Aqueous humor myocilin protein levels in normal, genetic carriers, and glaucoma Beagles

Objective The gene (myocilin: MYOC) has been attributed to be involved in over 6% of inherited types of human glaucoma, the highest correlation for any gene to date. This study determines myocilin protein levels in the aqueous humor (AH) of normal laboratory quality, genetic carrier (offspring of normal laboratory quality and POAG Beagles), and primary open angle glaucoma (POAG) Beagles. Materials and methods Eighteen dogs were used and classified as either normal, carrier or having mild, moderate or advanced POAG. A 0.1‐mL sample of AH was drawn from the anterior chamber of each dog in the study and frozen on dry ice. A modified Coomassie stain and Western blot, using a polyclonal rabbit antihuman myocilin antibody (Santa Cruz Biotechnologies, Santa Cruz, CA), was run on each sample to compare the myocilin levels. A purified human trabecular meshwork excreted myocilin protein sample was used as a control (Alcon Research Laboratories, Fort Worth, TX) and its band/densitometry measurement was defined as one unit of myocilin for comparisons. Results Comparisons of AH myocilin levels differed among normal laboratory quality, genetic carrier, and POAG Beagles at different stages of the disease. In the normal laboratory, Beagles the AH myocilin measured 0.817 ± 0.075 units (mean ± SEM); in the carrier Beagles the AH myocilin was 3.117 ± 0.290 units. As POAG progressed, myocilin protein levels also increased to 6.097 ± 0.810, 8.844 ± 1.079, and 17.228 ± 1.198 units in the early, moderate, and advanced forms, respectively. Overall comparisons between normal, carrier and all POAG Beagles combined showed significant differences (P < 0.0010). Individual comparisons between normal and carrier eyes showed significant differences (P < 0.0193). Comparisons between normal and all POAG eyes also showed significant differences (P < 0.0426). Conclusion This study shows myocilin protein is present in normal Beagles, markedly increased in POAG Beagles, and mildly increased in genetic carrier Beagles. There is a strong correlation between amounts of AH myocilin protein and the presence and severity of POAG. The exact role of AH myocilin levels in the genesis of ocular hypertension remains unresolved, but myocilin may adversely affect AH outflow.     

Detection of anti-lens crystallin antibody in dogs with and without cataracts

Objective To determine if antilens crystallin (ALC) serum and aqueous humor antibodies were present in normal dogs and dogs with cataracts, whether antibody incidence varied with stage of cataract, and whether antibody titer had a relationship to the presence of lens‐induced uveitis. Methods Serum and aqueous humor samples were obtained from normal dogs and dogs with cataracts. Lens crystallin was separated by SDS‐polyacrylamide gel electrophoresis (SDS‐PAGE), and antilens crystallin antibodies were detected by Western immunoblot analysis. An indirect ELISA using crystallin protein as antigen was also used to detect antilens crystallin antibodies in serum and aqueous humor. Test groups included normal, incipient, immature, mature, hypermature and diabetic cataract. Results SDS‐PAGE identified bands with molecular weights of lens crystallin subunits. Western immunoblotting demonstrated reaction between canine serum and these protein bands. The five canine serum samples that reacted with crystallin subunits on Western blots had corresponding reactivity on the ELISA. All aqueous humor samples (30) were negative. Serum ALC antibodies were detected in 59.3% (16/27) of controls, 66.7% (16/24) of incipients, 50.0% (10/20) of immatures, 37.9% (11/29) of matures, 28.6% (6/21) of hypermatures, and 26.7% (4/15) of diabetics. Serum ALC antibodies were detected in 43.1% (47/109) of all cataract samples. There was a statistically significant negative association between the presence (P = 0.004) and maturity (P = 0.004) of cataract and presence of ALC serum antibodies. In the immature and hypermature cataract groups, there was a statistically significant negative association between ALC serum antibody titer and severity of uveitis (95% confidence interval). Conclusions There is a negative association between the presence (P = 0.004) and maturity (P = 0.004) of cataract and presence of ALC serum antibodies.     

