Prof G Kumaramanickavel, MD Deputy Director of Research and Head Department of Genetics & Molecular Biology Vision Research Foundation Sankara Nethralaya, Chennai, India.
gkumarmvel@gmail.com
Summary of the scientific contributions:
Dr. G. Kumaramanickavel, has worked extensively in ophthalmic genomics for more than two decades and has contributed significantly to the progress in the area of ocular genetics both nationally and internationally. He had worked on both single gene (retinoblastoma, retinitis pigmentosa) and complex disorders (diabetic retinopathy and glaucoma), particularly in the area of gene mapping and was successful in identifying ocular genes, contributing significantly to the Human Genome Project, a global initiative.
He has intensively worked on the genetics of retinal dystrophies with the special mention to autosomal recessive childhood onset severe retinal dystrophy. He along with his team did a genome wide scan and identified the causative gene RPE65 and subsequently he was able to detect 5 pathogenic novel mutations in the gene. He detected a novel missense rhodopsin mutation in a family with recessive retinitis pigmentosa; this is of significance because this is the second report of its kind in the world literature.
Other than family linkage work on retinal dystrophies the he had worked on linkage of Oguchi disease and blepharophimosis (abnormal eyelid development disorder). He is currently involved in the gene mapping of complex disorders like glaucoma and diabetic retinopathy which is the major cause of adult blindness in India.
He had put inputs in the molecular pathology of diabetic retinopathy in which he found significant association of the disease with single nucleotide polymorphisms and the number of the microsatellite repeats in the candidate genes. His studies revealed that Tumor necrosis factor alpha and Ser82 allele in exon 3 of the Receptor for Advanced Glycation End product gene (RAGE) are the low risk alleles & Z-2 aldose reductase and iNOS (inducible nitric oxide synthase gene) are the high-risk alleles in diabetic retinopathy individuals in India.
He is working on the molecular genetics of various types of glaucoma (primary open angle glaucoma, angle closure glaucoma, normal tension glaucoma, congenital glaucoma, juvenile open angle glaucoma) and he and his group have identified novel mutation in the myocilin gene, which is the candidate gene for primary open angle glaucoma and also had reported the association of GSTM1 polymorphism with normal tension glaucoma; currently he is focused on gene mapping of angle closure glaucoma.
He had done fundamental work on the genetic causes of retinoblastoma (tumor of the retina and this disease occurs in the pediatric age group) like mutations, cytogenetic deletions, DNA methylation of RB1 promoter (epigenetic mechanism), loss of heterozygosity. He has initiated the process of establishing a diagnostic model for this disease, which would result in efficient molecular diagnosis and early detection of the disease, to assist clinical management in India.
His future objectives are formulating gene therapy for untreatable ocular diseases and to establish an ideal molecular diagnostic laboratory for genetic ocular diseases in the country.
Abstract: PURPOSE: In the Icelandic and Swedish populations, pseudoexfoliation syndrome (XFS) and pseudoexfoliation glaucoma (XFG) has been significantly associated with LOXL1 exon 1 polymorphisms - allele G of rs1048661 (R141L) and allele G of rs3825942 (G135D). In this study, we looked at the association of rs1048661 and rs3825942 in a southern Indian population. METHODS: Fifty-two cases with XFS (including XFG) and 97 matched controls that had thorough glaucoma evaluations were included in the study. Exon 1 of the LOXL1 gene with the single nucleotide polymorphisms (SNPs) were amplified and sequenced. For statistical significance, Pearson's Chi(2) test was performed. The HAPLOVIEW program v4.0 was used to determine the Hardy-Weinberg equilibrium and haplotype association. RESULTS: In our study population, there was a significant association of allele G of rs3825942 with XFS (p=0.0001) and genotype GG (p=0.000305) with XFS. CONCLUSIONS: Out of the two non-synonymous SNPs in exon 1 of the LOXL1 gene, rs3825942 has a significant association with XFS cases in the patients of the southern Indian population. To the best of our knowledge, this is the first Asian study replicating the European studies.
