Comparison of Retinal Structure Using Optical Coherence Tomography Angiography between Persons with and without HIV Infection.

HIV retinal microangiopathy is an important predictor for increased risk of mortality. Optical coherence tomography angiography (OCTA) can investigate microvascular changes resulting from retinal diseases. Study included 25 persons with HIV and 25 healthy persons. OCTA evaluated the vascularization of retinal layers, choriocapillary, and optic disk. HIV group had lower vessel flow density (VFD) in superficial plexus. No difference was observed in the deep plexus. VFD of the optic disk and peripapillary region showed no difference between the groups. HIV group showed a thinner retinal nerve fiber layer and smaller area of the optic disk rim. HIV infection is associated with VFD reduction in superficial retinal plexus, neural rim area reduction, and retinal nerve fiber layer thinning in individuals without microangiopathic alterations on fundus examination. Therefore, OCTA can find retinal changes before clinical evidence of retinopathy.

Optical coherence tomography angiography: microvascular alterations in diabetic eyes without diabetic retinopathy / Angiografia por tomografia de coerência óptica: alterações microvasculares em olhos diabéticos sem retinopatia diabética

ABSTRACT Purpose: To describe microvascular changes in the maculas of individuals with type 2 diabetes observed on optical coherence tomography angiography (OCTA) images. We compared the maculas of diabetic subjects without diabetic retinopathy with those of healthy subjects and correlated the findings with the clinical profiles of diabetic subjects. Methods: One eye each of 30 patients with diabetes and 30 healthy individuals were examined. The patients with diabetes underwent funduscopy, retinography, and fluorescein angiography to rule out retinopathy. All subjects underwent optical coherence tomography angiography of a macular area (6×6 mm2), and the foveal and parafoveal vascular densities were analyzed in the superficial and deep retinal vascular plexus. The foveal and parafoveal thicknesses, foveal avascular zone of the superficial plexus, and choriocapillaris flow area were also examined. The optical coherence tomography angiography results were compared between the two study groups and correlated with the following parameters: visual acuity, time since diabetes diagnosis, glycemic control, lipid profile, and renal function of patients with diabetes. Results: A minimal increase in the choriocapillaris flow area was observed in the patients with diabetes (mean area, 22.3 ± 4.6 mm2 in controls; 22.6 ± 3.9 mm2 in patients with diabetes) (p=0.017). No significant differences were observed between other optical coherence tomography angiography parameters analyzed in the two groups. Glycosylated hemoglobin and fasting blood glucose levels were significantly negatively correlated with the foveal vascular density of both plexuses; conversely, fasting blood glucose levels were positively correlated with the choriocapillaris flow area (p=0.034). The other clinical parameters were not correlated with the optical coherence tomography angiography findings. Conclusion: Optical coherence tomography angiography may not be the most appropriate tool for detecting preclinical changes in patients with diabetes, moreover, optical coherence tomography angiography; does not replace clinical examinations. Glycemic control should be the primary clinical parameter considered during retinopathy screening. Larger studies are necessary to confirm these findings.

RESUMO Objetivo: Descrever alterações microvasculares na mácula em diabéticos do tipo 2 sem retinopatia diabética e pacientes saudáveis, e correlacionar achados com perfil clínico nos diabéticos. Métodos: Foram incluídos 60 olhos de 30 diabéticos e 30 pacientes saudáveis. Diabéticos realizaram fundoscopia, retinografia® (CR2; Canon Inc., New York, New York, USA) e angiografia fluoresceínica® (TRC-50DXC; Topcon Inc., Tokyo, Japan) para descartar a presença de retinopatia. Os 60 pacientes realizaram a angiografia por tomografia de coerência óptica® (RTVue XR, Avanti, Optovue, Fremont, CA, USA) (área macular: 6 x 6 mm2) e foram analisados densidade vascular total, foveal e parafoveal no plexo capilar superficial e plexo capilar profundo, espessura foveal, espessura parafoveal, área da zona avascular da fóvea no plexo capilar superficial e área de fluxo da coriocapilar. Resultados da angiografia por tomografia de coerência óptica foram comparados entre os 2 grupos e correlacionados com acuidade visual, tempo de diabetes, controle glicêmico, perfil lipídico e função renal nos diabéticos. Resultados: Observou-se aumento mínimo da área de fluxo da coriocapilar nos diabéticos, média das áreas foi de 22,3 ± 4,6 mm2 no grupo controle e 22,6 ± 3,9 mm2 em diabéticos (p=0,017). Não foi observada diferença estatisticamente significante entre outras variáveis da angiografia por tomografia de coerência óptica analisadas nos dois grupos. Hemoglobina glicosilada e glicemia de jejum apresentaram correlação negativa estatisticamente significante com densidade vascular foveal de ambos os plexos e a glicemia de jejum se correlacionou positivamente com área de fluxo da coriocapilar (p=0,034). Outros dados clínicos avaliados não apresentaram correlação com achados da angiografia por tomografia de coerência óptica. Conclusão: Resultados sugerem que a angiografia por tomografia de coerência óptica pode não ser a melhor ferramenta na detecção de alterações pré-clínicas em diabéticos, não substituindo o exame clínico, e corroboram a ideia de que o controle glicêmico deve ser o principal parâmetro clínico a ser considerado na triagem da retinopatia. Estudos com amostras maiores são necessários para confirmar os achados.