Secondary glaucomas in the dog in North America

OBJECTIVE: To determine the prevalence of secondary glaucomas in dogs associated with cataract formation, lens luxation or displacement, cataract surgery, uveitis, hyphema and intraocular neoplasia. METHODS: Information was obtained from the Veterinary Medical Data Base (VMDB) from all veterinary medical teaching hospitals in North America from March 1964 to March 2003. Secondary glaucomas were diagnosed at the same examination or after the primary diagnosis was made, and included those associated with cataract formation, lens luxation, cataract surgery, uveitis of unknown cause, hyphema of unknown cause, and intraocular neoplasia. The data were evaluated by decade, breed, gender and age of presentation. RESULTS: A total of 1 592 831 dogs were presented, and 9695 canine secondary glaucomas. Secondary glaucoma associated with cataract formation represented 81% of all the canine secondary glaucomas. Breeds (n = 7890 dogs) predisposed to secondary glaucoma and cataracts had an overall prevalence of 0.5%, but nearly 20% of all the cataractous dogs developed secondary glaucoma in at least one eye. For the years 1994-2003, these breeds included the American Cocker Spaniel; Boston Terrier; Toy, Miniature and Standard Poodle; English Springer Spaniel; Bichon Frise; and Labrador Retriever. The other forms of secondary glaucoma occurred less frequently, and included those glaucomas with lens luxation or displacement (779 dogs; 12.0%), postcataract surgery (528 dogs; 5.1%), with uveitis from unknown cause (399 dogs; 7.1%), with hyphema from unknown cause (117 dogs; 7.3%), and with intraocular neoplasia (19 dogs; 3.5%). The risk of the secondary glaucomas from 1984 to 2002 was highest after the intracapsular lens extraction (ICLE), less in the extracapsular technique (ECLE), and lowest for the phacoemulsification/phacofragmentation method. CONCLUSION: Prevalence of the canine secondary glaucomas ranges from 0.25% (1964-1973), 0.46% (1974-1983), 0.79% (1984-1993), to 0.80% (1994-2003) and are as frequent as the primary or breed-related glaucomas during these same time periods.     

Antioxidant profile of cataractous English Cocker Spaniels

Cataracts have been attributed to oxidative injury in proteins and lipids. Primary defenses that directly protect the lens against oxidative damage include small molecule antioxidants (vitamin C, vitamin E, glutathione and carotenoids) and antioxidant enzymes (superoxide dismutase, catalase, and the glutathione enzyme systems – glutathione peroxidase, glutathione reductase and glucose-6-phosphate dehydrogenase). In humans, low plasmatic levels of vitamin C, vitamin E and carotenoids have been associated with a high risk of senile cataracts. Dogs are more prone to develop cataracts. A decrease in antioxidant defenses could be responsible for increased lens oxidation and cataract development. In this study we report the levels of erythrocytic enzymatic antioxidants (superoxide dismutase, catalase, glutathione peroxidase and glucose-6-phosphate dehydrogenase) and plasma vitamin C as well as malondialdehyde, in normal and cataractous English Cocker Spaniel dogs. Plasma vitamin C levels were consistently lower in cataractous dogs (20.17 μ m  ± 8.2 μ m ) when compared with normal dogs (24.1 μ m  ± 9.4 μ m ). These results indicate a possibly decreased synthesis in vitamin C, leading to lower aqueous humor levels of this vitamin. Considering that vitamin C levels in the aqueous humor may be responsible for lens antioxidant maintenance, and that these levels are obtained from plasma secretion through the ciliary epithelium, decreased plasma levels may indicate a decrease in the antioxidant capacity of the aqueous humor.     

Vascular endothelial growth factor in diabetic and nondiabetic canine cataract patients

Objective To measure vascular endothelial growth factor (VEGF) levels in aqueous humor, serum, and plasma in diabetic and nondiabetic cataractous dogs. Methods Canine VEGF was assayed in the plasma and serum of 32 dogs (20 diabetics; 12 nondiabetics) and aqueous humor in 57 eyes of those dogs (39 diabetic; 18 nondiabetic) undergoing phacoemulsification, using a commercial canine VEGF assay. Statistical analysis was performed using Fisher’s PLSD, t‐test, and regression analysis to compare values by diabetic status, duration of diabetes, age, weight, gender, left vs. right eye, and blood clarity. Results Plasma, but not serum or aqueous humor VEGF values of diabetics were significantly greater than nondiabetics (P = 0.019). Older nondiabetics (10–15 years) had higher plasma VEGF values than younger (0–5 and 5–10 years) dogs (P = 0.0002 and 0.0001, respectively). There was no significant difference in aqueous humor VEGF between left and right eyes in all patients. Serum and plasma, but not aqueous humor, VEGF values in females were significantly higher than males in both groups. Conclusion Similar to human diabetic patients, VEGF aqueous humor values in all dogs are significantly higher than blood values. Aqueous humor VEGF values in human diabetics are elevated and correlate with the severity of diabetic retinopathy. However, aqueous humor values of VEGF in diabetic dogs are not greater than nondiabetics and may serve to protect the dog against development of diabetic retinopathy.     

