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E Randy Craven,1 Vikas Chopra,2 Jeffrey L Goldberg,3 Kenneth M Marion,4 Xiaoming Chen,5 Cheng-Tao Chang,6 Michelle Y Chen6 1Johns Hopkins University School of Medicine, Baltimore, MD, USA; 2Doheny Eye Center UCLA, Pasadena, CA, USA; 3Byers Eye Institute at Stanford University, Palo Alto, CA, USA; 4Doheny Eye Institute, Los Angeles, CA, USA; 5West China Hospital Sichuan University, Sichuan, People’s Republic of China; 6Allergan, an AbbVie Company, Irvine, CA, USACorrespondence: E Randy Craven, 600 N. Wolfe Street, Baltimore, MD, 21287, USA, Tel +1 240-482-1100, Fax +1 240-482-1105, Email erandycraven@gmail.comPurpose: To quantitatively compare iridocorneal angle assessments using gonioscopy and anterior segment optical coherence tomography (AS-OCT).Patients: US and Chinese patients with open-angle glaucoma (OAG) and/or ocular hypertension (OHT).Methods: Analysis was pooled from 2 multicenter, noninterventional studies conducted in the US and China. Gonioscopy Shaffer grade and an AS-OCT method that approximates the angle width relative to local morphologic variations were compared by measuring the same iridocorneal angles. A third, separate, single-center, noninterventional study was conducted to verify results observed from the pooled analysis.Results: From the pooled studies, a total of 239 eyes were measured using Shaffer grade and AS-OCT. Of these, 6 were Shaffer grade 2, 37 in Shaffer grade 3, and 196 in Shaffer grade 4. There was a trend of increasing Shaffer grade with increasing AS-OCT angle width. Open iridocorneal angles, Shaffer grade ≥ 3, had a ∼ 98% sensitivity and 88% positive predictive value for identifying AS-OCT angle width ≥ 300 μm, using the AS-OCT method. To verify these results, a total of 28 right eyes were imaged for the third study. A trend of increasing Shaffer grade with increasing AS-OCT angle width was observed, and angles with Shaffer grade ≤ 2 had AS-OCT angle width < 300 μm.Conclusion: The AS-OCT method can determine the space in the anterior chamber and can potentially identify angles that are the appropriate size for certain glaucoma devices. Information gathered from AS-OCT can provide additional comprehensive and quantitative assessment to gonioscopy.Keywords: gonioscopy, anterior segment optical coherence tomography, angle assessment, open-angle glaucoma

Background: Domestic family violence (DFV) is a global health concern affecting one in three women worldwide. Women are vulnerable to DFV throughout their life; however, pregnancy introduces an increased risk of experiencing DFV for millions of women and birthing people., Methods: Routinely collected data from two hospitals in one local health district in New South Wales, Australia, were examined to determine the prevalence of DFV from 2010 to 2019. Demographics and outcome factors were compared by a reported history of DFV. Multivariable logistic regression was used to assess for predictors of DFV and to assess DFV as a predictor of adverse maternal and perinatal outcomes., Results: One percent of women (538/52,469) experienced DFV in the past year. Women experiencing domestic violence were more likely to be younger and have previous children, and had higher Edinburgh Depression Scores. These women were more likely to experience stillbirth (1.5% vs. 0.6%, p = 0.005). Maternal age < 25 years, cigarette smoking, alcohol use in pregnancy, mental health issues, and place of birth were associated with a recent history of DFV after adjusting for confounders. Recent DFV was associated with preterm birth and mental health issues but was not associated with admission to the neonatal nursery, small-for-gestational-age birthweight, or caesarean section after adjusting for confounders., Conclusion: There was a relationship between DFV and poorer health outcomes for both women and their babies. This study highlighted that stillbirth is high among the population of women who experience DFV when compared to women who do not experience DFV., (© 2024 The Author(s). Birth published by Wiley Periodicals LLC.)

Purpose: To identify optical coherence tomography (OCT) biomarkers, including thin and thick double-layer sign (DLS) for the progression from intermediate AMD (iAMD) to exudative macular neovascularization (MNV) over 24 months., Design: Retrospective cohort study., Methods: Setting: Retina consultants of Texas., Patient Population: 458 eyes of 458 subjects with iAMD in at least 1 eye with 24 months of follow-up data., Main Outcomes Measures: The following biomarkers were assessed at baseline: high central drusen volume (≥0.03 mm 3 ), intraretinal hyper-reflective foci (IHRF), subretinal drusenoid deposits, hyporeflective drusen cores, thick DLS, thin DLS, and central choroidal thickness. A binary logistic regression was computed to investigate the association between baseline OCT covariates and the conversion to exudative MNV within 24 months. In addition, fellow eye status was also included in the model., Results: During follow-up, 18.1% (83 of 458) of eyes with iAMD progressed to exudative MNV. Thick DLS, IHRF, and fellow eye exudative MNV were found to be independent predictors for the development of exudative MNV within 2 years. The baseline frequencies, odds ratios, 95% confidence intervals, and P values for these biomarkers were as follows: thick DLS (9.6%, 4.339, 2.178-8.644; P < .001), IHRF (36.0%, 2.340, 1.396-3.922; P = 0.001), and fellow eye exudative MNV (35.8%, 1.694, 1.012-2.837; P = .045)., Conclusions: Thick DLS, IHRF, and fellow eye exudative MNV were associated with an increased risk of progression from iAMD to exudative MNV. These biomarkers, which are readily identified by the review of OCT volume scans, may aid in risk prognostication for patients and for identifying patients for early intervention trials., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Purpose: To determine the frequency of multiple OCT biomarkers of intermediate age-related macular degeneration (iAMD) and their relationship with the development of complete retinal pigment epithelium and outer retinal atrophy (cRORA) after 2 years., Design: Retrospective cohort study., Participants: This retrospective analysis included 330 eyes of 330 consecutive patients with iAMD in ≥ 1 eye who had 24 months of follow-up data., Methods: Spectralis OCT volume scans (49 B-scans over 6 × 6 mm, automatic real time = 6, fovea-centered) at baseline were evaluated for the previously described iAMD biomarkers, including a high-central drusen volume (DV; ≥ 0.03 mm 3 ), intraretinal hyper-reflective foci (IHRF), subretinal drusenoid deposits (SDDs), hypo-reflective drusen cores (hDCs), and a thin or thick (multilayered) double-layer sign (DLS). The age-related macular degeneration (AMD) status in the fellow eye was also assessed and classified as normal or early AMD, iAMD, exudative macular neovascularization, or cRORA., Main Outcome Measures: Incidence of cRORA, odds ratio for demographics, and OCT features., Results: At month 24, 16.36% (54/330) of the iAMD eyes developed cRORA. Several baseline features, including high-central DV, IHRF, SDD, hDC, thin DLS, and cRORA in the fellow eye, were associated with a significantly greater risk for development of cRORA at 2 years. The odds ratio, 95% confidence interval, P value, and baseline frequencies of these biomarkers were DV (6.510, 2.467-17.176, P < 0.001, 49.1%), IHRF (12.763, 4.763-34.202, P < 0.001, 38.8%), SDD (2.307, 1.003-5.304, P = 0.049, 34.2%), hDC (3.012, 1.152-7.873, P = 0.024, 13.0%), thin DLS (4.517, 1.555-13.126, P = 0.006, 11.8%), and cRORA in the fellow eye (7.184, 1.938-26.623, P = 0.003, 8.2%)., Conclusions: In addition to the 4 previously reported factors that are present in a significant proportion of iAMD (DV, IHRF, hDC, and SDD), a thin DLS and cRORA in the fellow eye were associated with an increased risk of progression to cRORA over 2 years. These biomarkers may aid in prognostication, risk stratification, and selection of patients for clinical trials., Financial Disclosure(s): Proprietary or commercial disclosure may be found after the references., (Copyright © 2022 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

Subretinal injection (SRI) is a widely used technique in retinal research and can be used to deliver nucleic acids, small molecules, macromolecules, viruses, cells or biomaterials such as nanobeads. Here we describe how to undertake SRI of mice. This protocol was adapted from a technique initially described for larger animals. Although SRI is a common procedure in eye research laboratories, there is no published guidance on the best practices for determining what constitutes a 'successful' SRI. Optimal injections are required for reproducibility of the procedure and, when carried out suboptimally, can lead to erroneous conclusions. To address this issue, we propose a standardized protocol for SRI with 'procedure success' defined by follow-up examination of the retina and the retinal pigmented epithelium rather than solely via intraoperative endpoints. This protocol takes 7-14 d to complete, depending on the reagent delivered. We have found, by instituting a standardized training program, that trained ophthalmologists achieve reliable proficiency in this technique after ~350 practice injections. This technique can be used to gain insights into retinal physiology and disease pathogenesis and to test the efficacy of experimental compounds in the retina or retinal pigmented epithelium., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)

