Blood vessel maturation in human uveal melanoma: spatial distribution of neovessels and mature vasculature.

PURPOSE: The aims of this study are (1) to evaluate the spatial distribution of neovessels and mature vessels in human uveal melanoma tumors and (2) to determine whether vessel maturation is associated with the major indicators for poor prognosis. METHODS: Immunohistochemical analyses were performed on human tissue specimens from enucleated eyes (n = 14) to assess total vessels, neovessels, mature vessels, and cell proliferation. Tumor morphology was analyzed by hematoxylin and eosin and modified periodic acid-Schiff (PAS) staining.The spatial distribution of neovessels and mature vessels was analyzed by immunohistochemistry, and correlated with major indicators of poor prognosis (i.e., aggressive PAS patterns, epithelioid cytology, mitotic figures, extraocular extension, anterior tumor location, ciliary body involvement, large tumor size, cell proliferation, and angiogenic activity). RESULTS: Neovesseldensity was greater than mature vessel density in apical (p = 0.17), central (p = 0.036), and peripheral (p = 0.31) regions of the tumors, while mature vessel density was greater than neovessel density in basal areas of the tumor (p = 0.47). This pattern indicated that vessel maturation begins at the base of the tumor and later extends to the peripheral and apical regions. The difference between mature and neovessel densities for the apical (-0.8 +/- 1.9) and central areas (-0.8 +/- 1.3) of the tumor was significantly higher than the difference obtained for the basal area (0.3 +/- 1.6; p = 0.014 and p = 0.012, respectively), indicating a higher density of mature vessels compared to neovessels at the base. Statistical correlations were found between mature vessel density and tumor size (r = 0.48, p = 0.084), cell proliferation (r = 0.62, p = 0.042), and mitotic figures (r = 0.76, p = 0.001). CONCLUSIONS: Significant differences exist in the spatial distribution of mature versus neovessels in human uveal melanoma. Vessel maturation is associated with known clinical and pathologic indicators of poor prognosis (e.g., cell proliferation). Antiangiogenic therapy should be considered for the treatment of ocular malignancies; however, the results of this study indicate that blood vessel maturation heterogeneity may limit the efficacy of vessel targeting agents.

Histopathologic examination of failed grafts in descemet's stripping with automated endothelial keratoplasty.

PURPOSE: To study the histopathologic features of 19 corneal posterior lamellar grafts in eyes for which Descemet's stripping with automated endothelial keratoplasty (DSAEK) has failed. DESIGN: Retrospective case series with clinicopathologic correlation. PARTICIPANTS: Nineteen cases of DSAEK failures undergoing repeat DSAEK or penetrating keratoplasty. METHODS: The histopathologic results of posterior lamellar grafts (also termed DSAEK grafts), recipient corneas, or both from 19 cases of failed DSAEK were examined. MAIN OUTCOME MEASURES: Abnormalities in the DSAEK graft and in the interface between the recipient cornea and the DSAEK graft were assessed. RESULTS: Histopathologic features in 19 failed DSAEK grafts revealed attenuation of endothelial cells (16 cases) and presence in the graft-host interface of fibrocellular tissue (11 cases), retained Descemet's membrane (5 cases), epithelial ingrowth (4 cases), or a combination thereof. Four DSAEK grafts had full-thickness corneal layers at 1 edge. CONCLUSIONS: Presence of interface material, such as fibrocellular tissue, retained Descemet's membrane, and epithelial ingrowth, are potential causes of dislocation. Endothelial attenuation was the most common finding in failed grafts. Decentered DSAEK grafts with full-thickness corneal layers at 1 edge are a potential cause for epithelial ingrowth.

Optic nerve invasion from an amelanotic peripapillary choroidal melanoma.

PURPOSE: To report an unusual case of an amelanotic optic nerve head melanoma. PATIENTS/METHODS/RESULTS: This is a retrospective case report of an 81-year-old woman with a slowly enlarging amelanotic optic nerve head lesion. Histopathologic evaluation of the lesion after enucleation revealed a peripapillary choroidal melanoma. The lesion was highly atypical in that the patient's visual acuity was excellent despite the lesion's growth and the intraocular pressure did not differ from the fellow eye. The patient did have a history of primary open angle glaucoma, which may have rendered the optic nerve more susceptible to tumor invasion. CONCLUSION: Optic nerve invasion by choroidal melanoma is a rare event, which is typically associated with poor visual acuity, elevated intraocular pressure, and large necrotic lesions; however, invasive melanoma with optic nerve invasion may occur in the absence of these classic findings.

