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Diabetic retinopathy (DR) is the commonest cause of blindness in the working-age population of the developed world. The molecular pathophysiology of DR is complex, and a complete spatiotemporal model of the disease is still being elucidated. Recently, a role for angiopoietin (Ang) proteins in the pathophysiology of DR has been proposed by several research groups, and several aspects of Ang signalling are being explored as novel therapeutic strategies. Here, we review the role of the Ang proteins in two important forms of DR, diabetic macular oedema and proliferative diabetic retinopathy. The function of the Ang proteins in regulating blood vessel permeability and neovascularisation is discussed, and we also evaluate recent preclinical and clinical studies highlighting the potential benefits of modulating Ang signalling as a treatment for DR.

Purpose To compare the intraocular pressure (IOP)-lowering effect of latanoprostene bunod (LBN) 0.024% with timolol maleate 0.5% in subjects with open-angle glaucoma (OAG) or ocular hypertension (OHT). Design Prospective, randomized, double-masked, parallel-group, noninferiority clinical trial. Methods Adults with OAG or OHT from 46 clinical sites (United States and European Union) were randomized 2:1 to LBN instilled once daily (QD) in the evening and vehicle in the morning or timolol instilled twice a day (BID) for 3 months. IOP was measured at week 2, week 6, and month 3 (8 AM, 12 PM, and 4 PM each visit). Results A total of 387 subjects (LBN, n = 259; timolol, n = 128) completed the study. Analysis of covariance showed that mean IOP reduction with LBN was not only noninferior to timolol but significantly greater ( P ≤ .025) than timolol at all but the first time point in this study (week 2, 8 AM). Of LBN- and timolol-treated subjects, respectively, 31.0% and 18.5% ( P = .007) had their IOP reduced ≥25% from baseline, and 17.7% and 11.1% ( P = .084) had their IOP reduced to ≤18 mm Hg over all time points/visits in this study. Ocular treatment-emergent adverse events, while uncommon, appeared more frequently in the LBN group (all mild-moderate except 1 case of severe hyperemia). Conclusions LBN 0.024% QD in the evening was noninferior to timolol 0.5% BID over 3 months of treatment, with significantly greater IOP lowering in subjects with OAG or OHT at all but the earliest time point evaluated, and demonstrated a good safety profile.

Purpose of review Biological three-dimensional printing has received a lot of media attention over recent years with advances made in printing cellular structures, including skin and heart tissue for transplantation. Although limitations exist in creating functioning organs with this method, the hope has been raised that creating a functional retina to cure blindness is within reach. The present review provides an update on the advances made toward this goal. Recent findings It has recently been shown that two types of retinal cells, retinal ganglion cells and glial cells, can be successfully printed using a piezoelectric inkjet printer. Importantly, the cells remained viable and did not change certain phenotypic features as a result of the printing process. In addition, recent advances in the creation of complex and viable three-dimensional cellular structures have been made. Summary Some first promising steps toward the creation of a functional retina have been taken. It now needs to be investigated whether recent findings can be extended to other cells of the retina, including those derived from human tissue, and if a complex and viable retinal structure can be created through three-dimensional printing.

BACKGROUND The aim of the study is to measure the impact of age-related macular degeneration on vision-related activity limitation and preference-based status for glaucoma patients. DESIGN This was a cross-sectional study. PARTICIPANTS Two-hundred glaucoma patients of whom 73 had age-related macular degeneration were included in the research. METHODS Sociodemographic information, visual field parameters and visual acuity were collected. Age-related macular degeneration was scored using the Age-Related Eye Disease Study system. MAIN OUTCOME MEASURES The Rasch-analysed Glaucoma Activity Limitation-9 and the Visual Function Questionnaire Utility Index measured vision-related activity limitation and preference-based status, respectively. Regression models determined factors predictive of vision-related activity limitation and preference-based status. Differential item functioning compared Glaucoma Activity Limitation-9 item difficulty for those with and without age-related macular degeneration. RESULTS Mean age was 73.7 (±10.1) years. Lower better eye mean deviation (β: 1.42, 95% confidence interval: 1.24-1.63, P

The accumulation of chondroitin sulfate proteoglycans (CSPGs) in the glial scar following acute damage to the central nervous system (CNS) limits the regeneration of injured axons. Given the rich diversity of CSPG core proteins and patterns of GAG sulfation, identifying the composition of these CSPGs is essential for understanding their roles in injury and repair. Differential expression of core proteins and sulfation patterns have been characterized in the brain and spinal cord of mice and rats, but a comprehensive study of these changes following optic nerve injury has not yet been performed. Here, we show evidence that the composition of CSPGs in the optic nerve and retina following optic nerve crush (ONC) in mice and rats exhibits an increase in aggrecan, brevican, phosphacan, neurocan and versican, similar to changes following spinal cord injury. We also observe an increase in inhibitory 4-sulfated (4S) GAG chains, which suggests that the persistence of CSPGs in the glial scar opposes the growth of CNS axons, thereby contributing to the failure of regeneration and recovery of function.

Background Gout is a frequently occurring, complex rheumatologic form of inflammatory arthritis caused by the accumulation of serum uric acid (sUA) and deposition of uric acid crystals in the joints and tissues of the body. Hyperuricemia is also a significant independent risk factor for all-cause and cardiovascular morbidity and mortality and is associated with hypertension, diabetes, obesity, and osteoarthritis. However, patient adherence to prescribed urate-lowering therapies ranges from 20% to 70%, suggesting that other additional strategies, such as dietary intervention with specific, efficacious foods or beverages, may be necessary to mitigate the risk of arthritis, as well as other comorbidities. Tart cherry juice (TCJ) has been used for decades by some for gout based largely on anecdotal evidence of its efficacy and its antioxidant and anti-inflammatory properties. Objectives We designed this study to test the effect of TCJ on uricemia, lipidemia, glycemia, and inflammation in at-risk overweight and obese humans with a specific hypothesis that TCJ consumption would reduce sUA concentrations. Methods In this randomized, placebo-controlled crossover study, we recruited overweight and obese participants with body mass index (BMI) >25.0 kg/m2 (n = 26, 18 women/8 men, 41 ±11 y; BMI 31.3 ± 6.0; 12 obese, 14 overweight) to consume 240 mL/d (8 oz/d) of either TCJ or placebo beverage, for 4 wk each with a 4-wk intervening washout period followed by 4 wk of the alternate beverage. Results TCJ significantly reduced sUA concentration by 19.2% (P

Subclinical inflammation is frequently noted in chronic diseases such as diabetes, cardiovascular disease (CVD) and obesity. Accumulating epidemiological evidence demonstrates that diets rich in vegetables and fruits, e.g., cherries, may significantly reduce the risk of chronic disease, in part, via antioxidant and anti-inflammatory activities. In this randomized, placebo-controlled crossover study, we recruited 10 at-risk participants (38.1 ± 12.5 years; 8 females, 2 males) with BMI >25.0 kg m−2 (32.2 ± 4.6 kg m−2; 5 obese, 5 overweight) to consume 240 mL (8 ounces) daily of either 100% tart cherry juice (TCJ) or an alternate placebo beverage, for 4 weeks with a 2-week intervening washout period before switching to the alternate beverage for four weeks. Fasting blood samples were collected at the beginning and end of each arm for measurement of biomarkers of inflammation. The erythrocyte sedimentation rate (ESR), an indicator of chronic inflammation, was significantly (p < 0.05) lower with TCJ than with the placebo beverage, which increased ESR by 19%. Mean baseline hsCRP, an indicator of acute inflammation, was 7.0 ± 5.2 mg L−1 and consumption of TCJ did not affect hsCRP levels. The chemokine MCP-1 and cytokine TNF-alpha were lower in participants after consuming TCJ compared to those consuming the placebo beverage. Plasma IL-6 and IL-l0 were not different between treatments. Collectively, the data suggest that authentic 100% TCJ may reduce biomarkers of inflammation often noted in chronic disease and may be a preferable dietary selection compared to artificially flavored beverages with added sugars.