Progressive changes in ophthalmic blood velocities in Beagles with primary open angle glaucoma

Objective To measure changes in the ocular and orbital blood flow velocities by color Doppler imaging (CDI) in beagles with primary open angle glaucoma as the disease progressed from early to advanced stages. Methods CDI measurements were performed periodically on 13 glaucomatous Beagles during the nontreated mild, moderate and advanced stages of POAG over the course of 4 years. CDI was performed with the dogs lightly anesthetized (butorphanol 0.1 mg/kg IV, acepromazine maleate 0.02 mg/kg IV, and atropine sulfate 0.05 mg/kg) while the CD transducer was placed directly on the cornea anesthetized with 0.5% tetracaine hydrochloride. Intraocular pressure (IOP) by pneumatonography or TonoPen XL, heart rate and mean arterial blood pressure were measured at the beginning, middle and end of each study. The ophthalmic vessels examined included: external ophthalmic arteries and veins, long and short posterior ciliary arteries, anterior ciliary arteries and veins, primary retinal arteries, and vortex veins. Recordings of each vessel included peak systolic velocity (PSV), end diastolic velocity (EDV) and time averaged velocity (TAV), and when possible the resistive index (RI) and pulsatility index (PI) were computed. Results CDI abnormalities were present before intraocular pressure exceeded the normal range. As the animals aged, and the glaucoma progressed with higher levels of IOP, significant changes occurred in nearly all vessels, and generally included a major increase in RI (P < 0.001) and an increase in the PI (P < 0.001). Mean arterial blood pressure (105 ± 18 mmHg) and heart rate (118 ± 33/min) remained reasonably constant. The IOP gradually increased as the disease progressed (early and normotensive: 19.4 ± 3.9 mmHg; moderate: 29.7 ± 2 mmHg; and advanced: 44.5 ± 6 mmHg). The ocular veins seemed most influenced early on in the disease. Late in the disease, ocular venous blood flow could not be consistently demonstrated. An increase in the PI of ocular veins occurred in the moderately and severely affected glaucomatous Beagles. As the IOP increased, there were trends of increasing resistive index and pulsatility index in most arteries, and periods of marked decreased velocities of the vortex and external ophthalmic veins in severe cases. Conclusion CDI measurements in Beagles with primary open angle glaucoma during the course of 4 years indicate easily measurable and repeatable progressive blood flow abnormalities before the elevation of IOP and, thereafter, with gradually increased levels of IOP.     

Epidemiology of canine glaucoma presented to University of Zurich from 1995 to 2009. Part 1: Congenital and primary glaucoma (4 and 123 cases)

Objective To investigate the epidemiology of canine congenital and primary glaucoma in the cases presented to the University of Zurich, Vetsuisse Faculty (UZH) from 1995 to 2009. Methods Information was obtained from the computer database of patients examined by members of the UZH Ophthalmology Service, between January 1995 and August 2009. Congenital and primary glaucoma was diagnosed based on the age of onset, the lack of evidence of any antecedent eye conditions, and/or the presence and severity of iridocorneal angle defects. The data was evaluated for breed, gender and age at presentation. Results A total of 5984 dogs presented to the UZH Ophthalmology service between 1995 and 2009. Four dogs of different breed were diagnosed with congenital glaucoma and 123 dogs were diagnosed with primary glaucoma. For the primary glaucomas the overall male to female ratio (M:F) was 1:1.41 and the age of onset ranged from 0.12 to 18.3 years with a mean of 7.3 ± 3.6 years. Data suggested a predisposition for primary glaucoma in the Siberian Husky, Magyar Vizsla and Newfoundland from 2004 to 2009. Conclusion The report presents the epidemiology of canine congenital and primary glaucomas presented to the UZH from 1995 to 2009. A previous suspicion of predisposition for primary glaucoma in the Newfoundland dog (n = 6) and the Magyar Vizsla breed (n = 8) was confirmed.     

The relationship of cataract maturity to intraocular pressure in dogs

Objective To determine the distribution of intraocular pressure, as measured by applanation tonometry, in dogs with cataracts, and compare these tonometric results to the different stages of cataract formation (incipient, immature, mature, and hypermature). Animals studied Retrospection study of canine clinical patients (86 dogs). Procedures All records of dogs presented from 1991 to 1996 to the university veterinary medical teaching hospital for diagnosis of cataracts and evaluation for cataract surgery were reviewed. The tonometric measurements from the initial ophthalmic examination were selected in cataractous and nonglaucomatous eyes either receiving no topical or no systemic medications. The stage of cataracts was based on the degree of opacification, tapetal reflection, clinical vision, and visibility of the ocular fundus by indirect ophthalmoscopy. The distribution of tonometric results were grouped by the cataract maturity, and compared by anova and Tukey’s general linear tests. Results Intraocular pressure with incipient cataracts ranged from 9 to 17 mmHg (mean 12.7 ± 1.2 mmHg). Intraocular pressure with immature cataracts ranged from 3 to 27 mmHg (mean 13.6 ± 0.6 mmHg). For the mature cataracts, IOP ranged from 5 to 22 mmHg (mean 11.9 ± 0.7 mmHg). For the hypermature cataract group, IOP ranged from 4 to 23 mmHg (mean 10.8 ± 0.6 mmHg). Comparison of the tonometric results among the different stages of cataract formation indicated a significant difference (P = 0.0086) between only the immature and hypermature groups. Conclusions Intraocular pressure in lens‐induced uveitis (LIU) is lowered but the relationship to the stage of cataract maturity is less clear. Significant tonometric differences were present between the immature and hypermature cataract groups, but these differences are too small to be clinically useful. Decreased intraocular pressure of dogs with all stages of cataract formation suggests concurrent LIU during all stages of cataract formation, especially with the mature and hypermature stages. The average tonometric measurements in dogs with these cataracts were about two standard deviations below the mean IOP reported in normal dogs.     

