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Objective: Hemodialysis patients with chronic kidney disease often exhibit inflammation characterized by elevated levels of C-reactive protein, Interleukin 6 and tumor necrosis factor-alpha, and they are shown to be associated with cardiovascular impairment and enhanced renal failure. This study aims to assess the impact of fish oil intake on inflammation indicators in adult hemodialysis patients., Methods: From the inception to December 2023, the datasets Cochrane Central, Google Scholar, Science Direct, Embase, and Pubmed were examined. Two authors independently searched, selected, and screened the literature. The pooled results are represented by weighted mean difference (WMD) with 95% confidence intervals. To investigate the causes of heterogeneity, subgroup analysis was done. Sensitivity analysis was then used to evaluate the validity of the combined findings., Results: Thirteen randomized control trials studies were included. The pooled results showed that fish oil supplementation caused a significant reduction of the C-reactive protein level (WMD, -2.92 mg/L; 95% Confidence interval, -5.23, to -0.61; p  = 0.01; I 2  = 99%), especially in patients with baseline C-reactive protein ⩾5 mg/L (WMD, -4.39 mg/L; 95% Confidence interval, -5.93 to 2.85; p  < 0.00001; I 2  = 33%). Subgroup analyses showed that C-reactive protein baseline level (C-reactive protein <5 mg/L) was the main source of heterogeneity. Fish oil intake may not reduce the level of Interleukin 6 (WMD, -2.26; 95% Confidence interval: -19.61 to 15.09; p  = 0.80; I 2  = 93%), nor will it reduce the level of tumor necrosis factor-alpha (random model: WMD, -2.51; 95% Confidence interval: 6.08 to 1.06; p  = 0.17; I 2  = 98%)., Conclusion: Hemodialysis patients, especially those with C-reactive protein > 5 mg/L, responded to fish oil supplementation to reduce their C-reactive protein level; however, Interleukin 6 and tumor necrosis factor-alpha levels did not appear to be affected., Competing Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2024.)

Purpose: Strabismus (STBMS) is a multifactorial ocular disorder in children that leads to misalignment of the eyes. Insulin-like growth factor 1 ( IGF1 ) has been shown to be involved in the development of extraocular muscles and myopia; however, data are limited on the genetic associations of IGF1 with STBMS in Pakistan., Methods: Two hundred seventy-four STBMS cases and 272 unaffected controls were recruited, and their DNA was extracted. Two IGF1 single nucleotide polymorphisms, rs6214 and rs5742632, were genotyped using PCR-restriction fragment length polymorphism. Univariate logistic regression analysis was performed to determine the association of these single nucleotide polymorphisms with STBMS, and the results were adjusted for age and sex. In addition, 26 extraocular muscle tissues were collected from patients with STBMS undergoing squint correction surgery, along with 3 deceased control samples. IGF1 mRNA expression was measured by quantitative PCR; the Mann-Whitney U test was applied, and fold change was calculated. Logistic regression analysis was applied to determine the association of RNA expression and fold change with genotype., Results: Multivariate logistic regression analysis revealed that rs5742632 (odds ratio [95% confidence interval] = 1.05[1.01-1.06], p = 0.03) is associated with STBM. Moreover, rs6214 (1.03[1.01-1.05], p = 0.03) and rs5742632 (1.09[1.04-1.11], p = 0.04) were associated with exotropia. Statistically, no significant difference in IGF1 mRNA expression in the extraocular muscles between the STBMS cases and the controls was observed., Conclusions: IGF1 polymorphisms rs5742632 (A>G) and rs6214 (C>T) are plausible risk factors for the development of exotropia. However, the physiologic mechanism requires further evaluation., (Copyright © 2022 Molecular Vision.)

Background: Primary congenital glaucoma (PCG) is a heterogeneous rare recessively inherited disorder prevalent in regions with high consanguinity. Disease phenotype is associated with increased intra ocular pressure and is a major cause of childhood blindness. Sequence variations in Cytochrome P450 1B1 (CYP1B1) gene are a major cause of PCG. Current study was conducted to screen CYP1B1 gene in highly consanguineous PCG affected families from Pakistani population consistent with the autosomal recessive pattern of PCG inheritance., Methods: For this study, patients and controls (clinically unaffected individuals of each family) from 25 consanguineous families belonging to Punjab, Baluchistan and Khyber Pakhtunkhwa, Pakistan were recruited through ophthalmologists. DNA was isolated from collected blood samples. Genetic screening of CYP1B1 gene was done for all enrolled families. In-silico analysis was performed to identify and predict the potential disease-causing variations., Results: Pathogenicity screening revealed sequence variants segregating with disease phenotype in homozygous or compound heterozygous form in eleven out of 25 analyzed families. We identified a total of sixteen disease causing variants among which five frameshift i.e., c.629dup (p.Gly211Argfs*13), c.287dup (p.Leu97Alafs*127), c.662dup (p.Arg222Profs*2), c.758&#95;759insA (p.Val254Glyfs*73) and c.789dup (p.Leu264Alafs*63), two silent c.1314G>A, c.771T>G and six missense variations c.457C>G (p.Arg153Gly), c.516C>A (p.Ser172Arg), c.722T>A (p.Val241Glu), c.740T>A (p.Leu247Gln), c.1263T>A (p.Phe421Leu), and c.724G>C (p.Asp242His) are previously un reported. However two frameshift c.868dup (p.Arg290Profs*37), c.247del (p.Asp83Thrfs*12) and one missense variant c.732G>A (p.Met244Ile), is previously reported. Furthermore, six polymorphisms c.1347T>C, c.2244&#95;2245insT, c.355G>T, c.1294G>C, c.1358A>G and c.142C>G were also identified. In the intronic region, a novel silent polymorphism i.e., g.35710&#95;35711insT was found in homozygous state. All the newly detected disease-causing variants were negative in 96 ethnically matched controls., Conclusion: Among twenty-five screened families, eight families (PCG50, 52-54, 58, 59, 63 and 67) were segregating disease causing variants in recessive manner. Two families (PCG049 and PCG062) had compound heterozygosity. Our data confirms genetic heterogeneity of PCG in Pakistani population however we did not find molecular variants segregating with PCG in fifteen families in coding exons and intron-exon boundaries of CYP1B1 gene. Genetic counseling was provided to families to refrain from practicing consanguinity and perform premarital screening as a PCG control measure in upcoming generations., Competing Interests: The authors have declared that no competing interests exist.