BackgroundBleb needling is widely used to restore flow and lower intraocular pressure (IOP) in a failing trabeculectomy. We aimed to measure the safety and efficacy of needling in a large cohort and identify factors that were associated with success and failure.MethodsThis retrospective audit included all patients who underwent needling at Addenbrooke’s Hospital, Cambridge over a 10-year period. Data were available on 91 patients (98% of patients identified), including 191 needlings on 96 eyes. Success was defined as IOP below 21 mm Hg or 16 mm Hg or 13 mm Hg consistently, without reoperation or glaucoma medication. Risk factors for failure were assessed by Cox proportional hazard regression and Kaplan-Meier curves.ResultsSuccess defined as IOP ConclusionNeedling was most successful soon after trabeculectomy, but resuscitation of a long-failed trabeculectomy had lower likelihood of success. The safety and efficacy compare favourably with alternative treatment approaches.

The failure of mammalian CNS neurons to regenerate their axons derives from a combination of intrinsic deficits and extrinsic factors. Following injury, chondroitin sulfate proteoglycans (CSPGs) within the glial scar inhibit axonal regeneration, an action mediated by the sulfated glycosaminoglycan (GAG) chains of CSPGs, especially those with 4-sulfated (4S) sugars. Arylsulfatase B (ARSB) selectively cleaves 4S groups from the non-reducing ends of GAG chains without disrupting other, growth-permissive motifs. We demonstrate that ARSB is effective in reducing the inhibitory actions of CSPGs both in in vitro models of the glial scar and after optic nerve crush (ONC) in adult mice. ARSB is clinically approved for replacement therapy in patients with mucopolysaccharidosis VI and therefore represents an attractive candidate for translation to the human CNS.

Previous studies have demonstrated that intravitreal delivery of brain-derived neurotrophic factor (BDNF) by injection of recombinant protein or by gene therapy can alleviate retinal ganglion cell (RGC) loss after optic nerve injury. BDNF gene therapy can improve RGC survival in experimental models of glaucoma, the leading cause of irreversible blindness worldwide. However, the therapeutic efficacy of BDNF supplementation alone is time limited at least in part due to BDNF receptor downregulation. Tropomyosin-related receptor kinase-B (TrkB) downregulation has been reported in many neurological diseases including glaucoma, potentially limiting the effect of sustained or repeated BDNF delivery. Here, we characterize a novel adeno-associated virus (AAV) gene therapy (AAV2 TrkB-2A-mBDNF) that not only increases BDNF production but also improves long-term neuroprotective signaling by increasing expression of the BDNF receptor (TrkB) within the inner retina. This approach leads to significant and sustained elevation of survival signaling pathways ERK and AKT within RGCs over 6 months and avoids the receptor downregulation which we observe with treatment with AAV2 BDNF alone. We validate the neuroprotective efficacy of AAV2 TrkB-2A-mBDNF in a mouse model of optic nerve injury, where it outperforms conventional AAV2 BDNF or AAV2 TrkB therapy, before showing powerful proof of concept neuroprotection of RGCs and axons in a rat model of chronic intraocular pressure (IOP) elevation. We also show that there are no adverse effects of the vector on retinal structure or function as assessed by histology and electroretinography in young or aged animals. Further studies are underway to explore the potential of this vector as a candidate for progression into clinical studies to protect RGCs in patients with glaucoma and progressive visual loss despite conventional IOP-lowering treatment.

The eye is at the forefront of the application of gene therapy techniques to medicine. In the United States, a gene therapy treatment for Leber’s congenital amaurosis, a rare inherited retinal disease, recently became the first gene therapy to be approved by the FDA for the treatment of disease caused by mutations in a specific gene. Phase III clinical trials of gene therapy for other single-gene defect diseases of the retina and optic nerve are also currently underway. However, for optic nerve diseases not caused by single-gene defects, gene therapy strategies are likely to focus on slowing or preventing neuronal death through the expression of neuroprotective agents. In addition to these strategies, there has also been recent interest in the potential use of precise genome editing techniques to treat ocular disease. This review focuses on recent developments in gene therapy techniques for the treatment of glaucoma and Leber’s hereditary optic neuropathy (LHON). We discuss recent successes in clinical trials for the treatment of LHON using gene supplementation therapy, promising neuroprotective strategies that have been employed in animal models of glaucoma and the potential use of genome editing techniques in treating optic nerve disease.

PURPOSE: To establish the medium-term repeatability of the iPad perimetry app Melbourne Rapid Fields (MRF) compared to Humphrey Field Analyzer (HFA) 24-2 SITA-standard and SITA-fast programs. DESIGN: Multicenter longitudinal observational clinical study. METHODS: Sixty patients (stable glaucoma/ocular hypertension/glaucoma suspects) were recruited into a 6-month longitudinal clinical study with visits planned at baseline and at 2, 4, and 6 months. At each visit patients undertook visual field assessment using the MRF perimetry application and either HFA SITA-fast (n = 21) or SITA-standard (n = 39). The primary outcome measure was the association and repeatability of mean deviation (MD) for the MRF and HFA tests. Secondary measures were the point-wise threshold and repeatability for each test, as well as test time. RESULTS: MRF was similar to SITA-fast in speed and significantly faster than SITA-standard (MRF 4.6 ± 0.1 minutes vs SITA-fast 4.3 ± 0.2 minutes vs SITA-standard 6.2 ± 0.1 minutes, P < .001). Intraclass correlation coefficients (ICC) between MRF and SITA-fast for MD at the 4 visits ranged from 0.71 to 0.88. ICC values between MRF and SITA-standard for MD ranged from 0.81 to 0.90. Repeatability of MRF MD outcomes was excellent, with ICC for baseline and the 6-month visit being 0.98 (95% confidence interval: 0.96-0.99). In comparison, ICC at 6-month retest for SITA-fast was 0.95 and SITA-standard 0.93. Fewer points changed with the MRF, although for those that did, the MRF gave greater point-wise variability than did the SITA tests. CONCLUSIONS: MRF correlated strongly with HFA across 4 visits over a 6-month period, and has good test-retest reliability. MRF is suitable for monitoring visual fields in settings where conventional perimetry is not readily accessible.