Purpose: Subretinal injection (SRI) in mice is widely used in retinal research, yet the learning curve (LC) of this surgically challenging technique is unknown., Methods: To evaluate the LC for SRI in a murine model, we analyzed training data from three clinically trained ophthalmic surgeons from 2018 to 2020. Successful SRI was defined as either the absence of retinal pigment epithelium (RPE) degeneration after phosphate buffered saline injection or the presence of RPE degeneration after Alu RNA injection. Multivariable survival-time regression models were used to evaluate the association between surgeon experience and success rate, with adjustment for injection agents, and to calculate an approximate case number to achieve a 95% success rate. Cumulative sum (CUSUM) analyses were performed and plotted individually to monitor each surgeon's simultaneous performance., Results: Despite prior microsurgery experience, the combined average success rate of the first 50 cases in mice was only 27%. The predicted SRI success rate did not reach a plateau above 95% until approximately 364 prior cases. Using the 364 training cases as a cutoff point, the predicted probability of success for cases 1 to 364 was 65.38%, and for cases 365 to 455 it was 99.32% (P < 0.0001). CUSUM analysis showed an initial upward slope and then remained within the decision intervals with an acceptable success rate set at 95% in the late stage., Conclusions: This study demonstrates the complexity and substantial LC for successful SRI in mice with high confidence. A systematic training system could improve the reliability and reproducibility of SRI-related experiments and improve the interpretation of experimental results using this technique., Translational Relevance: Our prediction model and monitor system allow objective quantification of technical proficiency in the field of subretinal drug delivery and gene therapy for the first time, to the best of our knowledge.

Detection of microbial products by multiprotein complexes known as inflammasomes is pivotal to host defense against pathogens. Nucleotide-binding domain leucine-rich repeat (NLR) CARD domain containing 4 (NLRC4) forms an inflammasome in response to bacterial products; this requires their detection by NLR family apoptosis inhibitory proteins (NAIPs), with which NLRC4 physically associates. However, the mechanisms underlying sterile NLRC4 inflammasome activation, which is implicated in chronic noninfectious diseases, remain unknown. Here, we report that endogenous short interspersed nuclear element (SINE) RNAs, which promote atrophic macular degeneration (AMD) and systemic lupus erythematosus (SLE), induce NLRC4 inflammasome activation independent of NAIPs. We identify DDX17, a DExD/H box RNA helicase, as the sensor of SINE RNAs that licenses assembly of an inflammasome comprising NLRC4, NLR pyrin domain–containing protein 3, and apoptosis-associated speck-like protein–containing CARD and induces caspase-1 activation and cytokine release. Inhibiting DDX17-mediated NLRC4 inflammasome activation decreased interleukin-18 release in peripheral blood mononuclear cells of patients with SLE and prevented retinal degeneration in an animal model of AMD. Our findings uncover a previously unrecognized noncanonical NLRC4 inflammasome activated by endogenous retrotransposons and provide potential therapeutic targets for SINE RNA–driven diseases.

The atrophic form of age-related macular degeneration (dry AMD) affects nearly 200 million people worldwide. There is no Food and Drug Administration (FDA)-approved therapy for this disease, which is the leading cause of irreversible blindness among people over 50 y of age. Vision loss in dry AMD results from degeneration of the retinal pigmented epithelium (RPE). RPE cell death is driven in part by accumulation of Alu RNAs, which are noncoding transcripts of a human retrotransposon. Alu RNA induces RPE degeneration by activating the NLRP3-ASC inflammasome. We report that fluoxetine, an FDA-approved drug for treating clinical depression, binds NLRP3 in silico, in vitro, and in vivo and inhibits activation of the NLRP3-ASC inflammasome and inflammatory cytokine release in RPE cells and macrophages, two critical cell types in dry AMD. We also demonstrate that fluoxetine, unlike several other antidepressant drugs, reduces Alu RNA-induced RPE degeneration in mice. Finally, by analyzing two health insurance databases comprising more than 100 million Americans, we report a reduced hazard of developing dry AMD among patients with depression who were treated with fluoxetine. Collectively, these studies identify fluoxetine as a potential drug-repurposing candidate for dry AMD., Competing Interests: Competing interest statement: M.A., B.D.G., S.N., S.-b.W., I.A., and F.P. are named as inventors on patent applications on macular degeneration filed by the University of Virginia. S.S.S. has received research grants from Boehringer Ingelheim, Gilead Sciences, Portola Pharmaceuticals, and United Therapeutics unrelated to this work. S.R.S. has been a consultant for 4DMT, Allergan, Amgen, Centervue, Heidelberg, Roche/Genentech, Novartis, Optos, Regeneron, and Thrombogenics and has received research funding from Carl Zeiss Meditec, all unrelated to this work. B.D.G. is a co-founder of DiceRx.

Long interspersed nuclear element-1 (L1)–mediated reverse transcription (RT) of Alu RNA into cytoplasmic Alu complementary DNA (cDNA) has been implicated in retinal pigmented epithelium (RPE) degeneration. The mechanism of Alu cDNA–induced cytotoxicity and its relevance to human disease are unknown. Here we report that Alu cDNA is highly enriched in the RPE of human eyes with geographic atrophy, an untreatable form of age-related macular degeneration. We demonstrate that the DNA sensor cGAS engages Alu cDNA to induce cytosolic mitochondrial DNA escape, which amplifies cGAS activation, triggering RPE degeneration via the inflammasome. The L1-extinct rice rat was resistant to Alu RNA–induced Alu cDNA synthesis and RPE degeneration, which were enabled upon L1-RT overexpression. Nucleoside RT inhibitors (NRTIs), which inhibit both L1-RT and inflammasome activity, and NRTI derivatives (Kamuvudines) that inhibit inflammasome, but not RT, both block Alu cDNA toxicity, identifying inflammasome activation as the terminal effector of RPE degeneration.

Nonfibrillar amyloid-β oligomers (AβOs) are a major component of drusen, the sub-retinal pigmented epithelium (RPE) extracellular deposits characteristic of age-related macular degeneration (AMD), a common cause of global blindness. We report that AβOs induce RPE degeneration, a clinical hallmark of geographic atrophy (GA), a vision-threatening late stage of AMD that is currently untreatable. We demonstrate that AβOs induce activation of the NLRP3 inflammasome in the mouse RPE in vivo and that RPE expression of the purinergic ATP receptor P2RX7, an upstream mediator of NLRP3 inflammasome activation, is required for AβO-induced RPE degeneration. Two classes of small molecule inflammasome inhibitors-nucleoside reverse transcriptase inhibitors (NRTIs) and their antiretrovirally inert modified analog Kamuvudines-both inhibit AβOs-induced RPE degeneration. These findings crystallize the importance of P2RX7 and NLRP3 in a disease-relevant model of AMD and identify inflammasome inhibitors as potential treatments for GA.

Alu retroelements propagate via retrotransposition by hijacking long interspersed nuclear element-1 (L1) reverse transcriptase (RT) and endonuclease activities. Reverse transcription of Alu RNA into complementary DNA (cDNA) is presumed to occur exclusively in the nucleus at the genomic integration site. Whether Alu cDNA is synthesized independently of genomic integration is unknown. Alu RNA promotes retinal pigmented epithelium (RPE) death in geographic atrophy, an untreatable type of age-related macular degeneration. We report that Alu RNA-induced RPE degeneration is mediated via cytoplasmic L1-reverse-transcribed Alu cDNA independently of retrotransposition. Alu RNA did not induce cDNA production or RPE degeneration in L1-inhibited animals or human cells. Alu reverse transcription can be initiated in the cytoplasm via self-priming of Alu RNA. In four health insurance databases, use of nucleoside RT inhibitors was associated with reduced risk of developing atrophic macular degeneration (pooled adjusted hazard ratio, 0.616; 95% confidence interval, 0.493-0.770), thus identifying inhibitors of this Alu replication cycle shunt as potential therapies for a major cause of blindness., Competing Interests: Competing interest statement: J.A. is a co-founder of iVeena Holdings, iVeena Delivery Systems, and Inflammasome Therapeutics, and has been a consultant for Allergan, Biogen, Boehringer-Ingelheim, Immunovant, Janssen, Olix Pharmaceuticals, Retinal Solutions, and Saksin LifeSciences unrelated to this work. J.A., B.D.G., B.J.F., S.N., K.A., S.-b.W., I.A., M.A., F.P., N.K., and S.F. are named as inventors on patent applications on macular degeneration filed by the University of Virginia or the University of Kentucky. J.W.H. has received consulting fees from Celgene Corporation unrelated to this work. S.S.S. has received research grants from Boehringer Ingelheim, Gilead Sciences, Portola Pharmaceuticals, and United Therapeutics unrelated to this work. J.A. and B.D.G. are co-founders of DiceRx.

