The effect of exogenous ciliary neurotrophic factor on cell cycle and neural differentiation markers of in vitro model cells: New insights for future therapeutic approaches.

Retinoblastoma is known as childhood rare malignancy of the retina. Ciliary neurotrophic factor (CNTF) was previously found to reduce degeneration and promote retina survival. This work investigated the effects of CNTF supplementation on in-vitro model cells including retinoblastoma (Y79) and adipose-derived mesenchymal stem cells (AMSCs) viability, proliferation, gene expression and cell cycle. A drop of viability was detected in Y79 treated with CNTF in a dose-dependent manner (P < .05). However, the proliferation of AMSCs was increased at lower concentrations of CNTF (5 ng/mL), but declined in higher doses (50 and 100 ng/mL). The BrdU assay confirmed the MTT assay results. Cell cycle was arrested in both Y79 and AMSCs in the G0/G1 phase by CNTF treatment. A considerable down-regulation of Bcl2, CycD1 and N-Myc genes expression (P < .05) inversely, P15 and P21 genes up-regulation in treated Y79 cells was observed. Besides, stemness genes' transcription was reduced in AMSCs (P < .05), and levels of neuronal-specific markers such as neuron-specific enolase (NSE) and neuronal nuclei (NeuN) were increased (P < .05). The findings of this study suggest a promising potential of CNTF in terms of arresting Y79 retinoblastoma cells, and differentiation-inducing to AMSCs, which could be valuable for managing future innovative treatments targeting retinoblastoma. SIGNIFICANCE OF THE STUDY: We demonstrate that CNTF has the potential to reduce proliferation of Y79 cells and induce the cell cycle arrest of them. Also, down-regulation of oncogenes (such as N-Myc) while up-regulation of tumour suppressor genes (such as P21) was detected by exposure of Y79 cells to CNTF. Furthermore, we observed the cell cycle arrest, reduction of stemness gene and up-regulation of neural differentiation markers in AMSCs treated with CNTF. These results support the probable promising effects of CNTF for controlling retinoblastoma.

SVEP1 as a Genetic Modifier of TEK-Related Primary Congenital Glaucoma.

Purpose: Affecting children by age 3, primary congenital glaucoma (PCG) can cause debilitating vision loss by the developmental impairment of aqueous drainage resulting in high intraocular pressure (IOP), globe enlargement, and optic neuropathy. TEK haploinsufficiency accounts for 5% of PCG in diverse populations, with low penetrance explained by variable dysgenesis of Schlemm's canal (SC) in mice. We report eight families with TEK-related PCG, and provide evidence for SVEP1 as a disease modifier in family 8 with a higher penetrance and severity. Methods: Exome sequencing identified coding/splice site variants with an allele frequency less than 0.0001 (gnomAD). TEK variant effects were assayed in construct-transfected HEK293 cells via detection of autophosphorylated (active) TEK protein. An enucleated eye from an affected member of family 8 was examined via histology. SVEP1 expression in developing outflow tissues was detected by immunofluorescent staining of 7-day mouse anterior segments. SVEP1 stimulation of TEK expression in human umbilical vascular endothelial cells (HUVECs) was measured by TaqMan quantitative PCR. Results: Heterozygous TEK loss-of-function alleles were identified in eight PCG families, with parent-child disease transmission observed in two pedigrees. Family 8 exhibited greater disease penetrance and severity, histology revealed absence of SC in one eye, and SVEP1:p.R997C was identified in four of the five affected individuals. During SC development, SVEP1 is secreted by surrounding tissues. SVEP1:p.R997C abrogates stimulation of TEK expression by HUVECs. Conclusions: We provide further evidence for PCG caused by TEK haploinsufficiency, affirm autosomal dominant inheritance in two pedigrees, and propose SVEP1 as a modifier of TEK expression during SC development, affecting disease penetrance and severity.

Medical therapy versus trabeculectomy in patients with open-angle glaucoma.

