Influence of subretinal fluid in advanced stage retinopathy of prematurity on proangiogenic response and cell proliferation

Purpose The clinical phenotype of advanced stage retinopathy of prematurity (ROP, stages 4 and 5) cannot be replicated in an animal model. To dissect the molecular events that can lead up to advanced ROP, we examined subretinal fluid (SRF) and surgically dissected retrolental membranes from patients with advanced ROP to evaluate its influences on cell proliferation, angiogenic properties, and macrophage polarity. Methods: We compared our findings to SRF collected from patients with uncomplicated rhegmatogenous retinal detachment (RD) without proliferative vitreoretinopathy and surgically dissected epiretinal membrane from eyes with macular pucker. All subretinal fluid samples were equalized for protein. The angiogenic potential of SRF from ROP eyes was measured using a combination of capillary cord formation in a fibrin clot assay, and its proliferative effect was tested with a DNA synthesis of human retinal microvascular endothelial cells. Findings were compared with SRF collected from participants with uncomplicated rhegmatogenous RD without proliferative vitreoretinopathy. The ability of SRF to induce nitric oxide production was measured in vitro using murine J774A.1 macrophages. Cytokine profiles of SRF from ROP and RD eyes were measured using a multienzyme-linked immunosorbent assay (ELISA). Fluorescent immunohistochemistry of retrolental membranes from ROP was performed to detect the presence of leukocytes and the composition of tissue macrophages using markers for M1 and M2 differentiation. Results: The cytokine composition in SRF revealed that in ROP, not only were several proangiogenic factors were preferentially elevated but also the profile of proinflammatory factors was also increased compared to the RD eyes. SRF from ROP eyes supported cell proliferation and endothelial cord formation while SRF from RD eyes had inhibitory effects. SRF from eyes with ROP but not RD robustly induced nitric oxide production in macrophages. Furthermore, fluorescent immunostaining revealed a preponderance of M1 over M2 macrophages in retrolental fibrous membranes from ROP eyes. The cytokine profile and biologic properties of SRF in ROP promote a proangiogenic environment, which supports the maintenance and proliferation of fibrous membranes associated with advanced stages of ROP. In contrast, SRF from RD eyes exhibits a suppressive environment for endothelial cell proliferation and angiogenesis. Conclusions: Our investigation demonstrates that the microenvironment in advanced ROP eyes is proangiogenic and proinflammatory. These findings suggest that management of advanced ROP should not be limited to the surgical removal of the fibrovascular membranes and antiangiogenic therapy but also directed to anti-inflammatory therapy and to promote M2 activation over M1 activity

Measuring Vision With Temporally Modulated Stripes in Infants and Children with ROP

Purpose. To determine differences in preferential looking (PL) acuities using stationary and temporally modulated stripe patterns in patients with various stages of retinopathy of prematurity (ROP). Methods. We measured the PL acuities of 134 patients (ages 4 mo to 13 yr) with various stages of ROP. Patients were divided into six subgroups according to PL vision measured with stationary stripes: (1) equal to or better than 20/200 (n = 24); (2) worse than 20/200 to 20/400 (n = 10); (3) worse than 20/400 to 20/800 (n = 15); (4) worse than 20/800 to 20/1600 (n = 13); (5) worse than 20/1600 to 20/6400 (n = 26); and (6) worse than 20/6400 (n = 46; no stationary vision). Results. In the group with PL acuity equal to or better than 20/200, no difference in vision was apparent between the two methods. In patients with acuities worse than 20/200 to 20/400, the temporally modulated PL acuities were 0.23 octave better than the PL acuities measured with the stationary stripes. The difference increased to 0.86 and 1.12 octaves in the groups with visual acuities worse than 20/400 to 20/800 and worse than 20/800 to 20/1600, respectively. The difference in the group with PL acuities worse than 20/1600 to 20/6400 was 1.69 octaves. The 46 patients with no stationary vision detected only the temporally modulated stripes. Conclusions. The results suggest that the PL acuity difference between the temporally modulated and stationary stripes increases with visual impairment. Measuring PL acuity with temporally modulated stripes is an important addition to the evaluation of severely visually impaired subjects. Invest Ophthalmol Vis Sci. 1993;34:496-502. A he preferential looking (PL) test has been useful in clinical settings to evaluate vision in infants and young children. However, some patients with severe visual impairment cannot discriminate the lowest spatial freFrom. th