To evaluate current delivery of glaucoma care in Botswana; in particular, the service infrastructure available and glaucoma-related workload.A multi-center cross-sectional study was undertaken comprising government eye care institutions and ophthalmic personnel across Botswana. Data on human resources, equipment types and numbers, diagnostic criteria routinely used, treatments routinely provided, and new and repeat glaucoma consultations were obtained through quantitative and qualitative surveys.In 27 government eye care institutions there were two general ophthalmologists, neither of whom had a subspecialty interest in glaucoma, 64 ophthalmic nurses, two optometrists, one low vision therapist, one refractionist, and two equipment technicians. Only 8.5% of available ophthalmic human resources were taken up with provision of glaucoma care. About 1/3 of hospitals did not have tonometers, most primary hospitals lacked slit lamp biomicroscopes and most hospitals lacked sensitive diagnostic equipment. A diagnosis of glaucoma was made by either an ophthalmic nurse or an ophthalmologist, but only 10% of institutions could meet recommendations for follow-up assessment. Topical glaucoma medications were prescribed by almost all hospital clinics, usually by ophthalmic nurses. Drug choices were largely determined by local availability. Glaucoma surgery accounted for 0.8% of total eye operations. Glaucoma patients took up 8.5% of total clinic visits. The total number of glaucoma visits was highest in the two hospitals with ophthalmologists. New glaucoma cases took up 10.3% of total glaucoma visits.This study highlights the challenges faced in caring for glaucoma patients in Botswana; in particular, lack of professional human resources, equipment and availability of effective treatments.

Optic neuropathies such as glaucoma occur when retinal ganglion cells (RGCs) in the eye are injured. Strong evidence suggests mesenchymal stem cells (MSCs) could be a potential therapy to protect RGCs; however, little is known regarding their effect on the human retina. We, therefore, investigated if human MSCs (hMSCs), or platelet‐derived growth factor (PDGF) as produced by hMSC, could delay RGC death in a human retinal explant model of optic nerve injury. Our results showed hMSCs and the secreted growth factor PDGF‐AB could substantially reduce human RGC loss and apoptosis following axotomy. The neuroprotective pathways AKT, ERK, and STAT3 were activated in the retina shortly after treatments with labeling seen in the RGC layer. A dose dependent protective effect of PDGF‐AB was observed in human retinal explants but protection was not as substantial as that achieved by culturing hMSCs on the retina surface which resulted in RGC cell counts similar to those immediately post dissection. These results demonstrate that hMSCs and PDGF have strong neuroprotective action on human RGCs and may offer a translatable, therapeutic strategy to reduce degenerative visual loss. Stem Cells 2018;36:65–78

Background In the assessment of a pituitary mass, objective visual field testing represents a valuable means of evaluating mass effect, and thus in deciding whether surgical management is warranted. Case presentation In this vignette, we describe a 73 year-old lady who presented with a three-week history of frontal headache, and ‘blurriness’ in the left side of her vision, due to a WHO grade III anaplastic haemangiopericytoma compressing the optic chiasm. We report how timely investigations, including an iPad-based visual field test (Melbourne Rapid Field, (MRF)) conducted at the bedside aided swift and appropriate management of the patient. Conclusions We envisage such a test having a role in assessing bed-bound patients in hospital where access to formal visual field testing is difficult, or indeed in rapid testing of visual fields at the bedside to screen for post-operative complications, such as haematoma.

Background To assess the impact of cataract on quality of life (QoL) for patients with glaucoma. Design Cross-sectional study. Participants Two hundred forty-two patients with mild (n = 67), moderate (n = 80) or severe (n = 45) glaucoma and 50 controls (glaucoma suspects). Patients with cataract of any severity were included. Methods The following data were collected: sociodemographic information, phakic/pseudophakic status, cataract grade according to the Lens Opacities Classification System (LOCS) III, visual acuity and visual field test parameters. Glaucoma severity was stratified according to binocular visual field loss. Visually significant cataract was defined as: LOCS III criteria nuclear cataract ≥3/6.9, cortical cataract ≥3/5.9 or posterior subcapsular cataract ≥2/5.9. Main Outcome Measures Patients’ QoL was measured using Rasch-transformed scores from the Glaucoma Activity Limitation-9 (GAL-9) questionnaire. Multiple linear regression models were used to determine the association between cataract and GAL-9 (logit) score. Results Fifty-six (23.1%) patients had at least one visually significant cataract. At least one visually significant cataract (standardized coefficient [β] 1.19, 95% confidence interval 1.04–1.34, P = 0.011) and poor visual field index (better eye) (β 1.47, 95% confidence interval 1.36–1.88, P

Background With a higher life expectancy, there is an increased demand for hospital glaucoma services in the United Kingdom. Design The Cambridge community Optometry Glaucoma Scheme (COGS) was initiated in 2010, where new referrals for suspected glaucoma are evaluated by community optometrists with a special interest in glaucoma, with virtual electronic review and validation by a consultant ophthalmologist with special interest in glaucoma. Participants 1733 patients were evaluated by this scheme between 2010 and 2013. Methods Clinical assessment is performed by the optometrist at a remote site. Goldmann applanation tonometry, pachymetry, monoscopic colour optic disc photographs and automated Humphrey visual field testing are performed. A clinical decision is made as to whether a patient has glaucoma or is a suspect, and referred on or discharged as a false positive referral. The clinical findings, optic disc photographs and visual field test results are transmitted electronically for virtual review by a consultant ophthalmologist. Main Outcome Measures The number of false positive referrals from initial referral into the scheme. Results Of the patients, 46.6% were discharged at assessment and a further 5.7% were discharged following virtual review. Of the patients initially discharged, 2.8% were recalled following virtual review. Following assessment at the hospital, a further 10.5% were discharged after a single visit. Conclusions The COGS community-based glaucoma screening programme is a safe and effective way of evaluating glaucoma referrals in the community and reducing false-positive referrals for glaucoma into the hospital system.

Glaucoma is a leading cause of blindness that affects the dendrites, axons and cell bodies of retinal ganglion cells (RGCs). Lowering intraocular pressure is currently the mainstay of glaucoma treatment but fails to prevent visual loss in many patients. Dendrite changes in RGCs appear to be an early sign of glaucoma in experimental models and recent evidence suggests that changes in the inner plexiform layer of human glaucoma patients may be predictive of future visual loss. Strengthening dendritic connectivity has been shown to improve functional outcomes in a number of neurodegenerative diseases which share some pathological features with glaucoma. We therefore propose that better understanding of RGC dendrites could suggest new targets for glaucoma therapy, and might aid identification of patients who are most at risk of future visual loss and blindness.

