Radiation-Induced DNA Damage in Uveal Melanoma Is Influenced by Dose Delivery and Chromosome 3 Status.

Purpose: The purpose of this study was to analyze the extent of DNA breaks in primary uveal melanoma (UM) with regard to radiotherapy dose delivery (single-dose versus fractionated) and monosomy 3 status. Methods: A total of 54 patients with UM were included. Stereotactic radiotherapy (SRT) was performed in 23 patients, with 8 undergoing single-dose SRT (sdSRT) treatment and 15 receiving fractionated SRT (fSRT). DNA breaks in the enucleated or endoresected tumors were visualized by a TUNEL assay and quantified by measuring the TUNEL-positive area. Protein expression was analyzed by immunohistochemistry. Co-detection of chromosome 3 with proteins was performed by immuno-fluorescent in situ hybridization. Results: The amount of DNA breaks in the total irradiated group was increased by 2.7-fold (P < 0.001) compared to non-irradiated tissue. Tumors treated with fSRT were affected more severely, showing 2.1-fold more DNA damage (P = 0.007) compared to the cases after single (high) dose irradiation (sdSRT). Monosomy 3 tumors showed less DNA breaks compared to disomy 3 samples (P = 0.004). The presence of metastases after radiotherapy correlated with monosomy 3 and less DNA breaks compared to patients with non-metastatic cancer in the combined group with fSRT and sdSRT (P < 0.05). Conclusions: Fractionated irradiation led to more DNA damage than single-dose treatment in primary UM. As tumors with monosomy 3 showed less DNA breaks than those with disomy 3, this may indicate that they are less radiosensitive, which may influence the efficacy of irradiation.

Quercetin Impairs the Growth of Uveal Melanoma Cells by Interfering with Glucose Uptake and Metabolism.

Monosomy 3 in uveal melanoma (UM) increases the risk of lethal metastases, mainly in the liver, which serves as the major site for the storage of excessive glucose and the metabolization of the dietary flavonoid quercetin. Although primary UMs with monosomy 3 exhibit a higher potential for basal glucose uptake, it remains unknown as to whether glycolytic capacity is altered in such tumors. Herein, we initially analyzed the expression of n = 151 genes involved in glycolysis and its interconnected branch, the "pentose phosphate pathway (PPP)", in the UM cohort of The Cancer Genome Atlas Study and validated the differentially expressed genes in two independent cohorts. We also evaluated the effects of quercetin on the growth, survival, and glucose metabolism of the UM cell line 92.1. The rate-limiting glycolytic enzyme PFKP was overexpressed whereas the ZBTB20 gene (locus: 3q13.31) was downregulated in the patients with metastases in all cohorts. Quercetin was able to impair proliferation, viability, glucose uptake, glycolysis, ATP synthesis, and PPP rate-limiting enzyme activity while increasing oxidative stress. UMs with monosomy 3 display a stronger potential to utilize glucose for the generation of energy and biomass. Quercetin can prevent the growth of UM cells by interfering with glucose metabolism.

Comparison of Automated Thresholding Algorithms in Optical Coherence Tomography Angiography Image Analysis.

(1) Background: Calculation of vessel density in optical coherence tomography angiography (OCTA) images with thresholding algorithms varies in clinical routine. The ability to discriminate healthy from diseased eyes based on perfusion of the posterior pole is critical and may depend on the algorithm applied. This study assessed comparability, reliability, and ability in the discrimination of commonly used automated thresholding algorithms. (2) Methods: Vessel density in full retina and choriocapillaris slabs were calculated with five previously published automated thresholding algorithms (Default, Huang, ISODATA, Mean, and Otsu) for healthy and diseased eyes. The algorithms were investigated with LD-F2-analysis for intra-algorithm reliability, agreement, and the ability to discriminate between physiological and pathological conditions. (3) Results: LD-F2-analyses revealed significant differences in estimated vessel densities for the algorithms (p < 0.001). For full retina and choriocapillaris slabs, intra-algorithm values range from excellent to poor, depending on the applied algorithm; the inter-algorithm agreement was low. Discrimination was good for the full retina slabs, but poor when applied to the choriocapillaris slabs. The Mean algorithm demonstrated an overall good performance. (4) Conclusions: Automated threshold algorithms are not interchangeable. The ability for discrimination depends on the analyzed layer. Concerning the full retina slab, all of the five evaluated automated algorithms had an overall good ability for discrimination. When analyzing the choriocapillaris, it might be useful to consider another algorithm.