Abstract: PURPOSE: Leber congenital amaurosis (LCA) is one of the most common causes of hereditary blindness in infants. To date, mutations in 13 known genes and at two other loci have been implicated in LCA causation. An examination of the known genes highlights several processes which, when defective, cause LCA, including photoreceptor development and maintenance, phototransduction, vitamin A metabolism, and protein trafficking. In addition, it has been known for some time that defects in sensory cilia can cause syndromes involving hereditary blindness. More recently evidence has come to light that non-syndromic LCA can also be a "ciliopathy." METHODS: Here we present a homozygosity mapping analysis in a consanguineous sibship that led to the identification of a mutation in the recently discovered LCA5 gene. Homozygosity mapping was done using Affymetrix 10K Xba I Gene Chip and a 24.5cM region on chromosome 6 (6q12- q16.3) was identified to be significantly homozygous. The LCA5 gene on this region was sequenced and cDNA sequencing also done to characterize the mutation. RESULTS: A c.955G>A missense mutation in the last base of exon 6 causing disruption of the splice donor site was identified in both the affected sibs. Since there is a second consensus splice donor sequence 5 bp into the adjacent intron, this mutation results in a transcript with a 5 bp insertion of intronic sequence, leading to a frameshift and premature truncation. CONCLUSIONS: We report a missense mutation functionally altering the splice donor site and leading to a truncated protein. This is the second report of LCA5 mutations causing LCA. It may also be significant that one affected child died at eleven months of age due to asphyxia during sleep. To date the only phenotype unambiguously associated with mutations in this gene is LCA. However the LCA5 gene is known to be expressed in nasopharynx, trachea and lungs and was originally identified in the proteome of bronchial epithelium ciliary axonemes. The cause of death in this child may therefore imply that LCA5 mutations can in fact cause a wider spectrum of phenotypes including respiratory disease.
Abstract: BACKGROUND: Polymorphisms in vascular endothelial growth factor (VEGF) gene have been associated with diabetic retinopathy (DR) in various populations. A promoter polymorphism and a 3'UTR variation are studied for association with DR. MATERIALS AND METHODS: Type 2 diabetic patients with and without retinopathy were recruited. The -634C/G and 936C/T polymorphisms were genotyped by direct sequencing and their frequencies were analyzed using relevant statistical tests. RESULTS: No significant association was observed between genotypes, alleles and haplotypes of -634C/G and 936C/T polymorphisms and DR or its severity. However, C(-634)G genotype was found to increase the risk for DR in patients with microalbuminuria (OR: 8.9, 95% CI: 1.4, 58.3). CONCLUSION: Our study broadly suggests lack of association of VEGF gene polymorphisms with DR.
Abstract: Purpose: To compare the prevalence of refractive errors and factors associated with spectacle use in a rural and urban south Indian population. Materials and Methods: Four thousand eight hundred subjects (age> 39 years) each from rural and urban Tamil Nadu were enumerated for a population-based study. All participants underwent a complete ophthalmic evaluation including best-corrected visual acuity (BCVA), objective and subjective refraction. Out of 3924 rural responders 63.91% and out of 3850 urban responders 81.64% were phakic in the right eye with BCVA of 20/40 or better and were included in the study. Association of spectacle use and refractive errors with different parameters were analysed using logistic regression. Statistical Analysis: Chi square, t test, Chi square for trend and Pearson's correlation coefficient were used for analysis. Results: Spectacle use was significantly higher and positively associated with literacy and employment in the urban population. The age and gender-adjusted prevalence of emmetropia, myopia of spherical equivalent (SE) </=-0.50 diopter sphere (DS), high myopia (SE </=-5.00DS), hyperopia (SE> 0.50DS) and astigmatism ≤ 0.50 diopter cylinder (DC) were 46.8%, 31.0%, 4.3%, 17.9% and 60.4% respectively in the rural population and 29.0%, 17.6%, 1.5%, 51.9%, 59.1% respectively in the urban population. The prevalence of emmetropia decreased with age ( p p = 0.001) and were associated with nuclear sclerosis ( p = 0.001) in both populations. Hyperopia was commoner among women than men ( p = 0.001); was positively associated with diabetes mellitus ( p = 0.008) in the rural population and negatively with nuclear sclerosis ( p = 0.001) in both populations. Conclusion: Spectacle use was found to be significantly lower in the rural population. The pattern of refractive errors was significantly different between both populations.
Abstract: PURPOSE: To determine the relationship between tumor cell differentiation and age at diagnosis in retinoblastoma. METHODS: Medical records of 170 patients with retinoblastoma treated by enucleation were reviewed retrospectively. Age at diagnosis and histopathological features were analyzed. RESULTS: Well-differentiated tumors presented earlier than poorly differentiated tumors. The frequency of presentation was highest in the first year of age for well-differentiated tumors and in the third year of age for poorly differentiated tumors (P < .0001). Bilateral well-differentiated tumors presented earlier than bilateral poorly differentiated tumors. Similarly, unilateral well-differentiated tumors presented earlier than unilateral poorly differentiated tumors. CONCLUSIONS: Differentiated tumors present earlier than poorly differentiated tumors, irrespective of laterality in retinoblastoma.