The development of neuroprotective strategies to attenuate retinal ganglion cell death could lead to novel therapies for chronic optic neuropathies such as glaucoma. Intravitreal transplantation of mesenchymal stem cells slows retinal ganglion cell death in models of optic nerve injury, but the mechanism of action remains unclear. Here we characterized the neuroprotective effects of mesenchymal stem cells and mesenchymal stem cell-derived factors in organotypic retinal explant culture and an in vivo model of ocular hypertensive glaucoma. Co-culture of rat and human bone marrow-derived mesenchymal stem cells with retinal explants increased retinal ganglion cell survival, after 7 days ex vivo, by ∼2-fold and was associated with reduced apoptosis and increased nerve fibre layer and inner plexiform layer thicknesses. These effects were not demonstrated by co-culture with human or mouse fibroblasts. Conditioned media from mesenchymal stem cells conferred neuroprotection, suggesting that the neuroprotection is mediated, at least partly, by secreted factors. We compared the concentrations of 29 factors in human mesenchymal stem cell and fibroblast conditioned media, and identified 11 enriched in the mesenchymal stem cell secretome. Treatment of retinal explants with a cocktail of these factors conferred retinal ganglion cell neuroprotection, with factors from the platelet-derived growth factor family being the most potent. Blockade of platelet-derived growth factor signalling with neutralizing antibody or with small molecule inhibitors of platelet-derived growth factor receptor kinase or downstream phosphatidylinositol 3 kinase eliminated retinal ganglion cell neuroprotection conferred by mesenchymal stem cell co-culture. Intravitreal injection of platelet-derived growth factor -AA or -AB led to profound optic nerve neuroprotection in vivo following experimental induction of elevated intraocular pressure. These data demonstrate that mesenchymal stem cells secrete a number of neuroprotective proteins and suggest that platelet-derived growth factor secretion in particular may play an important role in mesenchymal stem cell-mediated retinal ganglion cell neuroprotection. Furthermore, platelet-derived growth factor may represent an independent target for achieving retinal ganglion cell neuroprotection.

Purpose To investigate whether modification of liver complement factor H (CFH) production, by alteration of liver CFH Y402H genotype through liver transplantation (LT), influences the development of age-related macular degeneration (AMD). Design Multicenter, cross-sectional study. Participants We recruited 223 Western European patients ≥55 years old who had undergone LT ≥5 years previously. Methods We determined AMD status using a standard grading system. Recipient CFH Y402H genotype was obtained from DNA extracted from recipient blood samples. Donor CFH Y402H genotype was inferred from recipient plasma CFH Y402H protein allotype, measured using enzyme-linked immunosorbent assays. This approach was verified by genotyping donor tissue from a subgroup of patients. Systemic complement activity was ascertained by measuring levels of plasma complement proteins using an enzyme-linked immunosorbent assay, including substrates (C3, C4), activation products (C3a, C4a, and terminal complement complex), and regulators (total CFH, C1 inhibitor). Main Outcome Measures We evaluated AMD status and recipient and donor CFH Y402H genotype. Results In LT patients, AMD was associated with recipient CFH Y402H genotype (P = 0.036; odds ratio [OR], 1.6; 95% confidence interval [CI], 1.0–2.4) but not with donor CFH Y402H genotype (P = 0.626), after controlling for age, sex, smoking status, and body mass index. Recipient plasma CFH Y402H protein allotype predicted donor CFH Y402H genotype with 100% accuracy (n = 49). Plasma complement protein or activation product levels were similar in LT patients with and without AMD. Compared with previously reported prevalence figures (Rotterdam Study), LT patients demonstrated a high prevalence of both AMD (64.6% vs 37.1%; OR, 3.09; P

Background To report for the first time acute transient corneal endothelial changes following selective laser trabeculoplasty. Design Observational case series at a tertiary referral ophthalmological centre in the United Kingdom (Addenbrooke's Hospital, Cambridge) Participants Ten consecutive routinely treated patients. Methods Baseline measurements of corneal specular microscopy, endothelial cell counts and in vivo confocal microscopy were performed prior to routine selective laser trabeculoplasty treatment. Repeat measurements were made approximately 1 h later and at a 6-week follow up. Main Outcome Measures Endothelial in vitro laser confocal microscopy, specular microscopic endothelial cell count, endothelial morphology, visual acuity and intraocular pressure. Results All patients had normal corneal endothelia prior to routine selective laser trabeculoplasty treatment, where 180 degrees of angle was treated. Approximately an hour afterwards, nearly all patients still had normal specular microscopy and cell counts (2237 ± 211 cells/mm2) but had subtle endothelial changes on slit-lamp examination. These changes were found diffusely across the cornea. The changes were not present at the 6-week follow up and cell counts remained unchanged (2278 ± 242 cells/mm2). Acuity remained unchanged throughout and the patients were not aware of these changes subjectively. Conclusions Selective laser trabeculoplasty appears to cause transient corneal endothelial changes in most patients that have no impact on cell count or visual acuity. Further work is required to elucidate the mechanism of this phenomenon and any long-term impact.

Retinal ganglion cell degeneration underlies several conditions which give rise to significant visual compromise, including glaucoma, hereditary optic neuropathies, ischaemic optic neuropathies, and demyelinating disease. In this review, we discuss the emerging strategies for neuroprotection specifically in the context of glaucoma, including pharmacological neuroprotection, mesenchymal stem cells, and gene therapy approaches. We highlight potential pitfalls that need to be considered when developing these strategies and outline future directions, including the prospects for clinical trials.

Background To investigate whether pre-operative intraocular pressure (IOP) predicts outcome of trabeculectomy surgery in patients with primary open angle glaucoma over a 3-year period of follow-up. Methods Retrospective cohort study, of a total of 61 patients (80 procedures) who had undergone trabeculectomy surgery after failed medical management at a single centre between 2000 and 2011. Patients were identified through surgical logbooks. A subsequent case note-review identified 61 patients (80 procedures) with primary open angle glaucoma (POAG). The primary outcome was success of trabeculectomy surgery, with failure defined as intraocular pressure (IOP) > 21 mmHg, ≤ 5 mmHg or not reduced by 20% at two consecutive follow-up visits 3-months post-operatively. Qualified success was defined as surgical success with the use of supplemental medical therapy. Secondary outcomes included visual acuity, Humphrey visual field MD, surgical complications and post-operative interventions. Results At 3 years, the odds ratio of failure was 0.93 per mmHg pre-operative IOP (95% C.I. 0.83-1.03, p = 0.15 Wald Χ2 test), and the odds ratio of failure or qualified success was 0.96 (95% C.I. 0.89-1.04, p = 0.35). The incidence of surgical complications showed an odds ratio of 1.02 per mmHg pre-operative IOP (95% C.I. 0.95-1.10, p = 0.55 Wald Χ2 test). The incidence of post-operative interventions showed an odds ratio of 1.01 per mmHg pre-operative IOP (95% C.I. 0.94-1.09, p = 0.80 Wald Χ2 test). Conclusions Pre-operative IOP does not predict success of trabeculectomy surgery in POAG patients during the first 3 years of follow-up. The incidence of surgical complications and post-operative interventions shows no association with pre-operative IOP.

Glaucoma is an optic neuropathy that is characterized by the progressive degeneration of the optic nerve, leading to visual impairment. Glaucoma is the main cause of irreversible blindness worldwide, but typically remains asymptomatic until very severe. Open-angle glaucoma comprises the majority of cases in the United States and western Europe, of which, primary open-angle glaucoma (POAG) is the most common type. By contrast, in China and other Asian countries, angle-closure glaucoma is highly prevalent. These two types of glaucoma are characterized based on the anatomic configuration of the aqueous humour outflow pathway. The pathophysiology of POAG is not well understood, but it is an optic neuropathy that is thought to be associated with intraocular pressure (IOP)-related damage to the optic nerve head and resultant loss of retinal ganglion cells (RGCs). POAG is generally diagnosed during routine eye examination, which includes fundoscopic evaluation and visual field assessment (using perimetry). An increase in IOP, measured by tonometry, is not essential for diagnosis. Management of POAG includes topical drug therapies and surgery to reduce IOP, although new therapies targeting neuroprotection of RGCs and axonal regeneration are under development.

