Generation of an EYS-associated retinitis pigmentosa patient-derived human pluripotent stem cell line (MUi038-A).

Mutations in the eyes shut homolog (EYS) gene are one of the common causes of autosomal recessive retinitis pigmentosa (RP). The lack of suitable animal models hampers progress understanding of the disease mechanism and drug development. This study reported the reprogramming of CD34+ hematopoietic stem/progenitor cells from a patient with compound heterozygous EYS mutations (c.6416 G > A and c.7228 + 1 G > A) into the induced pluripotent stem cell line, MUi038-A, using non-integrating vectors. The MUi038-A demonstrates pluripotency, tri-lineage differentiation potential, and a normal karyotype, offering a valuable model for studying the mechanism of EYS-related RP and new therapeutic development.

Occult retinal neovascularization following intravitreal bevacizumab and laser treatment for retinopathy of prematurity.

BACKGROUND: We present a patient with retinopathy of prematurity (ROP) who developed worsening plus disease after complete regression of stage 3 ROP. The use of fundus fluorescein angiography (FFA) aided the visualization of occult neovascularization that caused the disease progression. CASE PRESENTATION: The patient was at high risk for ROP due to low birth weight of 690 g and gestational age of 25 weeks. After the diagnosis of stage 3 ROP in zone I without plus disease, she was treated initially with bilateral intravitreal bevacizumab (IVB) and followed by laser photocoagulation 5 weeks later. Despite the resolution of ROP stage, the plus disease worsened. Neither systemic risk factors nor skip laser areas were observed. Hence, FFA was performed and subsequently identified occult neovascularization with active leakage. Additional IVB and laser treatment in the capillary dropout area inside vascularized retina were added. The plus disease improved but mild arteriolar tortuosity persisted. CONCLUSIONS: Worsening of plus disease after completion of laser ablation and IVB with complete regression of stage 3 ROP is rare. Systemic risk factors such as continuous oxygen therapy and cardiovascular disease should be ruled out. FFA aided in identifying occult neovascularization and prompted further treatment.

Validation of the postnatal growth and retinopathy of prematurity (G-ROP) screening criteria in a Thai cohort.

PURPOSE: To validate Postnatal Growth and Retinopathy of Prematurity (G-ROP) criteria for Thai infants. STUDY DESIGN: A retrospective review of infants receiving ROP screening during 2009-2020. METHODS: Baseline characteristics, clinical progression and final ROP outcomes were collected. G-ROP was applied to infants who met at least one of the following 6 criteria: birth weight (BW) below 1051 g, gestational age (GA) under 28 weeks, weight gain (WG) less than 120 g during postnatal day 10-19, WG less than 180 g during day 20-29, WG less than 170 g during day 30-39 and hydrocephalus. RESULTS: A total of 684 infants (boys, 53.4%) were included. Median (IQR) BW was 1200 (960-1470) grams and median GA was 30 (28-32) weeks. Prevalence of ROP was 26.6%, with 28 (4.1%) having type 1, 19 (2.8%) type 2 and, 135 (19.7%) having other ROP. Treatment was performed in 26 infants (3.8%). Sensitivity of G-ROP to include type 1, 2 or treatment-requiring ROP cases was 100% with 36.9% specificity, excluding 235 (34.4%) cases of unnecessary screening. To adjust for our setting of initial eye examination at 4 weeks' postnatal date, the last 2 criteria of G-ROP were replaced by the occurrence of grade 3 or 4 intraventricular hemorrhage (IVH). This modified G-ROP criteria yielded 100% sensitivity, 42.5% specificity and excluded 271 (39.6%) cases of unnecessary screening. CONCLUSION: G-ROP criteria can be applied to our hospital setting. Occurrence of IVH grade 3 or 4 was proposed as an alternative in modified G-ROP criteria.

A generation of human-induced pluripotent stem cell line (MUi032-A) from a Choroideremia disease patient carrying a hemizygous mutation on the CHM gene.