Impact of Silicone Oil Removal on Macular Perfusion.

(1) Background: Silicone oil (SO) can be used as an endotamponade during vitreoretinal surgery for retinal detachment. There is emerging evidence that SO filling of the vitreous cavity and its removal may impact macular perfusion. So far, studies have not focused on choroidal sublayer perfusion, yet. (2) Methods: Optical coherence tomography angiography was applied in 19 patients with SO endotamponade before and four weeks after removal of SO. (3) Results: Perfusion of choriocapillaris increased significantly after SO removal, while perfusion of Haller's and Sattler's layer decreased significantly. (4) Conclusions: Removal of SO impacts choroidal perfusion and leads to a perfusion shift within choroidal sublayers. This study underlines that it is worth to conduct larger prospective studies that evaluate the choroidal perfusion and its functional implications in more detail.

Case Report: GNAQ- and SF3B1 Mutations in an Aggressive Case of Relapsing Uveal Ring Melanoma.

The molecular mechanisms for uveal ring melanoma are still unclear until today. In this case report, we describe a patient with a malignant uveal melanoma with exudative retinal detachment that had been treated with plaque brachytherapy, resulting in successful tumor regression. After 1 year, a ring-shaped recurrence with extraocular extension appeared, and the eye required enucleation. Histological and molecular genetic analyses revealed an epithelioid-cell-type melanoma with complete circumferential involvement of the ciliary body and, so far, unreported GNAQ and SF3B1 mutations in ring melanoma. Therefore, this report gives new genetic background information on this ocular tumor usually leading to enucleation.

Specific Autoantibodies in Neovascular Age-Related Macular Degeneration: Evaluation of Morphological and Functional Progression over Five Years.

(1) Background: Altered levels of autoantibodies (aab) and their networks have been identified as biomarkers for various diseases. Neovascular age-related macular degeneration (nAMD) is a leading cause for central vision loss worldwide with highly variable inter- and intraindividual disease courses. Certain aab networks could help in daily routine to identify patients with a high disease activity who need to be visited and treated more regularly. (2) Methods: We analyzed levels of aab against Angiotensin II receptor type 1 (AT1-receptor), Protease-activated receptors (PAR1), vascular endothelial growth factor (VEGF) -A, VEGF-B, and VEGF-receptor 2 in sera of 164 nAMD patients. In a follow-up period of five years, we evaluated changes in functional and morphological characteristics. Using correlation analyses, multiple regression models, and receiver operator characteristics, we assessed whether the five aab have a clinical significance as biomarkers that correspond to the clinical properties. (3) Results: Neither the analyzed aab individually nor taken together as a network showed statistically significant results that would allow us to draw conclusions on the clinical five-year course in nAMD patients. (4) Conclusions: The five aab that we analyzed do not correspond to the clinical five-year course of nAMD patients. However, larger, prospective studies should reevaluate different and more aab to gain deeper insights.

Rare Case of Bilateral Diffuse Uveal Melanocytic Proliferation with Dermal and Mucosal Hyperpigmentations.

PURPOSE: The demonstration of a rare case of bilateral diffuse uveal melanocytic proliferation (BDUMP) due to a lung carcinoma with unusual dermal lesions. CASE DESCRIPTION: A 76-year-old man with painless bilateral vision loss revealed leopard or giraffe spot chorioretinopathy and bilateral serous retinal detachment. Ultrasound biomicroscopy revealed uveal swelling expanding into the anterior chamber angle. Dermal and mucosal lesions were present on the lip, breast, groin, scrotum, and penis. Screening analyses revealed a non-small cell lung carcinoma. CONCLUSIONS: The diagnosis of BDUMP, a rare paraneoplastic syndrome, was made. The ophthalmological diagnosis led to screening investigations and revealed the underlying malignant disease. Uncommonly, multiple dermal and mucosal lesions could be detected and were analyzed histopathologically.