Glaucoma is an optic neuropathy that is characterized by the progressive degeneration of the optic nerve, leading to visual impairment. Glaucoma is the main cause of irreversible blindness worldwide, but typically remains asymptomatic until very severe. Open-angle glaucoma comprises the majority of cases in the United States and western Europe, of which, primary open-angle glaucoma (POAG) is the most common type. By contrast, in China and other Asian countries, angle-closure glaucoma is highly prevalent. These two types of glaucoma are characterized based on the anatomic configuration of the aqueous humour outflow pathway. The pathophysiology of POAG is not well understood, but it is an optic neuropathy that is thought to be associated with intraocular pressure (IOP)-related damage to the optic nerve head and resultant loss of retinal ganglion cells (RGCs). POAG is generally diagnosed during routine eye examination, which includes fundoscopic evaluation and visual field assessment (using perimetry). An increase in IOP, measured by tonometry, is not essential for diagnosis. Management of POAG includes topical drug therapies and surgery to reduce IOP, although new therapies targeting neuroprotection of RGCs and axonal regeneration are under development.

This study provides ophthalmologists who manage uveitic glaucoma with important information on factors that can affect the success of surgical management of this challenging disease.This study examines surgical outcomes of trabeculectomy and glaucoma device implant (GDI) surgery for uveitic glaucoma, in particular the effect of uveitis activity on surgical outcomes.Retrospective chart review at a tertiary institution.Eighty-two cases with uveitic glaucoma (54 trabeculectomies and 28 (GDI) surgeries) performed between 1 December 2006 and 30 November 2014.Associations of factors with surgical outcomes were examined using univariate and multivariate analysis.Surgical outcomes as defined in Guidelines from World Glaucoma Association.Average follow up was 26.4 ± 21.5 months. Overall qualified success rate of the trabeculectomies was not statistically different from GDI, being 67% and 75%, respectively (P = 0.60). Primary and secondary GDI operations showed similar success rates. The most common postoperative complication was hypotony (~30%). Active uveitis at the time of operation was higher in trabeculectomy compared with GDI group (35% vs. 14%). Active uveitis at the time of surgery did not significantly increase risk of failure for trabeculectomies. Recurrence of uveitis was significantly associated with surgical failure in trabeculectomy group (odds ratio 4.8, P = 0.02) but not in GDI group.Surgical success rate of GDI was not significantly different from trabeculectomy for uveitic glaucoma in this study. Regular monitoring, early and prolonged intensive treatment of ocular inflammation is important for surgical success particularly following trabeculectomy.

PURPOSE: Platelet-derived growth factor (PDGF) promotes neuronal survival in experimental glaucoma and recruits glial cells that regulate synapses. We investigated the effects of intravitreal PDGF on the inflammatory milieu and retinal synapses in the presence of raised IOP. METHODS: Animals with laser-induced IOP elevation received intravitreal injections of either saline or 1.5 μg PDGF. At 7 days, a further intravitreal injection was administered so groups received "PDGF-saline" (n = 15), "PDGF-PDGF" (n = 13), or "saline-saline" (n = 20). Platelet-derived growth factor receptor activation was assessed after 2 weeks using Western blot for PI3 kinase. Immunohistochemistry was performed for markers of synapses in the inner plexiform layer (IPL): PSD-95, GluR1, SY38; RGCs: βIII-tubulin, and glial cells: Iba-1, CD45. Real-time quantitative polymerase chain reaction (qPCR) was performed for Arc, selp, MCP-1, IL-6, IL-10, and CX3CR1 (n = 13). RESULTS: A single injection of PDGF increased IPL synaptic density in high IOP eyes (PSD-95 = 8.65 ± 0.43, SY38 = 8.68 ± 0.51, GluR1 = 9.03 ± 0.60 puncta/μm3, P < 0.001) and expression of synaptic modulator Arc (6.92 ± 3.71-fold change/control, P < 0.05) in comparison with vehicle (PSD-95 = 4.59 ± 0.41, SY38 = 4.46 ± 0.38, GluR1 = 5.94 ± 0.50 puncta/μm3, Arc = 1.46 ± 0.31-fold/control). This was associated with more resident microglia (8.16 ± 1.34-fold change/control, P < 0.001) and infiltrating monocyte-derived macrophages in the retina as well as increased Selp expression (26.8 ± 14.12-fold change/control, P < 0.05). Optic nerve head (ONH) showed an increased microglia (saline = 1.44 ± 0.13 versus PDGF = 2.23 ± 0.18-fold change/control, P < 0.01) but not infiltrating macrophages. IL-10 expression was significantly increased in PDGF-treated eyes (5.43 ± 0.47-fold change/control, P < 0.05) relative to vehicle (2.51 ± 0.67-fold change/control). CONCLUSIONS: Platelet-derived growth factor increased microglial and monocyte-derived macrophage populations in the eye and protected intraretinal synapses from degeneration in our experimental glaucoma model.

Consumption of polyphenol-rich fruits, for example, tart cherries, is associated with a lower risk of cardiovascular disease and cancer. This is due, in large part, to the diverse myriad bioactive agents, that is, polyphenol anthocyanins, present in fruits. Anthocyanin-rich tart cherries purportedly modulate numerous cellular processes associated with oncogenesis such as apoptosis, cellular proliferation (CP), and cell cycle progression, although the effective concentrations eliciting these effects are unclear. We hypothesized that several dose-dependent effects over a large concentration range of 100% tart cherry juice (TCJ) would exist and affect these processes differentially with the potential for cellular protection and cellular death either by apoptosis or by necrosis. In this in vitro study, we tested the dose response of TCJ on CP and cell death in MCF-7 human breast cancer cells. TCJ was added at 0.03-30% (v/v) to cells and incubated overnight with the medium alone or with increasing TCJ. Bromodeoxyuridine incorporation was significantly reduced by 20% at ≥10% (v/v) TCJ and associated with necrosis, but was not different between the control and treatment groups at10% TCJ. MTT reduction was also significantly reduced by 27% and 80% at 10% and 30% TCJ, respectively, and associated with necrosis. Apoptosis, but not necrosis, was increased ∼63% at 3% TCJ (∼307 nM monomeric anthocyanins), yet significantly decreased (P.05) by 20% at 1% TCJ (920 nM) both of which were physiologically relevant concentrations of anthocyanins. The data support a biphasic effect on apoptosis and no effect on proliferation.

