Uveal spindle cell tumor of blue-eyed dogs: an immunohistochemical study.

Immunohistochemical techniques were used to investigate the origin of a spindle cell tumor in the anterior uveal tract of dogs and the influence of ultraviolet radiation on the development of this tumor. Thirteen tumors were identified from the 4,007 canine ocular samples examined at the Comparative Ocular Pathology Laboratory of Wisconsin between 1978 and 2005. Siberian Husky and Siberian Husky mix dogs were overrepresented (10/13 dogs, overall median age 10 years). Light microscopic evaluation (all dogs) and electron microscopy (2 dogs) were performed. Immunohistochemical staining included alpha-smooth muscle actin (SMA), vimentin, S-100, desmin, glial fibrillary acidic protein (GFAP), Melan A, microphthalmic transcription factor (MITF-1), protein gene product 9.5 (PGP 9.5), laminin, gadd45, p53, proliferating cell nuclear antigen (PCNA), anti-UVssDNA (antibody for detection of (6-4)-dipyrimidine photoproducts), and telomerase reverse transcriptase (TERT). All tumors occurred in the iris with or without ciliary body involvement and were composed of spindle cells arranged in fascicles and whorls (variable Antoni A and B behavior). All tumors were positive when immunostained for vimentin and S-100. Nine of 13 tumors exhibited GFAP immunopositivity. All tumors were negative for SMA, desmin, Melan A, and MITF-1. Tumors were variably positive for PGP 9.5, laminin, gadd45, p53, PCNA, anti-UVssDNA, and TERT. Electron microscopy revealed intermittent basal laminae between cells. These tumors are morphologically and immunohistochemically most consistent with schwannoma. The relationship between spindle cell tumors of the anterior uvea of dogs, altered neural crest, blue iris color, and ultraviolet radiation has not yet been fully elucidated.

Immunohistochemical properties of ocular adenomas, adenocarcinomas and medulloepitheliomas.

Ocular medulloepitheliomas, adenomas and adenocarcinomas share a common phenotype and originate from the optic cup neuroectoderm. This can make it very difficult to differentiate between these tumors histopathologically. Therefore, this study focused on identifying a combination of immunologic markers that might be used in the diagnosis of these tumors. These markers included AE1/AE3, CK7, CK20, and telomerase reverse transcriptase (TERT). Routine immunohistochemical staining was performed on 27 whole globes diagnosed with one of these tumors. The tumors that immunostained for TERT showed increasing immunoreactivity as the tumor types increased in aggressiveness. None of the tumor types were immunopositive for CK7. CK20 immunostaining was found in the adenomas but not in the adenocarcinomas or medulloepitheliomas. AE1/AE3 expression was present more consistently in the adenocarcinomas and less frequently in the adenomas. AE1/AE3 expression was present in only one of six medulloepitheliomas. Furthermore, CK20 and TERT showed inverse expression patterns, i.e. TERT increased in expression and CK20 decreased in expression with increasing aggressiveness. These results may be important diagnostic and prognostic indicators for these tumors.

Canine cataracts, diabetes mellitus and spontaneous lens capsule rupture: a retrospective study of 18 dogs.

OBJECTIVE: To describe the clinical presentation and surgical outcome of diabetic canine patients with cataracts and preoperative spontaneous lens capsule rupture. ANIMALS STUDIED: A total of 20 dogs and 40 eyes were included in the retrospective evaluation. The patients' ages ranged from 5 to 14 years (mean 8.5 years). RESULTS: All dogs had clinical diabetes mellitus, with the duration since diagnosis ranging from 30 to 240 days (mean 123 days). Cataracts were bilateral and noted to have been present for 14-112 days (mean 39 days). Of the 40 eyes affected with cataracts, 30 had a spontaneous rupture of the lens capsule prior to surgery. The capsular rupture was diagnosed on clinical examination in 28/30 eyes and was noted intraoperatively in 2/30. The location of the capsular rupture was equatorial in 29/30 and posterior in 1/30 eyes. Surgery was performed in 38/40 eyes, with one case lost to follow-up without surgical intervention. Prior to surgery, routine diagnostic ophthalmic examination, ocular ultrasound, electroretinography, and systemic evaluation were performed in all dogs. Surgical procedures included phacoemulsification in 28/40 eyes, with IOL placement performed in 20/28 eyes. Intrascleral prosthesis placement or enucleation was performed in 8/40 and 2/40 eyes, respectively, due to a significantly reduced ERG or secondary glaucoma. CONCLUSIONS: The duration of clinical follow-up (19/20 dogs) ranged from 1 to 36 months (mean 12.9 months). All eyes that had cataract surgery with or without IOL placement were sighted at the time of the last follow-up examination. Spontaneous lens capsule rupture associated with diabetes mellitus, cataract and rapid lens intumescence occurs in the dog. Early surgical intervention, prior to secondary complications of glaucoma and loss of retinal function, is associated with a favorable outcome.

Telomerase activity with concurrent loss of cell cycle regulation in feline post-traumatic ocular sarcomas.

Paraffin wax-embedded ocular globes of cats with post-traumatic ocular sarcomas were examined for the presence of TERT, the active subunit of telomerase. The latter is a ribonucleoprotein complex essential for immortalization and expressed by most malignant tumours, germ line cells, lens epithelial cells, and some stem cells. Due to the frequent loss of cell cycle control with the increased expression of telomerase activity, post-traumatic ocular sarcomas were also examined for loss of p16 expression and alterations in p53, the findings being related to mitotic score, tumour grade, and proliferating cell nuclear antigen. These sarcomas expressed telomerase at a high frequency (62.5%); in addition, the majority showed alterations in cell cycle control, as evaluated by lack of p16 immunolabelling (66.7%). Alterations in p53 were the sole mechanism by which cell cycle control was dysregulated in only two tumours expressing TERT (13%). These findings suggest that p16, and not p53, represents the primary mechanism by which post-traumatic ocular sarcomas that express telomerase activity escape cell cycle control.