Influence of Retinal Microsecond Pulse Laser Treatment in Central Serous Chorioretinopathy: A Short-Term Optical Coherence Tomography Angiography Study.

BACKGROUND: Central serous chorioretinopathy (CSC) is a common macular condition characterized by detachment of the neuroretina and is a frequent cause of central vision loss in adults. Among the various therapeutic strategies, subthreshold microsecond pulsed laser (SML) treatment has become a useful option. Despite the suggested involvement of choroidal circulatory disturbances in CSC, the effects of this treatment on macular microperfusion have not been fully evaluated yet. Herein, we report the impact of SML on retinal and choroidal microvascular flow using non-invasive optical coherence tomography (OCT) angiography (OCTA). METHODS: In this study, CSC patients with persistent subretinal fluid (SRF) with or without secondary choroidal neovascularization (CNV) were included (referred to as the pachychoroid neovasculopathy (PNV) group and the CSC group, respectively). SML was conducted using a yellow (577 nm) laser with a duty cycle of 10%, spot size of 200 µm and duration of 200 ms. Best corrected visual acuity (BCVA) as well as OCT and OCTA images were evaluated at baseline and 4 weeks after SML. OCTA parameters of interest included full retinal perfusion (FRP), choriocapillaris perfusion (CCP), Sattler's layer perfusion (SLP), and Haller's layer perfusion (HLP), which were evaluated longitudinally and compared to unaffected fellow eyes. RESULTS: 27 affected eyes and 17 fellow eyes from 27 patients were included. Before treatment, central retinal thickness (CRT) and subfoveal choroidal thickness (SFCT) of affected eyes were significantly larger than in fellow eyes. Four weeks after SML, CRT decreased significantly, whereas perfusion parameters did not change. In subgroup analyses, the CSC group showed a significant decrease in SFCT, whereas the PNV group did not despite the decrease in CRT. CONCLUSION: Our results suggest that the SML may affect the SFCT of the CSC, but not the PNV patients at least within four weeks following treatment. This effect seems to be independent of the change in choroidal perfusion measured with OCTA.

Association of Locus Coeruleus and Substantia Nigra Pathology With Cognitive and Motor Functions in Patients With Parkinson Disease.

OBJECTIVE: To investigate the contribution of substantia nigra (SN) and locus coeruleus (LC) pathology to clinical signs and symptoms in Parkinson disease (PD) by applying neuromelanin-weighted imaging. METHODS: Forty-seven patients with PD and 53 matched controls underwent motor assessment, a neuropsychological test battery, and neuromelanin-weighted MRI. Patients with PD were enrolled after fulfilling the criteria for clinically established PD as defined by the Movement Disorders Society Clinical Diagnostic Criteria. Two independent raters identified SN and LC and calculated the contrast-to-noise ratio (CNR). RESULTS: The intrarater reliability demonstrated good reliability between raters with an intraclass correlation coefficient of 0.88 (p < 0.001) and an interrater reliability of 0.80 (p < 0.001). Both SN and LC CNRs were lower in patients with PD (p ≤ 0.001) compared to controls. The CNR of SN but not of LC was strongly correlated with disease duration (p ≤ 0.001). Neuromelanin pathology of the pars compacta-containing dorsolateral SN correlated with Movement Disorders Society-sponsored version of the Unified Parkinson's Disease Rating Scale I, II, and III but not cognitive function. In contrast, neuromelanin pathology of LC was associated with cognitive function in all tested domains but not with motor impairment or activities of daily living. No such associations were present in controls. CONCLUSIONS: Neuromelanin imaging of the SN and LC is well-suited to map neurodegeneration in PD. Neuromelanin pathology of the SN correlates with motor dysfunction whereas LC pathology is related to cognitive impairment. Neuromelanin-weighted imaging of the LC could thus serve as an imaging marker of executive and other cognitive dysfunction in PD. CLASSIFICATION OF EVIDENCE: This study provides Class I evidence that neuromelanin-weighted imaging was associated with the severity of various signs and symptoms in patients with PD.

Evaluating Retinal and Choroidal Perfusion Changes after Isometric and Dynamic Activity Using Optical Coherence Tomography Angiography.