This study was undertaken to determine the performance of trabeculectomy surgery over a 20-year period and examine the associations between outcome and risk factors for trabeculectomy failure.Retrospective cohort study.A total of 234 patients (330 procedures) who had undergone trabeculectomy surgery at Addenbrooke's Hospital, Cambridge, United Kingdom, between January 1988 and December 1990.Patients were identified through surgical logbooks (n = 521 procedures on 380 patients); after this, a case-note review was undertaken, which identified 234 patients (330 procedures) who had available case notes.Surgical success was defined as "complete success" while intraocular pressure (IOP) remained21 mm Hg with no additional medication and as "qualified success" if those requiring additional topical medication were included. Functional success was defined if patients did not progress to legal blindness (visual acuity3/60 or visual field10 degrees).After 20 years, 57% were classified as complete success, 88% were classified as qualified success, and 15% had become blind. Those at risk of trabeculectomy failure were younger or had uveitic glaucoma. Those with pseudoexfoliation or aphakia were more likely to progress to blindness. Furthermore, those using 2 or more topical medications or with advanced visual field loss at the time of surgery were more at risk of both trabeculectomy failure and blindness.This study indicates that trabeculectomy survival at 20 years may be approximately 60% with no topical medication and approximately 90% with additional topical medication. Patient age, preoperative topical medication use, glaucoma type, and glaucoma severity will independently influence this outcome. Trabeculectomy surgery is therefore a long-term solution to IOP control.

Glaucoma, a leading cause of blindness, is a neurodegenerative disease characterized by progressive loss of retinal ganglion cell axons in the optic nerve and their cell bodies in the retina. Reactive retinal glial changes have been observed in glaucoma but the role of such glial changes in the pathogenesis of the condition remains unclear. In the present study we found that retinal ganglion cells in an experimental animal model of glaucoma have an increased axon regenerative potential. Regeneration of adult rat retinal ganglion cell axons after optic nerve crush was significantly increased in vivo when combined with intraocular pressure-induced experimental glaucoma. This enhanced axon regeneration response was correlated with a significant increase in activation of glial fibrillary acidic protein+retinal glia. Using a dissociated retinal ganglion cell culture model we showed that reducing the number of activated retinal glia with a glial specific toxin, α-Aminoadipic acid, significantly reduced the growth potential of retinal ganglion cells from glaucomatous rat eyes, suggesting that activated retinal glia mediate, at least in part, the growth promoting effect. This was shown to be mediated by both membrane-bound and soluble glial-derived factors. Neurotrophin and ciliary neurotrophic/leukemia inhibitory factor blockers did not affect the regenerative potential, excluding these growth factors as principal mediators of the enhanced growth response occurring in glaucomatous retinal cultures. These observations are the first to reveal that retinal ganglion cells from glaucomatous rat eyes have an enhanced regenerative capacity. Furthermore, our results suggest that activated retinal glia mediate at least part of this response. Further work to understand and enhance the regeneration-promoting effect of activated retinal glia is required to determine if this approach could be useful as part of a therapeutic strategy to encourage optic nerve regeneration in glaucoma.

Glaucoma is a progressive optic neuropathy and a major cause of visual impairment worldwide. Neuroprotective therapies for glaucoma aim to ameliorate retinal ganglion cell degeneration through direct or indirect action on these neurons. Neurotrophic factor (NTF) delivery is a key target for the development of potential neuroprotective glaucoma treatments. This article will critically summarize the evidence that NTF deprivation and/or dysfunction plays a role in the pathogenesis of glaucoma. Experimental support for the neuroprotective potential of NTF supplementation in animal models of glaucoma will be reviewed, in particular for brain-derived neurotrophic factor, ciliary neurotrophic factor, and glial cell line-derived neurotrophic factor. Finally, the challenges of clinical translation will be considered with an emphasis on the most promising NTF delivery strategies including slow-release drug delivery, gene therapy, and cell transplantation.

The Method of Standard Additions or The Standard Addition Method, often referred to by its acronym as just SAM, is a proverbial workhorse in both inorganic and organic quantitative analytical chemistry and in related disciplines such as geochemistry. Its advantage in mitigating the effects of matrix interferences compared with the calibration curve approach is well known and is one of its major benefits. It is presented in virtually all standard textbooks on analytical chemistry in varying degrees of complexity. Yet the story of how it originated, and by whom, is not well known. It is generally believed that it originated in the early 1950s, introduced by spectroscopists. We have determined that the priority of its use and discovery apparently belongs exclusively to Hans Hohn (1906– 1978), a mining chemist, dating from the exposition of the method in his 1937 book on polarography, Chemische Analysen mit dem Polarographen. How the method became established in other disciplines quite different from polarography is not completely clear. It is likely that it was rediscovered independently in at least two separate disciplines, X-ray fluorescence (XRF) and atomic spectroscopy, almost simultaneously in 1953, 16 years after Hohn’s original description.

Tau inclusions play a key role in the pathogenesis of tauopathies. Altered tau levels have been detected in retina and optic nerve of patients with glaucoma, suggesting the possibility of shared pathogenic mechanisms with tauopathies. Here we report that hyperphosphorylated transgenic tau accumulates in the nerve fibre layer and, from 2 months of age, aggregates into filamentous inclusions in retinal ganglion cells of human P301S tau transgenic mice. Axonopathy and accumulation of hyperphosphorylated tau in the nerve fibre layer preceded inclusion formation. Hyperphosphorylated tau and tau inclusions were also detected in cultured retinal explants from 5-month-old transgenic mice. Axonal outgrowth was similar in transgenic and wild-type retinal explants under basal conditions. However, when exposed to growth-promoting stimuli, axon elongation was enhanced in explants from wild-type but not transgenic mice, indicating that the presence of abnormal tau can impair stimulated axonal outgrowth. These findings suggest that the retina is a good model system for investigating tau-driven neurodegeneration and for assessing potential pharmacological modifiers for tauopathies.

High intraocular pressure induces glaucomatous degeneration of retinal ganglion cells (RGCs). The cellular mechanisms leading to activation of the apoptosis cascade are multidimensional and only partially understood. A small dynein subunit, the light chain DYNLL1 (synonym LC8-1, PIN) has recently been shown to be an important regulator of neuron proteins known to be involved in glaucomatous RGC death including NO synthases, the pro-apoptotic protein Bim and the dynein intermediate chain. Also, DYNLL1 is a regulator of mitochondria anchorage in axons, which is impaired in glaucoma. We investigated expression of DYNLL1 and 2 and its dynein binding partner dynein intermediate chain in a rat model of chronic glaucoma. Laser capture microdissection (LCM) allowed us to collect distinct cell layers and cell bodies from the retina to gain data highly specific for retinal ganglion cells. Glaucoma was induced in 23 rats by laser treatment to the aqueous outflow tract. RNA was extracted from LCM dissected ganglion cell layers (GCL) and 100 pooled RGCs per retina. Expression levels for 1, 2 and 4 week timepoints were analysed by quantitative PCR for DYNLL1 and 2, dynein intermediate chain and GFAP. DYNLL protein abundance in RGCs was quantified in immunostained retina sections. DYNLL gene 1 but not 2 was expressed in RGCs. In the glaucoma model DYNLL1 was strongly and persistently downregulated at all timepoints. DYNLL protein was significantly less abundant at the 4 week timepoint. In contrast, the motorprotein binding partner dynein intermediate chain 1 was more stably expressed. DYNLL2 was upregulated in glia cells at 2 weeks. Expression of DYNLL1, the only form of the dynein light chain expressed in RGCs, is downregulated persistently in glaucoma, while its binding partner dynein IC-1 is unchanged. The specific lack of DYNLL1 could have an impact on the function of their regulatory binding partners and contribute in several ways to neuron dysfunction and apoptosis.