Inverted papilloma of middle ear is an extremely rare lesion of the respiratory epithelium that normally occurs in the nasal cavity and paranasal sinuses. So far less than 17 cases were described in literature. A 45-year-old patient was admitted in our Department with hearing loss, otorrhea and pulsing tinnitus on the right ear. The clinical examination showed a granulation tissue on the right eardrum. No tumor formation was seen in the nasal cavity. The MRI showed a tissue formation in the tympanic cavity with an extension in the middle cranial fossa. A mastoidectomy with antrotomy and duraplasty was performed. The histological diagnosis was inverted papilloma of the middle ear. In a second step occurred an eradication of the tumor with a subtotal petrosectomy. The etiology of the inverted papilloma of the middle ear is unknown. Our case is so far the 18 nd case described.Our experience has shown that the eradication of the tumor with a subtotal petrosectomy resulted as reasonable procedure. A long-term follow-up is suggested in order to detect possible recurrence or malignant transformation., Competing Interests: The authors declare that they have no conflict of interest., (Thieme. All rights reserved.)

Purpose: To evaluate the effect of averaging en face OCT angiography (OCTA) images on quantitative measurements of the retinal microvasculature and their correlation to diabetic retinopathy (DR) disease severity., Design: Cross-sectional cohort study., Participants: One hundred five eyes (65 patients) comprising 28 eyes from 19 healthy, aged-matched control participants, 14 eyes from 9 diabetics without DR, and 63 eyes from 37 diabetics with varying levels of DR., Methods: Spectral-domain CIRRUS 5000 (Carl Zeiss Meditec, Dublin, CA) OCTA images with no macular edema or significant motion artifact were acquired 5 times with the 3 × 3-mm scan pattern. En face images of the superficial retinal layer (SRL) and deep retinal layer were registered and averaged. Vessel length density (VLD), perfusion density (PD), and foveal avascular zone (FAZ) parameters were measured on averaged versus single OCTA images., Main Outcome Measures: Univariate and multivariate linear regression correlated quantitative metrics to DR severity and best-corrected visual acuity (BCVA)., Results: Eighty-four eyes (55 patients) met the inclusion criteria. Almost uniformly, lower VLD and PD parameters were associated significantly with worse DR severity and BCVA. Multivariate linear regression for DR severity resulted in an R 2 value of 0.82 and 0.77 for single and averaged groups, respectively. No variables remained associated significantly with DR severity in multivariate analysis with single images, but in averaged images, increased superior SRL PD significantly predicted worse DR severity (coefficient, 52.7; P = 0.026). Multivariate linear regression for BCVA had an R 2 value of 0.42 and 0.47 for single and averaged groups, respectively. Foveal avascular zone size was not associated with DR severity when single OCTA images (P = 0.98) were considered, but was highly associated when using averaged images (coefficient, 6.18; P < 0.001). Foveal avascular zone size was predictive for logarithm of the minimum angle of resolution BCVA with averaged images (0.21; P = 0.004), but not with single images (P = 0.31)., Conclusions: Averaging of en face OCTA images improves the visualization of capillaries, particularly increasing the clarity of the FAZ borders, and therefore improves the correlation of vessel density and FAZ-specific parameters to DR severity and BCVA., (Copyright © 2020 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

Purpose: To identify baseline risk factors for macular atrophy (MA) development in HARBOR via a longitudinal assessment of monthly spectral-domain (SD)-OCT scans. Previous analyses of MA in HARBOR examined data from color fundus photography (CFP) and fluorescein angiography (FA)., Design: Retrospective, post hoc analysis of SD-OCT images from HARBOR (ClinicalTrials.gov identifier, NCT00891735), a phase 3, multicenter, prospective, randomized, double-blind, active treatment-controlled clinical trial., Participants: Patients (N = 1097) with subfoveal choroidal neovascularization secondary to neovascular age-related macular degeneration (nAMD) treated with intravitreal ranibizumab 0.5 mg monthly (n = 275), 0.5 mg pro re nata (PRN) after 3 loading doses (n = 275), 2.0 mg monthly (n = 274), or 2.0 mg PRN (n = 273)., Methods: Evaluable SD-OCT macular cube scans from patients with 24 months of follow-up (N = 941) were examined monthly from baseline to month 24 by masked reading center-trained graders. Atrophy diagnosis criteria were consistent with those proposed by the Classification of Atrophy Meetings (CAM) group: hypertransmission of light into the choroid, loss of retinal pigment epithelium, and loss of outer retinal layers. Multivariable proportional hazards regression was performed for time to atrophy development., Main Outcome Measures: Risk factors for MA as determined by time to MA development over 24 months of treatment., Results: Baseline risk factors for MA were confirmed from prior analyses that used CFP and FA data: absence of subretinal fluid, presence of intraretinal cysts, presence of Type 3 neovascularization, and presence of atrophy in the fellow eye. This analysis of SD-OCT data identified new baseline risk factors for MA: higher central drusen volume, lower choroidal thickness, presence of nascent atrophy, presence of reticular pseudodrusen, and increased central foveal thickness. Ranibizumab treatment regimen and dose level were not found to be risk factors for MA development., Conclusions: In this analysis of a major nAMD trial using CAM atrophy criteria, new baseline risk factors for MA development were identified using an SD-OCT dataset. Risk factors for MA development identified by prior analyses were confirmed. Monthly treatment with ranibizumab 0.5 mg was not found to be a risk factor for MA development over 24 months., (Copyright © 2020 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

Purpose: To evaluate retinal vessel quantity within various retinal structural layers using optical coherence tomography angiography (OCTA)., Methods: In this IRB-approved study, 22 normal eyes (from 22 subjects) were imaged using the Spectralis OCT2, with a 15 × 15 degree OCTA scan centered on fovea and two additional 15 × 5 degree OCTA scans, displaced temporally and nasally by 15 degrees along the fovea-Bruch's membrane opening (BMO) axis. Following projection artifact removal (PAR), vessel quantity (i.e., amount of flow signal) within each retinal nuclear and plexiform layer was assessed across the scan and was plotted as a vessel quantity profile over this fovea-BMO axis. Vessel quantity was correlated against the retinal layer thickness at the corresponding locations using the Spearman correlation., Results: For the nerve fiber layer (NFL), the vessel quantity was highest nasally and declined towards the fovea and was near zero temporal to the fovea with or without PAR. For all other retinal layers, the retinal vessel quantities were greatest in the parafoveal retina, peaking approximately 5 degrees from the foveal center. Before PAR, the parafoveal vessel quantity was highest in the inner plexiform layer (IPL). Following PAR, the vessel quantity in the IPL decreased but was relatively unchanged in the other layers. The vessel quantity correlated moderately well with retinal layer thickness (r = 0.432 to 0.511; P < 0.05 among the various layers)., Conclusions: Retinal vessel quantity varies significantly among the various structural layers, with significant regional variability. Projection artifact can significantly impact retinal vessel quantity in the deeper layers, but the effect appears to be most pronounced in the IPL.

Purpose: Azidothymidine (AZT), a nucleoside reverse transcriptase inhibitor, possesses anti-inflammatory and anti-angiogenic activity independent of its ability to inhibit reverse transcriptase. The aim of this study was to evaluate the efficacy of 5'-glucuronyl azidothymidine (GAZT), an antiretrovirally inert hepatic clinical metabolite of AZT, in mouse models of retinal pigment epithelium (RPE) degeneration and choroidal neovascularization (CNV), hallmark features of dry and wet age-related macular degeneration (AMD), respectively., Methods: RPE degeneration was induced in wild-type (WT) C57BL/6J mice by subretinal injection of Alu RNA. RPE degeneration was assessed by fundus photography and confocal microscopy of zonula occludens-1-stained RPE flat mounts. Choroidal neovascularization was induced by laser injury in WT mice, and CNV volume was measured by confocal microscopy. AZT and GAZT were delivered by intravitreous injections. Inflammasome activation was monitored by western blotting for caspase-1 and by ELISA for IL-1β in Alu RNA-treated bone marrow-derived macrophages (BMDMs)., Results: GAZT inhibited Alu RNA-induced RPE degeneration and laser-induced CNV. GAZT also reduced Alu RNA-induced caspase-1 activation and IL-1β release in BMDMs., Conclusions: GAZT possesses dual anti-inflammatory and anti-angiogenic properties and could be a viable treatment option for both forms of AMD.