Background: Congenital cataract is causing one-third of blindness worldwide. Congenital cataract is heterogeneous in its inheritance patterns. The current study is aimed to explore the unknown genetic causes underlying congenital cataracts., Methods: Blood samples from affected and normal individuals of n = 25 Pakistani families identified with congenital cataracts were collected. Genomic DNA was extracted and Sanger sequencing was performed to identify novel pathogenic variants in the FYCO1 (MIM#607182) gene. Later structural bioinformatics tools and molecular dynamics simulations were performed to analyze the impact of these variants on protein structure and function., Results: Sanger sequencing resulted in the identification of a novel splice site mutation (NM&#95;024513.3: c.3151-29&#95;3151-7del) segregating in an autosomal recessive manner. This novel variant was confirmed to be absent in the n = 300 population controls. Further, bioinformatics tools revealed the formation of a mutant protein with a loss of the Znf domain. In addition, we also found a previously known (c.4127 T > C; p.Leu1376Pro) mutation in four families. We also report a novel heterozygous variant (c.3419G > A; p.Arg1140Gln) in another family., Conclusions: In conclusion, we report a novel deletion (NM&#95;024513.3: c.3151-29&#95;3151-7del) in one family and a frequent homozygous missense mutation (c.4127 T > C; p.Leu1376Pro) in four Pakistani families. The current research highlights the importance of autophagy in lens development and maintaining its transparency., (© 2022 The Authors. Molecular Genetics & Genomic Medicine published by Wiley Periodicals LLC.)

Background: Off-label medication usage (OLMU) is prevalent in the treatment of various diseases, including female reproductive health issues (FRHIs). However, there is a paucity of literature on the perspective of health professionals on this subject. The purpose of the current study was to assess gynecologists/obstetricians' knowledge, attitude and practice toward OLMU in the treatment of FRHIs., Methods: The current cross-sectional study was conducted in September and October 2021, at five tertiary care hospitals (two public and three private sector), different clinics and maternity homes in a metropolitan city of Karachi, Pakistan. The target population was gynecologists, obstetricians and physicians/residents working in the ob/gyn department in various hospitals and clinical settings of Karachi., Results: The overall response rate was 77.1%. The mean age of the study respondents was 36.1 ± 7.7 years; n = 85 (55.9%) respondents were working in primary patient care. The majorly reported OLMU by the respondents were clomiphene citrate in unexplained infertility ( n = 66; 43.4%), metformin to improve cycle regularity in females with polycystic ovary syndrome (PCOS) ( n = 59; 38.8%) and letrozole to induce ovulation ( n = 31; 20.4%). The majorly stated categories of OLMU were at a different dose ( n = 95; 62.5%) and at different indications than approved to treat ( n = 89; 58.5%). It was reported by the majority of the respondents ( n = 95; 62.5%) that they do not follow any guidelines or regulations for OLMU in their work setting; however, the response was statistically varied with the working organization (CI 2.14-2.93; p = 0.037) and practice area (CI 2.85-4.32; p = 0.0001) of respondents., Conclusions: The present study revealed that the respondents were well-familiar with the practice of OLMU in the treatment of FRHIs. They expressed their concerns about decreasing such practices by being involved in collective decision-making procedures, and they were inclined to accept initiatives aimed at ensuring drug safety in patients., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer SI declared a past co-authorship with the author HR to the handling editor., (Copyright © 2022 Shakeel, Iffat, Qamar, Nesar, Butt, Siddiqui, Rehman and Rehman.)

This is a rare case of development of bi-lateral chylous pleural effusion (containing parenteral nutrition material) along with pneumomediastinum due to punctured left subclavian vein following insertion of a peripherally inserted central venous catheter (PICC) line. Parenteral nutrition is usually preferred for patients unable to tolerate enteral feeding. Due to hypertonicity of the total parenteral nutrition material, it is usually administered via internal jugular or subclavian vein which have a rapid blood flow; therefore, resulting in adequate mixing. Literature studies are yet to clearly explain the communications between two pleural cavities; therefore, development of bilateral pleural effusions in association with pneumomediastinum makes this case more intriguing. We present the journey of a 43-year-old woman who required insertion of bilateral chest drains, followed by sternotomy and repair of the left subclavian vein after she was found in hypoxic respiratory failure 2 days following insertion of PICC line into her left subclavian vein., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2021. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Purpose: Axenfeld-Rieger syndrome (ARS) is a rare autosomal dominant disorder that affects the anterior segment of the eye. The aim of this study was to examine the PITX2 gene to identify possible novel mutations in Pakistani and Mexican families affected by the ARS phenotype., Methods: Three unrelated probands with a diagnosis of ARS were recruited for this study. Genomic DNA was isolated from the peripheral blood of the probands and their family members. Polymerase chain reaction and Sanger sequencing were used for the analysis of coding exons and the flanking intronic regions of the PITX2 gene. Bioinformatics tools and database (VarSome, Provean, and MutationTaster, SIFT, PolyPhen-2, and HOPE) were evaluated to explore missense variants., Results: We identified novel heterozygous variations in the PITX2 gene that segregated with the ARS phenotype within the families. The variant NM&#95;153426.2(PITX2):c.226G > T or p.(Ala76Ser) and the mutation NM&#95;153426.2(PITX2):c.455G > A or p.(Cys152Tyr) were identified in two Pakistani pedigrees, and the mutation NM&#95;153426.2(PITX2):c.242&#95;265del or p.(Lys81&#95;Gln88del), segregated in a Mexican family., Conclusion: Our study extends the spectrum of PITX2 mutations in individuals with ARS, enabling an improved diagnosis of this rare but serious syndrome., (© 2020 The Authors. Molecular Genetics & Genomic Medicine published by Wiley Periodicals, Inc.)