Optical coherence tomography angiography: microvascular alterations in diabetic eyes without diabetic retinopathy.

PURPOSE: To describe microvascular changes in the maculas of individuals with type 2 diabetes observed on optical coherence tomography angiography (OCTA) images. We compared the maculas of diabetic subjects without diabetic retinopathy with those of healthy subjects and correlated the findings with the clinical profiles of diabetic subjects. METHODS: One eye each of 30 patients with diabetes and 30 healthy individuals were examined. The patients with diabetes underwent funduscopy, retinography, and fluorescein angiography to rule out retinopathy. All subjects underwent optical coherence tomography angiography of a macular area (6×6 mm2), and the foveal and parafoveal vascular densities were analyzed in the superficial and deep retinal vascular plexus. The foveal and parafoveal thicknesses, foveal avascular zone of the superficial plexus, and choriocapillaris flow area were also examined. The optical coherence tomography angiography results were compared between the two study groups and correlated with the following parameters: visual acuity, time since diabetes diagnosis, glycemic control, lipid profile, and renal function of patients with diabetes. RESULTS: A minimal increase in the choriocapillaris flow area was observed in the patients with diabetes (mean area, 22.3 ± 4.6 mm2 in controls; 22.6 ± 3.9 mm2 in patients with diabetes) (p=0.017). No significant differences were observed between other optical coherence tomography angiography parameters analyzed in the two groups. Glycosylated hemoglobin and fasting blood glucose levels were significantly negatively correlated with the foveal vascular density of both plexuses; conversely, fasting blood glucose levels were positively correlated with the choriocapillaris flow area (p=0.034). The other clinical parameters were not correlated with the optical coherence tomography angiography findings. CONCLUSION: Optical coherence tomography angiography may not be the most appropriate tool for detecting preclinical changes in patients with diabetes, moreover, optical coherence tomography angiography; does not replace clinical examinations. Glycemic control should be the primary clinical parameter considered during retinopathy screening. Larger studies are necessary to confirm these findings.

Evaluation of the retinal vasculature of patients with branch retinal vein occlusion using optical coherence tomography angiography / Avaliação da vasculatura da retina em pacientes com oclusão de ramo da veia central da retina utilizando a angiografia por tomografia de coerência óptica

ABSTRACT Purpose: To evaluate vascular density in super­ficial and deep capillary plexuses of the retina, measured using optical coherence tomography angiography in patients with branch retinal vein occlusion. Affected eyes were compared with the contralateral eye of the same patient and both were compared with normal eyes. Methods: A cross-sectional study including 16 previously untreated patients with branch retinal vein occlusion. Patients with poor quality examinations, bilateral disease, high refractive error, or any other retinal or choroidal disease were excluded. A total of 31 patients without eye disease were also selected as a comparison group. All participants underwent five optical coherence tomography angiographies, and only those with at least two good quality examinations were selected. The Kruskal-Wallis, Wilcoxon signed-rank, and Mann-Whitney U tests were used for the statistical analysis. Results: Vascular density was lower in affected eyes compared with contralateral eyes: whole density (p=0.020 for capillary plexuses superficial; p=0.049 for deep capillary plexuses) and parafoveal density (p=0.020 for capillary plexuses superficial; p=0.011 for deep capillary plexuses). Vascular density was also lower in affected eyes compared with normal eyes: whole density (p<0.001 for capillary plexuses superficial and deep) and parafoveal density (p<0.001 for capillary plexuses superficial and deep). Whole density (p=0.001 for capillary plexuses superficial and deep) and parafoveal density (p=0.001 for capillary plexuses superficial; p<0.001 for deep capillary plexuses) were both lower in the contralateral eyes compared with normal eyes. Following adjustment for arterial hypertension, this difference was no longer observed. Conclusions: Vascular density in capillary plexuses and deep capillary plexuses was lower in the eyes affected by branch retinal vein occlusion. Furthermore, the lower vascular density noted in the contralateral eyes indicates that changes most likely occurred in these eyes prior to the appearance of any clinically detectable alterations, reflecting the early signs of hypertensive retinopathy.