The effects of oxidative stress on telomerase activity and other stress-related proteins in lens epithelial cells.

Telomerase is a ribonucleoprotein complex responsible for maintaining the ends of chromosomes and for repair of DNA strand breaks. While telomerase activity is generally found in cells that have unlimited proliferative potential such as neoplastic cells, germline cells and some stem cells, lens epithelial cells (though not highly proliferative) have telomerase activity. Our previous studies indicated that lens epithelial cells express high levels of telomerase despite their low proliferative potential, thus we hypothesized that telomerase expression protects lens epithelial cells from oxidative stress. We also determined levels of the stress proteins gadd45 and p16 and the stress and proliferation-related protein, proliferating cell nuclear antigen (PCNA). In acute studies, lenses were exposed to TBHP for 0-120 min. In recovery studies, lenses were exposed to TBHP for 1 hr, then allowed to recover for up to 18 hr. In acute studies, telomerase activity was increased, p16 initially decreased then normalized, PCNA levels did not change significantly even in the overnight recovery groups, and gadd45 was decreased in some TBHP exposed groups. In recovery studies, telomerase activity was increased in all groups, gadd45 decreased then became elevated, and p16 levels were decreased at later recovery times. PCNA levels remained constant during the studies, indicating that there was no change in proliferation. These studies showed that elevated telomerase activity did not correlate with increased proliferation in lens epithelial cells; instead, increased telomerase activity was associated with increased levels of the stress-related protein gadd45 only in the later recovery times. These findings support the hypothesis that telomerase plays a protective rather than a proliferative role in lens epithelial cells.

Telomerase activity and related properties of normal canine lymph node and canine lymphoma.

Telomerase activity (TA) and the expression of p16(INK4), telomerase reverse-transcriptase catalytic subunit (TERT) and proliferating cell nuclear antigen (PCNA) were analysed in lymph nodes from clinically normal dogs and from dogs with lymphoma. Telomere lengths were measured in 12 histologically normal lymph nodes. These data were related to the overall survival time of the lymphoma patients given chemotherapy, in an effort to identify prognostic significance of the measured variables. There was no significant difference between TA of normal lymph nodes (n = 16) and lymphoma lymph nodes (n = 6). PCNA expression was significantly higher in lymphoma (n = 30) than in normal lymph nodes (n = 10), but TERT expression was not. Expression of p16(INK4) was not significantly different between normal and lymphoma lymph nodes. TA and p16(INK4) expression were inversely correlated within the normal lymph nodes studied. Telomere lengths in normal lymph nodes were consistent with previous studies. No variables examined had any correlation with survival of the lymphoma patients given chemotherapy. The role of p16(INK4) in the regulation of TA warrants further investigation.

Orbital fibroma in a horse.

An 11-year-old American Quarterhorse gelding presented for moderate periorbital swelling and exophthalmia of the left eye. The menace response, and direct and consensual pupillary light reflexes were absent in the left eye. Conjunctival hyperemia, blepharedema, a mydriatic pupil, resistance to retropulsion, and an increased intraocular pressure were present. A soft-tissue mass could be palpated in the left retrobulbar space by pressing onto the orbit over the supraorbital fossa. Incomplete surgical resection of the mass was performed and histopathologic evaluation was consistent with a fibroma. Normal pupillary light reflexes and vision returned following surgery. The mass has not recurred 14 months after surgery.

Immunohistochemical analysis of lens epithelial-derived membranes following cataract extraction in the dog.

The objective of the study was to characterize the morphologic and immunohistochemical features of lens epithelial-derived proliferative membranes from the anterior segment of canine globes. These features were correlated with those previously identified for diseases resulting from lens epithelial cell (LEC) proliferation including posterior capsular opacification, traumatic subcapsular cataract, and subcapsular plaques associated with hypermature cataracts. Sixteen canine globes were removed as a result of glaucoma or other complications following cataract extraction. Light microscopic and immunohistochemical analysis was performed on sections from formalin-fixed, paraffin-embedded globes. The tissues were stained with a variety of antibodies for cellular markers for LECs, growth factors or other cellular constituents relevant to cellular metaplasia and proliferation. The membranes were composed of monolayers or multilayers of spindle-shaped cells on the external surfaces of the anterior and posterior lens capsule, ciliary processes, iris leaflets, and iridocorneal angle, and they could be seen extending from an obvious monolayer of LEC within the capsular sac. Variably, scattered pigment cells, presumably of uveal origin, were concurrently present. Cellular components of the membranes stained positive for vimentin, transforming growth factor-beta, basic fibroblast growth factor, and smooth muscle actin. An amorphous eosinophilic extracellular matrix consisting predominately of collagen was associated with the membranes. Proliferative anterior segment membranes following cataract surgery were morphologically and immunohistochemically similar to cellular and matrix components of posterior capsular opacification and capsular plaques seen with hypermature cataracts, both of which result from metaplasia and proliferation of LEC. The presence of these LEC-derived membranes in association with secondary glaucoma suggests that exuberant proliferation of LEC outside the confines of the lens capsular sac may cause pathologic alterations in the eye following cataract surgery in the dog.