Importance: Early diagnosis of retinoblastoma, the most common intraocular cancer, can save both a child's life and vision. However, anecdotal evidence suggests that many children across the world are diagnosed late. To our knowledge, the clinical presentation of retinoblastoma has never been assessed on a global scale., Objectives: To report the retinoblastoma stage at diagnosis in patients across the world during a single year, to investigate associations between clinical variables and national income level, and to investigate risk factors for advanced disease at diagnosis., Design, Setting, and Participants: A total of 278 retinoblastoma treatment centers were recruited from June 2017 through December 2018 to participate in a cross-sectional analysis of treatment-naive patients with retinoblastoma who were diagnosed in 2017., Main Outcomes and Measures: Age at presentation, proportion of familial history of retinoblastoma, and tumor stage and metastasis., Results: The cohort included 4351 new patients from 153 countries; the median age at diagnosis was 30.5 (interquartile range, 18.3-45.9) months, and 1976 patients (45.4%) were female. Most patients (n = 3685 [84.7%]) were from low- and middle-income countries (LMICs). Globally, the most common indication for referral was leukocoria (n = 2638 [62.8%]), followed by strabismus (n = 429 [10.2%]) and proptosis (n = 309 [7.4%]). Patients from high-income countries (HICs) were diagnosed at a median age of 14.1 months, with 656 of 666 (98.5%) patients having intraocular retinoblastoma and 2 (0.3%) having metastasis. Patients from low-income countries were diagnosed at a median age of 30.5 months, with 256 of 521 (49.1%) having extraocular retinoblastoma and 94 of 498 (18.9%) having metastasis. Lower national income level was associated with older presentation age, higher proportion of locally advanced disease and distant metastasis, and smaller proportion of familial history of retinoblastoma. Advanced disease at diagnosis was more common in LMICs even after adjusting for age (odds ratio for low-income countries vs upper-middle-income countries and HICs, 17.92 [95% CI, 12.94-24.80], and for lower-middle-income countries vs upper-middle-income countries and HICs, 5.74 [95% CI, 4.30-7.68])., Conclusions and Relevance: This study is estimated to have included more than half of all new retinoblastoma cases worldwide in 2017. Children from LMICs, where the main global retinoblastoma burden lies, presented at an older age with more advanced disease and demonstrated a smaller proportion of familial history of retinoblastoma, likely because many do not reach a childbearing age. Given that retinoblastoma is curable, these data are concerning and mandate intervention at national and international levels. Further studies are needed to investigate factors, other than age at presentation, that may be associated with advanced disease in LMICs.

Primary congenital glaucoma (PCG) causes blindness in early age. It has an autosomal recessive pattern of inheritance, hence is more prevalent in populations with frequent consanguineous marriages that occur in the Pakistani population. Mutations in the CYP1B1 gene are commonly associated with PCG. The aim of the present study was to identify genetic mutations in the CYP1B1 gene in PCG cases belonging to 38 Pakistani families. DNA was extracted using blood samples collected from all enrolled patients, their available unaffected family members and controls. Direct sequencing of the CYP1B1 gene revealed a novel 3' splice acceptor site causative variant segregating in an autosomal recessive manner in a large consanguineous family with four PCG-affected individuals. The novel variant was not detected in 93 ethnically matched controls. Furthermore, four already reported mutations, including p.G61E, p.R355X, p.R368H, and p.R390H were also detected in patients belonging to nine different families. All identified causative variants were evaluated by computational programs, that is, SIFT, PolyPhen-2, and MutationTaster. Pathogenicity of the novel splice site variant identified in this study was analyzed by Human Splicing Finder and MaxEntScan. Ten out of 38 families with PCG had the disease due to CYP1B1 mutations, suggesting CYP1B1 was contributing to PCG in these Pakistani patients. Identification of this novel 3' splice acceptor site variant in intron 2 is the first report for the CYP1B1 gene contributing to genetic heterogeneity of disease., (© 2018 Japanese Teratology Society.)

Purpose: Brittle cornea syndrome (BCS) is a rare recessive disorder affecting connective tissues, most prominently in the eye. Pathogenic mutations causing BCS have been identified in PRDM5 and ZNF469 genes. This study investigates the genetic cause of BCS in a large, consanguineous Pakistani family with 4 affected and 3 unaffected individuals., Methods: The coding region and exon-intron splice junctions of PRDM5 and ZNF469 genes were amplified by polymerase chain reaction, and bidirectional Sanger sequencing was performed to find the pathogenic change responsible for causing the disease in the family., Results: A novel homozygous duplication c.9831dupC (p.Arg3278GlnfsX197) in the ZNF469 gene was identified, which was found to be co-segregating with the disease in the family., Conclusions: This is the first report of a ZNF469 homozygous mutation causing a BCS phenotype in a consanguineous Pakistani family. Our data extend the mutation spectrum of ZNF469 variants implicated in BCS.

