Poor prognosis of elderly individuals >80 years of age with acute retinal necrosis

Purpose: To report the clinical features and prognosis of acute retinal necrosis (ARN) in elderly (>80 years of age) individuals. Methods: Six consecutive patients with unilateral ARN who attended the Department of Ophthalmology at Yamaguchi University Hospital between 2014 and 2015 were retrospectively reviewed. Clinical characteristics, causative virus, time from symptom onset to physician visit, visual acuity at presentation and final visit, and treatment were evaluated and compared between the three elderly and three middle-aged (<80 years) patients. Results: Varicella zoster virus (VZV) DNA was detected in aqueous humor by the polymerase chain reaction in all six cases. The mean ± SD time between symptom onset and medical attention was 18.0 ± 8.7 and 8.3 ± 1.5 days in the elderly and middle-aged groups, respectively. All patients were treated with intravenous aciclovir, oral prednisolone, and a nonsteroidal anti-inflammatory drug, and five of the six patients also received oral valaciclovir and underwent vitrectomy. The final best corrected visual acuity of the affected eye was worse for the elderly patients (20/400, hand motion, and light perception negative) than for the middle-aged patients (20/15, 20/50, and 20/25). Conclusions and importance: ARN in the elderly individuals of the present study was caused by VZV infection and associated with a poorer visual prognosis compared with that of middle-aged patients. A delay in the onset of antiviral treatment might contribute to the poor prognosis of elderly patients with ARN

Attenuation of choroidal neovascularization by dietary intake of ω-3 long-chain polyunsaturated fatty acids and lutein in mice

Dietary ω-3 long-chain polyunsaturated fatty acids (LCPUFAs) and lutein each protect against age-related macular degeneration (AMD). We here examined the effects of ω-3 LCPUFAs and lutein supplementation in a mouse model of AMD. Mice were assigned to four groups: (1) a control group fed an ω-3 LCPUFA–free diet, (2) a lutein group fed an ω-3 LCPUFA–free diet with oral administration of lutein, (3) an ω-3 group fed an ω-3 LCPUFA–supplemented diet, and (4) an ω-3 + lutein group fed an ω-3 LCPUFA–supplemented diet with oral administration of lutein. Mice were fed the defined diets beginning 2 weeks before, and received lutein with an oral gavage needle beginning 1 week before, induction of choroidal neovascularization (CNV) by laser photocoagulation. The area of CNV measured in choroidal flat-mount preparations was significantly reduced in mice fed ω-3 LCPUFAs or lutein compared with those in the control group, and it was reduced in an additive manner in those receiving both ω-3 LCPUFAs and lutein. The concentrations of various inflammatory mediators in the retina or choroid were reduced in mice fed ω-3 LCPUFAs or lutein, but no additive effect was apparent. The generation of reactive oxygen species (ROS) in chorioretinal lesions revealed by dihydroethidium staining as well as the expression of NADPH oxidase 4 (Nox4) in the retina revealed by immunohistofluorescence and immunoblot analyses were attenuated by ω-3 LCPUFAs and lutein in a synergistic manner. Our results thus show that dietary intake of ω-3 LCPUFAs and lutein attenuated CNV in an additive manner and in association with suppression of inflammatory mediator production, ROS generation, and Nox4 expression. Dietary supplementation with both ω-3 LCPUFAs and lutein warrants further study as a means to protect against AMD

Attenuation of choroidal neovascularization by dietary intake of ω-3 long-chain polyunsaturated fatty acids and lutein in mice