Lens morphometry determined by B-mode ultrasonography of the normal and cataractous canine lens

Purpose To determine axial lens thickness, anterior chamber depth and axial globe length in canine eyes with normal lenses and in eyes with immature, mature, congenital, posterior polar and diabetic cataract. Methods B‐mode ultrasonography was performed in 50 normal dogs and, as a prephacoemulsification screening procedure, in 100 dogs with cataract. Axial B‐mode ultrasonograms were used to determine lens thickness, anterior chamber depth and globe diameter. Statistical comparisons between groups were made by analysis of variance and multivariate analysis, with a significance level of P < 0.05. Results Axial globe lengths were not statistically significantly different between groups apart from the smaller globes in younger dogs with congenital cataract. Axial lens thickness in diabetics (8.4 ± 0.9 mm) was statistically significantly different from the lens thickness in normal eyes (6.7 ± 1.0 mm), eyes with immature cataract (6.4 ± 0.8 mm) and eyes with mature cataract (7.4 ± 0.9 mm) although these groups, while varying in thickness, were not statistically significantly different from each other. Anterior chamber depth was statistically significantly reduced in eyes with diabetic cataract (2.9 ± 0.1 mm) from that in normal eyes (3.8 ± 0.1 mm), eyes with immature cataract (3.5 ± 0.1 mm) and eyes with mature cataract (3.2 ± 0.6 mm) although these groups, while varying in chamber depth, were not statistically significantly different from each other. Conclusions Lenses with diabetic cataracts were significantly increased in axial thickness compared to lenses in other eyes, although lenses with mature cataracts showed a trend towards increased axial thickness and immature cataracts demonstrated a trend towards reduced thickness. While previous studies on cataract pathobiology have suggested a reduction in lens thickness in immature cataract through lens protein loss and an increase in thickness in mature cataracts through intumescence, this study is the first to document these changes in the canine lens.     

Prevalence of the breed-related glaucomas in pure-bred dogs in North America

Objective To determine the prevalence of the breed‐related glaucomas in pure‐bred dogs presented to the veterinary medical teaching hospitals in North America that participate in the Veterinary Medical Data Base (VMDB). Materials and methods In this retrospective study, age of first diagnosis, breed, and gender data for all breeds of dogs were collected from the VMDB with the clinical diagnosis of primary glaucoma (glaucoma‐NOS) at 5–10 year intervals from 1964 to 2002. The prevalence for each breed (affected dogs compared to all dogs of each breed), any changes over the 38 years, and any gender differences for these glaucomas were determined. Results The prevalence of the primary breed‐related glaucomas has gradually increased from 0.29% (1964–1973); 0.46% (1974–1983); 0.76% (1984–1993); to 0.89% (1994–2002). Breeds that consistently featured among the highest 10 for glaucoma prevalence from four different periods (1964 to 2002) included American Cocker Spaniel, Basset Hound, Wire Fox Terrier, and Boston Terrier. During the last observation period (1994–2002), 22 different breeds had 1% or higher prevalence of the glaucomas. The highest prevalence of glaucomas in 1994–2002 by breed included: American Cocker Spaniel (5.52%); Basset Hound (5.44%); Chow Chow (4.70%); Shar‐Pei (4.40%); Boston Terrier (2.88%); Wire Fox Terrier (2.28%); Norwegian ElkHound (1.98%); Siberian Husky (1.88%); Cairn Terrier (1.82%); and Miniature Poodle (1.68%). A predominance of females with glaucoma occurred in the American Cocker Spaniel, Basset Hound, Cairn Terrier, Chow Chow, English Cocker Spaniel, Samoyed, and perhaps the Siberian Husky, and a predominance of males in the Australian Cattle dog and St Bernard. Age affected the time for first presentation of the glaucomas in the pure‐bred dog. In the majority of breeds the glaucomas were presented for initial diagnosis in dogs between 4 and 10 years of age. Conclusion Breed‐related glaucomas in pure‐bred dogs are frequently presented to the veterinary medical teaching hospitals in North America. The prevalence of the breed‐related glaucomas in the dog appears similar to humans, and in some breeds exceeds that in humans. In many breeds the high prevalence of the glaucomas suggests a genetic basis.     