Purpose: Previous studies of macular atrophy (MA) in HARBOR analyzed color fundus photography and fluorescein angiography image data. This study performed a longitudinal assessment of monthly spectral-domain (SD) OCT scans to determine MA prevalence, incidence, and progression in HARBOR., Design: Post hoc analysis of SD OCT images from HARBOR (ClincalTrials.gov identifier, NCT00891735), a phase 3 multicenter, prospective, randomized, double-blind, active treatment-controlled clinical trial., Participants: Patients (n = 1097) with subfoveal choroidal neovascularization secondary to neovascular age-related macular degeneration (nAMD) treated with ranibizumab 0.5 mg monthly (n = 275), 0.5 mg pro re nata (PRN) after 3 loading doses (n = 275), 2.0 mg monthly (n = 274), or 2.0 mg PRN (n = 273)., Methods: Evaluable SD OCT macular cube scans from patients with 24 months of follow-up (N = 941) were examined by masked reading center-trained graders monthly from baseline to month 24. Atrophy diagnosis criteria were consistent with those proposed by the Classification of Atrophy Meetings (CAM) group: hypertransmission of light into the choroid, retinal pigment epithelium loss, and loss of outer retinal layers. Macular atrophy was considered Definite if all 3 criteria were met and Questionable if 2 were met. Study arms were compared for time to MA detection (log-rank test) and enlargement rates (time × arm interaction test)., Main Outcome Measures: Prevalence, incidence, and enlargement rates of MA., Results: At baseline, imbalance in MA rates across ranibizumab arms was evident (0.5 mg monthly, 19.1%; 0.5 mg PRN, 16.1%; 2.0 mg monthly, 10.1%; 2.0 mg PRN, 10.5%). At month 24, new MA development rates in eyes without baseline MA were similar between ranibizumab doses (0.5 mg, 25.9%; 2.0 mg, 25.4%) and treatment regimens (monthly, 26.4%; PRN, 25.0%). No significant differences in enlargement rate of new atrophy area (P = 0.479, square-root transformed) or time to detection of new MA (P = 0.997) were evident among study arms., Conclusions: In this analysis of a major nAMD trial using CAM atrophy criteria, no differences were observed in incidence or progression rates of new MA among study arms, ranibizumab doses, or treatment regimens. Monthly versus PRN treatment did not influence MA incidence or progression., (Copyright © 2019 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

Purpose: To evaluate the relationship between OCT features and progression to late age related-macular degeneration (AMD) in the fellow eyes of patients enrolled in the Study of Ranibizumab Administered Monthly or on an As-needed Basis in Patients With Subfoveal Neovascular AMD (HARBOR) (ClinicalTrials.gov identifier, NCT00891735)., Design: Post hoc analysis of a phase 3 multicenter, prospective, randomized, double-masked, active treatment-controlled clinical trial., Participants: Evaluable patients (n = 501) with macular neovascularization (MNV) secondary to neovascular AMD and early or intermediate AMD in the fellow eye., Methods: Volume OCT scans from 501 fellow eyes of 501 patients with MNV were reviewed. Baseline OCT features that were assessed included intraretinal hypereflective foci (IHRF), hyporeflective foci (hRF) within drusenoid lesions (DLs), subretinal drusenoid deposits (SDDs), and drusen volume (DV) of 0.03 mm 3 or more. OCT images obtained at months 6, 12, 18, and 24 were graded by masked graders for late AMD (defined as MNV, complete retinal pigment epithelium and photoreceptor atrophy [cRORA], or both). Participant demographic characteristics (age, gender, and smoke exposure) and baseline OCT features were correlated with progression to late AMD., Main Outcome Measures: Incidence of late AMD, hazard ratio (HR) for demographics, and OCT risk factors., Results: At month 24, 33.13% of eyes (166/501) demonstrated late AMD: 20.96% (105/501) demonstrated cRORA, whereas 12.18% (61/501) demonstrated MNV. Baseline demographic factors were not associated significantly with development of late AMD, whereas significant associations were identified for all OCT features. Intraretinal hypereflective foci had an HR of 5.21 (95% confidence interval [CI], 3.29-8.26), hRF within DLs had an HR of 2.42 (95% CI, 1.74-3.38), SDD had an HR of 1.95 (95% CI, 1.34-2.82), and DV of 0.03 mm 3 or more had an HR of 1.46 (95% CI, 1.03-2.07). The correlation remained significant when considering only the progression to cRORA and MNV alone, except for DV, which was not associated significantly with progression to MNV., Conclusions: We confirmed that 4 previously reported OCT risk factors were associated with progression to late AMD in the fellow eyes of patients newly diagnosed with MNV. Although outcomes of more than 2 years were not evaluated, these findings may help to identify high-risk AMD patients., (Copyright © 2019 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

Purpose: To investigate fluorescence lifetime imaging ophthalmoscopy (FLIO) findings in preclinical Alzheimer's disease (AD)., Methods: This prospective, observational study enrolled patients with early AD undergoing Alzheimer's biomarker analysis and matched controls. Alzheimer-associated parameters (β-amyloid [Aβ], total tau in cerebrospinal fluid [CSF], Mini-Mental Status Examination [MMSE], etc.), risk factor-associated data (body mass index [BMI], hypertension, lipid profile, etc.), ganglion cell layer plus inner plexiform layer (GCIPL) thickness in structural optical coherence tomography (OCT), OCT angiography data, and FLIO-derived parameters (τ m , τ 1 , τ 2 , and τ 3 ) in short and long spectral channels (SSC and LSC) were compared and correlated between the two groups. Additional analyses were performed separately within subgroups of phakic and pseudophakic., Results: A total of 28 eyes from 15 subjects (8 control and 7 AD) were included in this analysis. In FLIO parameters, τ m in AD group showed longer lifetimes compared to the controls in phakic subjects (593.9 ± 93.3, 454.4 ± 38.6 ps; 475.0 ± 71.6, 394.1 ± 28.2  ps in SSC and LSC of AD and control groups, respectively, p = 0.036 and 0.024). Aβ, tau in CSF, and GCIPL thickness correlated with τ m in the LSC for phakic subjects (r = -0.611 to 0.562, p < 0.05 for all), but only the GCIPL thickness showed a correlation with FLIO parameters in pseudophakic subjects (r = -0.893 to -0.795, p < 0.001 for all)., Conclusion: FLIO-derived parameters appear to correlate with Aβ, tau levels in the CSF, and GCIPL thickness on OCT in AD patients. If these findings can be validated in future longitudinal studies, FLIO may prove to be useful as a simple, non-invasive diagnostic tool for AD.

Purpose: To evaluate the utility of swept-source (SS) optical coherence tomography (OCT) to objectively analyze the degree of anterior chamber (AC) inflammation., Methods: Thirty-eight eyes of 32 patients with uveitis and 20 control eyes were enrolled. SS OCT B-scans were obtained, and the number of cells in the B-scans was counted using two methods: (1) manual grading by Point Picker plug-in of Image J ( http://bigwww.epfl.ch/thevenaz/pointpicker/ ) and (2) automated grading by the Image J Particle Analysis algorithm ( http://imagej.net/Particle&#95;Analysis ). The automated and manual AC cell counts were correlated with the Standardization of Uveitis Nomenclature score., Results: The average numbers of AC inflammatory cells counted by the automated method were 8 ± 4.0, 18 ± 3.0, 42 ± 14.0, 81 ± 32.0, 117 ± 57.0, and 275 ± 67.0 cells/mm 2 for grades 0, 0.5 + , 1 + , 2 + , 3 + , and 4 + , respectively. For the same clinical categories, the average manual cell counts were 6 ± 4.0, 18 ± 3.0, 34 ± 14.0, 72 ± 32.0, 92 ± 43.0, and 168 ± 65.0 cells/mm 2 , respectively. Zero cells were detected in the AC of healthy eyes. The automated and manual methods were highly correlated (R = 0.98, p < 0.001) and showed good correlation with the clinical grading (R = 0.88, p < 0.001). A mean AC particle size of 117.4 ± 108.8 μm was obtained by the automated method., Conclusions: Quantification of the AC cells imaged by SS AS-OCT shows good correlation with categorical clinical severity assessments in uveitis eyes. This approach may provide a more objective method for monitoring uveitis and response to uveitis therapy.