RESUMO Objetivo: Avaliar a densidade vascular do plexo capilar superficial e profundo da retina, usando angiografia por tomografia de coerência óptica em pacientes com oclusão de ramo da veia central da retina, comparando o olho afetado com o contralateral do mesmo paciente e ambos com olhos normais. Métodos: Estudo transversal. Incluídos dezesseis pacientes com oclusão de ramo da veia central da retina sem tratamento prévio. Pacientes com exames de baixa qualidade, altas ametropias, outras patologias de retina ou coróide foram excluídos. Para comparação, trinta e um pacientes sem doença ocular foram selecionados. Todos foram submetidos a cinco exames angiografia por tomografia de coerência óptica, apenas aqueles com pelo menos dois exames de boa qualidade permaneceram no estudo. Os testes Kruskal-Wallis, Wilcoxon, e Mann-Whitney foram utilizados. Resultados: Densidades vasculares mais baixas do plexo capilar superficial e plexo capilar profundo foram observadas quando olhos com oclusão de ramo da veia central da retina foram comparados com os contralaterais: densidade total (p=0,02 para plexo capilar superficial, p=0,049 para plexo capilar profundo), densidade parafoveal (p=0,02 para plexo capilar superficial, p=0,011 para plexo capilar profundo). Comparando olhos acometidos com olhos normais, também foram observadas densidades vasculares mais baixas de plexo capilar superficial e plexo capilar profundo: densidade total (ambos com p<0,001) e densidade parafoveal (ambos com p<0,001). Quando os olhos contralaterais foram comparados aos normais, tanto a densidade total do plexo capilar superficial e plexo capilar profundo (ambos com p=0,001) quanto a densidade parafoveal (plexo capilar superficial com p=0,001, plexo capilar profundo com p<0,001) foram menores. Ao se realizar uma subanálise, minimizando o fator hipertensão arterial, esta diferença não se manteve. Conclusões: Densidades vasculares mais baixas do plexo capilar superficial e do plexo capilar profundo foram observadas em olhos com oclusão de ramo da veia central da retina. Além disso, a presença de densidades vasculares mais baixas nos olhos contralaterais mostra que já existem altera­ções nesses olhos antes das alterações clínicas, devido a al­terações inicias da retinopatia hipertensiva.

Acetylsalicylic Acid Exerts Potent Antitumor and Antiangiogenic Effects in Cutaneous and Uveal Melanoma Cell Lines.

INTRODUCTION: Acetylsalicylic acid (ASA) has been investigated for a potential anticancer role in several cancers, such as colorectal, ovarian, and endometrial cancer. Moreover, ASA has been shown to abrogate various processes that contribute to tumor growth and progression. OBJECTIVE: The aim of this study was to evaluate the effects of ASA on cutaneous melanoma (CM) and uveal melanoma (UM). METHODS: Human CM and UM cells were treated with 5 mM ASA and assessed for changes in cellular functions. Antiangiogenic effects of ASA were determined using an ELISA-based assay for 10 proangiogenic cytokines, and then validated by Western blot. Finally, proteomic analysis of ASA-treated cells was performed to elucidate the changes that may be responsible for ASA-mediated effects in melanoma cells. RESULTS: Treatment with ASA significantly inhibited the proliferation, invasion, and migration capabilities, and caused a significant decrease in angiogenin and PIGF secretion in both CM and UM. Mass spectrometry revealed 179 protein changes associated with ASA in the CM and UM cell lines. CONCLUSIONS: These results suggest that ASA may be effective as an adjuvant therapy in metastatic CM and UM. Future studies are needed to determine the regulating targets that are responsible for the antitumor effects of ASA.

Evaluation of the retinal vasculature of patients with branch retinal vein occlusion using optical coherence tomography angiography.

PURPOSE: To evaluate vascular density in super-ficial and deep capillary plexuses of the retina, measured using optical coherence tomography angiography in patients with branch retinal vein occlusion. Affected eyes were compared with the contralateral eye of the same patient and both were compared with normal eyes. METHODS: A cross-sectional study including 16 previously untreated patients with branch retinal vein occlusion. Patients with poor quality examinations, bilateral disease, high refractive error, or any other retinal or choroidal disease were excluded. A total of 31 patients without eye disease were also selected as a comparison group. All participants underwent five optical coherence tomography angiographies, and only those with at least two good quality examinations were selected. The Kruskal-Wallis, Wilcoxon signed-rank, and Mann-Whitney U tests were used for the statistical analysis. RESULTS: Vascular density was lower in affected eyes compared with contralateral eyes: whole density (p=0.020 for capillary plexuses superficial; p=0.049 for deep capillary plexuses) and parafoveal density (p=0.020 for capillary plexuses superficial; p=0.011 for deep capillary plexuses). Vascular density was also lower in affected eyes compared with normal eyes: whole density (p<0.001 for capillary plexuses superficial and deep) and parafoveal density (p<0.001 for capillary plexuses superficial and deep). Whole density (p=0.001 for capillary plexuses superficial and deep) and parafoveal density (p=0.001 for capillary plexuses superficial; p<0.001 for deep capillary plexuses) were both lower in the contralateral eyes compared with normal eyes. Following adjustment for arterial hypertension, this difference was no longer observed. CONCLUSIONS: Vascular density in capillary plexuses and deep capillary plexuses was lower in the eyes affected by branch retinal vein occlusion. Furthermore, the lower vascular density noted in the contralateral eyes indicates that changes most likely occurred in these eyes prior to the appearance of any clinically detectable alterations, reflecting the early signs of hypertensive retinopathy.