Regular consumption of fruits and vegetables is strongly associated with a reduced risk of cardiovascular disease (CVD). This effect occurs, in part, because of the plethora of bioactive agents in foods and their subsequent function as antioxidants. Ergothioneine (ERT), a novel antioxidant, is present in edible mushrooms and is not synthesized, but is accumulated, by humans through diet. In this study, we tested whether ERT, a bioactive agent, could interrupt pro-inflammatory induction of adhesion molecule expression associated with atherogenesis. Human aortic endothelial cells (HAECs) were incubated with increasing concentrations of ERT (0.01-10.0 mM) overnight (16 hours) followed by incubation with medium alone or with the pro-inflammatory cytokine interleukin (IL)-1β (5 ng/mL) for 6 hours to induce expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and endothelial-leukocyte adhesion molecule (ELAM-1 or E-selectin). ERT at 0.1-0.3 mM significantly (P .05) reduced VCAM-1, ICAM-1, and E-selectin expression up to 41%. VCAM-1 was suppressed to the greatest extent followed by E-selectin and then ICAM-1. We next tested if binding of preloaded U937 human monocytes to HAECs would be inhibited. U937 binding to HAECs was significantly reduced in IL-1β-stimulated HAECs preincubated with 1 and 3 mM ERT. Unstimulated monolayers demonstrated marginal, but significant, reductions. ERT was not toxic to HAECs at any concentration used. These data provide evidence that ERT found in commonly consumed dietary mushrooms can protect against events observed in atherogenesis, suggesting increased dietary intake of edible mushrooms would be a prudent medicinal means of reducing CVD risk.

Chondroitinase ABC (ChABC) is a bacterial enzyme that can enhance plasticity following injury to the central nervous system (CNS) by degrading the glycosaminoglycan (GAG) side chains of proteoglycans. CNS lesions treated with ChABC often show enhanced axonal sprouting and improved functional recovery and there is therefore much interest in the potential use of ChABC as a clinical treatment in humans. When highly concentrated fibrin gel containing ChABC was implanted adjacent to a spinal cord lesion, bioactive ChABC was detectable in the spinal cord for at least three weeks. Nearly six times more bioactive ChABC was detected in the spinal cord 3 weeks after injury when the fibrin delivery system was used vs. an intraspinal injection of ChABC (61+/-30 mU vs. 11+/-4 mU). Furthermore, 3 weeks after injury the level of inhibitory GAG found in injured spinal cord treated with the delivery system was 37% lower than the level of GAG in spinal cord treated with an injection of ChABC. When using the delivery system, 24.4% of the initial ChABC dose could still be detected in the lesion after 3 weeks, compared to just 4.4% when using an intraspinal injection of ChABC.

High resolution in vivo retinal imaging in rodents is becoming increasingly important in eye research. Development of suitable imaging devices currently requires many lengthy animal procedures. We present an ex vivo rat model eye with fluorescently labelled retinal ganglion cells (RGC) and nerve fibre bundles that reduces the need for animal procedures while preserving key properties of the living rat eye. Optical aberrations and scattering of four model eyes and eight live rat eyes were quantified using a Shack-Hartmann sensor. Fluorescent images from RGCs were obtained using a prototype scanning laser ophthalmoscope. The wavefront aberration root mean square value without defocus did not significantly differ between model and living eyes. Higher order aberrations were slightly higher but RGC image quality was comparable to published in vivo work. Overall, the model allows a large reduction in number and duration of animal procedures required to develop new in vivo retinal imaging devices.

Retinal pigment epithelial cell malfunction is a causative feature of age-related macular degeneration, and transplantation of new retinal pigment epithelial cells is an attractive strategy to prevent further progression and visual loss. However, transplants have shown limited efficacy, mainly because transplanted cells fail to adhere and migrate onto pathological Bruch's membrane. Adhesion to Bruch's membrane is integrin-mediated. Ageing of Bruch's membrane leads to a decline in integrin ligands and, added to this, wet age-related macular degeneration leads to upregulation of anti-adhesive molecules such as tenascin-C. We have therefore investigated whether manipulation of integrin function in retinal pigment epithelial cells can restore their adhesion and migration on wet age-related macular degeneration-damaged Bruch's membrane. Using spontaneously immortalized human retinal pigment epithelial cells (adult retinal pigment epithelium-19), we show that adhesion and migration on the Bruch's membrane components is integrin-dependent and enhanced by integrin-activating agents manganese and TS2/16. These allowed cells to adhere and migrate on low concentrations of ligand, as would be found in aged Bruch's membrane. We next developed a method for stripping cells from Bruch's membrane so that adhesion and migration assays can be performed on its surface. Integrin activation had a moderate effect on enhancing retinal pigmented epithelial cell adhesion and migration on normal human and rat Bruch's membrane. However, on Bruch's membrane prepared from human wet age-related macular degeneration-affected eyes, adhesion was lower and integrin activation had a much greater effect. A candidate molecule for preventing retinal pigmented epithelial interaction with age-related macular degeneration-affected Bruch's membrane is tenascin-C which we confirm is present at high levels in wet age-related macular degeneration membrane. We show that tenascin-C is anti-adhesive for retinal pigmented epithelial cells, but after integrin activation, they can adhere and migrate on it using alphaVbeta3 integrin. Alternatively, we find that transduction of retinal pigmented epithelial cells with alpha9 integrin, a tenascin-C-binding integrin, led to a large increase in alpha9beta1-mediated adhesion and migration on tenascin-C. Both expression of alpha9 integrin and integrin activation greatly enhanced the ability of retinal pigment epithelial cells to adhere to tenascin-rich wet age-related macular degeneration-affected Bruch's membranes. Our results suggest that manipulation of retinal pigment epithelial cell integrins through integrin activating strategies, or expression of new integrins such as alpha9, could be effective in improving the efficacy of retinal pigment epithelial cell transplantation in wet age-related macular degeneration-affected eyes.

Hereditary mucoepithelial dysplasia is a rare desmosomal gap junction abnormality known to produce keratitis and cataracts in addition to multiple systemic problems. It generally carries a poor visual prognosis due to corneal scarring, amblyopia, and side effects of the treatment. The authors present a sporadic case of hereditary mucoepithelial dysplasia with an unusually good visual outcome and suggest that timely and appropriate intervention can improve the visual prognosis of this rare disorder. [J Pediatr Ophthalmol Strabismus 2010;47:e1–e4.]

Retinal degenerative diseases are the leading cause of incurable blindness worldwide. Furthermore, existing pharmacological and surgical interventions are only partially effective in halting disease progression, thus adjunctive neuroprotective strategies are desperately needed to preserve vision. Stem cells appear to possess inherent neuroprotective abilities, at least in part by providing neurotrophic support to injured neurons. Advances in stem cell biology offer the hope of new therapies for a broad range of neurodegenerative conditions, including those of the retina. Experimental cell-mediated therapies also hint at the tantalizing possibility of achieving retinal neuronal replacement and regeneration, once cells are lost to the disease process. This article summarizes the latest advances in cell therapies for neuroprotection and regeneration in neurodegenerative pathologies of both the inner and outer retina.