Background: Depression in congestive heart failure (CHF) patients can increase morbidity and mortality. Given the ever-rising prevalence of CHF patients with depression, it is vital that we understand the predictors of depression in these patients to identify and better manage these patients. The main objective of this study was to evaluate the frequency and predictors of depression in CHF patients., Methods: A cross-sectional study was conducted in a tertiary care hospital. Patients with a diagnosis of CHF for more than 6 months based on signs and left ventricular ejection fraction <40% were included. Patients were interviewed with the Patient Health Questionnaire-9 (PHQ-9) consisting of nine items in line with the Diagnostic and Statistical Manual (DSM) - IV criteria to assess depression. Each item was scored from 0 to 3, and a PHQ-9 score of 10 or greater suggested clinical depression. Data were analyzed on SPSS, v22, and a p < 0.05 was considered significant., Results: Of 170 participants, 102 (60%) had depression. Among these 102 patients, 42% (n = 43) had mild depression, and the rest (n = 59) had moderate-to-severe depression. Predictors of depression were New York Heart Association stage 3 or 4 (p = 0.001), previous myocardial infarction (p = 0.001), living without a partner (p = 0.001), lack of a joint family system (p = 0.001), sedentary lifestyle (p = 0.001), aged 70 years or more (p = 0.01), and having been admitted in a hospital at least once in the past two months (p = 0.002)., Conclusion: Depression is common among patients with CHF. It is associated with multiple factors and needs to be addressed and targeted urgently., (Copyright © 2018 Cardiological Society of India. Published by Elsevier B.V. All rights reserved.)

Glaucoma is the cause of irreversible blindness worldwide. Mutations in six genes have been associated with juvenile- and adult-onset familial primary open angle glaucoma (POAG) prior to this report but they explain only a small proportion of the genetic load. The aim of the study is to identify the novel genetic cause of the POAG in the families with adult-onset glaucoma. Whole exome sequencing (WES) was performed on DNA of two affected individuals, and predicted pathogenic variants were evaluated for segregation in four affected and three unaffected Dutch family members by Sanger sequencing. We identified a pathogenic variant (p.Val956Gly) in the PRPF8 gene, which segregates with the disease in Dutch family. Targeted Sanger sequencing of PRPF8 in a panel of 40 POAG families (18 Pakistani and 22 Dutch) revealed two additional nonsynonymous variants (p.Pro13Leu and p.Met25Thr), which segregate with the disease in two other Pakistani families. Both variants were then analyzed in a case-control cohort consisting of Pakistani 320 POAG cases and 250 matched controls. The p.Pro13Leu and p.Met25Thr variants were identified in 14 and 20 cases, respectively, while they were not detected in controls (p values 0.0004 and 0.0001, respectively). Previously, PRPF8 mutations have been associated with autosomal dominant retinitis pigmentosa (RP). The PRPF8 variants associated with POAG are located at the N-terminus, while all RP-associated mutations cluster at the C-terminus, dictating a clear genotype-phenotype correlation.

Objective: To evaluate the state of refraction in children with bilateral congenital nasolacrimal duct obstruction (CNLDO)., Study Design: Descriptive study., Place and Duration of Study: Al-Shifa Trust Eye Hospital, Rawalpindi, Pakistan, from April 2014 to April 2016., Methodology: Children with bilateral CNLDO were studied. Patients' refractive status were evaluated by performing cycloplegic refraction, followed by appropriate management plan. The refractive errors of both eyes were noted and compared for any significant anisometropia., Results: One hundred and seventeen (n=117) children with median age (IQR) of 32 (12) months having bilateral CNLDO were enrolled. Children with bilateral CNLDO had insignificant interocular difference in terms of spherical equivalent (SE) and cylindrical refractive errors (p>0.05). The rate of the anisometropia (>1 D difference between the two eyes) was 5.98% (n=7) in children with bilateral CNLDO., Conclusion: Performing cycloplegic refraction routinely in patients with bilateral CNLDO is not as urgent as compared to ones with unilateral CNLDO. Further, avoidance of early surgical intervention in children with bilateral CNLDO will spare the parents from the emotional trauma and positively influence the health economics worldwide.

Congenital cataract is a clinically and genetically heterogeneous disease. The present study was undertaken to find the genetic cause of congenital cataract families. DNA samples of a large consanguineous Pakistani family were genotyped with a high resolution single nucleotide polymorphism Illumina microarray. Homozygosity mapping identified a homozygous region of 4.4 Mb encompassing the gene GJA3 . Sanger sequence analysis of the GJA3 gene revealed a novel homozygous variant c.950dup p.(His318ProfsX8) segregating in an autosomal recessive (AR) manner. The previously known mode of inheritance for GJA3 gene mutations in cataract was autosomal dominant (AD) only. The screening of additional probands ( n = 41) of cataract families revealed a previously known mutation c.56C>T p.(Thr19Met) in GJA3 gene. In addition, sequencing of the exon-intron boundaries of the GJA8 gene in 41 cataract probands revealed two additional mutations: a novel c.53C>T p.(Ser18Phe) and a known c.175C>G p.(Pro59Ala) mutation, both co-segregating with the disease phenotype in an AD manner. All these mutations are predicted to be pathogenic by in silico analysis and were absent in the control databases. In conclusion, results of the current study enhance our understanding of the genetic basis of cataract, and identified the involvement of the GJA3 in the disease etiology in both AR and AD manners.

Objective: The objective of the study was to evaluate the refractive status and thereby assess anisometropia in children with unilateral congenital nasolacrimal duct obstruction (CNLDO)., Study Design: This study design was a descriptive cross-sectional study., Place and Duration: this study was conducted at the Department of Pediatric Ophthalmology and Strabismology, Al-Shifa Trust Eye Hospital, Rawalpindi; from August 2013 to July 2014., Methodology: This study assessed consecutive children with unilateral CNLDO. Cycloplegic refraction on all children with CNLDO was performed followed by appropriate intervention. Refractive errors of the affected and normal eyes were compared., Results: One hundred and twenty-four children with a mean age of 29.69 ± 21.12 months (range, 2 months to 8 years) were studied. Based on spherical equivalent (SE), hypermetropia was more common in the affected eyes ( P < 0.001). Anisometropia of >1.5 diopters (D) was present in n = 17 (13.7%). Interocular difference was significant for spherical error and SE ( P < 0.001) but not cylindrical errors., Conclusion: Unilateral CNLDO is associated with statistically significant anisometropia, especially anisohypermetropia which has amblyogenic potential. It is vital to perform cycloplegic refraction routinely and counsel parents regarding prognosis and regular follow-ups., Competing Interests: The authors declare that there are no conflicts of interests of this paper.