Pediatric nasopharyngeal carcinoma with retinal metastasis.

PURPOSE: To report an unusual case of nasopharyngeal carcinoma (NPC) metastatic to the lungs and retina. To our knowledge, this is the first report of retinal metastasis from NPC. DESIGN: Observational case report. METHODS: We studied the case of a 15-year-old boy who had been recently diagnosed with metastatic NPC and was referred for evaluation of a suspicious retinal lesion in the right eye. RESULTS: Clinical examination and fundus photography showed an amelanotic, 1-disk-area infiltration in the retina along the inferotemporal arcade that was suggestive of metastatic disease. The lesion was small and irregularly shaped by echographic examination, and a nodular area of retinal thickening was seen by optical coherence tomography. Pathologic analysis of lung biopsy and nasopharynx biopsy specimens revealed undifferentiated NPC. CONCLUSIONS: Retinal metastases need to be considered in the differential diagnosis of patients presenting with NPC and vision changes.

Photodynamic therapy and vitrectomy for a large optic nerve hemangioma with neovascularization and retinal detachment: a clinicopathologic correlation.

PURPOSE: To report the clinicopathologic correlation of a young man with a von Hippel-Lindau disease-associated peripapillary hemangioblastoma and its satisfactory response to a combination of photodynamic therapy (PDT) and vitrectomy. DESIGN: Clinicopathologic correlation. METHODS: We studied the case of a 14-year-old boy with an optic nerve mass and large inferior exudative retinal detachment complicated by a significant tractional component from extensive secondary neovascularization over the lesion. RESULTS: A juxtapapillary hemangioblastoma with secondary neovascularization was documented by clinical examination, fundus photography, and optical coherence tomography. A von Hippel-Lindau gene mutation was detected. The patient responded satisfactorily to a combination of PDT and vitrectomy. CONCLUSIONS: A staged approach to treatment of peripapillary hemangioblastoma with a combination of PDT and vitrectomy may be favorable to therapy with one modality.

LOC387715/HTRA1 and complement factor H variants in patients with age-related macular degeneration seen at the mayo clinic.

PURPOSE: To confirm association of the complement factor H allelic variant (CFH Y402H) and the LOC387715/HTRA1 (LOC387715 A69S) risk alleles with age-related macular degeneration (AMD). STUDY POPULATION: Study of 89 Caucasian patients with neovascular (exudative) AMD and 232 Caucasian controls. METHODS: The Y402H variant of CFH gene and A69S variant of LOC387715/HTRA1 gene locus were examined. RESULTS: For CFH, the odds ratio for the homozygous variant was 4.97 (CI 2.52 to 9.79). For LOC387715/HTRA1 the odds ratio for the homozygous risk variant was 7.75 (CI 3.46 to 17.35). The odds ratio for heterozygous carriers was 3.35 (CI 1.91 to 5.90).

The effect of L-thyroxine supplementation in a neonatal rat model of ROP.

PURPOSE: The role of L-thyroxine in retinopathy of prematurity (ROP) is controversial. Recent animal studies suggest both high and low levels of serum thyroxine (exogenous supplementation and pharmacologic inhibition) are associated with preretinal neovascularization (NV) or retinal vascular retardation, a precursor of NV. To address this controversy, we studied L-thyroxine supplementation in an animal model of ROP. METHODS: Five hundred newborn Sprague-Dawley rats were raised in 20 expanded litters of 25, under conditions of fluctuating high and low oxygen and high carbon dioxide, to induce preretinal neovascularization. Rats received either 3 days of intraperitoneal T4, 7 days of T4 or saline control. Doses of T4 ranged from 0.005 microg/g to 0.5 microg/g. Retinae from left eyes were dissected, flat-mounted, and ADPase-stained. The presence and severity of NV, retinal vascular area, and retinal vascular density were scored in a masked manner. RESULTS: The incidence of NV was similar in rats receiving either 3 days of T4 or 7 days of T4 and saline controls (55% and 43% NV in 3-day experiments [0.05 microg g-1 day-1 and 0.5 microg g-1 day-1] compared with 51% in saline controls, p = 0.49; 52% and 38% in 7-day experiments [0.005 microg g-1 day-1 and 0.05 microg g-1 day-1], p = 0.22). Retinal vascular area and vessel density were also similar to saline controls. CONCLUSIONS: Systemic T4 supplementation does not increase, or decrease, the incidence or severity of preretinal neovascularization in an animal model of ROP, despite its positive effect on overall animal growth. Further work is needed to elucidate the potential role of premature infant hypothyroidism in the pathogenesis of ROP.