Choroideremia (CHM) is a monogenic, X-linked inherited retinal disease caused by mutations in the CHM gene. CHM patients develop progressive loss of vision due to degeneration of cell layers in the retina. In this report, the human-induced pluripotent stem cell, MUi032-A, was generated from CD34+ hematopoietic stem/progenitor cells of a male CHM patient by co-electroporation of non-integration episomal vectors containing OCT4/shp53, Sox-2/KLF4, and L-MYC/LIN-28. The MUi032-A showed normal karyotype and a hemizygous c.715C > T mutation. They expressed pluripotency markers and differentiated into cells derived from three germ layers. This cell line may be useful for disease mechanisms and gene therapy studies.

Effect of surgical simulation training on the complication rate of resident-performed phacoemulsification.

OBJECTIVE: To study the effect of additional training with ophthalmic surgical simulation on the intraoperative complication rates of phacoemulsification performed by residents. METHODS AND MATERIALS: This was a retrospective study of phacoemulsification surgeries performed by third-year residents at Siriraj Hospital. The operations were classified into two groups according to the experience of the surgeon in simulation training, that is, trained vs untrained. The main outcome was the total rate of complications. Other outcomes, including posterior capsule rupture, anterior capsulorhexis tearing, zonular dehiscence, retaining of lens material and intraocular lens (IOL) implantation methods, were also analysed. RESULTS: In total, 2971 operations were performed, comprising 1656 operations by 21 residents in the trained group, and 1315 by 20 residents in the untrained group. The total rate of complications in the simulator-trained group was lower than in the untrained group (13.6% vs 17.3%, p=0.005). Only the rate of retaining lens material showed a statistically significantly reduction (p<0.001); however, the rates of posterior capsule rupture, anterior capsulorhexis tearing and zonular dehiscence were not significantly different (p=0.08, 0.17, 0.23, respectively). The IOL implantation methods and surgical aphakia rate between the two groups were similar (p=0.44). In the subgroup analysis, the posterior capsule rupture rate in the first half of all cases performed by the residents was lower in the trained group (8.8% vs 12.4%, p=0.02). CONCLUSION: Ophthalmic simulation training reduces the total rate of complications of resident-performed phacoemulsification. It also shortens the learning curve for cataract surgery training, as indicated by the decreased posterior capsule rupture rate in the initial cases of cataract surgery.

Autoantibody profiles and clinical association in Thai patients with autoimmune retinopathy.

Autoimmune retinopathy (AIR) is a rare immune-mediated inflammation of the retina. The autoantibodies against retinal proteins and glycolytic enzymes were reported to be involved in the pathogenesis. This retrospective cohort study assessed the antiretinal autoantibody profiles and their association with clinical outcomes of AIR patients in Thailand. We included 44 patients, 75% were females, with the overall median age of onset of 48 (17-74, IQR 40-55.5) years. Common clinical presentations were nyctalopia (65.9%), blurred vision (52.3%), constricted visual field (43.2%), and nonrecordable electroretinography (65.9%). Underlying malignancy and autoimmune diseases were found in 2 and 12 female patients, respectively. We found 41 autoantibodies, with anti-α-enolase (65.9%) showing the highest prevalence, followed by anti-CAII (43.2%), anti-aldolase (40.9%), and anti-GAPDH (36.4%). Anti-aldolase was associated with male gender (P = 0.012, OR 7.11, 95% CI 1.54-32.91). Anti-CAII showed significant association with age of onset (P = 0.025, 95% CI - 17.28 to - 1.24), while anti-α-enolase (P = 0.002, OR 4.37, 95% CI 1.83-10.37) and anti-GAPDH (P = 0.001, OR 1.87, 95% CI 1.32-2.64) were significantly associated with nonrecordable electroretinography. Association between the antibody profiles and clinical outcomes may be used to direct and adjust the treatment plans and provide insights in the pathogenesis of AIR.

Intravitreal autologous mesenchymal stem cell transplantation: a non-randomized phase I clinical trial in patients with retinitis pigmentosa.