Dietary ω-3 long-chain polyunsaturated fatty acids (LCPUFAs) and lutein each protect against age-related macular degeneration (AMD). We here examined the effects of ω-3 LCPUFAs and lutein supplementation in a mouse model of AMD. Mice were assigned to four groups: (1) a control group fed an ω-3 LCPUFA–free diet, (2) a lutein group fed an ω-3 LCPUFA–free diet with oral administration of lutein, (3) an ω-3 group fed an ω-3 LCPUFA–supplemented diet, and (4) an ω-3 + lutein group fed an ω-3 LCPUFA–supplemented diet with oral administration of lutein. Mice were fed the defined diets beginning 2 weeks before, and received lutein with an oral gavage needle beginning 1 week before, induction of choroidal neovascularization (CNV) by laser photocoagulation. The area of CNV measured in choroidal flat-mount preparations was significantly reduced in mice fed ω-3 LCPUFAs or lutein compared with those in the control group, and it was reduced in an additive manner in those receiving both ω-3 LCPUFAs and lutein. The concentrations of various inflammatory mediators in the retina or choroid were reduced in mice fed ω-3 LCPUFAs or lutein, but no additive effect was apparent. The generation of reactive oxygen species (ROS) in chorioretinal lesions revealed by dihydroethidium staining as well as the expression of NADPH oxidase 4 (Nox4) in the retina revealed by immunohistofluorescence and immunoblot analyses were attenuated by ω-3 LCPUFAs and lutein in a synergistic manner. Our results thus show that dietary intake of ω-3 LCPUFAs and lutein attenuated CNV in an additive manner and in association with suppression of inflammatory mediator production, ROS generation, and Nox4 expression. Dietary supplementation with both ω-3 LCPUFAs and lutein warrants further study as a means to protect against AMD

Inhibitory effect of nintedanib on VEGF secretion in retinal pigment epithelial cells induced by exposure to a necrotic cell lysate.

Necrosis is a form of cell death that results in rupture of the plasma membrane and the release of cellular contents, and it can give rise to sterile inflammation in the retina and other tissues. The secretion of vascular endothelial growth factor (VEGF) by retinal pigment epithelial (RPE) cells contributes to retinal homeostasis as well as to pathological angiogenesis. We have now examined the effect of a necrotic cell lysate prepared from human RPE cells (NLR) on the release of VEGF by healthy RPE cells. We found that NLR markedly increased the release of VEGF from RPE cells and that this effect was attenuated by nintedanib, a multiple receptor tyrosine kinase inhibitor, whereas it was unaffected by inhibitors of NF-κB signaling or of caspase-1. NLR also induced the phosphorylation of extracellular signal-regulated kinase (Erk) and signal transducer and activator of transcription 3 (Stat3) in a manner sensitive to inhibition by nintedanib, although inhibitors of Erk and Stat3 signaling pathways did not affect NLR-induced VEGF secretion. In addition, nintedanib attenuated the development of choroidal neovascularization in mice. Our results have thus shown that a necrotic lysate of RPE cells induced VEGF secretion from healthy RPE cells and that this effect was mediated by receptor tyrosine kinase signaling. They therefore suggest that VEGF secretion by healthy RPE cells is a potential therapeutic target for retinal diseases associated with sterile inflammation and pathological angiogenesis

Dendritic cells mediate the anti-inflammatory action of omega-3 long-chain polyunsaturated fatty acids in experimental autoimmune uveitis.

We previously showed that dietary omega (ω)-3 long-chain polyunsaturated fatty acids (LCPUFAs) suppress inflammation in mice with experimental autoimmune uveitis (EAU). We have now investigated the role of antigen presenting cells (APCs) in this action of ω-3 LCPUFAs. C57BL/6 mice were fed a diet supplemented with ω-3 or ω-6 LCPUFAs for 2 weeks, after which splenocytes were isolated from the mice and cocultured with CD4+ T cells isolated from mice with EAU induced by injection of a human interphotoreceptor retinoid-binding protein peptide together with complete Freund's adjuvant. The proliferation of and production of interferon-γ and interleukin-17 by T cells from EAU mice in vitro were attenuated in the presence of splenocytes from ω-3 LCPUFA-fed mice as compared with those from mice fed ω-6 LCPUFAs. Splenocyte fractionation by magnetic-activated cell sorting revealed that, among APCs, dendritic cells (DCs) were the target of ω-3 LCPUFAs. Adoptive transfer of DCs from mice fed ω-3 LCPUFAs attenuated disease progression in EAU mice as well as the production of pro-inflammatory cytokines by T cells isolated from these latter animals. The proliferation of T cells from control Balb/c mice was also attenuated in the presence of DCs from ω-3 LCPUFA-fed mice as compared with those from ω-6 LCPUFA-fed mice. Furthermore, T cell proliferation in such a mixed lymphocyte reaction was inhibited by prior exposure of DCs from mice fed an ω-6 LCPUFA diet to ω-3 LCPUFAs in vitro. Our results thus suggest that DCs mediate the anti-inflammatory action of dietary ω-3 LCPUFAs in EAU

Effects of ω-3 LCPUFAs and lutein on Nox4 expression in the choroid-retina.