PURPOSE: To compare therapeutic outcomes between trabeculectomy and medical therapy in patients with open-angle glaucoma. METHODS: In the present retrospective comparative study, the medical charts of 284 patients (eyes) newly diagnosed with open-angle glaucoma who had received conventional medications (n=188) or undergone fornix-based trabeculectomy (n=96) at a teaching eye hospital were reviewed. RESULTS: At a mean follow-up of 6.6 years, post-treatment changes in intraocular pressure (IOP), visual field (VF), best spectacle-corrected visual acuity (BSCVA), and number of required drugs were significantly more favorable in the surgical group (P<0.001 for all comparisons). However, the frequency of clinically desirable IOP (≤21 mmHg) at the endpoint was comparable between the surgical and medical groups (87.2% vs. 82.3%; P=0.26). The rate of conversion to surgical therapy was 34% in the medical group. A greater baseline requirement for anti-glaucoma drugs (two or more) was the only independent predictor of treatment failure in the present study. CONCLUSIONS: Although more severe cases naturally receive trabeculectomy, the surgical approach had greater efficacy than conventional medical therapy in patients with open-angle glaucoma. An initial requirement for two or more anti-glaucoma drugs may predict failure of medical therapy.

Medical therapy versus trabeculectomy in patients with open-angle glaucoma / Tratamento clínico versus trabeculectomia em pacientes com glaucoma de ângulo aberto

ABSTRACT Purpose: To compare therapeutic outcomes between trabeculectomy and medical therapy in patients with open-angle glaucoma. Methods: In the present retrospective comparative study, the medical charts of 284 patients (eyes) newly diagnosed with open-angle glaucoma who had received conventional medications (n=188) or undergone fornix-based trabeculectomy (n=96) at a teaching eye hospital were reviewed. Results: At a mean follow-up of 6.6 years, post-treatment changes in intraocular pressure (IOP), visual field (VF), best spectacle-corrected visual acuity (BSCVA), and number of required drugs were significantly more favorable in the surgical group (P<0.001 for all comparisons). However, the frequency of clinically desirable IOP (≤21 mmHg) at the endpoint was comparable between the surgical and medical groups (87.2% vs. 82.3%; P=0.26). The rate of conversion to surgical therapy was 34% in the medical group. A greater baseline requirement for anti-glaucoma drugs (two or more) was the only independent predictor of treatment failure in the present study. Conclusions: Although more severe cases naturally receive trabeculectomy, the surgical approach had greater efficacy than conventional medical therapy in patients with open-angle glaucoma. An initial requirement for two or more anti-glaucoma drugs may predict failure of medical therapy.

RESUMO Objetivo: Comparar o resultado terapêutico de trabeculectomia versus terapia médica em pacientes com glaucoma de ângulo aberto. Método: Neste estudo comparativo retrospectivo, prontuários médicos de 284 pacientes (olhos), de um hospital de ensino oftalmológico, com diagnóstico recente de glaucoma de ângulo aberto que receberam medicamentos convencionais (n=188) ou foram submetidos a trabeculectomia de base fórnice (n=96) foram revisados. Resultados: Com seguimento médio de 6,6 anos, as mudanças pós-tratamento da pressão intraocular (PIO), campo visual (VF), melhor acuidade visual corrigida por óculos (BSCVA), e o número de medicações necessárias foram significativamente mais favorável ao grupo cirúrgica (p<0,001 para todas as comparações). No entanto, os grupos foram comparáveis para a frequência de PIO clinicamente desejável (≤21 mmHg) na visita final (87,2% vs. 82,3%, respectivamente; p=0,26). A taxa de conversão para o tratamento cirúrgico foi de 34% no grupo médico e a necessidade inicial de mais drogas antiglaucomatosas (2 ou mais) foi o único preditor independente desta conversão. Conclusões: Embora os casos mais graves de glaucoma são naturalmente designados o grupo de trabeculectomia, esta abordagem cirúrgica se mostrou mais eficaz do que a terapia médica convencional em pacientes com glaucoma de ângulo aberto. Uma necessidade inicial de 2 ou mais medicações antiglaucomatosas pode prever a falha em terapia médica.