Transplantation of stem or progenitor cells is an attractive new approach for treating neurodegenerative conditions of the central nervous system, which aims to protect or replace neurons and improve function. Proof of principle has already been shown in the retina that photoreceptors may be replaced by transplantation of neural progenitor cells. However, the task of retinal ganglion cell replacement is much more complex, as new cells will need to establish complex connections within the retina and also extend axons to precise targets in the brain. Although progress has been made in this field, it is likely that neuroprotective clinical applications will be established more quickly. Our laboratory has focused on the intraocular transplantation of cells to treat inner retinal disease, either by neuronal replacement or neuroprotection of existing cells. We have investigated the efficacy and effects of transplanting a variety of cell types, including human Müller stem cells (MIO-M1), oligodendrocyte precursor cells (OPCs), and bone marrow-derived mesenchymal stromal cells (MSCs) in a rat model of experimentally induced glaucoma. We also have developed and characterized a novel in vitro organotypic retinal explant culture system for exploring the methods of enhancing the efficacy of cell transplantation for the inner retina. In this review, we discuss the potentially beneficial effects of intraocular cell injections, identify current shortcomings of retinal stem cell therapy, and suggest directions for future research.

BACKGROUND: The Folin–Ciocalteu method is routinely used to calculate total polyphenol concentrations (TPP) in foodproducts. However, the use of inappropriate standards such as gallic acid (GA) may give inaccurate estimates of TPP content.A method for the production of a pomegranate ( Punica granatum L. ‘Wonderful’) standard was developed and validated toavoid this problem. The PPS was produced by solid phase (C18) isolation of polyphenols from POMx powder (a nutritionalsupplement) and used for TPP analysis.RESULTS: TPPcontentofPOMxwas > 90%usingPPS,whereastheGAstandardunderestimatedTPPbyupto30%.Compositionalanalysis confirmed a balance of about 9% non-polyphenolic compounds. Mass spectrometry analysis corroborated polyphenolcomposition and confirmed pomegranate-specific peaks, i.e., punicalagin and punicalin.CONCLUSION: Results support the use of pomegranate-specific standards to accurately determine TPP. c 2008 Society of Chemical IndustryKeywords: pomegranate; Folin–Ciocalteu; hydrolyzable tannin; ellagitannin; polyphenol; gallic acid

To quantify ciliary body position variability in glaucoma patients in order to determine if scleral transillumination should be used routinely to guide contact probe placement during cyclodestructive procedures.This was a prospective experimental study with human subjects. One hundred consecutive glaucoma patients attending a glaucoma clinic were recruited between June and November 2006. A fibre optic light source was used to identify ciliary body position by transscleral transillumination. The distance between the ciliary body and corneoscleral limbus was measured in the superior, temporal, inferior, and nasal quadrants of both eyes using surgical callipers.The anterior boundary of the ciliary body was located 1.5-5 mm posterior to the corneoscleral limbus. The ciliary body was located significantly more posteriorly in the superior and inferior quadrants compared to the nasal and temporal quadrants. (P0.001). Ciliary body position was significantly correlated with the mean sphere of the refractive error in phakic patients (r(2)=0.052; P=0.047). The ciliary body tended to be located most posteriorly in myopic eyes.The distance between the corneoscleral limbus and the ciliary body as identified by transillumination varies significantly in different quadrants of the eye. Considerable variability was also observed between different patients with glaucoma. Ciliary body transillumination on patients undergoing cyclodestructive procedures may have implications for optimal probe placement and therapeutic outcome.

Diseases such as age-related macular degeneration (AMD) affect the retinal pigment epithelium (RPE) and lead to the death of the epithelial cells and ultimately blindness. RPE transplantation is currently a major focus of eye research and clinical trials using human stem cell-derived RPE cells are ongoing. However, it remains to be established to which extent the source of RPE cells for transplantation affects their therapeutic efficacy and this needs to be explored in animal models. Autotransplantation of RPE cells has attractions as a therapy, but existing protocols to isolate adult RPE cells from rodents are technically difficult, time-consuming, have a low yield and are not optimized for long-term cell culturing. Here, we report a newly devised protocol which facilitates reliable and simple isolation and culture of RPE cells from adult rats. Incubation of a whole rat eyeball in 20 U/ml papain solution for 50 minutes yielded 4 x 104 viable RPE cells. These cells were hexagonal and pigmented upon culture. Using immunostaining, we demonstrated that the cells expressed RPE cell-specific marker proteins including cytokeratin 18 and RPE65, similar to RPE cells in vivo. Additionally, the cells were able to produce and secrete Bruch’s membrane matrix components similar to in vivo situation. Similarly, the cultured RPE cells adhered to isolated Bruch’s membrane as has previously been reported. Therefore, the protocol described in this article provides an efficient method for the rapid and easy isolation of high quantities of adult rat RPE cells. This provides a reliable platform for studying the therapeutic targets, testing the effects of drugs in a preclinical setup and to perform in vitro and in vivo transplantation experiments to study retinal diseases.

Over the last decade, a large number of research articles have been published demonstrating regeneration and/or neuroprotection of retinal ganglion cells following manipulation of specific genetic and molecular targets. Interestingly, of the targets that have been identified to promote repair following visual system damage, many are genes known to be mutated in different types of cancer. This review explores recent literature on the potential for modulating cancer genes as a therapeutic strategy for visual system repair and looks at the potential clinical challenges associated with implementing this type of therapy. We also discuss signalling mechanisms that have been implicated in cancer and consider how similar mechanisms may improve axonal regeneration in the optic nerve.

The aim of the study is to measure the impact of age-related macular degeneration on vision-related activity limitation and preference-based status for glaucoma patients.This was a cross-sectional study.Two-hundred glaucoma patients of whom 73 had age-related macular degeneration were included in the research.Sociodemographic information, visual field parameters and visual acuity were collected. Age-related macular degeneration was scored using the Age-Related Eye Disease Study system.The Rasch-analysed Glaucoma Activity Limitation-9 and the Visual Function Questionnaire Utility Index measured vision-related activity limitation and preference-based status, respectively. Regression models determined factors predictive of vision-related activity limitation and preference-based status. Differential item functioning compared Glaucoma Activity Limitation-9 item difficulty for those with and without age-related macular degeneration.Mean age was 73.7 (±10.1) years. Lower better eye mean deviation (β: 1.42, 95% confidence interval: 1.24-1.63, P 0.001) and age-related macular degeneration (β: 1.26 95% confidence interval: 1.10-1.44, P = 0.001) were independently associated with worse vision-related activity limitation. Worse eye visual acuity (β: 0.978, 95% confidence interval: 0.961-0.996, P = 0.018), high risk age-related macular degeneration (β: 0.981, 95% confidence interval: 0.965-0.998, P = 0.028) and severe glaucoma (β: 0.982, 95% confidence interval: 0.966-0.998, P = 0.032) were independently associated with worse preference-based status. Glaucoma patients with age-related macular degeneration found using stairs, walking on uneven ground and judging distances of foot to step/curb significantly more difficult than those without age-related macular degeneration.Vision-related activity limitation and preference-based status are negatively impacted by severe glaucoma and age-related macular degeneration. Patients with both conditions perceive increased difficulty walking safely compared with patients with glaucoma alone.