Effects of ranibizumab and amfenac on the functional abilities and radiosensitivity of uveal melanoma cells / Efeitos do ranibizumabe e do amfenac nas habilidades funcionais e na radiossensibilidade de células de melanoma uveal

ABSTRACT Purpose: To evaluate the effects of ranibizumab and amfenac in human uveal melanoma cell lines and to explore the ability of these compounds to sensitize uveal melanoma cells to radiation therapy. Methods: The 92.1 human uveal melanoma cell line was cultured and subjected to the proposed treatment (ranibizumab, amfenac, and a combination of both). Proliferation, migration, and invasion assays of the 92.1 uveal melanoma cell line were assessed after pretreatment with ranibizumab (125 mg/mL), amfenac (150 nM), or a combination of both. In addition, proliferation rates were assessed after treatment with ranibizumab and amfenac, and the cells were subsequently exposed to various radiation doses (0, 4, and 8 Gy). Results: Proliferation assay: cells treated with a combination of ranibizumab and amfenac had lower proliferation rates than controls (p=0.016) and than those treated with only ranibizumab (p=0.033). Migration assay: a significantly lower migration rate was observed in cells treated with amfenac than the control (p=0.014) and than those treated with ranibizumab (p=0.044). Invasion assay: there were no significant differences among the studied groups. Irradiation exposure: in the 4 Gy dose group, there were no significant differences among any groups. In the 8 Gy dose group, treatment with ranibizumab, amfenac, and their combination prior to application of the 8 Gy radiation led to a marked reduction in proliferation rates (p=0.009, p=0.01, and p=0.034, respectively) compared with controls. Conclusion: Combination of ranibizumab and amfenac reduced the proliferation rate of uveal melanoma cells; however, only amfenac monotherapy significantly decreased cell migration. The radiosensitivity of the 92.1 uveal melanoma cell line increased following the administration of ranibizumab, amfenac, and their combination. Further investigation is warranted to determine if this is a viable pretreatment strategy to render large tumors amenable to radiotherapy.

RESUMO Objetivo: Avaliar os efeitos do ranibizumabe em associação com o amfenac nas células de melanoma uveal humano e explorar a capacidade desses compostos em sensibilizar as células de melanoma uveal à radioterapia. Métodos: Células de melanoma uveal humano do tipo 92.1 foram cultivadas e submetidas ao tratamento proposto (ranibizumabe, amfenac e a combinação de ambos). Ensaios de proliferação, migração e invasão com as células de melanoma uveal do tipo 92.1 foram avaliados após tratamento com ranibizumabe (125 mg/ml), amfenac (150 nM) e a combinação de ambos. Além disso, as taxas de proliferação foram avaliadas após tratamento com ranibizumabe e amfenac com subsequente exposição das células a diferentes doses de radiação (0 Gy, 4 Gy e 8 Gy). Resultados: Ensaio de proliferação: células tratadas com ranibizumabe e amfenac combinados apresentaram taxas de proliferação inferiores em comparação ao grupo controle (p=0,016), do que as tratadas apenas com ranibizumabe (p=0,033). Ensaio de migração: foi observada uma taxa de migração significativamente mais baixa nas células tratadas com amfenac do que no grupo controle (p=0,014) e do que nas tratadas com ranibizumabe (p=0,044). Ensaio de invasão: não houve diferenças significativas entre os grupos estudados. Exposição à irradiação: no grupo da dose de 4 Gy, não houve diferença significante entre os grupos. No grupo da dose de 8 Gy, o tratamento com ranibizumabe, afenac e sua combinação antes da aplicação da radiação de 8 Gy levou a uma redução acentuada nas taxas de proliferação (p=0,009, p=0,01 e p=0,034, respectivamente) em comparação aos grupos controle. Conclusão: A combinação de ranibizumabe e amfenac reduziu a taxa de proliferação das células de melanoma uveal; no entanto, apenas o amfenac diminuiu significativamente a migração celular. A radiossensibilidade das células de melanoma uveal do tipo 92.1 aumentou após a administração de ranibizumabe, amfenac e sua combinação. Mais investigações são necessárias para determinar se esta é uma estratégia de pré-tratamento viável para tornar grandes tumores passíveis de radioterapia.

Effects of ranibizumab and amfenac on the functional abilities and radiosensitivity of uveal melanoma cells.