Optical coherence tomography angiography (OCTA) is a non-invasive tool for imaging and quantifying the retinal and choroidal perfusion state in vivo. This study aimed to evaluate the acute effects of isometric and dynamic exercise on retinal and choroidal sublayer perfusion using OCTA. A pilot study was conducted on young, healthy participants, each of whom performed a specific isometric exercise on the first day and a dynamic exercise the day after. At baseline and immediately after the exercise, heart rate (HR), mean arterial pressure (MAP), superficial capillary plexus perfusion (SCPP), deep capillary plexus perfusion (DCPP), choriocapillaris perfusion (CCP), Sattlers's layer perfusion (SLP), and Haller's layer perfusion (HLP) were recorded. A total of 34 eyes of 34 subjects with a mean age of 32.35 ± 7.87 years were included. HR as well as MAP increased significantly after both types of exercise. Both SCPP and DCPP did not show any significant alteration due to isometric or dynamic exercise. After performing dynamic exercise, CCP, SLP, as well as HLP significantly increased. Changes in MAP correlated significantly with changes in HLP after the dynamic activity. OCTA-based analysis in healthy adults following physical activity demonstrated a constant retinal perfusion, supporting the theory of autoregulatory mechanisms. Dynamic exercise, as opposed to isometric activity, significantly changed choroidal perfusion. OCTA imaging may represent a novel and sensitive tool to expand the diagnostic spectrum in the field of sports medicine.

Three-year clinical and optical coherence tomography follow-up after stereotactic radiotherapy for neovascular age-related macular degeneration.

PURPOSE: The long-term clinical outcome of adjuvant stereotactic radiotherapy (SRT) in neovascular age-related macular degeneration (nAMD) patients was evaluated. METHODS: This case-control study included patients with unilateral nAMD, who underwent SRT complementary to standard anti-VEGF treatment. Only patients with monthly follow-up over at least three years were considered. Number of intravitreal injections, visual acuity (VA), central retinal thickness (CRT), and subfoveal choroidal thickness (SFCT) were evaluated and compared to baseline as well as to an age- and gender-matched control group, who received anti-VEGF monotherapy. RESULTS: Twenty patients were irradiated and had complete follow-up. Cumulatively, SRT patients needed significantly less injections than non-irradiated ones over three years (14 vs. 18, p â€‹= â€‹0.014), while median VA did not show statistically significant changes (0.4 logMAR at baseline to 0.65 logMAR at final follow-up, p â€‹= â€‹0.061). CRT remained steady, but SFCT showed a continuous thinning of almost 50 â€‹µm (p â€‹= â€‹0.031) in irradiated patients over three years. Multiple linear regression analysis revealed that SFCT and VA at time of irradiation are significant prognostic factors of VA change in SRT patients over the following three years (F(2,17) â€‹= â€‹23.946, p<0.001, R2 of 0.738). CONCLUSIONS: SRT significantly reduced the cumulative anti-VEGF treatment burden over three years, however, this was mainly driven by the results of the first year after irradiation. A thinner SFCT at time of irradiation was associated with poorer visual outcome. While further research and investigation are warranted to elucidate the underlying pathogenesis, SFCT could be a potential biomarker when evaluating a patient's suitability for SRT.

Correlation of retinal and choroidal microvascular impairment in systemic sclerosis.

PURPOSE: To investigate the correlation between retinal and choroidal microperfusion in patients with systemic sclerosis (SSc) using optical coherence tomography angiography (OCTA). METHODS: In this cross-sectional study SSc patients without clinical evidence of ocular involvement and healthy, age- and sex-matched volunteers were recruited. Participants underwent specific rheumatological and ophthalmological examinations, including optical coherence tomography (OCT) and OCTA. Retinal and choroidal thicknesses as well as perfusion of the retina and the choroidal sublayers were evaluated. RESULTS: A total of 15 SSc patients (30 eyes) with a median disease duration of 60 months and 15 matched, healthy controls (30 eyes) were recruited. OCT data revealed a significantly lower macular volume, as well as Sattler's layer and Haller's layer thickness in SSc patients compared to controls. In OCTA analysis, the perfusion of both retinal plexus as well as Sattler's and Haller's layer were significantly reduced in the SSc group. Patients with a disease duration of more than 60 months showed a statistically significant positive correlation between retinal and choroidal malperfusion, while those with a shorter disease duration did not. CONCLUSION: OCTA analysis confirmed impairment of retinal and choroidal microperfusion in SSc patients, supporting the hypothesis of wide spreading vascular injury. In early stages, either the retinal or the choroidal perfusion seems to be involved, while later on, vascular impairment affects both tissues alike. Both, retinal and choroidal examinations should be considered as soon as the diagnosis of SSc is made, to avoid missing out on early alterations.