Purpose: Obtaining slit-lamp gonioscopic images requires a high level of expertise and can be time consuming. The images typically are limited to 4 quadrants with limited resolution and can be affected by different lighting conditions. Our goal was to study the ability of the novel imaging system to quickly and easily document the full 360 degrees of the anterior chamber angle in a standardized manner., Design: Cross-sectional cohort study., Subjects: Eighty-four eyes from 50 subjects were included in this study, including 26 eyes from 17 normal participants and 58 eyes from 33 patients diagnosed with glaucoma., Methods: A newly introduced, commercially available gonioscope (GS-1, NIDEK Co, Gamagori, Japan) was utilized to perform semiautomated imaging of the full 360-degree iridocorneal angle on participants under an institutional review board-approved study. Topical proparacaine was administered to the eye to anesthetize the eye and imaging was completed using the 16-mirror, machine-attached gonioscopic lens coupled with lubricating ophthalmic gel ointment. The first step of the 2-step image acquisition process is analogous to manual gonioscopy, whereas the second step is performed by the instrument in an automated fashion. First, the instrument is manually moved forward by the operator to allow the machine-attached lens to make contact with the patient's corneal surface (time = 30 seconds). Second, the instrument automatically achieves fine focus on the iridocorneal angle and takes 16 sequential high-resolution photographs at multiple different focal planes (time = less than 30 seconds)., Main Outcome Measures: Successful qualitative imaging of anterior chamber iridocorneal angle in normal and glaucomatous eyes including visualization of various implanted devices within angle before and after surgery in less than 1 minute per eye., Results: In this pilot study, the full 360 degrees of iridocorneal angle of 84 eyes from 50 normal participants or glaucoma patients were successfully imaged using NIDEK GS-1 in under 60 seconds. All eyes were able to be imaged in 1 sitting, and we were able to image the full spectrum of angle from narrow/closed to fully open. The 360-degree views of the images were able to be displayed in 3 available formats (16-section display, a circular display, or a linear display). Successful imaging was also achieved of surgical devices and postsurgical conditions including CyPass, iStent, XEN, Baerveldt tube shunt, angle recession with iridodialysis, angle neovascularization, pigment dispersion, posttrabeculectomy sclerostomy and surgical iridectomy, and post-laser peripheral iridotomy., Conclusion: The newly introduced, semiautomated imaging system was able to document the full 360-degree iridocorneal angle views in normal and glaucoma eyes in less than 1 minute, which may allow longitudinal evaluation in a standardized manner. Furthermore, this system allows the ability to easily evaluate post-angle surgery changes and assess positions of implanted devices in the anterior chamber angle., (Copyright © 2019 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

Purpose: We evaluate the repeatability of fluorescence lifetime imaging ophthalmoscopy (FLIO) in normal subjects with mydriasis and explore factors that influence FLIO imaging., Method: Thirty-two healthy participants (63 eyes) were enrolled in this prospective study. The Heidelberg Engineering FLIO system uses a 473 nm blue laser light and the emitted fluorescence is detected in two wavelength channels, short and long spectral channels (SSC, LSC). The mean fluorescence lifetime (τ m ) values were computed for the entire scan area as well as in five regions of interest (ROI, 1 × 1 mm) at the fovea and superior, nasal, inferior, and temporal portions of the macula. Intraclass correlation coefficients (ICC) and coefficients of variation (CV) were used to assess the repeatability. Age, macular thickness, and vascular density also were correlated with τ m ., Results: The repeatability was good for both channels (ICC, 0.956∼0.995; CV, 9∼16%). The τ m for the entire scan was 367.8 ± 58.1 picoseconds (ps) in SSC and 322.5 ± 34.0 ps in LSC. τ m was the shortest in the fovea and significantly shorter in the temporal region compared to other regions. τ m was positively correlated with age ( r = 0.588 for SSC and r = 0.584 for LSC, P = 0.000) and retinal thickness ( r = 0.298 for SSC and r = 0.322 for LSC, P = 0.000), and negatively correlated with vascular density ( r = -0.112, P = 0.055 for SSC and r = -0.119, P = 0.040 for LSC)., Conclusion: Repeatable fluorescence lifetime values can be obtained with FLIO, but the lifetimes are affected by age, retinal thickness, vessel density, and macular location., Translational Relevance: Establishing repeatability of FLIO can introduce fluorescence lifetime imaging technique, which is used in basic science for analysis of excitation and emission wavelength spectrum of fixed and living cells into clinical practice.

Purpose: Anterior-segment OCT (AS-OCT) platforms provide noninvasive and quantifiable evaluations of anterior chamber angle (ACA) anatomy. Although these ACA measurements have been validated with high repeatability and reproducibility, there is a paucity of data previously published regarding the technique errors and artifacts (TEA) that may occur during imaging of the ACA. The purpose of this study was to report on the types and frequency of imaging TEA that are seen in ACA evaluation of a large cohort of patients with open-angle glaucoma., Design: Retrospective, cross-sectional design., Participants: Images were obtained with AS-OCT from 5082 eyes of 2541 patients with glaucoma and were reviewed at the Doheny Image Reading Center (DIRC)., Methods: Images had been captured using AS-OCT devices from 4 different manufacturers (Zeiss Cirrus [Oberkochen, Germany], Heidelberg Spectralis [Heidelberg, Germany], Optovue RTVue [Fremont, CA], or Zeiss Visante). All images were analyzed by 2 independent and certified DIRC image readers for angle opening distance under Schwalbe's line (SL-AOD), and then re-reviewed by a DIRC-certified principal investigator to adjudicate nonconsensus measurements. Images were defined to have TEA when image readers were unable to confidently measure the SL-AOD., Main Outcome Measures: Type and frequency of TEA in the AS-OCT image., Results: Of 5082 eyes reviewed, 208 images (4.09%) were found to have TEA that restricted proper visualization of irido-corneal angle structures and measurements of SL-AOD. The most common TEA was that the image readers were unable to identify Schwalbe's line (74% of TEA images). Other common TEA included poor visualization of the anterior iris surface (15%), misaligned scan location (11%), irregular corneal irregularities (5%), and illumination-related pupillary abnormalities (4%)., Conclusions: This large-scale study presents the types and frequency of TEA found in AS-OCT-derived angle images. More than 95% of the images reviewed were usable for the quantification of irido-corneal angle measurements. Knowing the common types of TEA may help in enhancing the training of both human image readers and in the design of automated algorithms to improve image acquisition and image interpretation parameters. This will become especially useful as AS-OCT becomes more widely used in clinical practice for anterior chamber assessment., (Copyright © 2019 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

Purpose: To evaluate corneal epithelial thickness (CET) and corneal thickness (CT) in healthy eyes using spectral domain optical coherence tomography., Methods: Thirty-six healthy eyes were imaged using the Cirrus high-definition (HD)-optical coherence tomography device. The average CET and CT were assessed using Cirrus Review Software within predefined concentric corneal ring-shaped zones. Specific regions of CET (superior, inferior, temporal, nasal, superonasal, inferotemporal, superotemporal, and inferonasal) were also assessed. The difference between zones was compared between males and females., Results: The average CET was 48.3, 47.1, 46.1, and 45.8 μm in the 4 concentric zones (0-2, 2-5, 5-7, and 7-9 mm), respectively (P < 0.001). The average CT was 533.5, 550.8, and 579.4 μm in the 3 zones (0-2, 2-5, and 5-7 mm), respectively (P < 0.001). There was no statistically significant correlation between CET and CT in any of the measured zones. Males had thicker corneas than did females in each of the 3 CT zones (P < 0.05), but CET did not differ significantly. The CET superonasal-inferotemporal in 2.0 to 5.0 mm and CET superotemporal-inferonasal in 5.0- to 7.0-mm zones were significantly thinner in males than in females (-1.15 vs. 0.9 μm, -3.5 vs. -1.9 μm), respectively (P < 0.05)., Conclusions: Optical coherence tomography-based analysis of CET reveals that it is thinner in the periphery, whereas the total corneal thickness is greater. Although total CT seems to be influenced by sex, CET is not. Regional and sex-based variations in CT may need to be considered when assessing corneal and epithelial alterations in the setting of disease.