Anterior chamber to frontal sinus shunt for the diversion of aqueous humor: a pilot study in four normal dogs

Silicone tubing was used to divert aqueous humor from the anterior chamber of the right eye to the rostral compartment of the right frontal sinus in four clinically normal mixed-breed dogs. Biomicroscopic examination, and pneumoapplanation tonometry and tonography, completed for up to 18 weeks postoperatively, confirmed gonioimplant function in all four cases. The dogs were euthanized at 6, 8, 16 and 18 weeks postoperatively. Gonioimplant patency was further confirmed by postmortem examination of the globes, implants and frontal sinuses. Gross and light microscopic examinations revealed iridal attachments to the implant ( n  = 4), mild anterior uveitis ( n  = 3), anterior subcapsular cataracts ( n  = 4), and focal corneal ( n  = 3) and scleral ( n  = 3) scarring in the operated globes. Light microscopic examination of frontal sinus specimens revealed mild lymphocytic proliferation and fibrosis immediately adjacent to the implant entrance site. There were no bacteria detected on aerobic or anaerobic cultures of the frontal sinuses or light microscopic examination of the globes or frontal sinuses. Results indicate that the frontal sinus shunting of aqueous humor is a safe and effective means of extraorbital aqueous diversion with potential applicability in the management of glaucoma.     

Cataracts in the Bichon Frise

Purpose To determine the clinical characteristics of possible inherited cataract in the Bichon Frise breed. These characteristics include the relative frequency, gender effects, site of first cataract formation, age of onset, relationship of age to cataract maturity, and other concurrent pre‐ and postoperative ophthalmic diseases. Methods Four different populations of Bichon Frise were examined. They included: (1) referred patients of the University of Florida (VMTH; 1990–2000); (2) patients from other universities and large institutions (VMDB; 1970–2000); (3) patients from the Canine Eye Registry Foundation (CERF; 1970–2000); and (4) special patients recruited from eye clinics and ACVO specialty practices (1995–2001). Blood samples were obtained from many patients for future DNA analyzes. Statistical comparisons between groups were by general linear and anova analyzes, and P < 0.05 was considered significant. Results The four populations of cataractous and total Bichon Frise dogs included: (1) UF‐VMTH: 57 cataractous dogs; (2) VMDB: 406 dogs (28%) with cataracts; total dogs ? 1407; (3) CERF: 505 cataractous dogs (6%); total dogs ? 8222; and (4) ACVO: 223 cataractous dogs (57%); total dogs ? 391. In each population group, gender did not affect cataract distribution. Dogs between 2 and 8 years of age were most frequently affected, and initial cataract involvement affected equally the anterior and posterior cortices. Immature cataracts occurred more often in younger dogs, and hypermature cataracts were more frequently diagnosed in older dogs. Dogs with early cataracts were encountered more frequently in the CERF population. Pre‐ and postoperative retinal detachments (RD) were not infrequently diagnosed in the UF‐VMTH and VMDB groups. RD in the UF‐VMTH and VMDB groups occurred in 33% and 13% of the patients, respectively. Conclusions Cataracts were first encountered in CERF and VMDB data between 1975 and 1979 and have increased since this time. As expected, both similarities and differences between the four different populations of Bichon Frise were encountered. Both sexes of dogs are equally affected. The anterior and posterior cortical areas of the lens were first involved and dogs 2–8 years of age are most frequently affected. Cataracts affected younger dogs in the CERF group and older cataractous dogs in the other populations. Cataract formation appears to be inherited in the Bichon Frise dog. The frequency of pre‐ and postoperative retinal detachments present higher risks for cataract surgery in this breed.     

Enhanced expression of cyclooxygenase-2 in glaucomatous dog eyes

Objective Cyclooxygenase‐2 (COX‐2)‐derived prostaglandins (PGs) are shown to play important pathophysiologic roles in various disease states. Recently, the effectiveness of topical PGs in reducing intraocular pressure (IOP) has stimulated further interest in the physiologic function of COX‐2 and PGs in normal and glaucomatous eyes. Therefore, we investigated the cell‐type distribution and expression of COX‐2 in normal and glaucomatous dog eyes. Procedures Using isoform‐specific antibodies, we immunohistochemically evaluated COX‐2 expression in formalin‐fixed and paraffin‐embedded normal (n = 5) and glaucomatous (n = 17) dog eyes. Results In the normal eyes, only minimal COX‐2 immunoreactivity was observed in the ciliary epithelium. In the glaucomatous eyes, COX‐2 expression was further observed in the cornea and corneoscleral limbus. In the cornea, moderate to strong COX‐2 expression was observed in all corneal layers (epithelium, stromal cells and endothelium), with the greatest expression present in the epithelial layer. In the corneoscleral limbus area, COX‐2 immunoreactivity was noted in the stromal cells of sclera, trabecular meshwork and endothelial cells of the angular aqueous plexus. Conclusions Increased expression of COX‐2 in dog glaucomatous eyes suggests that COX‐2‐derived PGs may have a potential role in the pathogenesis of canine glaucoma.     