PURPOSE: To evaluate the effects of ranibizumab and amfenac in human uveal melanoma cell lines and to explore the ability of these compounds to sensitize uveal melanoma cells to radiation therapy. METHODS: The 92.1 human uveal melanoma cell line was cultured and subjected to the proposed treatment (ranibizumab, amfenac, and a combination of both). Proliferation, migration, and invasion assays of the 92.1 uveal melanoma cell line were assessed after pretreatment with ranibizumab (125 mg/mL), amfenac (150 nM), or a combination of both. In addition, proliferation rates were assessed after treatment with ranibizumab and amfenac, and the cells were subsequently exposed to various radiation doses (0, 4, and 8 Gy). RESULTS: Proliferation assay: cells treated with a combination of ranibizumab and amfenac had lower proliferation rates than controls (p=0.016) and than those treated with only ranibizumab (p=0.033). Migration assay: a significantly lower migration rate was observed in cells treated with amfenac than the control (p=0.014) and than those treated with ranibizumab (p=0.044). Invasion assay: there were no significant differences among the studied groups. Irradiation exposure: in the 4 Gy dose group, there were no significant differences among any groups. In the 8 Gy dose group, treatment with ranibizumab, amfenac, and their combination prior to application of the 8 Gy radiation led to a marked reduction in proliferation rates (p=0.009, p=0.01, and p=0.034, respectively) compared with controls. CONCLUSION: Combination of ranibizumab and amfenac reduced the proliferation rate of uveal melanoma cells; however, only amfenac monotherapy significantly decreased cell migration. The radiosensitivity of the 92.1 uveal melanoma cell line increased following the administration of ranibizumab, amfenac, and their combination. Further investigation is warranted to determine if this is a viable pretreatment strategy to render large tumors amenable to radiotherapy.

Zika and the Eye: Pieces of a Puzzle.

Zika virus (ZIKV) is an arbovirus mainly transmitted to humans by mosquitoes from Aedes genus. Other ways of transmission include the perinatal and sexual routes, blood transfusion, and laboratory exposure. Although the first human cases were registered in 1952 in African countries, outbreaks were only reported since 2007, when entire Pacific islands were affected. In March 2015, the first cases of ZIKV acute infection were notified in Brazil and, to date, 48 countries and territories in the Americas have confirmed local mosquito-borne transmission of ZIKV. Until 2015, ZIKV infection was thought to only cause asymptomatic or mild exanthematous febrile infections. However, after explosive ZIKV outbreaks in Polynesia and Latin American countries, it was confirmed that ZIKV could also lead to Guillain-Barré syndrome and congenital birth abnormalities. These abnormalities, which can include neurologic, ophthalmologic, audiologic, and skeletal findings, are now considered congenital Zika syndrome (CZS). Brain abnormalities in CZS include cerebral calcifications, malformations of cortical development, ventriculomegaly, lissencephaly, hypoplasia of the cerebellum and brainstem. The ocular findings, which are present in up to 70% of infants with CZS, include iris coloboma, lens subluxation, cataract, congenital glaucoma, and especially posterior segment findings. Loss of retinal pigment epithelium, the presence of a thin choroid, a perivascular choroidal inflammatory infiltrate, and atrophic changes within the optic nerve were seen in histologic analyses of eyes from deceased fetuses. To date, there is no ZIKV licensed vaccines or antiviral therapies are available for treatment. Preventive measures include individual protection from mosquito bites, control of mosquito populations and the use of barriers measures such as condoms during sexual intercourse or sexual abstinence for couples either at risk or after confirmed infection. A literature review based on studies that analyzed ocular findings in mothers and infants with CZS, with or without microcephaly, was conducted and a theoretical pathophysiologic explanation for ZIKV-ocular abnormalities was formulated.

Blue-light filtering alters angiogenic signaling in human retinal pigmented epithelial cells culture model.

BACKGROUND: Light exposure and more specifically the spectrum of blue light contribute to the oxidative stress in Age-related macular degeneration (AMD). The purpose of the study was to establish whether blue light filtering could modify proangiogenic signaling produced by retinal pigmented epithelial (RPE) cells under different conditions simulating risk factors for AMD. METHODS: Three experiments were carried out in order to expose ARPE-19 cells to white light for 48 h with and without blue light-blocking filters (BLF) in different conditions. In each experiment one group was exposed to light with no BLF protection, a second group was exposed to light with BLF protection, and a control group was not exposed to light. The ARPE-19 cells used in each experiment prior to light exposure were cultured for 24 h as follows: Experiment 1) Normoxia, Experiment 2) Hypoxia, and Experiment 3) Lutein supplemented media in normoxia. The media of all groups was harvested after light exposure for sandwich ELISA-based assays to quantify 10 pro-angiogenic cytokines. RESULTS: A significant decrease in angiogenin secretion levels and a significant increase in bFGF were observed following light exposure, compared to dark conditions, in both normoxia and hypoxia conditions. With the addition of a blue light-blocking filter in normoxia, a significant increase in angiogenin levels was observed. Although statistical significance was not achieved, blue light filters reduce light-induced secretion of bFGF and VEGF to near normal levels. This trend is also observed when ARPE-19 cells are grown under hypoxic conditions and when pre-treated with lutein prior to exposure to experimental conditions. CONCLUSIONS: Following light exposure, there is a decrease in angiogenin secretion by ARPE-19 cells, which was abrogated with a blue light - blocking filter. Our findings support the position that blue light filtering affects the secretion of angiogenic factors by retinal pigmented epithelial cells under normoxic, hypoxic, and lutein-pretreated conditions in a similar manner.