Risk factors for late presentation of chronic glaucoma in an Iranian population.

PURPOSE: To identify risk factors for late presentation in chronic glaucoma. MATERIALS AND METHODS: In a hospital-based case-control study, 312 newly diagnosed consecutive patients with chronic glaucoma were recruited to study in Nikookari Hospital in Tabriz - Iran. Cases were defined as patients presenting with absolute field loss within 5° of fixation or a cup to disc ratio >0.8 in one or both eyes (188 patients). Controls were patients with no absolute field loss within 20° in either eye but otherwise glaucomatous field loss and a cup to disc ratio >0.5 or a difference of 0.2 or greater between the discs (124 patents). RESULTS: In logistic regression model, there was a significant association between late presentation and male gender, lower education status, pseudoexfoliative glaucoma, high intraocular pressure (IOP) in the first presentation and no eye exam in last 2 years. Patients with pseudoexfoliative glaucoma had 1.5 times more odd for late presentation (1.52 confidence interval [CI]: 1.14-2.24 P = 0.004). Patients that visited by an ophthalmologist in the last 2 years had 0.5 less likely to present with advanced glaucoma (0.54 CI: 0.32-0.92 P = 0.025). Higher IOP in initial examination was associated with advanced glaucoma. We estimated 1.08 increase in odd (CI: 1.03-1.04 P = 0.001) per increase in 1 mmHg in IOPs above 25 mmHg. CONCLUSION: Our study shows that several risk factors present for late presentation in chronic glaucoma. Our results suggest planning screening programs in high-risk individuals for early detection of glaucoma and prevention of blindness.

Customized Clinical Practice Guidelines for Management of Adult Cataract in Iran.

PURPOSE: To customize clinical practice guidelines (CPGs) for cataract management in the Iranian population. METHODS: First, four CPGs (American Academy of Ophthalmology 2006 and 2011, Royal College of Ophthalmologists 2010, and Canadian Ophthalmological Society 2008) were selected from a number of available CPGs in the literature for cataract management. All recommendations of these guidelines, together with their references, were studied. Each recommendation was summarized in 4 tables. The first table showed the recommendation itself in clinical question components format along with its level of evidence. The second table contained structured abstracts of supporting articles related to the clinical question with their levels of evidence. The third table included the customized recommendation of the internal group respecting its clinical advantage, cost, and complications. In the fourth table, the internal group their recommendations from 1 to 9 based on the customizing capability of the recommendation (applicability, acceptability, external validity). Finally, customized recommendations were sent one month prior to a consensus session to faculty members of all universities across the country asking for their comments on recommendations. RESULTS: The agreed recommendations were accepted as conclusive while those with no agreement were discussed at the consensus session. Finally, all customized recommendations were codified as 80 recommendations along with their sources and levels of evidence for the Iranian population. CONCLUSION: Customization of CPGs for management of adult cataract for the Iranian population seems to be useful for standardization of referral, diagnosis and treatment of patients.

Revisiting corneal topography for the diagnosis of keratoconus: use of Rabinowitz's KISA% index.