Objective: To observe the types of tumor regression after treatment, and identify the common pattern of regression in our patients., Study Design: Descriptive study., Place and Duration of Study: Department of Pediatric Ophthalmology and Strabismus, Al-Shifa Trust Eye Hospital, Rawalpindi, Pakistan, from October 2011 to October 2014., Methodology: Children with unilateral and bilateral retinoblastoma were included in the study. Patients were referred to Pakistan Institute of Medical Sciences, Islamabad, for chemotherapy. After every cycle of chemotherapy, dilated fundus examination under anesthesia was performed to record response of the treatment. Regression patterns were recorded on RetCam II., Results: Seventy-four tumors were included in the study. Out of 74 tumors, 3 were ICRB group A tumors, 43 were ICRB group B tumors, 14 tumors belonged to ICRB group C, and remaining 14 were ICRB group D tumors. Type IV regression was seen in 39.1% (n=29) tumors, type II in 29.7% (n=22), type III in 25.6% (n=19), and type I in 5.4% (n=4). All group A tumors (100%) showed type IV regression. Seventeen (39.5%) group B tumors showed type IV regression. In group C, 5 tumors (35.7%) showed type II regression and 5 tumors (35.7%) showed type IV regression. In group D, 6 tumors (42.9%) regressed to type II non-calcified remnants., Conclusion: The response and success of the focal and systemic treatment, as judged by the appearance of different patterns of tumor regression, varies with the ICRB grouping of the tumor.

Background: Anterior segment dysgenesis (ASD) disorders are a group of clinically and genetically heterogeneous phenotypes in which frequently cornea, iris, and lens are affected. This study aimed to identify novel mutations in PAX6, PITX2 and FOXC1 in families with anterior segment dysgenesis disorders., Methods: We studied 14 Pakistani and one Mexican family with Axenfeld Rieger syndrome (ARS; n = 10) or aniridia (n = 5). All affected and unaffected family members underwent full ophthalmologic and general examinations. Total genomic DNA was isolated from peripheral blood. PCR and Sanger sequencing were performed for the exons and intron-exon boundaries of the FOXC1, PAX6, and PITX2 genes., Results: Mutations were identified in five of the 15 probands; four variants were novel and one variant was described previously. A novel de novo variant (c.225C>A; p.Tyr75*) was identified in the PAX6 gene in two unrelated probands with aniridia. In addition, a known variant (c.649C>T; p.Arg217*) in PAX6 segregated in a family with aniridia. In the FOXC1 gene, a novel heterozygous variant (c.454T>C; p.Trp152Arg) segregated with the disease in a Mexican family with ARS. A novel homozygous variant (c.92&#95;100del; p.Ala31&#95;Ala33del) in the FOXC1 gene segregated in a Pakistani family with ARS and congenital glaucoma., Conclusions: Our study expands the mutation spectrum of the PAX6 and FOXC1 genes in individuals with anterior segment dysgenesis disorders. In addition, our study suggests that FOXC1 mutations, besides typical autosomal dominant ARS, can also cause ARS with congenital glaucoma through an autosomal recessive inheritance pattern. Our results thus expand the disease spectrum of FOXC1, and may lead to a better understanding of the role of FOXC1 in development.

Background: Primary congenital glaucoma (PCG) is the most common form of glaucoma in children. PCG occurs due to the developmental defects in the trabecular meshwork and anterior chamber of the eye. The purpose of this study is to identify the causative genetic variants in three families with developmental and primary congenital glaucoma (PCG) with a recessive inheritance pattern., Methods: DNA samples were obtained from consanguineous families of Pakistani ancestry. The CYP1B1 gene was sequenced in the affected probands by conventional Sanger DNA sequencing. Whole exome sequencing (WES) was performed in DNA samples of four individuals belonging to three different CYP1B1-negative families. Variants identified by WES were validated by Sanger sequencing., Results: WES identified potentially causative novel mutations in the latent transforming growth factor beta binding protein 2 (LTBP2) gene in two PCG families. In the first family a novel missense mutation (c.4934G>A; p.Arg1645Glu) co-segregates with the disease phenotype, and in the second family a novel frameshift mutation (c.4031&#95;4032insA; p.Asp1345Glyfs*6) was identified. In a third family with developmental glaucoma a novel mutation (c.3496G>A; p.Gly1166Arg) was identified in the PXDN gene, which segregates with the disease., Conclusions: We identified three novel mutations in glaucoma families using WES; two in the LTBP2 gene and one in the PXDN gene. The results will not only enhance our current understanding of the genetic basis of glaucoma, but may also contribute to a better understanding of the diverse phenotypic consequences caused by mutations in these genes.

Methicillin-resistant Staphylococcus aureus is one of the toughest organisms to treat, especially in cases where intraocular surgery is contemplated, because the risks are aggravated. Conjunctival swab culture and sensitivity tests are significant when there is history of recent hospitalization. In this report, an infant with successful cataract surgery after elimination of the organism is presented., (Copyright 2016, SLACK Incorporated.)