Evaluating Retinal and Choroidal Perfusion Changes After Ocular Massage of Healthy Eyes Using Optical Coherence Tomography Angiography.

Background and objectives: Ocular massage (OM) is used as a treatment option for acute retinal artery occlusion, under the assumption that it induces vessel dilatation and enhances perfusion. Since evidence of ocular perfusion alteration due to OM is lacking, we investigate the impact of OM on the hemodynamics of the posterior pole in healthy eyes in a noninvasive fashion by using optical coherence tomography angiography (OCTA). Materials and Methods: A prospective study was conducted on healthy volunteers, each of whom underwent measurements of intraocular pressure (IOP), central macular thickness (CMT), subfoveal choroidal thickness (SFCT), radial peripapillary capillary perfusion (RPCP), superficial capillary plexus perfusion (SCPP), deep capillary plexus perfusion (DCPP), choriocapillaris perfusion (CCP), Sattler's layer perfusion (SLP) and Haller's layer perfusion (HLP) before and after OM. OM was performed for 2 min, consisting of 10-s turns of compression and decompression of the globe. Results: A total of 21 eyes from 21 participants (median age 29) were included. After OM, IOP significantly declined (p < 0.001), while SFCT (p < 0.005), SCPP (p < 0.001), DCPP (p = 0.004) and CCP (p = 0.008) significantly increased. CMT, RPCP, SLP and HLP did not show any significant alteration due to OM. Changes in SCPP correlated positively with changes in CCP and vice versa. Conclusions: OCTA-based analysis in healthy adults following OM demonstrated a significant increase of retinal perfusion values, assumed to be due to failure of autoregulatory mechanisms. These findings may indicate a positive effect of OM as a treatment option for patients with acute retinal artery occlusion.

Ocular Blood Flow Changes Impact Visual Acuity Gain after Surgical Treatment for Idiopathic Epiretinal Membrane.

BACKGROUND: Idiopathic epiretinal membrane (iERM) is a common eye disease, which can be treated by surgical removal of the fibrotic tissue. Morphological outcome is generally evaluated by optical coherence tomography (OCT). Here, we investigate the impact of surgery on hemodynamics of the posterior pole, using OCT angiography (OCTA). METHODS: Patients with unilateral iERM and indication for treatment were included. OCT and OCTA images of the posterior pole were obtained preoperatively and 3-months after 23G vitrectomy with membrane peeling. Parameters of interest included full retinal perfusion, choriocapillaris perfusion (CCP), Sattler's layer perfusion (SLP), and Haller's layer perfusion, which were evaluated longitudinally and also compared to unaffected fellow eyes. Using these parameters, multiple regression analyses were used to predict visual outcomes. RESULTS: Sixty-three iERM eyes were recruited, which initially showed a significant bigger central retinal thickness (p < 0.001) and total macular volume (TMV) (p < 0.001) compared to fellow eyes, while perfusion parameters were alike. Three months later, treated eyes had a statistically significant thicker subfoveal choroid (p = 0.006) and showed an increase of CCP (p = 0.003), while SLP decreased (p = 0.014). Lower preoperative TMV and higher SLP were associated with better postoperative visual outcome. CONCLUSION: In this OCTA study, iERM itself does not affect the choroidal circulation. However, preoperative choroidal circulation will be a biomarker to know the influence on the choroidal circulation after ERM surgery and may be considered as a predictor for visual outcome.

Evaluating diurnal variations in retinal perfusion using optical coherence tomography angiography.