BACKGROUND: Retinitis pigmentosa (RP) is a progressive inherited retinal disease with great interest for finding effective treatment modalities. Stem cell-based therapy is one of the promising candidates. We aimed to investigate the safety, feasibility, and short-term efficacy of intravitreal injection of bone marrow-derived mesenchymal stem cells (BM-MSCs) in participants with advanced stage RP. METHODS: This non-randomized phase I clinical trial enrolled 14 participants, categorized into three groups based on a single dose intravitreal BM-MSC injection of 1 × 106, 5 × 106, or 1 × 107 cells. We evaluated signs of inflammation and other adverse events (AEs). We also assessed the best corrected visual acuity (BCVA), visual field (VF), central subfield thickness (CST), and subjective experiences. RESULTS: During the 12-month period, we noticed several mild and transient AEs. Interestingly, we found statistically significant improvements in the BCVA compared to baseline, although they returned to the baseline at 12 months. The VF and CST were stable, indicating no remarkable disease progression. We followed 12 participants beyond the study period, ranging from 1.5 to 7 years, and observed one severe but manageable AE at year 3. CONCLUSION: Intravitreal injection of BM-MSCs appears to be safe and potentially effective. All adverse events during the 12-month period required observation without any intervention. For the long-term follow-up, only one participant needed surgical treatment for a serious adverse event and the vision was restored. An enrollment of larger number of participants with less advanced RP and long-term follow-up is required to evaluate the safety and efficacy of this intervention. TRIAL REGISTRATION: ClinicalTrials.gov, NCT01531348 . Registered on February 10, 2012.

Whole Exome Sequencing in Eight Thai Patients With Leber Congenital Amaurosis Reveals Mutations in the CTNNA1 and CYP4V2 Genes.

Purpose: Our goal was to describe the clinical and molecular genetic findings in Thai patients with Leber congenital amaurosis (LCA). Methods: Whole exome sequencing (WES) was performed in eight unrelated patients. All genes responsible for inherited retinal diseases (IRDs) based on RetNet were selected for analysis. Potentially causative variants were filtered through a bioinformatics pipeline and validated using Sanger sequencing. Segregation analysis of the causative genes was performed in family members when available. Results: Eleven deleterious variants, six nonsense and five missense, were identified in seven genes: four LCA-associated genes (CEP290, IQCB1, NMNAT1, and RPGRIP1), one gene responsible for syndromic LCA (ALMS1), and two IRDs-related genes (CTNNA1 and CYP4V2). Clinical reassessment supported the diagnosis of syndromic LCA in those patients harboring potentially pathogenic variants in the ALMS1. Interestingly, two causative genes, CTNNA1 and CYP4V2, previously reported to cause butterfly-shaped pigment dystrophy (BSPD) and Bietti's crystalline dystrophy (BCD), respectively, were detected in two other patients. These two patients developed rapid and severe visual loss in contrast to BSPD and BCD patients in previous studies. The results of this study demonstrate that causative variants identified in the CTNNA1 and CYP4V2 genes are also associated with LCA. Conclusions: This is the first report describing the molecular genetics and clinical manifestations of Thai patients with LCA. The present study expands the spectrum of LCA-associated genes, which is a benefit for molecular diagnosis. The identification of mutations in the CTNNA1 and CYP4V2 genes requires further elucidation in larger cohorts with LCA.

Optic atrophy after anti-vascular endothelial growth factor injection in diabetic patients with proliferative diabetic retinopathy.

OBJECTIVE: To study the prevalence of optic atrophy in patients with proliferative diabetic retinopathy (PDR) who underwent intravitreal bevacizumab injection and risk factors associated with optic atrophy. MATERIAL AND METHOD: A retrospective case control study enrolled 269 cases (394 eyes) of patients with PDR, in which 166 cases (219 eyes) received intravitreal bevacizumab injection. Associated factors such as type of DM, hemoglobin A1c level, hypertension, hypercholesterolemia, chronic kidney disease, previous intravitreal surgery retinal detachment, and vitreous hemorrhage were recorded. Criteria for diagnosis of optic atrophy were decreased visual acuity, pale optic disc and decreased nerve fiber layer thickness, which was measured by Stratus optical coherence tomography (OCT). The association between intravitreal bevacizumab injection and optic atrophy was analyzed by multiple logistic regression. RESULTS: Two hundred sixty nine patients with PDR, consisting of 166 patients with intravitreal bevacizumab injection and 103 cases without bevacizumab injection. Optic atrophy was found in 11.4% (25/219 eyes) and 8% (14/175 eyes) respectively. There was no evidence that intravitreal bevacizumab injection and associated systemic diseases were related to optic atrophy. The risk factor that was related to optic atrophy was previous intravitreal surgery (adjusted odds ratio (OR), 2.57 [95% CI, 1.13, 5.84], p = 0.024). CONCLUSION: Anti-VEGF (bevacizumab) does not increase the risk of optic atrophy. The ophthalmologists should be aware of subsequent optic atrophy development in patients with PDR who undergo surgical intervention.