Purpose: To evaluate and compare the refractive state in children diagnosed as having unilateral or bilateral congenital nasolacrimal duct obstruction (CNLDO). This study also compares how the laterality of CNLDO affects the refractive state of the patients., Methods: This descriptive cross-sectional study includes consecutive children with unilateral and bilateral CNLDO over a period of 1 year. Cycloplegic refraction was performed on each child who presented with CNLDO followed by appropriate plans for treatment. The refractive errors of patients with unilateral and bilateral CNLDO were compared., Results: One hundred sixty-one patients with unilateral CNLDO (mean age: 29 ± 19.93 months) and 46 with bilateral CNLDO (mean age: 30 ± 16.21 months) were enrolled from August 2013 to July 2014. The rate of the anisometropia (≥ 1 diopters [D] difference between the two eyes) was 13.7% (n = 22) and 8.6% (n = 4) in patients with unilateral and bilateral CNLDO, respectively. Interocular difference was significant in terms of spherical equivalent (P < .01) but not cylindrical refractive error in patients with unilateral CNLDO. Patients with bilateral CNLDO had insignificant interocular differences in terms of spherical equivalent and cylindrical refractive errors (P > .05)., Conclusions: Unilateral CNLDO is associated with statistically significant anisometropia compared to bilateral CNLDO, which predisposes children with unilateral CNLDO to amblyopia. It is vital to perform cycloplegic refraction routinely and counsel parents regarding regular follow-ups. [J Pediatr Ophthalmol Strabismus. 2016;53(3):168-172.]., (Copyright 2016, SLACK Incorporated.)

Axenfeld-Rieger syndrome (ARS) is a disorder affecting the anterior segment of the eye, often leading to secondary glaucoma and several systemic malformations. It is inherited in an autosomal dominant fashion that has been associated with genetic defects in PITX2 and FOXC1. Known genes CYP1b1, PITX2, and FOXC1 were excluded by Sanger sequencing. The purpose of current study is to identify the underlying genetic causes in ARS family by whole exome sequencing (WES). WES was performed for affected proband of family, and variants were prioritized based on in silico analyses. Segregation analysis of candidate variants was performed in family members. A novel heterozygous PRDM5 missense variant (c.877A>G; p.Lys293Glu) was found to segregate with the disease in an autosomal dominant fashion. The novel missense variant was absent from population-matched controls, the Exome Variant Server, and an in-house exome variant database. The Lys293Glu variant is predicted to be pathogenic and affects a lysine residue that is conserved in different species. Variants in the PRDM5 gene were previously identified in anterior segment defects, i.e., autosomal recessive brittle cornea syndrome and keratoconus. The results of this study suggest that genetic variants in PRDM5 can lead to various syndromic and nonsyndromic disorders affecting the anterior segment of the eye.

Background: Recently nonsynonymous coding variants in the ankyrin repeats and suppressor of cytokine signaling box-containing protein 10 (ASB10) gene were found to be associated with primary open angle glaucoma (POAG) in cohorts from Oregon and Germany, but this finding was not confirmed in an independent cohort from Iowa. The aim of the current study was to assess the role of ASB10 gene variants in Pakistani glaucoma patients., Methods: Sanger sequencing of the coding exons and splice junctions of the ASB10 gene was performed in 30 probands of multiplex POAG families, 208 sporadic POAG patients and 151 healthy controls from Pakistan. Genotypic associations of individual variants with POAG were analyzed with the Fisher's exact or Chi-square test., Results: In total 24 variants were identified in POAG probands and sporadic patients, including 11 novel variants and 13 known variants. 13 of the variants were nonsynonymous, 6 were synonymous, and 5 were intronic. Three nonsynonymous variants (p.Arg49Cys, p.Arg237Gly, p.Arg453Cys) identified in the probands were not segregating in the respective families. This is not surprising since glaucoma is a multifactorial disease, and multiple factors are likely to be involved in the disease manifestation in these families. However a nonsynonymous variant, p.Arg453Cys (rs3800791), was found in 6 sporadic POAG patients but not in controls, suggesting that it infers increased risk for the disease. In addition, one synonymous variant was found to be associated with sporadic POAG: p.Ala290Ala and the association of the variant with POAG remained significant after correction for multiple testing (uncorrected p-value 0.002, corrected p-value 0.047). The cumulative burden of rare, nonsynonymous variants was significantly higher in sporadic POAG patients compared to control individuals (p-value 0.000006)., Conclusions: Variants in ASB10 were found to be significantly associated with sporadic POAG in the Pakistani population. This supports previous findings that sequence variants in the ASB10 gene may act as a risk factor for glaucoma.

Purpose: Photoreceptor neuronal degenerations are common, incurable causes of human blindness affecting 1 in 2000 patients worldwide. Only half of all patients are associated with known mutations in over 250 disease genes, prompting our research program to identify the remaining new genes. Most retinal degenerations are restricted to the retina, but photoreceptor degenerations can also be found in a wide variety of systemic diseases. We identified an X-linked family from Sri Lanka with a severe choroidal degeneration and postulated a new disease entity. Because of phenotypic overlaps with Bietti's crystalline dystrophy, which was recently found to have systemic features, we hypothesized that a systemic disease may be present in this new disease as well., Methods: For phenotyping, we performed detailed eye exams with in vivo retinal imaging by optical coherence tomography. For genotyping, we performed whole exome sequencing, followed by Sanger sequencing confirmations and cosegregation. Systemic investigations included electron microscopy studies of peripheral blood cells in patients and in normal controls and detailed fatty acid profiles (both plasma and red blood cell [RBC] membranes). Fatty acid levels were compared to normal controls, and only values two standard deviations above or below normal controls were further evaluated., Results: The family segregated a REP1 mutation, suggesting choroideremia (CHM). We then found crystals in peripheral blood lymphocytes and discovered significant plasma fatty acid abnormalities and RBC membrane abnormalities (i.e., elevated plasmalogens). To replicate our discoveries, we expanded the cohort to nine CHM patients, genotyped them for REP1 mutations, and found the same abnormalities (crystals and fatty acid abnormalities) in all patients., Conclusions: Previously, CHM was thought to be restricted to the retina. We show, to our knowledge for the first time, that CHM is a systemic condition with prominent crystals in lymphocytes and significant fatty acid abnormalities.