Abstract: OBJECTIVE: To estimate the prevalence and risk factors of primary open-angle glaucoma (POAG) in an urban population and compare the same with that of our published rural population data in southern India. DESIGN: Population-based cross-sectional study. PARTICIPANTS: Four thousand eight hundred subjects 40 years or older were selected using a multistage random cluster sampling procedure in Chennai city. INTERVENTION: Three thousand eight hundred fifty (80.2%) subjects underwent a complete ophthalmic examination, including applanation tonometry, gonioscopy, pachymetry, optic disc photography, and automated perimetry. MAIN OUTCOME MEASURES: Glaucoma was diagnosed using the International Society of Geographical and Epidemiological Ophthalmology Classification. RESULTS: The distribution of intraocular pressure (IOP) and vertical cup-to-disc ratio (VCDR) was obtained from the right eye of the 2532 subjects with normal suprathreshold visual fields. Mean IOP was 16.17+/-3.74 mmHg (97.5th and 99.5th percentiles, 24 mmHg and 30 mmHg). The mean VCDR was 0.43+/-0.17 (97.5th and 99.5th percentiles, 0.7 and 0.8). One hundred thirty-five (64 men, 71 women) subjects had POAG (3.51%; 95% confidence interval [CI], 3.04-4.0). Primary open-angle glaucoma subjects (58.4+/-11.3 years) were older (P<0.0001) than the study population (54.8+/-10.6 years). One hundred twenty-seven (94%) subjects were diagnosed to have POAG for the first time. Two subjects (1.5%) were bilaterally blind, and 3 (3.3%) were unilaterally blind due to POAG. The urban population prevalence was more than that of the rural population (1.62%; 95% CI, 1.4%-1.8%; P<0.0001). In both populations, increasing IOP (per millimeter of mercury) and older age were associated with the disease. There was no association with gender, myopia, systemic hypertension, diabetes, or central corneal thickness. CONCLUSIONS: The prevalence of POAG in a > or =40-year-old south Indian urban population was 3.51%, higher than that of the rural population. The prevalence increased with age, and >90% were not aware of the disease.
Abstract: OBJECTIVE: To determine the prevalence of primary angle-closure glaucoma (PACG), primary angle closure (PAC), and PAC suspect (PACS) in an urban population, and to compare prevalence and associated risk factors with a rural population. DESIGN: Population-based cross-sectional study. PARTICIPANTS: Four thousand eight hundred subjects 40 years or older were selected from Chennai city using multistage random cluster sampling. INTERVENTION: All subjects had a complete ophthalmic examination that included logarithm of the minimum angle of resolution visual acuity, applanation tonometry, gonioscopy, grading of lens opacities, dilated fundus examination, optic disc photography, and visual fields. MAIN OUTCOME MEASURES: Glaucoma was diagnosed using the International Society of Geographical and Epidemiological Ophthalmology classification. RESULTS: Three thousand eight hundred fifty (80.2%) responded; 34 subjects (17 female, 17 male) had PACG (0.88%; 95% confidence interval [CI], 0.60-1.16). The mean intraocular pressure (IOP) was 26.0+/-14.9 mmHg. Five subjects (14.7%) had been previously diagnosed to have glaucoma, 1 of whom had undergone glaucoma surgery and 2 of whom had been diagnosed to have open-angle glaucoma. Two subjects (5.9%) were bilaterally and 3 subjects (8.8%) were unilaterally blind. One hundred six subjects (2.75%; 95% CI, 2.01-3.49) were diagnosed to have PAC (62 female, 44 male). Thirty-nine subjects (36.8%) had presenting IOP > 24 mmHg, 83 (78.3%) had peripheral anterior synechiae, and 16 (15.1%) had both. Two hundred seventy-eight subjects (7.24%; 95% CI, 6.38-8.02) had PACS (183 female, 95 male). Prevalences of PACG and PACS were similar in the urban and rural populations. Primary angle closure prevalence was higher in the urban population (P<0.0001). Primary angle closure and PACG were positively associated with increasing age and IOP in both populations and were more common in rural women (odds ratio [OR], 4.3; 95% CI, 2.2-8.3). Association with hyperopia was seen only in the urban population (OR, 2.0; 95% CI, 1.4-2.8). CONCLUSIONS: Prevalences of PACG and PACS were similar in the rural and urban populations; PAC was more common in the urban population. In both groups, the disease was asymptomatic. Poor detection rates were probably due to lack of gonioscopy as a routine part of an eye examination.
Abstract: PURPOSE: To determine the frequency of pathogenic mutations in the gene encoding RPE65 in patients from India with Leber congenital amaurosis (LCA). METHODS: The coding sequence of all 14 exons and the adjacent flanking intron sequences of the RPE65 gene were directly sequenced in 60 unrelated Indian LCA patients. Bioinformatics tool was used to study the structural changes of the mutant protein. RESULTS: Three sequence variants were found; two missense and one isocoding change. Of two missense changes, one was a putative polymorphism (N321K) and the other was a novel missense, disease causing change that alters proline to leucine at codon 470 (P470L) in one LCA patient. RPE65 mutations contribute to 1.7% of LCA in our population. CONCLUSIONS: Mutations in the RPE65 gene are rare in patients with LCA and hence genes other than could be mainly responsible for causing LCA in India.