Visual outcome after cataract surgery complicated by posterior capsule rupture.

BACKGROUND: Posterior capsule rupture is one of the most common intra-operative complications of cataract surgery and may lead to other sequelae that affect visual outcome. OBJECTIVE: To determine the visual outcome and sequelae at 1 year after cataract surgery complicated by posterior capsule rupture and factors related to poor visual outcome. MATERIAL AND METHOD: Retrospective chart reviews of the patients who underwent cataract surgery complicated by posterior capsule rupture in Siriraj Hospital between January 2006 and December 2009 were performed. Data collected included demographic data, underlying systemic diseases, pre-existing ophthalmic diseases, type of cataract, type of operation, vitrectomy methods, type of intraocular lens (IOL) implantation, pre-operative and post-operative visual acuity, subsequent complications and management. RESULTS: There were 525 cases that received cataract surgery complicated by posterior capsule rupture. After excluding 280 cases with the follow-up period of shorter than 1 year 245 eyes of 242 patients were studied. These comprised 111 males and 131 females. The mean age was 69.3 years, ranged from 40 to 92 years. The mean logarithm of the minimum angle of resolution (logMAR) best-corrected visual acuity (BCVA) at 1 year was 0.43 (median 0.24). The proportion of patients who had BCVA of 0.3 logMAR or better was 64.5%. After excluding eyes with pre-existing diseases, 72.9% got this level of BCVA. Vitrectomy was required in 75.5%; which consisted of anterior vitrectomy (68.6%) and posterior vitrectomy (6.9%). Primary intra-ocular lens insertion was performed in 87.4%, mostly in the ciliary sulcus (75.5%). Subsequent complications occurred in 13.9%, which included secondary glaucoma (4.9%), cystoid macular edema (2.4o), endophthalmitis (1.6%), rhegmatogenous retinal detachment (1.2%o), IOL displacement (1.2%), uveitis (1.2%), corneal decompensation (0.8%) and fibrous ingrowth (0.4%). The factors associated with the poor visual outcome worse than 0.3 logMAR were pre-existing ocular diseases, incision requiring more than 2 stitches, posterior vitrectomy and subsequent complications (p < 0.05). CONCLUSION: Most of the patients had favorable visual outcome after cataract surgery complicated by posterior capsule rupture. Meticulous vitrectomy to prevent subsequent complications and foldable IOL insertion to minimize the wound size are recommended.

Selective ophthalmic arterial infusion of chemotherapeutic drugs for recurrent retinoblastoma.

Introduced in 1988 by Kaneko and colleagues, selective ophthalmic arterial infusion of chemotherapeutic drug has recently gained more interest among retinoblastoma experts worldwide. The report showed that the procedure could be repeated up to 12 treatments without serious side effects. We report a 4-year-old girl with bilateral retinoblastoma. The left eye was enucleated for the group E disease. The right eye started with 3 retinal tumors (group C) was treated with systemic chemotherapy plus local therapy. Seven months after the last cycle of chemotherapy, the tumor recurred close to the fovea. Systemic chemotherapy was reinitiated without success. To avoid aggressive cryotherapy and external-beam radiotherapy, selective ophthalmic arterial infusion of chemotherapeutic drugs was performed for 15 sessions. The tumor responded partially without evidence of drug-induced retinal toxicity by the electroretinogram. Minor irregularities of the inner wall of supraclinoid portion of the internal carotid artery were observed only at the sixth session. Narrowing of the vascular lumen occurred on the last 2 sessions. We demonstrate that this technique when performed repeatedly could result in the anatomic changes of the small blood vessel. Careful follow-up is necessary for early detection of any serious consequences.