Objective: To evaluate the mean changes in Central Corneal Thickness (CCT) and Endothelial Cell Count (ECC) in eyes after pediatric cataract surgery with foldable intraocular lens using scleral tunnel incision micro-surgical technique., Study Design: Qausi experimental study., Place and Duration of Study: Department of Pediatric Ophthalmology and Strabismus, Al-Shifa Trust Eye Hospital, Rawalpindi, from May 2011 to March 2012., Methodology: Fifty-two eyes of 37 children with pediatric cataract were included in the study. Extracapsular Cataract Extraction (ECE) with foldable Intra Ocular Lens (IOL) implantation using sclera tunnel incision was performed in all children. Endothelial Cell Count (ECC) and Central Corneal Thickness (CCT) were recorded before surgery and 1 month, 3 months and 6 months after surgery and the effect of currently practiced surgical technique on ECC and CCT was evaluated., Results: The mean age at the time of surgery was 8.8 ±2.7 years (range: 4 to 15 years). The postoperative ECC and CCT were significantly different from the pre-operative values. Mean pre-operative ECC was 3175.3 ±218.4 cell/mm2 and in first postoperative month the mean ECC was 3113.4 ±210.8 cell/mm2(p<0.0001). In the 3rd and 6th month postoperative means ECC were 3052 ±202.5 cell/mm2(p<0.0001) and 3015 ±190.6 cell/mm2(p<0.0001), respectively. The mean cell loss at first postoperative month was 1.95% and at 3rd and 6th postoperative month were 3.9% and 5.05%, respectively. Mean pre-operative CCT was 514 ±49.9 µm and first postoperative mean CCT after 1 month was 524.1 ±25 µm (p = 0.084). After the 3rd and 6th months postoperative, mean CCT were 527.3 ±24.6 µm, and 530 ±24.5 µm, respectively. Third and 6th months postoperative means were significantly higher than baseline CCT, p = 0.024 and 0.007, respectively., Conclusion: Endothelial cell loss with closed chamber micro-surgical technique using scleral tunnel incision is within acceptable limits and within the range of normal ECC in children.

Fraser's Syndrome (FS) is a rare autosomal recessive disorder with a spectrum of malformations. The most consistent features are Cryptophthalmos (CO), syndactyly, genitourinary tract abnormalities, laryngeal and tracheal anomalies, craniofacial dysmorphism, malformations of the ear and nose, orofacial clefting and musculoskeletal defects. FS is genetically heterogeneous; so far mutations in FRAS1, FREM2 and GRIP1 genes have been linked to FS. FS can be diagnosed on clinical examination, pre-natal ultrasound or perinatal autopsy. We present a case of a 3 months old child born to consanguineous healthy parents with bilateral complete CO, unilateral microphthalmia, hypertelorism, syndactyly (hands and feet bilaterally), ambiguous genitalia with cryptorchidism and an umbilical hernia. We also present the criteria for diagnosing FS and the significant features on pre-natal ultrasonography. Around 200 case reports of patients with FS and CO have been published. To our knowledge, this is the first reported case of FS in Pakistan.

This study presents a comparative account of biochemical responses evoked by a high-zinc-accumulating (HZnG) and a low-zinc-accumulating (LZnG) genotype of chickpea (Cicer arietinum L.) under conditions of zinc deficiency (0 mg ZnSO4 kg(-1) soil) and on exogenous zinc supply (2.5, 5, and 10 mg ZnSO4 kg(-1) soil), at different growth stages (pre-flowering, flowering, and post-flowering) of the crop. Both the genotypes exhibited significant variation in contents of chlorophyll, protein, nitrate, and leghemoglobin, and in the activity of enzymes (nitrate reductase, superoxide dismutase, and carbonic anhydrase), under zinc-limiting condition. The HZnG maintained a significantly higher level of biochemical parameters at deficient zinc levels, while LZnG was more affected by Zn deficiency (with reduced biochemical attributes), compared to HZnG. However, application of soil zinc alleviated the Zn-induced disturbance and improved the biochemical parameters in both genotypes. The differential biochemical response of plants to Zn deficiency indicated existence of genotypic variation in efficient utilization of the available Zn. The HZnG was more effective in overcoming Zn deficiency than the LZnG. This study suggests that metal-dependent biochemical parameters can be used as reliable indices for selection of genotypes with a better Zn utilization capacity, i.e., the Zn-efficient genotypes.

Background: Homozygosity mapping has facilitated the identification of the genetic causes underlying inherited diseases, particularly in consanguineous families with multiple affected individuals. This knowledge has also resulted in a mutation dataset that can be used in a cost and time effective manner to screen frequent population-specific genetic variations associated with diseases such as inherited retinal disease (IRD)., Methods: We genetically screened 13 families from a cohort of 81 Pakistani IRD families diagnosed with Leber congenital amaurosis (LCA), retinitis pigmentosa (RP), congenital stationary night blindness (CSNB), or cone dystrophy (CD). We employed genome-wide single nucleotide polymorphism (SNP) array analysis to identify homozygous regions shared by affected individuals and performed Sanger sequencing of IRD-associated genes located in the sizeable homozygous regions. In addition, based on population specific mutation data we performed targeted Sanger sequencing (TSS) of frequent variants in AIPL1, CEP290, CRB1, GUCY2D, LCA5, RPGRIP1 and TULP1, in probands from 28 LCA families., Results: Homozygosity mapping and Sanger sequencing of IRD-associated genes revealed the underlying mutations in 10 families. TSS revealed causative variants in three families. In these 13 families four novel mutations were identified in CNGA1, CNGB1, GUCY2D, and RPGRIP1., Conclusions: Homozygosity mapping and TSS revealed the underlying genetic cause in 13 IRD families, which is useful for genetic counseling as well as therapeutic interventions that are likely to become available in the near future.