Fluorescence lifetime imaging ophthalmoscopy (FLIO) is a novel technique that measures in vivo autofluorescence intensity decay over time of endogenous fluorophores in the retina. The Heidelberg Engineering FLIO system was used to obtain two 30 degree scans centered on the fovea of both eyes. The FLIO system uses a 473nm blue scanning laser light source and the emitted fluorescence is detected in two wavelengths channels, short and long spectral channels (SSC, LSC). Since the mydriatic status influence the FLIO result, the impact of mydriasis on FLIO need to be clarified. In this prospective, observational study, the impact of mydriasis on measurements from fluorescence lifetime imaging ophthalmoscope (FLIO) images in normal subjects were evaluated. 12 healthy participants (24 eyes) were volunteered and all subjects were scanned twice and the mean fluorescence lifetime (τm) values were computed with dilation and without dilation on different days. Intraclass correlation coefficients (ICC) and coefficients of variation (CV) were calculated from the measured τm in dilated, nondilated and between the dilated and non-dilated setting. Test duration was also compared and correlated with lifetimes in both settings. Repeatability was excellent for both the dilation and non-dilation settings (ICC; 0.967-0.996; 0.926-0.986, respectively). The agreement between the dilation and non-dilation settings, however, were lower (ICC; 0.688-0.970). The τm in the non-dilation setting was significantly longer than in the dilation setting for the SSC (P<0.05). The FLIO test duration in the non-dilation setting was significantly longer than with dilation for the SSC (P <0.05). Although good repeatability in τm measurements between imaging sessions were observed both with and without dilation, the agreement was not as good when comparing dilated with non-dilated measurements. Since FLIO without mydriasis results in longer τm in the SSC and takes a longer time for image acquisition, maximal dilation is recommended for FLIO testing., Competing Interests: SRS serves as a consultant for Centervue, Optos and Carl Zeiss Meditec. SRS has access to research instruments provided by Heidelberg Engineering, Topcon Medical Systems, Optos, Carl Zeiss Meditec, Nidek, and Centervue. There are no patents, products in development or marketed products associated with this research to declare. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Objectives: To determine the repeatability and reproducibility of Central Corneal Thickness (CCT) measurements using two different anterior segment imaging modalities, including those obtained with the new anterior segment lens attachments for the Cirrus 5000 HD-OCT., Methods: A total of 32 eyes from 16 normal volunteers (8 male, 8 female) were enrolled in this prospective study. CCT was measured by the same examiner using the Cirrus 5000 HD-OCT and Pentacam HR. The results of CCT obtained by each method were averaged and compared using t-test analysis. The agreement between the measurement methods was evaluated. Coefficient of Repeatability (CoR) and Intra-Class Correlation Coefficient (ICC) were computed., Results: The mean measurements taken with the Cirrus OCT anterior chamber lens (CCT AC ), HD cornea lens (CCT HDC ) and pachymetry scans (CCT Pach ) were 545.35 ± 31.02, 537.87 ± 26.82, and 532.04 ± 29.82 µm, respectively. The mean CCT obtained with the Pentacam (CCT Pent ) was 545.51 ± 30.71 µm. CCT Pent were significantly higher than CCT HDC and CCT Pach ( p < 0.0001). In contrast, the CCT Pent and CCT AC were similar ( p =0.87). CCT, as evaluated by the two different instruments, showed excellent correlation ( r > 0.98, p < 0.0001) with an ICC > 0.99 (95% CI, 0.97 - 0.99). CoR was the highest for CCT Pach (3.7 ± 1.4, 95% CI (3.0- 4.6))., Conclusion: CCT measurements from the Cirrus OCT using the new anterior segment lens attachments and the Pentacam HR are highly correlated. This should allow the use of a standardized correction factor if necessary to inter-relate the measurements between the two devices.

Geographic atrophy is a blinding form of age-related macular degeneration characterized by retinal pigmented epithelium (RPE) death; the RPE also exhibits DICER1 deficiency, resultant accumulation of endogenous Alu-retroelement RNA, and NLRP3-inflammasome activation. How the inflammasome is activated in this untreatable disease is largely unknown. Here we demonstrate that RPE degeneration in human-cell-culture and mouse models is driven by a noncanonical-inflammasome pathway that activates caspase-4 (caspase-11 in mice) and caspase-1, and requires cyclic GMP-AMP synthase (cGAS)-dependent interferon-β production and gasdermin D-dependent interleukin-18 secretion. Decreased DICER1 levels or Alu-RNA accumulation triggers cytosolic escape of mitochondrial DNA, which engages cGAS. Moreover, caspase-4, gasdermin D, interferon-β, and cGAS levels were elevated in the RPE in human eyes with geographic atrophy. Collectively, these data highlight an unexpected role of cGAS in responding to mobile-element transcripts, reveal cGAS-driven interferon signaling as a conduit for mitochondrial-damage-induced inflammasome activation, expand the immune-sensing repertoire of cGAS and caspase-4 to noninfectious human disease, and identify new potential targets for treatment of a major cause of blindness.

Purpose: To test the effect of ambient illumination scaling on the reproducibility and reliability anterior chamber metrics using the Visante time domain optical coherence tomography (TD-OCT) instrument., Materials and Methods: The inferior irido-corneal angles of 25 normal, healthy eyes were imaged twice with the Zeiss Visante TD-OCT under five strictly controlled ambient light conditions (foot candles (fc) measured with a light meter at camera/eye interface). Each eye was imaged 10 times totaling 250 assessments. Angle opening distance (AOD500/750), trabecular iris space area (TISA500/750), and scleral spur (SS) angle were graded twice by masked, trained graders at the Doheny Imaging Reading Center using the Visante's intrinsic tools. Lighting effects on measurements, intra-/inter-grader and acquisition analyses, and Bland-Altman plots were computed using Statistical Package for Social Science (SPSS Inc. version 18.0, Armonk, NY)., Results: With a near linear relationship of angle metrics to lights levels (R 2 = 0.8-0.95), the analysis examines the differences from the brightest to darkest light levels. Decreasing ambient light levels from 1.0 to 0.0 fc decreased the average AOD500 measurement from 407 ± 136 µm to 315 ± 114 µm (mean percent difference (MPD) 29%, p < 0.001), AOD750 from 587 ± 184 µm to 496 ± 155 µm (MPD 18%, p < 0.001), TISA500 from 136 ± 43 µm 2 to 101 ± 37 µm 2 (MPD 35%, p < 0.001), TISA750 from 269 ± 81 µm 2 to 212 ± 68 µm 2 (MPD 27%, p < 0.001), and SS angle from 38.3% ± 9% to 32.1% ± 9% (MPD 19%, p < 0.001). Intra-/inter-grader results showed good reproducibility for each grader (MPD = 0.7-3%; coefficient of variation (CV) = 3.2-8.3%; R 2 = 0.8-0.95; p < 0.001 for all metrics) and between graders (MPD = 1.4-5.9%; CV = 6.7-14.2%; R 2 = 0.81-0.89; Pearson Correlation Coefficient (PCC) = 0.8-0.97 (p<0.001)). Bland-Altman plots did not demonstrate any apparent bias, with similar repeatability and agreement., Conclusions: The results of this study show the high sensitivity of the anterior chamber to changes in the illumination. The slight decrease in light had a corresponding large decrease in Anterior Chamber Angle (ACA) metrics. With clinical diagnoses and treatments of eye diseases relying on these angle measurements, these findings emphasize the importance of strictly controlling light conditions in order to obtain reproducible measurements of anterior chamber geometry.

Mutations in the ganglioside-induced differentiation associated protein 1 (GDAP1) cause severe peripheral motor and sensory neuropathies called Charcot-Marie-Tooth disease. GDAP1 expression induces fission of mitochondria and peroxisomes by a currently elusive mechanism, while disease causing mutations in GDAP1 impede the protein's role in mitochondrial dynamics. In silico analysis reveals sequence similarities of GDAP1 to glutathione S-transferases (GSTs). However, a proof of GST activity and its possible impact on membrane dynamics are lacking to date. Using recombinant protein, we demonstrate for the first time theta-class-like GST activity for GDAP1, and it's activity being regulated by the C-terminal hydrophobic domain 1 (HD1) of GDAP1 in an autoinhibitory manner. Moreover, we show that the HD1 amphipathic pattern is required to induce membrane dynamics by GDAP1. As both, fission and GST activities of GDAP1, are critically dependent on HD1, we propose that GDAP1 undergoes a molecular switch, turning from a pro-fission active to an auto-inhibited inactive conformation.

Background: Evaluate the impact of variations in ambient lighting conditions on the reproducibility/reliability of Schwalbe's Line (SL)-based anterior chamber angle (ACA) metrics using spectral domain optical coherence tomography (SD-OCT)., Design: Images were taken at Doheny Eye Centers-UCLA clinic, which were randomized, masked and graded twice by dual reading centre graders., Participants: Twenty-five normal/healthy participants with open angles, Methods: Inferior angles were imaged using Cirrus SD-OCT under five light levels (foot-candles (fc) measured at camera-eye interface with Sper light-meter) with the instrument's corneal illumination function set to default level of 50 (CIdef) and low 5 (CIlow). Each eye was imaged 20 times, totaling 500 assessments., Main Outcome Measures: SL-angle-opening-distance (SL-AOD) and SL-trabecular-iris-space-area (SL-TISA) measured using custom ImageJ software. Intra-/inter-grader variability analyses were completed using Statistical-Package-for-Social-Science and Bland-Altman plots demonstrated limits of agreement for comparisons., Results: Light level demonstrated a linear relationship with angle size, thus differences from highest to lowest light levels were compared. Decreasing light from 1.0 → 0.0fc at CIdef decreased SL-AOD from 394 µm +/- 137 µm → 356 µm +/- 137 µm (mean percent difference (MPD) = 10.71%,P < 0.001) and SL-TISA from 297 µm(2)  +/- 114 µm(2)  → 261 µm(2)  +/- 109 µm(2) (MPD = 13.7%, P < 0.001). Decreasing from 1.0 → 0.0fc at CIlow decreased SL-AOD from 366 µm +/- 136 µm → 329 µm +/- 122 µm (MPD = 10.9%, P < 0.001) and SL-TISA from 271 µm(2)  +/- 113 µm(2)  → 234 µm(2)  +/- 98 µm(2) (MPD = 15.8%, P < 0.001). There was 7.9/11.4% (both P < 0.001) difference for SL-AOD/SL-TISA between CIdef → CIlow at 1.0fc, and 7.7/9.4% (both P < 0.001) difference at 0.0fc. Intra-/inter-grader results showed high reproducibility for all metrics (MPD = 0.33-4.4%; CV = 0.96-1.36; PCC = 0.93-0.95(P < 0.001); R2 = 0.94-0.98). Bland-Altman plots did not demonstrate bias, with repeat-ability and agreement among measurements., Conclusions: Using Cirrus SD-OCT, we found that SL-based ACA morphometrics are exquisitely sensitive to changes in ambient illumination and also corneal illumination by the OCT instrument. Consistently imaging in the darkest room possible is recommended (≤0.2 fce)., (© 2016 Royal Australian and New Zealand College of Ophthalmologists.)