Cyclooxygenase-2 expression in retinoblastoma: an immunohistochemical analysis.

PURPOSE: Increased level of cyclooxygenase-2 (COX-2) plays a significant role in the pathogenesis of cancers. High expression of COX-2 has been demonstrated in several cancer types including retinoblastoma. However, the in vivo study did not confirm the anti-proliferative effect of COX-2 inhibitor, celecoxib, on a murine transgenic retinoblastoma model. We, therefore, aim to investigate COX-2 expression in paraffin-embedded retinoblastoma specimens in a larger study group. METHODS: We reviewed 55 retinoblastoma specimens obtained during 1995 to 2005. Clinical and histopathological data were recorded. Immunohistochemical evaluation of COX-2 expression was performed using a rabbit monoclonal antibody to human cyclooxygenase-2. RESULTS: Forty-four of 55 specimens (80%) showed negative immunoreactivity for COX-2 expression. For the 11 specimens (20%, 95% CI = 11.6-32.4%) with positive COX-2, all immunostainings were less than 50% of tumor area. Demographic data and treatment details were available in 53 specimens. Enucleation was performed as a primary treatment in 43 specimens (81%). Other treatments, mainly systemic chemotherapy, were given prior to enucleation in 10 specimens (19%). There was no statistical difference in COX-2 expression between the specimens identified as primary and secondary enucleation (p = 0.66). Regarding the histopathological findings, there were no significant differences between COX-2 negative and COX-2 positive groups. CONCLUSIONS: It appears that COX-2 is not overexpressed in our retinoblastoma specimens, which is different from previous studies. This conflicting data reduces the possibility of introducing Cox-2 inhibitors in the treatment of retinoblastoma.

Prognostic factors and treatment outcomes of retinoblastoma in pediatric patients: a single-institution study.

PURPOSE: Since 1997, our institute has used neoadjuvant chemotherapy for intraocular retinoblastoma. However, some of the patients showed signs of recurrence months to years later. We therefore attempted to determine the prognostic factors of treatment outcomes and survival in our patients. METHODS: We reviewed 90 patients treated from 1997 to 2006. The following information was recorded: demographic and ophthalmological data, clinical classification, histopathological data, and treatment methods and outcomes. RESULTS: Enucleation was avoided in two of 57 eyes in the unilateral group. Sixteen of 54 eyes in the bilateral group were salvaged by systemic chemotherapy with local treatment. There was no difference in histopathological findings between the two groups. Nine of 57 patients in the unilateral group demonstrated poor outcomes, compared with four of 27 in the bilateral group. Significant poor prognostic factors for survival were optic nerve head invasion, orbital involvement, abnormal results on bone marrow aspiration, lumbar puncture, bone scan, and group E or F classification. CONCLUSIONS: The 15% mortality rate in our patients is slightly higher than that reported in developed countries. Delayed diagnosis and treatment is the main cause of avoidable deaths. Improving education of both clinicians and parents would increase survival rates in this potentially fatal disease.

Mutation analysis of the VMD2 gene in thai families with best macular dystrophy.

BACKGROUND: To identify genetic mutations of the VMD2 gene in two Thai families with Best macular dystrophy. MATERIALS AND METHODS: Ophthalmic examination including best-corrected visual acuity (BCVA), dilated fundus examination and fundus photography, and electro-oculography (EOG) was performed in two probands and their family members. Mutation screening of the VMD2 gene was performed by single-strand conformation polymorphism (SSCP) analysis followed by direct DNA sequencing of the abnormal exons. RESULTS: The 58-year-old male proband demonstrated typical egg yolk-like macular lesion in both eyes. Mutational screening of VMD2 identified a band shift in exon 7, which was confirmed by direct DNA sequencing to be a G to A transition at position 724 bp. This novel missense mutation resulted in the change of an amino acid valine to methionine and was responsible for the abnormal Arden ratio in the proband's daughter. The second male proband age 25 had a characteristic egg yolk-like macular lesion in the left eye and a scrambled egg appearance in the right. Mutation analysis identified a C to T transition at position 652 bp in exon 6. This reported missense mutation led to an amino acid substitution of cysteine for arginine. The mutation was documented in the maternal grandmother, the mother, as well as the elder sister of the proband. CONCLUSIONS: The Val-242-Met mutation is associated with a late-onset visual disturbance and the Arg-218-Cys mutation was associated with marked intra-familial clinical variability of expression. Presymptomatic testing will be available to the family members at risk with high accuracy.