BACKGROUND: Optical coherence tomography angiography (OCTA) is a non-invasive tool for imaging and quantifying the retinal and choroidal vasculature as well as perfusion state in healthy eyes. Choroidal perfusion is subject to diurnal variation, showing lowest perfusion in the morning and highest in the afternoon. In this index study, OCTA was used to investigate diurnal changes of the retinal perfusion in healthy adult eyes and to identify impacting factors since retinal perfusion is known to be mainly determined by autoregulatory mechanisms. METHODS: A prospective study was conducted on healthy volunteers, each of whom underwent repeated measurements of mean arterial pressure (MAP), intraocular pressure (IOP), macular volume (MV), subfoveal choroidal thickness (SFCT), foveal avascular zone (FAZ) and retinal perfusion of the superficial capillary plexus (SCP), deep capillary plexus (DCP) and full retina (FR) slab at 7 a.m. and 4 p.m. Possible influence of MAP or IOP on the retinal perfusion was evaluated. RESULTS: A total of 22 eyes of 22 participants (mean age 55.91 ± 14.84) were analysed. Significant diurnal changes from 7 a.m. to 4 p.m. were observed for MAP (p < 0.001) and SFCT (p = 0.017). The perfusion of SCP, DCP and FR as well as the size of the FAZ and the MV did not show significant fluctuation during the day. No significant correlation between MAP or IOP and retinal perfusion values were detectable. CONCLUSION: OCTA-based analysis of the retina in healthy adults demonstrated a steady perfusion of both plexus during the day, independently of changes in MAP. These findings support the theory of autoregulatory mechanisms of the retinal blood flow.

Mapping diurnal variations in choroidal sublayer perfusion in patients with idiopathic epiretinal membrane: an optical coherence tomography angiography study.

BACKGROUND: Optical coherence tomography angiography (OCTA) is a non-invasive tool for imaging and quantifying the choroidal vasculature and perfusion state. In this index study, OCTA was used to investigate diurnal changes in choroidal sublayer perfusion in eyes with idiopathic epiretinal membrane (ERM) and to identify impacting factors. METHODS: A prospective study was conducted on volunteers with symptomatic ERM, each of whom underwent repeated measurements of subfoveal choroidal thickness (SFCT) using enhanced-depth imaging optical coherence tomography and perfusion of choroidal vascular sublayers using OCTA at 7 a.m., 12 p.m., 4 p.m., and 8 p.m. Possible interactions between diurnal variations and other factors, such as gender and age, were evaluated. RESULTS: A total of 21 eyes of 21 participants (mean age 72.43 ± 7.06 years) were analysed. A significant pattern of diurnal variation was observed for SFCT (p = 0.008) as well as perfusion of Haller's layer (HLP, p = 0.001). SFCT and HLP both demonstrated a quadratic relation to time of the day, decreasing from morning to afternoon, before increasing again in the evening. No significant differences with regard to gender or age were detectable. CONCLUSION: OCTA-based analysis of choroidal sublayer perfusion demonstrated significant diurnal variations in patients with symptomatic ERM, which are quite different from changes reported in healthy eyes. Therefore, it is important to account for time of day, when comparing longitudinal OCTA data.

Subfoveal choriocapillaris, Sattler's and Haller's layer thickness predict clinical response to stereotactic radiotherapy in neovascular age-related macular degeneration patients.

PURPOSE: To evaluate the significance of choroidal substructure analysis in predicting the clinical response to adjuvant stereotactic radiotherapy (SRT) in neovascular age-related macular degeneration (nAMD) patients. METHODS: Patients with nAMD, who underwent SRT (baseline) in addition to common intravitreal injections (IVIs) and subsequently had at least 12 months of complete follow-up, were enrolled. In a post hoc analysis, optical coherence tomography (OCT) data were reviewed, and subfoveal choroidal thickness (CT) as well as the thickness of choroidal substructures, such as choriocapillaris (CC), Sattler's layer (SL), and Haller's layer (HL), was measured to determine if these influenced SRT efficacy. RESULTS: A total of 35 eyes of 35 patients were included. While each of the 4 choroidal metrics significantly forecasts the clinical response to SRT, combining them all together produced the most reliable prediction model. CONCLUSION: In terms of clinical response to SRT in nAMD patients, choroidal substructure analysis does improve the quality of the prediction model when combined with subfoveal CT.