Purpose: To compare anterior chamber angle parameters based on the location of Schwalbe line (SL) from 2 spectral domain optical coherence tomography (SD-OCT) instruments and to measure their reproducibility., Methods: Forty-two eyes from 21 normal, healthy participants underwent imaging of the inferior irido-corneal angle with the Spectralis and Cirrus SD-OCT under tightly controlled low-light conditions. SL-angle opening distance (SL-AOD) and SL-trabecular iris space area (SL-TISA) were measured by masked, certified graders at the Doheny Imaging Reading Center using customized grading software. Interinstrument and intrainstrument, as well as interobserver and intraobserver reproducibility of SL-AOD and SL-TISA measurements were evaluated by intraclass correlation coefficients (ICCs) and Bland-Altman plots with limits of agreement (LoA)., Results: The mean SL-AOD was 0.662±0.191 mm in Spectralis and 0.677±0.213 mm in Cirrus. The mean SL-TISA was 0.250±0.073 mm in Spectralis and 0.256±0.082 mm in Cirrus. The agreement for intrainstrument (ICCs>0.979), intragrader (ICCs>0.992), and intergrader (ICCs>0.929) was excellent. Excellent agreement between the 2 devices was also documented with a mean difference of -0.016 (LoA -0.125 to 0.092) mm for SL-AOD and -0.007 (LoA -0.056 to 0.043) mm in SL-TISA., Conclusions: Both SD-OCTs provided comparable measurements and permitted calculation of SL-based angle metrics. There was excellent interinstrument and intrainstrument and intraobserver and interobserver reproducibility for Spectralis and Cirrus SD-OCTs, suggesting true interchangeability between SD-OCT devices. This has the potential to lead to development of standardized grading assessments and quantification of angle parameters that would be valid across various SD-OCT devices.

Purpose. To compare the effects of physiologic versus pharmacologic pupil dilation on anterior chamber angle (ACA) measurements obtained with spectral domain optical coherence tomography (SD-OCT). Methods. Forty eyes from 20 healthy, phakic individuals with open angles underwent anterior segment OCT imaging under 3 pupillary states: (1) pupil constricted under standard room lighting, (2) physiologic mydriasis in a darkened room, and (3) postpharmacologic mydriasis. Inferior angle Schwalbe's line-angle opening distance (SL-AOD) and SL-trabecular-iris-space area (SL-TISA) were computed for each eye and pupillary condition by masked, certified Reading Center graders using customized grading software. Results. SL-AOD and SL-TISA under pupillary constriction to room light were 0.87 ± 0.31 mm and 0.33 ± 0.14 mm(2), respectively; decreased to 0.75 ± 0.29 mm (P < 0.01) and 0.29 ± 0.13 mm(2)  (P < 0.01), respectively, under physiologic mydriasis; and increased to 0.90 ± 0.38 mm (P < 0.01) and 0.34 ± 0.17 mm(2)  (P = 0.06) under pharmacologic mydriasis compared to baseline. Conclusions. Using SD-OCT imaging, pharmacologic mydriasis yielded the widest angle opening, whereas physiologic mydriasis yielded the most angle narrowing in normal individuals with open iridocorneal angles. Accounting for the state of the pupil and standardizing the lighting condition would appear to be of importance for future studies of the angle.

Background: Recombinant human thyrotropin (rhTSH; Thyrogen®) is approved for use in a 0.9 mg dose/day for 2 consecutive days for diagnosis and treatment of differentiated thyroid cancer. It is recommended that it be injected immediately after reconstitution in the distilled water diluent supplied by the manufacturer. However, Thyrogen has been used off-label in doses less than the standard 0.9 mg dose for stimulation of radioiodine uptake in the treatment of multinodular goiter. To determine whether the biologic activity of Thyrogen can be preserved after dilution, we designed experiments to assess the biologic stability of Thyrogen under different durations and temperatures of storage., Methods: rhTSH was diluted in 1% bovine serum albumin in phosphate-buffered saline to a concentration of 0.9 mg /mL and further diluted to 0.1 mg/mL. Aliquots of 0.5 mL were stored at room temperature, 4°C, -11°C, and -60°C for various lengths of time. In addition, rhTSH aliquots were also subjected to incubation for 1 hour at 50°C and to 10 cycles of freezing in dry ice alternating with thawing at 37°C. Bioassays were performed in FRTL-5 cells. rhTSH was added to the media at a final concentration of either 5 ng/mL or 20 ng/mL, and the cells were then incubated for 48 hours. Potency was assessed by measurement of ¹²⁵I-iodide uptake in comparison to cells treated with perchlorate to block iodide uptake. Samples were immunoassayed at day 185 of storage., Results: Samples stored at 4°C, -11°C, -60°C, and room temperature retained activity after storage periods of up to 204 days. Samples subjected to 10 freeze-thaw cycles or heated to 50°C for 1 hour retained full biologic activity. Immunoassay at day 185 showed no difference in immunoactivity in relation to the storage condition., Conclusion: rhTSH kept at 4°C, -11°C, -60°C, and room temperature maintained good biologic potency for more than 6 months of storage when tested in vitro, indicating that the biologic activity is very stable. However, altered sialylation occurring during storage could have altered the half-life of rhTSH. Nevertheless, the data provide reassurance that storage in the cold for a few months does not result in significant loss of biologic activity.

Background: Sunitinib malate (Sutent, Pfizer, Inc.; SU11248) is a selective, multitargeted inhibitor of receptor tyrosine kinases and has been shown to inhibit receptors for VEGF, PDGF, KIT, FLT3, and RET. The objective of this study was to determine the effects of sunitinib on signal transduction pathways and on gene expression of iodide-metabolizing proteins in papillary cancer cells with the RET/PTC1 rearrangement., Methods: We investigated the effects of sunitinib on cell growth, signal transduction pathways, and thyroid-specific gene expression in papillary thyroid cancer (PTC) cell lines that had the RET/PTC1 rearrangement., Results: Sunitinib inhibited proliferation of RET/PTC1 subclones in a time- and dose-related manner. The mean 50% lethal concentration in the RET/PTC1 subclones was 1.81 microM. Incubation of RET/PTC1 cells with 1 microM sunitinib inhibited their migration potential and transformed their morphology. Sunitinib inhibited RET autophosphorylation at Y1062 and the activation of signal transducer and activator of transcription 3 by blocking Y705 phosphorylation. Sunitinib caused cell cycle arrest in the G0/G1 phase and dephosphorylation of retinoblastoma protein, but did not induce apoptosis. Western blot analysis of the p38, MEK/ERK, and SAPK/JNK mitogen-activated protein kinase signal transduction pathways showed that sunitinib blocked ERK 1/2 and JNK phosphorylation in the cytoplasm. Sunitinib treatment of RET/PTC1 cell lines, in combination, with forskolin induced expression of the sodium (Na)/iodide (I) symporter (NIS) and the transcription factors that bind the NIS upstream enhancer. Mechanistically, the inhibition of both MEK/ERK and SAPK/JNK cytoplasmic pathways individually and in combination caused an increase in NIS gene expression., Conclusion: Sunitinib appears to target the cytosolic MEK/ERK and SAPK/JNK pathways in the RET/PTC1 cell lines, suggesting that blocking these pathways is at least part of the mechanism by which sunitinib inhibits cell proliferation and causes stimulation of NIS gene expression in RET/PTC1 cells.