Comparative Doppler imaging of the ophthalmic vasculature in normal Beagles and Beagles with inherited primary open-angle glaucoma

The objective of this study was to compare orbital and ocular vasculature velocity, measured by Doppler imaging, in normal Beagles and Beagles with inherited primary open-angle glaucoma. Eight normal Beagles and 13 Beagles with different stages of primary open-angle glaucoma were evaluated twice with a 2–4-week period between measurements. Doppler imaging was performed with the dogs anesthetized, and the Doppler transducer applied directly on the corneal surface. The majority of the orbital vasculature (external ethmoidal artery; internal ophthalmic artery and vein; and external ophthalmic artery and vein) and ocular blood vessels (anterior ciliary artery and veins; long posterior ciliary arteries; short posterior ciliary arteries; primary retinal arteries; and the vortex veins) were identified and Doppler blood velocity parameters were determined. The glaucomatous dogs demonstrated significant differences in the Doppler velocity parameters of several orbital vessels (external ethmoidal, external ophthalmic, and internal ophthalmic arteries), and several ocular vessels (anterior ciliary, short posterior ciliary, and long posterior ciliary arteries). These differences included decreased blood velocities, and increased pulsatility and resistive indexes. The Doppler blood flow velocities of the primary retinal arteries were unchanged between the normal and glaucomatous dogs. In the glaucomatous dogs, the Doppler imaging suggests increased vascular resistance downstream in both the orbital and ocular vasculature. These blood velocity parameter changes may be primary or secondary, and may offer therapeutic opportunities to increase perfusion, prolong the retina and optic nerve head function, and maintain vision in the canine glaucomas.     

Quantitation of uveoscleral outflow in normotensive and glaucomatous Beagles by 3H-labeled dextran

In uveoscleral outflow, aqueous humor leaves the anterior chamber and passes caudally through the trabecular meshwork and the sclerociliary cleft to enter the supraciliary and suprachoroidal spaces. The fluid is then absorbed by choroidal and scleral circulations. Using 3H-labeled dextran, uveoscleral outflow was quantitated in normotensive and glaucomatous Beagles under general anesthesia. The intrascleral plexus was isolated and 3H-labeled dextran was injected into the anterior chamber. Intrascleral plexus contents were sampled every 5 minutes over a 30- to 60-minute period. The eyes were enucleated, sectioned, and prepared for scintillation counting. Uveoscleral outflow accounted for 15% and 3% of the total aqueous humor outflow in the normotensive dogs and in the advanced glaucomatous dogs, respectively. In the advanced glaucomatous Beagle, conventional and uveoscleral outflow pathways were reduced and contributed to the etiopathogenesis of glaucoma.     

Effect of different dose schedules of latanoprost on intraocular pressure and pupil size in the glaucomatous Beagle

