different arrays in an 8 array x 1900 element format. Glaucoma filtration surgery was performed on one eye of six rabbits. After 14 days, tissue was harvested from the conjunctiva and Tenon's capsule of both the surgically treated and untreated control eyes. Total RNA from each sample was labeled with cyanine dyes and hybridized to our custom microarrays. RESULTS: Of the 3,154 total probes present on the two arrays, 2,522 had a signal value above the background. The expression of 315 genes was significantly altered by glaucoma filtration surgery. Genes whose expression was altered included proteins associated with inflammatory response, defense response, and proteins involved in synthesis of the extracellular matrix. CONCLUSIONS: The results of this rabbit microarray study are consistent with those from other wound healing studies, indicating that this array can provide valid information on broad patterns of gene expression. This is the first microarray available for rabbit studies and is a valuable tool that can be used to study molecular events in the eye.

Surgical management of a malacic corneal ulcer in a greater one-horned Asian rhinoceros (Rhinoceros unicornis) using a free island tarsoconjunctival graft.

A greater one-horned Asian rhinoceros (Rhinoceros unicornis) presented for presumed ocular trauma to the left eye, with secondary bacterial infection, resulting in severe and progressive corneal ulceration. Following a poor response to medical therapy, the animal was anesthetized for further examination, and a bulbar conjunctival pedicle graft performed. This graft failed by 48-h postsurgery as a result of self-trauma. The animal was re-anesthetized, and a free island tarsoconjunctival graft performed. This second procedure was successful, resulting in globe preservation, cosmesis, and functional vision in the affected eye.

Microarray analysis of the failure of filtering blebs in a rat model of glaucoma filtering surgery.

PURPOSE: To generate data concerning changes in levels of protein expression associated with wound healing and bleb failure in a rat model of glaucoma filtration surgery (GFS), and to identify factors that may play a role in this process. METHODS: Of 36 Sprague-Dawley rats, GFS was performed on 27 by introducing a silicone cannula through a scleral tunnel under a conjunctival flap, resulting in aqueous-filtering blebs that failed over 8 to 13 days. The additional nine rats were used as the nonsurgical control. Nine blebs were harvested at each of days 0, 2, 5, and 12 and pooled, yielding three replicates of three blebs per time point. RNA was extracted, labeled, and hybridized to 230A rat GeneChip arrays (Affymetrix, Santa Clara, CA). RESULTS: Of the 15,924 probe sets/genes present on the array, 923 genes were indicated to have a significant treatment effect at P < 0.005. Eight gene expression clusters were identified that could be broadly classified into three basic patterns. These were an increase on day 2, a decrease on day 2, or an increase on either day 5 or 12. The greatest change occurred between days 0 and 2. The most heavily populated functional categories included growth factors, structural proteins, and matrix metalloproteinases. CONCLUSIONS: This study represents the first large-scale gene expression analysis after GFS. Expression patterns for known mediators of the bleb scarring process, including transforming growth factor-beta, connective tissue growth factor, and matrix metalloproteinases, were confirmed, and a number of mediators not previously associated with this process were identified.

Expression of connective tissue growth factor after glaucoma filtration surgery in a rabbit model.

PURPOSE: Connective tissue growth factor (CTGF) appears to play a significant role in mediating fibrosis in several tissues. To gain further understanding of the role of CTGF in the scar formation that occurs after glaucoma filtering surgery (GFS), experiments were performed in a rabbit model. METHODS: . Three experiments were performed: (1) CTGF and transforming growth factor (TGF)-beta expression were measured quantitatively after GFS, using ELISA. (2) After GFS conjunctival bleb tissues were immunostained for the presence of CTGF and TGF-beta. (3) Exogenous CTGF was injected into mitomycin-C (MMC)-treated filtering blebs and the scaring response compared to TGF-beta and physiological saline-injected blebs. RESULTS: CTGF and TGF-beta were expressed maximally by day 5 after surgery and were both shown to be present in the bleb tissues after GFS. The addition of exogenous CTGF and TGF-beta increased the rate of failure of GFS blebs. CONCLUSIONS: These data support the hypothesis that CTGF plays an important role in scarring and wound contracture after GFS. Inhibition of CTGF synthesis or its action may help prevent bleb failure and improve long-term GFS outcomes.