Quantitative Assessment of Microstructural Changes of the Retina in Infants With Congenital Zika Syndrome.

Importance: A better pathophysiologic understanding of the neurodevelopmental abnormalities observed in neonates exposed in utero to Zika virus (ZIKV) is needed to develop treatments. The retina as an extension of the diencephalon accessible to in vivo microcopy with spectral-domain optical coherence tomography (SD-OCT) can provide an insight into the pathophysiology of congenital Zika syndrome (CZS). Objective: To quantify the microstructural changes of the retina in CZS and compare these changes with those of cobalamin C (cblC) deficiency, a disease with potential retinal maldevelopment. Design, Setting, and Participants: This case series included 8 infants with CZS and 8 individuals with cblC deficiency. All patients underwent ophthalmologic evaluation at 2 university teaching hospitals and SD-OCT imaging in at least 1 eye. Patients with cblC deficiency were homozygous or compound heterozygotes for mutations in the methylmalonic aciduria and homocystinuria type C (MMACHC) gene. Data were collected from January 1 to March 17, 2016, for patients with CZS and from May 4, 2015, to April 23, 2016, for patients with cblC deficiency. Main Outcomes and Measures: The SD-OCT cross-sections were segmented using automatic segmentation algorithms embedded in the SD-OCT systems. Each retinal layer thickness was measured at critical eccentricities using the position of the signal peaks and troughs on longitudinal reflectivity profiles. Results: Eight infants with CZS (5 girls and 3 boys; age range, 3-5 months) and 8 patients with cblC deficiency (3 girls and 5 boys; age range, 4 months to 15 years) were included in the analysis. All 8 patients with CZS had foveal abnormalities in the analyzed eyes (8 eyes), including discontinuities of the ellipsoid zone, thinning of the central retina with increased backscatter, and severe structural disorganization, with 3 eyes showing macular pseudocolobomas. Pericentral retina with normal lamination showed a thinned (<30% of normal thickness) ganglion cell layer (GCL) that colocalized in 7 of 8 eyes with a normal photoreceptor layer. The inner nuclear layer was normal or had borderline thinning. The central retinal degeneration was similar to that of cblC deficiency. Conclusions and Relevance: Congenital Zika syndrome showed a central retinal degeneration with severe GCL loss, borderline inner nuclear layer thinning, and less prominent photoreceptor loss. The findings provide the first, to date, in vivo evidence in humans for possible retinal maldevelopment with a predilection for retinal GCL loss in CZS, consistent with a murine model of the disease and suggestive of in utero depletion of this neuronal population as a consequence of Zika virus infection.

Pretreatment of RPE Cells with Lutein Can Mitigate Bevacizumab-Induced Increases in Angiogenin and bFGF.

PURPOSE: To determine whether pretreatment of retinal pigmented epithelial (RPE) cells with lutein can affect the response of cells to bevacizumab therapy. METHODS: One human RPE cell line (ARPE-19) was used for all experiments. The cells were treated with lutein in different concentrations (0.01, 0.1, 1, 10, or 100 µg/ml). After 24 h, all plates were treated with bevacizumab (0.25 mg/ml). Media were harvested 24 h later for sandwich ELISA-based angiogenesis arrays. A Quantibody Human Angiogenesis Array was used in order to quantify the secretion of the following 10 proangiogenic cytokines: angiogenin, ANG2, EGF, bFGF, HB-EGF, PDGF-BB, leptin, PIGF, HGF and VEGF. RESULTS: Treatment with bevacizumab alone led to a significant decrease in VEGF, as well as a significant increase in angiogenin and bFGF. Pretreatment with 0.1 and 1.0 µg/ml of lutein led to significant decreases in both bFGF and angiogenin following treatment with bevacizumab compared to bevacizumab treatment alone. Lutein alone did not modify the secretion of proangiogenic cytokines. CONCLUSIONS: Pretreatment of human RPE cells in culture with specific doses of lutein prior to bevacizumab treatment mitigated the increase in bFGF and angiogenin caused by bevacizumab monotherapy.

Optical Coherence Tomography of Retinal Lesions in Infants With Congenital Zika Syndrome.