Objective To evaluate the changes in intraocular pressure and pupil size in glaucomatous dogs after instillation of 0.005% latanoprost (Xalatan, Pharmacia and Upjohn, Kalamazoo, MI, USA) once in the morning, or once in the evening, or twice daily in five‐day multiple‐dose studies. Animals studied Eight Beagles with the moderate stage of inherited primary open‐angle glaucoma. Procedures Applanation tonometry (IOP) and pupil size (PS) measurements were obtained at 8 am, 10 am, 12 noon, 2 pm, and 4 pm in eight glaucoma dogs. Methylcellulose (0.5% as placebo) was instilled in the control eye, and 0.005% latanoprost was instilled in the opposite drug eye. Control and drug eyes were selected using a random table. For these three studies, 0.5% methylcellulose and 0.005% latanoprost were instilled the second through the fifth days with instillations in the morning (8.30 am), or evening (8 pm), or twice daily (8.30 am and 8 pm). Statistical comparisons between drug groups included control, placebo, and treated (0.005% latanoprost) eyes for three multiple‐dose studies. Results In the 8‐am latanoprost study, the mean ± SEM diurnal declines in IOP for the placebo and drug eyes for the first day were 6.5 ± 3.6 mmHg and 8.4 ± 4.0 mmHg, respectively. The mean ± SEM diurnal changes in IOP after 0.005% latanoprost at 8 am once daily for the next four days were 23.3 ± 5.0 mmHg, 25.4 ± 2.1 mmHg, 25.7 ± 1.7 mmHg, and 26.1 ± 1.7 mmHg, respectively, and were significantly different from the control eye. A significant miosis also occurred starting 2 h postdrug instillation, and the resultant mean ± SD pupil size was 1.0 ± 0.1 mm. In the first day of the second latanoprost study, the mean ± SEM diurnal changes in the placebo and drug eye IOPs were 11.6 ± 3.8 mmHg, and 12.0 ± 4.4 mmHg, respectively. For the following four days with latanoprost instilled at 8 pm, the mean ± SEM diurnal changes in IOP in the drug eyes were 24.9 ± 2.1 mmHg, 22.4 ± 1.8 mmHg, 21.6 ± 1.9 mmHg, and 26.6 ± 2.2 mmHg, respectively. Compared to the fellow placebo eyes, the diurnal changes in IOP were significantly different. Significant changes in pupil size were similar to the IOP changes, with miosis throughout the day and return to baseline pupil size the following morning before drug instillation. In the last study, the mean ± SEM diurnal changes in IOP for the placebo and drug eyes for the first day were 6.6 ± 2.1 mmHg and 9.4 ± 2.8 mmHg, respectively. For the four subsequent days with latanoprost instilled twice daily, the mean ± SEM diurnal IOP changes were 19.6 ± 1.5 mmHg, 19.1 ± 1.4 mmHg, 19.9 ± 1.7 mmHg, and 20.3 ± 0.7 mmHg, respectively, and were significantly different from the placebo eyes. The mean changes in PS were 3.1 ± 0.7 mm. Conclusion 0.005% latanoprost instilled once daily (am or pm) as well as twice daily produces significant decreases in IOP and PS in the glaucomatous Beagle. The evening instillation of 0.005% latanoprost produced less daily fluctuations in IOP than when the drug was instilled in the morning. 0.005% latanoprost instilled twice daily produced the greatest decline in IOP with the least daily fluctuations, but longer duration miosis.     

Epidemiology of canine glaucoma presented to University of Zurich from 1995 to 2009. Part 2: secondary glaucoma (217 cases)

Objective To investigate the epidemiology of canine secondary glaucomas in the cases presented to the University of Zurich, Vetsuisse Faculty (UZH) from 1995 to 2009 focusing on possible risk factors for developing secondary glaucoma in this population of dogs. Methods Information was obtained from the computer database of patients examined by members of the UZH Ophthalmology Service, between January 1995 and August 2009. Secondary glaucoma was diagnosed based on the presence of antecedent eye conditions. The data was evaluated for breed, gender, age at presentation, and for antecedent eye conditions known to cause glaucoma including anterior uveitis of unknown cause (AU), lens luxation (LL), intraocular surgery (SX), intraocular neoplasia (IN), unspecified trauma to the globe (T), ocular melanosis (OM), hypermature cataract (PY), hyphema (HY), and six other less frequent conditions. Results A total of 217 dogs were diagnosed with secondary glaucoma from 1995 to 2009. The age of the dogs with secondary glaucoma ranged between 88 days and 19 years (mean 7.7 ± 3.6 years). Data suggested a predisposition for secondary glaucoma in the Cairn Terrier and the Jack Russell Terrier breeds from 2004 to 2009. Common causes of secondary glaucoma from 1995 to 2009 were AU (23.0%), LL (22.6%), SX (13.4%), IN (10.6%), T (8.3%), OM and PY (both 6.9%) and HY (3.23%). Conclusion The report presents the epidemiology of secondary glaucomas presented to UZH from 1995 to 2009. Fourteen risk factors were recorded for secondary glaucoma. This is the first paper documenting OM in the Swiss Cairn Terrier dog population.     

Investigation of serum concentrations and immunohistochemical localization of α1-acid glycoprotein in tumor dogs

The purpose of this study was to measure the dynamics of serum α1-acid glycoprotein (AGP) concentration in dogs with various tumors, and to investigate the localization of AGP in some tissues using immunohistochemical staining. Sera were obtained from 171 dogs bearing tumors of various types. Serum AGP concentration was measured by single radial immunodiffusion. Tumors occurring in the liver and spleen were also investigated immunohistochemically using anti-canine AGP antibody. Mean serum AGP levels were 749 ± 602 mg/L in dogs with carcinoma (n = 39), 1,014 ± 971 mg/L with sarcoma (n = 18), and 887 ± 935 mg/L with round cell tumors (n = 46), all significantly higher than serum AGP level in healthy dogs (n = 137, 364 ± 106 mg/L). Mean serum AGP levels were significantly higher than in healthy dogs in complex mammary gland carcinoma (n = 5, 876 ± 721 mg/L), malignant melanoma (n = 7, 1,010 ± 821 mg/L), and hepatocellular carcinoma (n = 5, 936 ± 741 mg/L) among carcinomas, hemangiosarcoma (n = 5, 1,740 ± 1,323 mg/L) among sarcomas, and lymphoma (n = 19, 1,072 ± 965 mg/L) and histiocytic tumor (n = 6, 1,800 ± 1,387 mg/L) among round cell tumors. In an immunohistochemical investigation of AGP localization, both weak and strong staining for anti-AGP antibody were seen in hepatic tissue in dogs with primary non-tumorous lesions originating in the spleen (hematoma) and elevated serum AGP, but all tumor tissue in the spleen was negative. Among dogs with primary tumor lesions of the spleen (hemangiosarcoma) and elevated serum AGP levels, both weak, moderate and strong staining for anti-AGP antibody were seen in hepatic tissue, while strong positive staining was apparent in all tumorous tissue from the spleen. In primary tumor lesions in the liver (hepatocellular carcinoma), both moderate and strong staining for anti-AGP antibody were seen in normal hepatic tissue, and both weak, moderate and strong staining were seen in tumor tissues of the liver. AGP levels thus appear to be elevated in dogs with carcinomas, sarcomas, and round cell tumors. With some of these malignant tumors, localization of AGP in tumor tissue was seen.     