Objective: To determine the clinical manifestations and results of current treatment for patients with retinoblastoma (Rb) in a tertiary care eye hospital in the north west of Pakistan., Study Design: Case series., Place and Duration of Study: Al-Shifa Trust Eye Hospital, Rawalpindi, Pakistan, from January 2006 and December 2009., Methodology: The data of 139 patients diagnosed as having retinoblastoma was collected. Gender, age at diagnosis, laterality, presenting sign, classification of tumour, treatment modality and outcome were noted., Results: The mean age of presentation in this patients ranged from 6 to 50 months (mean: 24.05 ± 10.74 months). The most common presenting sign was leucocoria in 78 eyes (44.1%). One hundred and one (72.7%) patients had unilateral retinoblastoma. Using the International Classification of Retinoblastoma (ICRB), 135 (76.3%) eyes were placed in group-E. one hundred and twenty four (77.5%) eyes were enucleated or exenterated while globe preservation was achieved by chemoreduction and/or focal therapy in the rest of the treated eyes (n = 36, 22.5%). Twenty three (16.5%) cases were lost to follow-up before one year. Ninety two (66.2%) patients survived, being free of tumour, at least one year after the completion of treatment., Conclusion: Most children with Rb showed an advanced stage of tumour at the time of diagnosis. Measures to improve the rate of globe preservation and patient survival by early diagnosis and intervention are the need of the hour.

Purpose: To evaluate the visual outcome and corneal endothelial cell density after Artisan aphakic intraocular lens (IOL) implantation (Ophtec, Groningen, the Netherlands) in pediatric eyes with subluxated lenses., Methods: Artisan aphakic IOLs were implanted in 18 eyes of 11 children with subluxated lenses. Idiopathic subluxations and ectopia lentis due to Marfan syndrome were included, whereas subluxations due to trauma or buphthalmos were excluded. Best-corrected visual acuity (BCVA) and endothelial cell density were monitored. Mean postoperative BCVA and endothelial cell density at last follow-up visit were calculated., Results: The age of children at the time of Artisan aphakic IOL implantation ranged from 8 to 16 years (mean: 11.58 ± 2.9 years). Mean follow-up was 9.12 ± 4.30 months. Mean postoperative logarithm of the minimum angle of resolution BCVA was 0.26 ± 0.13 (P = .001) and mean postoperative endothelial cell density was 2,860 ± 435 cells/mm(2) (P = .000). Mean endothelial cell loss was 17.1%., Conclusion: Artisan aphakic IOL implantation is a safe surgical choice in the management of ectopia lentis in the pediatric age group. It has minimal complications and is less traumatic to pediatric eyes. However, long-term follow-up of these children is required.[J Pediatr Ophthalmol Strabismus 2013;50(3):178-182.]., (Copyright 2013, SLACK Incorporated.)

Purpose: To study effects of Artisan iris fixated intraocular lens (IOL) on central corneal thickness (CCT) and intraocular pressure (IOP) in pediatric eyes with crystalline subluxated lenses., Materials and Methods: The study included 17 eyes undergoing Artisan aphakic IOL implantation after lensectomy for subluxated crystalline lenses. CCT and IOP measurements were recorded pre-operatively and post-operatively taking the mean of 4 post-operative visits. Patients were divided into Group A (n = 8) including patients with lensectomy and iris fixation of Artisan IOL as a primary procedure and Group B (n = 9) including patients in which lensectomy was carried out as a primary surgery and Artisan IOL fixation as a secondary procedure., Results: Children ranged in age from 08 years to 16 years, mean 11.59 ± 2.96 years. Follow-up period ranged from 7 months to 16 months, mean 11.24 months ± 4.27. Mean pre-operative and post-operative IOP in Group A was 14.88 ± 2.80 and 14.16 ± 0.59 respectively (P = 0.528). In Group B it was 12.44 ± 2.79 and 14.44 ± 1.15 respectively (P = 0.080). Mean pre-operative and post-operative CCT in Group A was 529.13 ± 24.23 and 529.87 ± 17.46 respectively (P = 0.674). In Group B it was 567.33 ± 29.13 and 568.83 ± 25.69 respectively (P = 0.859)., Conclusions: Primary and secondary Artisan aphakic IOL implantation did not cause any significant changes in corneal thickness or IOP during the follow-up period.

Objective: To evaluate the outcome of nasolacrimal intubation as a primary treatment of congenital nasolacrimal duct obstruction (NLDO) in children up to 4 years of age., Methods: During the 3 years period from July 2008 to June 2011, in the Paediatric Ophthalmology Department. Alshifa Trust Eye Hospital, Rawalpindi, 65 eyes of 59 children, aged 12 to 48 months with congenital NLDO and no prior Nasolacrimal duct surgery were enrolled. After written informed consent, all nasolacrimal intubations using olive tip silicon tube were performed under general anaesthesia. The planned tube retention was at least 3 months. The study outcome visit was timed 1 month after tube removal and treatment success was analyzed. Intra operative and post operative complications were also noted. Data were analyzed by SPSS 16. Frequencies and percentages were calculated for categorical variables. Mean +/- SD were computed for age and duration of intubation. Chi-square test was used to compare proportion of outcomes in different age groups and duration., Results: The overall success rate of Nasolacrimal intubation as a primary treatment of congenital NLDO was 89% in children between 12 to 48 months age (mean 25.8 +/- 9.8 months). The success was 92% in children under 2 years of age (P < 0.0001) and 90% in children between 2-3 years of age (P < 0.0001). The procedure remained less successful in children between 3-4 years of age (P < 0.2860) as compared to children under 3 years of age. The success rate was consistently high (92.3%) when the tube was left in situ for more than 3-6 months (P < 0.0001)., Conclusion: Nasolacrimal intubation with Olive tip silicon intubation tube is a successful procedure as a primary treatment of Nasolacrimal duct obstruction in children under 4 years of age.