<p>(<b>A</b>) Representative images of Nox4 immunofluorescence (blue) in RPE-choroid sections prepared from mice of the four experimental groups at 7 days after laser photocoagulation. Red ovals enclose the lesion area. Scale bar, 100 μm. (<b>B</b>) The amount of Nox4 in the retina isolated from mice of the four experimental groups at 7 days after laser photocoagulation was determined by densitometric scanning of immunoblots. Data were normalized by the abundance of β-actin and are means ± SEM of triplicate determinations for a pooled sample. **<i>P</i> < 0.01, ***<i>P</i> < 0.001 versus control group. (<b>C</b>) Representative immunoblot of Nox4 and β-actin.</p

Effects of ω-3 LCPUFAs and lutein on the development of CNV.

<p>(A) The area of CNV at 7 days after laser photocoagulation was determined by staining of choroidal flat-mount preparations with Alexa Fluor 488–conjugated <i>G</i>. <i>simplicifolia</i> isolectin B4 for mice in the control group (<i>n</i> = 35 lesions), the ω-3 group (<i>n</i> = 23 lesions), the lutein group (<i>n</i> = 34 lesions), and the ω-3 + lutein group (<i>n</i> = 33 lesions). Data are means ± SEM. *<i>P</i> < 0.05, ***<i>P</i> < 0.001, ****<i>P</i> < 0.0001 (Tukey’s multiple comparison test). NS, not significant. The data shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0196037#pone.0196037.g002" target="_blank">Fig 2A</a> are from one of the three experiments performed. (<b>B</b>) Representative staining of CNV lesions for mice in the four groups. Scale bar, 100 μm.</p

Effects of ω-3 LCPUFAs and lutein on the concentrations of representative proinflammatory cytokines and chemokines in the choroid.

<p>The concentrations of IL-1β (<b>A</b>), IL-12 (p40) (<b>B</b>), TNF-α (<b>C</b>), G-CSF (<b>D</b>), MCP-1 (<b>E</b>), MIP-1α (<b>F</b>), MIP-1β (<b>G</b>), and RANTES (<b>H</b>) in the choroid of mice in the four experimental groups were determined with a multiplex assay at 7 days after CNV induction. Data are means ± SEM of triplicate determinations for a pooled sample and are the same as those shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0196037#pone.0196037.s002" target="_blank">S2 Fig</a>. *<i>P</i> < 0.05, **<i>P</i> < 0.01, ***<i>P</i> < 0.001, ****<i>P</i> < 0.0001 versus the control group (Dunnett’s test). ND, not detected.</p

Experimental design.

<p>Mice maintained on a normal diet up to 6 weeks of age were then fed a diet either supplemented with ω-3 LCPUFAs (<i>n</i> = 80) or free of ω-3 LCPUFAs (<i>n</i> = 80) beginning 2 weeks before the induction of CNV. Half of the mice on each diet received lutein daily via an oral gavage needle beginning 1 week before the induction of CNV. The animals were killed 7 days after laser photocoagulation for measurement of CNV size (<i>n</i> = 15 for each group [<i>n</i> = 5 for each of three separate experiments), serum fatty acid and lutein concentrations (<i>n</i> = 5 for each group from among the 15 mice examined for measurement of CNV size, and one sample was used for the calibration of gas chromatography), and cytokine and chemokine concentrations in the retina and choroid (<i>n</i> = 5 for each group) as well as for immunoblot analysis of Nox4 (<i>n</i> = 5 for each group) and immunohistofluorescence analysis of Nox4 (<i>n</i> = 5 for each group). For detection of ROS (<i>n</i> = 10 for each group [<i>n</i> = 5 for each of two separate experiments]), the animals were killed 5 days after laser photocoagulation.</p

Effects of ω-3 LCPUFAs and lutein on ROS levels in RPE-choroid sections.

<p>(<b>A</b>) RPE-choroid sections prepared from mice in the four experimental groups at 5 days after laser photocoagulation were stained with DHE for detection of ROS. In lower panels, phase image was taken of corresponding DHE-stained images in upper panel, respectively. Scale bar, 100 μm. (<b>B</b>) Integrated density for DHE staining in the CNV area was measured with Image J software. Data are means ± SEM for four determinations of leasion of each experimental group. *<i>P</i> < 0.05 versus the control group (Dunnett’s test).</p