In vitro analysis of antiangiogenic activity of fungi isolated from clinical cases of equine keratomycosis.

OBJECTIVE: The goal of this project was to explore the possibility that fungal organisms produce metabolites that inhibit angiogenesis. Procedures Fungal cultures were obtained from cases of keratomycosis, grown in Sabouraud's dextrose broth, and sterile filtered for use in experiments. The Matrigel assay was used to screen the filtrate samples for antiangiogenic activity. Matrigel is a basement membrane matrix that supports the differentiation of human umbilical vein endothelial (HUVE) cells into a capillary-like network of tubules. HUVE cells were cultured using standard techniques and passaged at confluence, with all cells being used at passage 3-6. HUVE cells (40 000 cells) were pipetted into each well of a 24-well tissue-culture plate coated with Matrigel. An aliquot of fungal media filtrate was added to each well and the plates allowed to incubate for 18 h, at which time they were evaluated for tubule formation. RESULTS: Two fungal isolates showed inhibition of tubule formation. The addition of 100, 200 and 400 &mgr;L of the fungal media filtrate from the first isolate (Fusarium sp. 99A34574) produced a consistent and dose-dependent inhibition of tubule formation. The second isolate (Aspergillus sp. 271599) did not show inhibition of tubule formation with 100 or 200 &mgr;L added to the wells, however, it did show inhibition at 400 &mgr;L/well. The remaining three isolates did not cause inhibition at any concentration. CONCLUSIONS: Our findings suggest that certain fungal organisms produce metabolites that inhibit tubule formation in vitro, and that these metabolites may play a significant role in altering the host vascular response to fungal infections of the cornea.

Fungal flora of the healthy camelid conjunctival sac.

Swab specimens for fungal isolation were collected from the healthy conjunctival sacs of 3 species of captive camelids (Lama glama, L guanicoe, L pacos) and llama-guanaco hybrids. Fungi were collected from over half the animals in winter (53%) and summer (56%). Fungal species of 10 genera were isolated. In both seasons, Aspergillus was the most commonly isolated genus; at least 9 species of Aspergillus were found. The fungal organisms isolated were similar to those found in healthy eyes of other domestic animals and may represent a random seeding from the environment where they are ubiquitous.

Bacterial and mycoplasmal flora of the healthy camelid conjunctival sac.

Healthy conjunctival sacs of 88 animals of 3 species of captive camelids (Lama glama, Lama guanicoe, Lama pacos) and llama-guanaco hybrids were sampled for bacterial and mycoplasmal flora. Mycoplasmas were not isolated from any animal. Eleven genera of bacteria were isolated. The most frequent isolates were Staphylococcus epidermidis and Pseudomonas spp. Nine varieties of Pseudomonas were found, which represented at least 3 Pseudomonas species. Many of the bacterial isolates (especially the pseudomonads) are potential pathogens in the eyes of these camelids.

Blindness in a dog with IgA-forming myeloma.

A 9-year-old dog with a 2-month history of weight loss and a 1-week history of blindness had an IgA-forming myeloma. Seemingly, the blindness was a result of bilaterally detached retinas. The dog also had leukopenia, anemia, hypoalbuminemia, hyperglobulinemia, and proteinuria as well as lytic lesions in the cervical portion of the spine and high IgA concentrations in serum and urine. Evaluation of aspirates from the subretinal spaces revealed lymphocytes in a proteinaceous fluid. Histologically, retinal lesions consisted of vascular endothelial cell damage and intraretinal cysts and hemorrhages.

Ocular manifestations of a metastatic pulmonary adenocarcinoma in a cat.

A primary pulmonary bronchogenic adenocarcinoma originating from an intramediastinal accessory lung was diagnosed in a 14.5-year-old cat. The cat had been admitted because of a cloudy right eye. Physical examination revealed a thin cat with severe iritis, aqueous flare, and a fibrin clot in the anterior chamber of the right eye. Right fundic examination revealed bullous retinal detachment superior to the optic disc. Euthanasia and necropsy were requested when FeLV test results were positive. Metastatic neoplastic cells similar to those of the primary tumor were detected in the choroid, ciliary body, and ciliary processes of the right eye. Ciliary and iridic stromal necrosis attributable to neoplastic embolization of uveal vessels had led to severe uveitis. Foci of metastasis were also in the heart, kidney, and cerebral meninges.

Xylazine-induced mydriasis: possible involvement of a central postsynaptic regulation of parasympathetic tone.

Intravenous injection of xylazine (0.01-1 mg/kg) produced a dose-dependent mydriasis associated with a depression of tonic ciliary nerve activity in anesthetized cats. Xylazine-induced mydriasis was apparent in the sympathectomized iris but was absent in the parasympathectomized, physostigmine-treated iris. Epinephrine (30 micrograms/kg, i.v.) produced a slightly greater mydriasis in the sympathectomized iris than in the parasympathectomized, physostigmine-treated iris. The alpha 2-adrenergic blocking agent, yohimbine (0.5 mg/kg, i.v.) antagonized the pupillary dilation and reversed the depression of ciliary nerve activity induced by xylazine administration. In rats pretreated with reserpine (7.5 mg/kg, s.c., 20 h) and alpha-methyl-p-tyrosine (250 mg/kg, i.p., 5 h), intravenous injection of xylazine (0.01-1 mg/kg) resulted in mydriasis of similar magnitude as control animals. However, xylazine induced bradycardia in the control group but not in the pretreated animals. The results suggest that pupillary dilation produced by i.v. xylazine is primarily the result of a central inhibition of parasympathetic tone to the iris. It also appears that xylazine produces this effect via postsynaptic alpha 2-adrenergic mechanisms, while it produces bradycardia through a presynaptic alpha 2-adrenergic mechanism.

Xylazine-induced mydriasis in rats and its antagonism by alpha-adrenergic blocking agents.

Pupillary response to xylazine (10-300 micrograms/kg, i.v.) norepinephrine (1-30 micrograms/kg. i.v.) and atropine (3-100 micrograms/kg, i.v.) were observed in rats anaesthetized with pentobarbital. Xylazine caused a dose-dependent mydriasis which was antagonized by a selective alpha 2-adrenergic blocking agent, yohimbine (2.5 mg/kg, i.v.). was less effective in antagonizing this effect of xylazine. A selective alpha 1-adrenergic blocking agent, prazosin (2.5 mg/kg, i.v.) was ineffective in reducing the xylazine-induced mydriasis. In contrast, both phentolamine and prazosin blocked the pupillary dilation produced by norepinephrine, while yohimbine was much less effective in antagonizing norepinephrine-induced mydriasis. Atropine also induced a dose-dependent mydriasis which was not affected by yohimbine pretreatment. The present study suggests that the mydriatic effect of xylazine in the rat is mediated by an adrenergic mechanism, possibly by stimulating the alpha 2-adrenergic receptors in the iris and CNS.