The effect of L-thyroxine supplementation on retinal vascular development in neonatal rats.

PURPOSE: Thyroxine (T4) plays a role in neuroretinal maturation, but little is known regarding its role in retinal vascularization. The neonatal rat retina is incompletely vascularized at birth, providing a model for the human premature infant retina and for retinopathy of prematurity (ROP). We hypothesized that T4 supplementation would accelerate vascular development of normal neonatal rat retina. METHODS: Two hundred twenty Sprague-Dawley rats were raised in litters of 10 in room air and received either 0.05 microg/g, 0.5 microg/g, or 1.0 microg/g of intraperitoneal T4 or saline control beginning on day 1 of life for 3 days, 7 days, or 3 days followed by 4 days recovery. Rats were sacrificed on either day 4 or day 8 of life. Left eyes were fixed, retinae dissected and ADPase-stained. Flat mounted retinae were digitized and total retinal areas and retinal vascular density were evaluated in a masked manner. Serum T4, thyroid stimulating hormone (TSH), and insulin-like growth factor-1 (IGF-1) were measured at each time point. RESULTS: Retinal vascular density was reduced in animals receiving daily 1 microg/g T4 compared with saline controls after 3 days of T4 (16.8 +/- 1.4 vessels/mm vs. 18.3 +/- 1.3 vessels/mm, p = 0.04) and 7 days of T4 (14.4 +/- 1.3 vessels/mm vs. 16.8 +/- 1.1 vessels/mm, p < 0.0006). However, retinal vascular density returned to normal after 3 days of treatment and 4 days of recovery. Vascularized retinal area was reduced in animals receiving 1 microg/g T4 for 3 days followed by 4 days recovery compared with saline controls (85 +/- 6% vs. 92 +/- 3%, p = 0.002). At lower doses of T4 (0.05 microg/g and 0.5 microg/g for 3 or 7 days) and at 1 microg/g T4 for 7 days, there was no effect on vascularized retinal area. Serum T4 levels were increased, with corresponding TSH suppression, after T4 treatment for 3 or 7 days. Serum IGF-1 levels were unaffected by T4 supplementation. CONCLUSIONS: Systemic T4 supplementation at 1 microg/g per day was detrimental to retinal vascular development in neonatal animals. If these effects are paralleled in human neonates, T4 supplementation might increase, rather than decrease, the risk of developing ROP. Further work on the role of T4 in the pathogenesis of ROP is warranted.

The GLH proteins, Caenorhabditis elegans P granule components, associate with CSN-5 and KGB-1, proteins necessary for fertility, and with ZYX-1, a predicted cytoskeletal protein.

The GLH proteins belong to a family of four germline RNA helicases in Caenorhabditis elegans. These putative ATP-dependent enzymes localize to the P granules, which are nonmembranous complexes of protein and RNA exclusively found in the cytoplasm of all C. elegans germ cells and germ cell precursors. To determine what proteins the GLHs bind, C. elegans cDNA libraries were screened by the yeast two-hybrid method, using GLHs as bait. Three interacting proteins, CSN-5, KGB-1, and ZYX-1, were identified and further characterized. GST pull-down assays independently established that these proteins bind GLHs. CSN-5 is closely related to the subunit 5 protein of COP9 signalosomes, conserved multiprotein complexes of plants and animals. RNA interference (RNAi) with csn-5 results in sterile worms with small gonads and no oocytes, a defect essentially identical to that produced by RNAi with a combination of glh-1 and glh-4. KGB-1 is a putative JNK MAP kinase that GLHs bind. A kgb-1 deletion strain has a temperature-sensitive, sterile phenotype characterized by the absence of mature oocytes and the presence of trapped, immature oocytes that have undergone endoreplication. ZYX-1 is a LIM domain protein most like vertebrate Zyxin, a cytoskeletal adaptor protein. In C. elegans, while zyx-1 appears to be a single copy gene, neither RNAi depletion nor a zyx-1 deletion strain results in an obvious phenotype. These three conserved proteins are the first members in each of their families reported to associate with germline helicases. Similar to the loss of GLH-1 and GLH-4, loss of either CSN-5 or KGB-1 causes oogenesis to cease, but does not affect the initial assembly of P granules.