Background Uveal melanoma (UM) is the most common intraocular tumor in adults. Despite good primary tumor control, up to 50% of patients develop metastasis, which is lethal. UM often presents asymptomatically and is usually diagnosed by clinical examination and imaging, making it one of the few cancer types diagnosed without a biopsy. Hence, alternative diagnostic tools are needed. Circulating tumor DNA (ctDNA) has shown potential as a liquid biopsy target for cancer screening and monitoring. The aim of this study was to evaluate the feasibility and clinical utility of ctDNA detection in UM using specific UM gene mutations. Methods We used the highly sensitive digital droplet PCR (ddPCR) assay to quantify UM driver mutations (GNAQ, GNA11, PLCβ4 and CYSTLR2) in cell-free DNA (cfDNA). cfDNA was analyzed in six well established human UM cell lines with known mutational status. cfDNA was analyzed in the blood and aqueous humor of an UM rabbit model and in the blood of patients. Rabbits were inoculated with human UM cells into the suprachoroidal space, and mutated ctDNA was quantified from longitudinal peripheral blood and aqueous humor draws. Blood clinical specimens were obtained from primary UM patients (n = 14), patients presenting with choroidal nevi (n = 16) and healthy individuals (n = 15). Results The in vitro model validated the specificity and accuracy of ddPCR to detect mutated cfDNA from UM cell supernatant. In the rabbit model, plasma and aqueous humor levels of ctDNA correlated with tumor growth. Notably, the detection of ctDNA preceded clinical detection of the intraocular tumor. In human specimens, while we did not detect any trace of ctDNA in healthy controls, we detected ctDNA in all UM patients. We observed that UM patients had significantly higher levels of ctDNA than patients with nevi, with a strong correlation between ctDNA levels and malignancy. Noteworthy, in patients with nevi, the levels of ctDNA highly correlated with the presence of clinical risk factors. Conclusions We report, for the first time, compelling evidence from in vitro assays, and in vivo animal model and clinical specimens for the potential of mutated ctDNA as a biomarker of UM progression. These findings pave the way towards the implementation of a liquid biopsy to detect and monitor UM tumors.

Background/Aims: To evaluate and compare cardiovascular effects (blood pressure and heart rate) of phenylephrine 2.5% versus phenylephrine 10%, and compare pupil diameter before and after instillation of eyedrops. Methods: A total of 58 patients scheduled for funduscopy were dilated with either phenylephrine 2.5% and tropicamide 1% (group 0, n = 29 patients, 58 eyes) or phenylephrine 10% and tropicamide 1% (group 1, n = 29 patients, 58 eyes). Only one drop per eye of each drug was administered. In both groups, pupil diameter, blood pressure and heart rate were measured before and 40 min after eyedrop instillation. Results: We did not observe significant changes in blood pressure or heart rate. Mean pupil diameter in group 0, before the instillation of eyedrops, was 3.51 mm and, in group 1, 2.66 mm. After medication, mean values were 7.38 mm in group 0 and 7.42 mm in group 1. Mean variation was 3.87 mm in group 0 and 4.76 mm in group 1 (p < 0.001). Conclusion: Our results corroborate the finding that one single drop of either 2.5 or 10% phenylephrine is safe and, when 1% tropicamide is combined, satisfactory pupil dilation is achieved.

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.

BackgroundThe influence of multiple gestation on the occurrence of retinopathy of prematurity (ROP) is still not completely understood.ObjectivesTo verify the incidence of any stage of ROP and threshold ROP in singletons and in multiple gestation among preterm infants.MethodsThis was an institutional, prospective, and descriptive cohort study, which included preterm newborns with birth weight (BW) of 1500 g or less and/or gestational age (GA) of 32 weeks or less, as admitted to the neonatal units at Laranjeiras and Amparo Hospitals in Rio de Janeiro, Brazil, between January 2001 and July 2005, and whom remained hospitalized for at least 28 days. There were no exclusion criteria. Patients were divided into two groups: Group 1 included multiples; and Group 2 consisted of singletons.ResultsA total of 159 infants that remained in neonatal unit care for at least 28 days were included in this study. Group 1 comprised 56 (35%) multiples; and Group 2 comprised 103 (65%) singletons. Mean BW was 1072 g ± 272 and 1089 g ± 282 in Groups 1 and 2, respectively (analysis of variance [ANOVA] P > 0.05). Mean GA among multiple gestation (Group 1) was 29 weeks ± 2.1; and 29 weeks ± 2.4 among singletons (Group 2) (ANOVA P > 0.05). Days in oxygen therapy ranged from 0 to 188 days. Median among Group 1 was 15 days, while median in Group 2 was 10 days (Kruskal–Wallis P > 0.05). Any stage ROP was detected in 66 (41.5%) of the whole cohort comprising 159 babies. Among the 56 multiples, 30 (53.6%) achieved any stage ROP, and among 103 singletons, 36 (35%) achieved any stage ROP (Chi-square test P < 0.05). Threshold ROP occurred in 12 (7.5%) of the 159 patients included. Three (5.3%) patients from Group 1 and nine (8.7%) patients in Group 2 reached threshold ROP needing laser treatment (Fisher’s exact test P > 0.05).ConclusionThis study showed higher frequency of any stage of ROP in twins and triplets but not regarding threshold disease. Because of the relatively small number of patients in this sample, other studies are necessary to determine if gemelarity plays a role in the occurrence of ROP.

When an appropriate drug combination is used, diabetic patients can achieve mydriasis as satisfactory as non-diabetic patients, allowing adequate fundus examination and/or retinopathy treatment.