IMPORTANCE: Zika virus (ZIKV) can cause severe changes in the retina and choroid that may result in marked visual impairment in infants with congenital Zika syndrome (CZS), the term created for a variety of anomalies associated with intrauterine ZIKV infection. OBJECTIVE: To evaluate the affected retinal layers in infants with CZS and associated retinal abnormalities using optical coherence tomography (OCT). DESIGN, SETTING, AND PARTICIPANTS: This cross-sectional, consecutive case series included 8 infants (age range, 3.0-5.1 months) with CZS. Optical coherence tomographic images were obtained in the affected eyes of 7 infants with CZS who had undergone previous ophthalmologic examinations on March 17, 2016, and in 1 infant on January 1, 2016. An IgM antibody-capture enzyme-linked immunosorbent assay for ZIKV was performed on the cerebrospinal fluid samples of 7 of the 8 infants (88%), and other congenital infections were ruled out. MAIN OUTCOMES AND MEASURES: Observation of retinal and choroidal findings in the OCT images. RESULTS: Among the 8 infants included in the study (3 male; 5 female; mean [SD] age at examination, 4.1 [0.7] months), 7 who underwent cerebrospinal fluid analysis for ZIKV had positive findings for IgM antibodies. Eleven of the 16 eyes (69%) of the 8 infants had retinal alterations and OCT imaging was performed in 9 (82%) of them. Optical coherence tomography was also performed in 1 unaffected eye. The main OCT findings in the affected eyes included discontinuation of the ellipsoid zone and hyperreflectivity underlying the retinal pigment epithelium in 9 eyes (100%), retinal thinning in 8 eyes (89%), choroidal thinning in 7 eyes (78%), and colobomatouslike excavation involving the neurosensory retina, retinal pigment epithelium, and choroid in 4 eyes (44%). CONCLUSIONS AND RELEVANCE: Zika virus can cause severe damage to the retina, including the internal and external layers, and the choroid. The colobomatouslike finding seen in the OCT images relate to the excavated chorioretinal scar observed clinically.

High frequency of squamous intraepithelial neoplasia in pterygium related to low ultraviolet light exposure.

PURPOSE: The aim of this study was to determine the frequency of clinically unsuspected ocular surface squamous neoplasia (OSSN) in cases of biopsied pterygium (PT). METHODS: We reviewed 15,016 cases presented at the Henry C. Witelson Ocular Pathology Laboratory during the period 1993-2013. All cases with a clinical diagnosis of PT were included. Histopathological diagnoses were reviewed and demographic data were retrieved from histopathological request forms. All cases associated with OSSN were re-evaluated independently by two ocular pathologists. The classification of OSSN in PT cases was made based on the Armed Forces Institute of Pathology (AFIP) recommendations. RESULTS: Two hundred and fifteen cases were diagnosed clinically as PT (1.43%) and 54% were from male patients. The average age at diagnosis was 53.4 ± 15.5 years. OSSN was identified in five cases (2.33%), and four of these cases were from female patients (80%). The average age of patients with PT and OSSN was similar to PT patients without OSSN (P > 0.05). Cases with OSSN were diagnosed as conjunctival intraepithelial neoplasia (CIN) I (60%), CIN II (20%), and CIN III (20%). There was complete agreement between the two pathologists (100%). CONCLUSIONS: The relatively high rate of dysplasia in a low ultraviolet light index area challenges the main cause of this disease in our population, a hypothesis that should be evaluated in future studies. We suggest that all PT samples should be sent for histopathological evaluation even in areas with low ultraviolet light index.

Pericyte Status in Routinely Discarded Vitrectomy Samples May Be an Early Marker of Diabetic Retinopathy.

BACKGROUND: Diabetic retinopathy (DR) is the leading cause of blindness among the working-age population. The earliest morphological manifestation of the disease is pericyte loss, as shown by animal models. AIMS: The purpose of this study was to evaluate the presence of pericytes in vitreous samples (VS) from diabetic and nondiabetic patients. METHODS: VS from 125 patients with and without diabetes were analyzed. Thirty-three of the VS contained blood vessels and were therefore included in further analysis. Pericyte status was evaluated using α-smooth muscle actin and quantified using the following scoring system: total loss (3), >50% loss (2), <50% loss (1), and no loss (0). RESULTS: Of the 33 VS, 29 samples were from patients with diabetes and 4 from nondiabetic patients. Six diabetic cases had a score of 1, 8 diabetic cases had a score of 2, and 15 cases had a score of 3. A positive correlation between glycemia levels and pericyte loss was observed (p = 0.0016; Spearman's r = 0.61). Moreover, all nondiabetic cases had a score of 0 (sensitivity and specificity = 100%). CONCLUSION: Pericyte loss in VS might be a sensitive and specific marker of DR that correlates with glycemia levels. Furthermore, VS, which are currently discarded, may contain valuable information for diabetic management.

Usefulness of Discarded Vitreous Samples from Routine Vitrectomy.

Purpose. To describe the histopathological features of vitreous samples obtained after vitrectomy surgery from diabetic and nondiabetic patients. Methods. Vitreous specimens from 137 patients who underwent vitrectomy for different clinical conditions were analysed. All samples were centrifuged and each resulting pellet was fixed and processed as part of routine paraffin section histopathology. The histopathological features were categorized in a semiquantitative fashion. The samples from diabetic and nondiabetic patients were compared. Results. The 125 included patients (58 diabetic, 60% males) were aged 64.2 ± 13.9 years. The presence of hemorrhage, inflammatory cells, and histiocytes was significantly higher in the diabetic group (P < 0.001, P = 0.028, and P = 0.016, resp.), showing more vessels (P < 0.001) and ghost vessels (P = 0.049). The presence of inflammatory cells was the feature with the highest sensitivity for detecting diabetes mellitus (98%) and also the highest negative predictive value (89%). In the multivariate analysis, three variables emerged as independent significant predictors of diabetes in vitrectomy samples: hemorrhage, endothelial-lined vessels, and age (P < 0.001, P < 0.001, and P = 0.019, resp.). Conclusions. Different histopathological features can be found in vitreous samples from diabetic patients. Analysis of vitrectomy samples may serve as a tool for diabetes management.