Introduction: University Centre of Woundhealing, Odense University Hospital (OUH) has conducted a prevalence study of pressure ulcers. The purpose of this article is to present the results of the study., Material and Methods: All patients above the age of 15 years who were in hospital in week 5-6 2008 were included in the study. Six wound nurses and two doctors from the University Centre of Woundhealing undertook the examination. The following were recorded: length of stay, gender, age, number and grade of pressure ulcer(s). The pressure ulcer relief of the patients and documentation in the hospital journals were also recorded., Results: A total of 591 inpatients were included. In all, 164 patients were excluded and 427 patients from the project group were examined. 192 patients were found with pressure ulcers of grade 0-IV, corresponding to a prevalence of 32.5%. If grade 0 pressure ulcers were not included, the prevalence was 17.3%., Conclusion: The study showed a high prevalence. There was an overall insufficient pressure ulcer relief of patients and missing documentation in hospital journals.

Introduction: Pressure sores are a major problem for patients as well as society in general. Immobilised patients are especially at risk. This group of patients with pressure sores should be hospitalised to perform surgical revision of the wound and reconstruction using a flap. Such surgery demands extensive postoperative relief of the flap. The University Centre for Wound Healing at Odense University Hospital has tested the effects of a reduction of the formerly recommended relief period from three to two weeks., Material and Methods: In this article we report results covering all patients who have undergone surgery and reconstruction of pressure sores during the period from 1st October 2001 to 1st November 2008. The results are divided into two periods: the period before and the period after the introduction of the reduced relief period. A total of 80 patients were included; 34 in the first period and 46 in the second period., Results: We achieved a considerable reduction in median length of stay from 38 to 27 days with no increase in surgical or complication frequency. Furthermore, the share of fully healed remained unchanged., Conclusion: We believe that there is no risk in shortening the immobile postoperative relief phase following reconstruction of pressure wounds in immobilised patients.

Mutations in the GDAP1 gene lead to recessively or dominantly inherited peripheral neuropathies (Charcot-Marie-Tooth disease; CMT). Here, we demonstrate that GDAP1 is a mitochondrial fission factor whose activity is dependent on the fission factors Drp1 and Fis1. Unlike other mitochondrial fission factors, GDAP1 overexpression or knockdown does not influence the susceptibility of cells to apoptotic stimuli. Recessively inherited CMT-associated forms of GDAP1 (rmGDAP1s) have reduced fission activity, whereas dominantly inherited forms (dmGDAP1s) interfere with mitochondrial fusion. Only the expression of dmGDAP1s increases the production of ROS, leads to uneven mitochondrial transmembrane potentials, and enhances the susceptibility to apoptotic stimuli. Taken together, our results indicate that wild-type GDAP1 promotes fission without increasing the risk of apoptosis. In CMT, recessive GDAP1 mutations are associated with reduced fission activity, while dominant mutations impair mitochondrial fusion and cause mitochondrial damage. Thus, different cellular mechanisms that disturb mitochondrial dynamics underlie the similar clinical manifestations caused by GDAP1 mutations, depending on the mode of inheritance.

Background: Sunitinib, a multitargeted vascular endothelial growth factor and receptor tyrosine kinase inhibitor, causes hypothyroidism in patients who take it for treatment of cancer. Although the pathophysiologic mechanism of the hypothyroidism is unclear, it has been claimed that it is due to inhibition of iodide uptake., Methods: To evaluate the pathologic mechanism of induction of the hypothyroidism, we studied the effect of sunitinib on FRTL-5 rat thyroid cells. We measured the effect of sunitinib on cell growth, (125)I-iodide uptake and efflux, TSH receptor (TSH-R), and sodium-iodide symporter (NIS) message., Results: At 48 hours, sunitinib caused a dose-related inhibition of growth with LC(50) of 14.6 muM, but there was no apparent inhibition of growth at 24 hours at concentrations of 0.1-25 microM. Preincubation with sunitinib did not impair the response to TSH, indicating that it did not affect the TSH-R. Incubation with sunitinib for 24 hours caused a dose-related increase of (125)I-iodide uptake and did not reduce iodide efflux or NIS mRNA expression., Conclusion: The data indicate that sunitinib is unlikely to cause hypothyroidism by inhibition of iodide uptake.

The lack of Na(+)/I(-) symporter (NIS) gene expression in some thyroid cancer patients has been a major hurdle that limits the efficacy of standard radioactive iodide therapy. The molecular mechanism that contributes to low NIS expression is not well understood. Activated NIS gene expression is stimulated by thyroid-stimulating hormone-mediated cAMP/protein kinase A signaling through a NIS upstream enhancer (NUE). The cAMP pathway is also stimulated by forskolin. In the current work, we studied the mechanism of transcriptional activation of NIS in normal thyroid cells and thyroid cancer cells. We identified the cAMP response element modulator (CREM) activator as a new component of the transcription complex that is important for NIS gene expression. The CREM complex is seen in the normal thyroid cells and BRAF (V600E) thyroid cancer cells (BHP 17-10) but is missing in rearranged in transformation/papillary thyroid carcinoma-1 rearrangement thyroid cancer cells (BHP 2-7). This complex is believed to be responsible for the loss of NUE activity and reduced NIS expression in the BHP 2-7 cell line. In BHP 2-7 cells, forskolin stimulated the thyroid-specific transcription factor Pax 8, but CREM activator mRNA did not increase, and this produced a small increase in NUE activity. Ectopic expression of CREM activator enhanced activity of the NUE, indicating that CREM is an essential regulator of NIS gene expression.

The postmortem diagnosis of shaken baby syndrome, a severe form of child abuse, may be difficult, especially when no other visible signs of significant trauma are obvious. An important finding in shaken baby syndrome is subdural haemorrhage, typically originating from ruptured cerebral bridging veins. Since these are difficult to detect at autopsy, we have developed a special postmortem computed tomographic (PMCT) method to demonstrate the intracranial vein system in infants. This method is minimally invasive and can be carried out conveniently and quickly on clinical computed tomography (CT) systems. Firstly, a precontrast CT is made of the infant's head, to document the original state. Secondly, contrast fluid is injected manually via fontanel puncture into the superior sagittal sinus, followed by a repeat CT scan. This allows the depiction of even very small vessels of the deep and superficial cerebral veins, especially the bridging veins, without damaging them. Ruptures appear as extravasation of contrast medium, which helps to locate them at autopsy and examine them histologically, whenever necessary.

6 years after introduction of the police ammunition QD-P.E.P. (9 mm caliber), 9 bullet wounds are presented from forensic investigations along with the results of experimental shots at pig carcasses. The evaluation reveals the characteristic pattern of such bullet wounds: a skin defect which is almost as large as the caliber and has a sharp-edged border thus looking like it has been punched out; the contusion ring ("abrasion ring") is conspicuously narrow. These findings can be explained by the shape and design of the projectile. In regular police work immediate incapacitation has not always been achieved, depending on the entry location of the bullet. Experimental shots at pig carcasses showed that in the case of shots at extremities, the residual energy of the exiting bullet may be sufficient to generate another penetrating bullet wound.

At autopsy, visualization of lesions of the bridging veins, a frequent source of subdural bleeding, is difficult due to their anatomical localization. On the other hand their demonstration is of great importance for the assignment to a chronologically defined trauma. For this reason a postmortem method using computed tomography was developed to visualize the intracranial venous system by means of X-ray contrast media. In subdural bleedings, in which the skull had not been opened up, ruptured vessels could be accurately localized with this method, so that targeted dissection was possible during the subsequent autopsy.

Ecdysone agonists are hormonally active insect growth regulators that disrupt development of pest insects and have potential for development as insecticides. Their effects have been particularly well-studied in Lepidoptera and Coleoptera, but significantly less is known about their effects on dipterans, particularly aquatic species. The potency of three ecdysone agonists on larvae of 3 mosquito species, Aedes aegypti, Anopheles gambiae, and Culex quinquefasciatus, was examined. Anopheles gambiae was the most susceptible species and Ae. aegypti was the most resistant species to the effects of the three compounds tested. Potency, in descending order, was RH-2485 > RH-5992 > RH-5849. Dose-response relationships were determined for the three agonists; RH-2485 was found to be the most effective endocrine disruptor against all three species. The observed biological effects of these compounds were similar to those reported for other insects, and mosquitoes initiated molting and apolysis but did not complete a molt. In some cases, mosquito larvae synthesized a new cuticle that appeared to be normally sclerotized but the larvae failed to ecdyse and shed the exuvium. These compounds may prove to be valuable insect growth regulators for control of mosquitoes to decrease the frequency of pathogen transmission to humans. Prospects for using these compounds to control mosquitoes in the field are discussed, along with possible impacts on non-target arthropods in mosquito habitats., ((c) 2004 Wiley-Liss, Inc.)