Reduction of endophthalmitis rate after cataract surgery with preoperative 5% povidone-iodine.

OBJECTIVE: Postoperative endophthalmitis reflects in part quality and safety aspects of cataract surgery. Preoperative 5% povidone-iodine was introduced as a quality improvement effort. This study evaluated the effect of this additional measure on the occurrence of endophthalmitis after cataract surgery. METHOD: Topical 5% povidone-iodine solution was applied onto the ocular surface just prior to transferring the patient into the operative theater for cataract surgery. Other prophylactic measures were allowed to continue as before. Patients developing postoperative intraocular inflammation and undergoing intravitreal antibiotic injection were included as occurrences of endophthalmitis. Alteration in endophthalmitis rate was analyzed using a 'p control chart' of a statistical process control method. The incidence in the povidone-iodine-receiving group was compared to those before implementation and concurrent nonreceiving groups. RESULT: The postoperative endophthalmitis rate showed a significant reduction after introduction of povidone-iodine. A year before, 9 of 3,052 eyes developed endophthalmitis (0.294%). After introduction, this occurred in 4 of 4,089 eyes receiving povidone-iodine (0.097%) and 1 of 502 nonreceiving eyes (0.199%) in the following 16 months. Despite the apparent lower rate, comparison between groups was not statistically significant. Moderate to severe but tolerable eye irritation after application was reported in 6.6%. No other adverse events were detected. CONCLUSION: Topical preoperative 5% povidone-iodine contributed an additional effect to the reduction of the postoperative endophthalmitis rate after cataract surgery. This measure was rather safe to apply as a prophylaxis against endophthalmitis in cataract surgery.

Surgical techniques of cataract surgery and subsequent postoperative endophthalmitis.

OBJECTIVES: To compare the incidence and characteristics of patients with endophthalmitis after extracapsular cataract extraction (ECCE) to those after phacoemulsification MATERIAL AND METHOD: Records of patients receiving intravitreal antibiotic injection to treat endophthalmitis after cataract surgery between Jan 2001 and Dec 2004 were reviewed. Demographic data and other characteristics including associated diseases, details of cataract surgical procedure and intraoperative complication, onset of endophthalmitis after cataract surgery, presenting symptoms and signs of endophthalmitis, how endophthalmitis was managed, causative organisms, duration of hospitalization and results of treatment were collected. This information was compared between those of endophthalmitis patients after ECCE and those after phacoemulsification. RESULTS: There were 5 cases who developed endophthalmitis after ECCE and 31 cases after phacoemulsification. The incidence was 0.365% after ECCE and 0.279% after phacoemulsification (p = 0.589). Visual acuity (VA) before cataract surgery in ECCE group was worse than the phacoemulsification group (median VA: counting fingers vs 6/36, p = 0.001). Median onset of endophthalmitis was 8 days after ECCE and 6 days after phacoemulsification. Presenting symptoms and signs were similar. Causative agents were identified in 4 (80%) and 14 (45%) cases in the ECCE and phacoemulsification groups respectively. Gram-positive bacteria were the major cause of infection in both groups. Endophthalmitis caused by citrobacter sp. in ECCE group and enterococcus or streptococcus sps. the phacoemulsification in the group ended up with enucleation or no light perception. CONCLUSION: The present study has not demonstrated an apparent difference between endophthalmitis after ECCE and those after phacoemulsification. Endophthalmitis after either procedure can be managed as the same condition.