Polyethylene Glycol-Based Synthetic Hydrogel Sealant for Closing Vitrectomy Wounds: An In Vivo and Histological Study

Purpose We conducted an in vivo study using Dutch pigmented rabbit eyes to test the usefulness of polyethylene glycol (PEG) sealant for the closure of sutureless sclerotomies in microincisional vitrectomy surgery (MIVS). Methods: Three-port, 23-gauge vitrectomy was performed on rabbit eyes. After air leakage was confirmed by the application of 0.625% povidone–iodine at the sclerotomy site, PEG sealant was subconjunctivally injected using a 27-gauge needle through conjunctival incisions to cover the sclerotomy wounds, following which it was polymerized by the application of xenon light for 60 seconds. Ophthalmological examinations and intraocular pressure measurements were conducted the day before and 1, 3, 5, and 7 days after surgery. The eyes were enucleated for histological evaluation 7 days after surgery. Results: PEG sealant was rapidly polymerized by the application of xenon light after subconjunctival injection, and it firmly sealed the sclerotomies without air leakage, as confirmed by povidone–iodine dropping, in all cases. Conjunctival and scleral wounds closed with PEG sealant were successfully attached and remained intact till the end of the follow-up period. There was no sign of postoperative hypotony or infection in any eye, and no adverse effects of PEG sealant were found. In histological examination, linear scar formation and eosinophilic staining of collagen fibers were observed at the sclerotomy sites, while the sclerotomy tunnels appeared tightly closed. Conclusions: PEG sealant can be useful for the closure of sutureless 23-gauge vitrectomy incisions in rabbits. Translational Relevance The PEG sealant may become an effective option for closing vitrectomy incisions including pediatric cases

Open-source Software for Developing Anthropomorphic Spoken Dialog Agents

An architecture for highly-interactive human-like spoken-dialog agent is discussed in this paper. In order to easily integrate the modules of different characteristics including speech recognizer, speech synthesizer, facial-image synthesizer and dialog controller, each module is modeled as a virtual machine that has a simple common interface and is connected to each other through a broker (communication manager). The agent system under development is supported by the IPA and it will be publicly available as a software toolkit this year

Search for dark matter produced in association with bottom or top quarks in √s = 13 TeV pp collisions with the ATLAS detector

A search for weakly interacting massive particle dark matter produced in association with bottom or top quarks is presented. Final states containing third-generation quarks and miss- ing transverse momentum are considered. The analysis uses 36.1 fb−1 of proton–proton collision data recorded by the ATLAS experiment at √s = 13 TeV in 2015 and 2016. No significant excess of events above the estimated backgrounds is observed. The results are in- terpreted in the framework of simplified models of spin-0 dark-matter mediators. For colour- neutral spin-0 mediators produced in association with top quarks and decaying into a pair of dark-matter particles, mediator masses below 50 GeV are excluded assuming a dark-matter candidate mass of 1 GeV and unitary couplings. For scalar and pseudoscalar mediators produced in association with bottom quarks, the search sets limits on the production cross- section of 300 times the predicted rate for mediators with masses between 10 and 50 GeV and assuming a dark-matter mass of 1 GeV and unitary coupling. Constraints on colour- charged scalar simplified models are also presented. Assuming a dark-matter particle mass of 35 GeV, mediator particles with mass below 1.1 TeV are excluded for couplings yielding a dark-matter relic density consistent with measurements