BACKGROUND: Numerous videokeratographic methods are used in the diagnosis of keratoconus. The purpose of this study was to compare the sensitivity and specificity of the KISA% index with the keratometry (K) value, inferior-superior (I-S) value, relative skewing of the steepest radial axes (SRAX), and keratometric astigmatism (AST) indices in 25 patients presenting with bilateral keratoconus. METHODS: Twenty-five patients presenting with bilateral keratoconus were enrolled. Fifty eyes from 25 healthy individuals without corneal abnormalities were used as controls. The clinical diagnosis of keratoconus was made by observation of scissors reflex at retinoscopy and characteristic slit lamp findings. Corneal topographic analysis was performed in each eye to calculate the K value, I-S value, AST, and SRAX indices. RESULTS: The keratoconus percentage index (KISA%) was significantly more sensitive and specific than the other indices examined. Furthermore, it was significantly better at predicting positive and negative results than the other indices included in the study. CONCLUSION: The current research suggests that KISA% could be used to detect/diagnose keratoconus but further studies are required to confirm the specificity and sensitivity of KISA% for the detection of early-stage disease and keratoconus suspects.

Aqueous humor hepcidin prohormone levels in patients with primary open angle glaucoma.

PURPOSE: This study was designed to evaluate the levels of Interleukin-6 (IL6) and Hepcidin prohormone (Hep) in the serum and aqueous humor of patients with primary open-angle glaucoma (POAG), as well as those with senile cataract as a control group. METHODS: Levels of IL6 and Hep were measured by enzyme-linked immunosorbent assay (ELISA) methods in serum and aqueous humor aspirates taken from 45 patients (POAG=20, Control=25) during anterior segment surgery. RESULTS: The mean aqueous humor Hep concentration in eyes with POAG was significantly higher than that controls (34.55±23.01 ng/ml versus 20.82± 24.63 ng/ml, p=0.04). There was no significant difference between the serum Hep concentration of POAG and the control group (359.46±113.03 ng/ml versus 287.36±130.53 ng/ml, p=0.08). There was also no significant difference between either serum (6.18±5.22 versus 10.84±3.37, pg/ml p=0.112) or aqueous humor (4.39±3.06 versus 5.79±2.63, pg/ml p=0.14) IL6 concentrations of POAG and the control groups. No significant correlations were found between aqueous humor and serum Hep and IL6 levels in either POAG or the control groups (p>0.05). CONCLUSIONS: The aqueous humor Hep level may exist locally and independent from the IL6 increase in patients with POAG, suggesting that Hep might represent a bridge protein between local inflammation and the consequent loss of retinal ganglion cells.

Outcomes of penetrating keratoplasty in keratoconus.

PURPOSE: Keratoconus (KCN) is one of the most common indications of corneal transplantation in Iran. This study was conducted to determine the outcomes of penetrating keratoplasty (PK) for KCN in patients operated in a private practice setting from 1994 to 2001. METHODS: This longitudinal retrospective study included 164 eyes of 164 patients. Variables included preoperative keratometry, trephination and suturing techniques, donor-recipient disparity, surgical complications, immunologic rejection, graft clarity, postoperative spherical and cylindrical refractive error, keratometry, uncorrected and best spectacle-corrected visual acuity, suture management, and the results of keratorefractive procedures. RESULTS: Patients were followed for a mean period of 33.5 months. Mean postoperative best spectacle-corrected visual acuity (BSCVA) at last follow-up was 0.14 +/- 0.11 LogMAR (20/25); mean spherical error and mean corneal astigmatism were -0.61 +/- 2.6 and 3.4 +/- 1.8 D, respectively. Final visual outcomes were not significantly correlated with trephination and suturing techniques or severity of the ectasia. Although donor-recipient disparity did not affect final astigmatism, more myopic shift was observed with greater disparity, but this finding was not of statistical significance. Overall, 26.8% of the patients required keratorefractive surgery, which resulted in 2.9 D reduction in corneal astigmatism. Immunologic graft rejection occurred in 28% of cases; however, all episodes responded to medical management, and none resulted in graft failure. CONCLUSION: Penetrating keratoplasty is a safe and effective procedure with remarkable optical and visual outcomes for patients with keratoconus who are contact lens intolerant or have unacceptable corrected visual acuity. Neither severity of the disorder nor trephination and suturing techniques significantly affects final visual outcomes. Less graft-recipient disparity (0.25 versus 0.50 mm) seems to induce less myopic shift.