Analysis of HSP90 Expression Is Valuable in the Differential Diagnosis of Ocular Surface Squamous Lesions.

OBJECTIVES: The aim of this study was to evaluate heat shock protein 90 (HSP90) expression in squamous lesions (SLs) and to assess its diagnostic value for different lesions within the SL spectrum. METHODS: A total of 70 conjunctival SLs, including 19 papillomas, 22 cases of conjunctival intraepithelial neoplasia (ConINs) I, 11 cases of ConIN II, six cases of ConIN III, and 12 squamous carcinomas (sqCAs), were evaluated using the German immunoreactive score against HSP90. RESULTS: Cytoplasmic HSP90 expression differed between low- and high-grade lesions (P < .001). Among high-grade lesions, the nuclear HSP90 score was higher in the ConIN III-sqCA group than in the ConIN II group (P = .0162). A percentage of total thickness staining of less than 73% differentiated between ConIN III and sqCA. CONCLUSIONS: The expression of HSP90 is particularly useful to differentiate low-grade from high-grade lesions of the conjunctiva. HSP90 may play an important role in the malignant transformation of SLs and could be a new target for therapy.

Sebaceous adenomas of the eyelid and Muir-Torre Syndrome.

BACKGROUND/AIMS: Sebaceous adenomas (SAs) are rare, benign sebaceous gland tumours of the eyelid. SAs may be associated with primary internal malignancies. This association is known as Muir-Torre Syndrome (MTS). The purpose of this study was to approximate the prevalence of SAs, to determine the reliability of the clinical diagnosis of SAs and to demonstrate immunohistochemical staining of DNA mismatch repair proteins mutL homologue 1 (MLH1) and mutS homologue 2 (MSH2) for a case of MTS. METHODS: We reviewed the histopathology reports from all eyelid specimens collected between 1993 and 2013 at the Henry C Witelson Ocular Pathology Laboratory to determine the proportion of SAs. For the SAs identified on histopathology, we looked at patient charts to see what diagnosis was originally suspected on clinical examination. Immunohistochemical staining for MLH1 and MSH2 was performed on all SAs to screen for MTS. RESULTS: Of the 5884 eyelid specimens collected, 9 were SAs (6 women, 3 men; 42-72 years old). The diagnosis of SA was suspected clinically in only one of the nine cases based on the gross appearance of the eyelid lesion. Immunohistochemistry revealed one SA case with positive MLH1 expression and negative MSH2 expression. These findings prompted systemic work-up and this patient was diagnosed with MTS after discovery of a colon adenocarcinoma T2M0N0. CONCLUSIONS: The diagnosis of eyelid SA is rare. The importance of this benign eyelid tumour stems from its association with internal malignancies in MTS. Immunohistochemical staining of mismatch repair proteins MLH1 and MSH2 is a valid and accessible strategy for investigating MTS in patients with SAs.

Diagnosing Pathological Prognostic Factors in Retinoblastoma: Correlation between Traditional Microscopy and Digital Slides.

OBJECTIVE: It was the aim of this study to determine the diagnostic accuracy of high-risk prognostic factors and morphological characteristics of retinoblastomas using digital whole slide images (WSI) generated by a scanner. METHODS: Forty-seven H&E sections of glass slides with high-risk morphological features of retinoblastoma were analyzed. Slides were scanned as WSI and reviewed. The results were compared with those obtained after reviewing the slides using a regular microscope as the gold standard. McNemar's test (MT), the percentage of agreement (POA), and sensitivity (S) and specificity (Sp) were evaluated between WSI and conventional microscopy. RESULTS: There were no differences with respect to multicentricity, growth type, rosette formation, choroidal invasion, anterior chamber invasion, extraocular extension, scleral extension, optic nerve invasion, necrosis, or Azzopardi effect between WSI analysis and light microscopy (MT, p = 1.0; POA = 100%; S = 100%, and Sp = 100%). Discordance was found in 1 case where calcification could not be found using WSI (MT, p = 1.00; POA = 97.9%; S = 100%, and Sp = 97.8%). CONCLUSION: To the best of our knowledge, this is the first report using digital pathology (WSI) to evaluate prognostic factors in eyes containing retinoblastomas. Using WSI, the pathologist was able to detect high-risk morphological features in retinoblastoma. To date, WSI is an important tool, in particular for ophthalmic pathologists examining enucleation and exenteration specimens.