Renal resistive index in 55 dogs with degenerative mitral valve disease

Background

Azotemia occurs frequently in dogs with degenerative mitral valve disease (DMVD). It could indicate changes in renal hemodynamics.

Hypothesis/Objectives

To assess the renal resistive index (RI) in dogs with DMVD, and the statistical link between heart failure class, azotemia, echo‐Doppler parameters, several plasma variables, and RI.

Animals

Fifty‐five dogs with naturally occurring DVMD were used (ISACHC class 1 [n = 28], 2 [n = 19], and 3 [n = 8]).

Methods

Observational, blinded study, performed under standardized conditions. Physical examination, renal ultrasonography, and echo‐Doppler examinations were performed in awake dogs. The RI of the renal, interlobar, and arcuate arteries were measured. Plasma creatinine, urea, and N‐terminal pro‐B‐type natriuretic peptide concentrations (NT‐proBNP) were determined. Statistical links between variables and RI were tested by means of a general linear model.

Results

Although the RI of renal and arcuate arteries were unaffected by ISACHC class, the left interlobar RI increased (P < .001) from 0.62 ± 0.05 (mean ± SD) in class 1 to 0.76 ± 0.08 in class 3. It was also higher (P < .001) in azotemic (0.74 ± 0.08) than in non‐azotemic (0.62 ± 0.05) dogs. Similar findings were observed for right interlobar RI. Univariate analysis showed a positive statistical link between NT‐proBNP (P = .002), urea (P < .001), creatinine (P = .002), urea‐to‐creatinine ratio (P < .001), left atrium‐to‐aorta ratio (P < .001), regurgitation fraction (P < .001), systolic pulmonary arterial pressure (P < .001), shortening fraction (P = .035), and RI.

Conclusion and Clinical Importance

In dogs with DMVD, interlobar RI increases with heart failure severity and azotemia but a cause and effect relationship remains to be established.     

The effects of moxifloxacin ophthalmic solution 0.5% or gatifloxacin ophthalmic solution 0.3% treatment on corneal wound healing in pigmented rabbits following anterior keratectomy

Purpose These studies examined corneal healing rates, Type‐IV collagen and zonula occludens membrane‐associated protein (ZO‐1) expression, as well as aqueous PGE₂ and IL‐1β concentrations in pigmented rabbits treated with either moxifloxacin 0.5%, gatifloxacin 0.3% or BSS^® following anterior keratectomy. Methods Anterior keratectomy surgery was followed by topical administration with commercial ophthalmic formulations of either moxifloxacin or gatifloxacin or BSS^® (TID for 96 h). Images of the fluorescein‐stained healing corneas were analyzed for wound area. At 48 or 96 h following surgery, aqueous humor samples were collected and analyzed for the inflammatory mediators PGE₂ and IL‐1β using an ELISA. The corneas were subsequently evaluated using both scanning and transmission electron microscopy. In a second parallel study, corneas were evaluated at both 48 and 96 h for Type‐IV collagen and ZO‐1 expression using immunohistochemistry. Results Fluorescein‐stained corneal images at 96 h postsurgery demonstrated that 90% ± 8% re‐epithelialization for moxifloxacin, 81% ± 14% for gatifloxacin, and 88 ± 6% for BSS^® (P > 0.05). PGE₂ levels in the aqueous humor of fluoroquinolone treated eyes were reduced at 48 h compared to BSS^® treated eyes. IL‐1β was undetectable in all samples. No differences in Type‐IV collagen or ZO‐1 expression were observed between any treatment groups. There were no differences between groups in histological appearance or in ultrastructural healing processes. Conclusions These studies demonstrated that the commercial ophthalmic formulations of moxifloxacin and gatifloxacin were similar to each other in their effects on the levels of aqueous humor PGE₂ and rates of corneal wound re‐epithelialization.