Abstract: Retinoblastoma has contributed much to the understanding of cancer. It provided the classic 'two-hit model' for oncogenesis and helped to identify the first tumor suppressor gene RB1. Thirty years since then, the search for additional events underlying disease progression continues. Phenotypic variations in retinoblastoma offer numerous clues to disease pathogenesis. Understanding their molecular biological basis will provide insight into mechanisms underlying tumor progression. These not fully understood genetic and stochastic events play a major role in uncontrolled retinal precursor cell proliferation. Comparative genomic hybridization and gene expression studies have facilitated probing of genes controlling basic events in cellular development, i.e. proliferation, differentiation and apoptosis. Research to determine the cell of origin that underlies the evolution of retinoblastoma can lead to understanding of the stochastic events underlying the genesis of this cancer, which currently remains unclear. In this review, we discuss the recent developments in retinoblastoma and describe how they are beginning to shape a new and revised picture of retinoblastoma pathogenesis and progression.
Abstract: The endothelial (posterior) corneal dystrophies, which result from primary endothelial dysfunction, include Fuchs endothelial corneal dystrophy (FECD), posterior polymorphous corneal dystrophy (PPCD) and congenital hereditary endothelial dystrophy (CHED). Mutations in SLC4A11 gene have been recently identified in patients with recessive CHED (CHED2). In this study, we show that heterozygous mutations in the SLC4A11 gene also cause late-onset FECD. Four heterozygous mutations [three missense mutations (E399K, G709E and T754M) and one deletion mutation (c.99-100delTC)] absent in ethnically matched controls were identified in a screen of 89 FECD patients. Missense mutations involved amino acid residues showing high interspecies conservation, indicating that mutations at these sites would be deleterious. Accordingly, immunoblot analysis, biochemical assay of cell surface localization and confocal immunolocalization showed that missense proteins encoded by the mutants were defective in localization to the cell surface. Our data suggests that SLC4A11 haploinsufficiency and gradual accumulation of the aberrant misfolded protein may play a role in FECD pathology and that reduced levels of SLC4A11 influence the long-term viability of the neural crest derived corneal endothelial cells.
Abstract: PURPOSE: To investigate the genetic basis of autosomal recessive retinal degeneration in a large consanguineous family from Pakistan. METHODS: Ophthalmic examinations were conducted on family members to establish their diagnosis. Genomic DNA extracted from peripheral blood was used for homozygosity mapping to discover the chromosomal region that harbors the defective gene. Direct sequence analysis and restriction enzyme digestion were used to identify and confirm the defect in the gene. RESULTS: There were three affected siblings in the family, each with limited peripheral vision and impaired visual acuity. We established linkage to a region on chromosome 2 that encompasses the RP26 locus. Upon sequencing the ceramide kinase-like (CERKL) gene, which is mutated in the original RP26 family, we identified a C>A transversion in exon 2 (c.316C>A) that substitutes an arginine residue with a serine (p.R106S) in the conserved nuclear localization signal sequence (KLKRR) of the protein. This mutation segregated with retinal degeneration in the Pakistani family and was not observed in the DNA of 174 ethnically matched unaffected controls. CONCLUSIONS: This is the third reported mutation in CERKL causing retinal degeneration but is the first report to show that a single amino acid change in CERKL, rather than a null mutation, can cause retinal disease. Although the function of CERKL is still unknown, the mutation described herein confirms that the nuclear localization signal sequence is important in the physiologic function of the protein.
Abstract: INTRODUCTION: Diabetes mellitus, particularly type II, is a major public health concern worldwide. While the occurrence of diabetic retinopathy cannot be prevented, with the provision of knowledge to sufferers, sight-threatening complications can be minimized. Purpose: To report the results of a KAP (Knowledge, Attitude and Practice) study among a rural population in two areas: diabetes mellitus (DM) and diabetic retinopathy (DR). The level of knowledge was evaluated for both DM and DR; however, the influence of knowledge on practices and attitude was evaluated in only the DR group. METHODS: In rural areas, 145 awareness meetings on DM and DR were conducted attended 28 347 individuals. Using systematic random sampling, the data were collected from every 14th individual. In total, 1938 individuals from a rural population were numbered for gaining their responses to the KAP questionnaire. Univariate and multiple regression analyses were performed to identify independent risk factors related to the knowledge of the disease and influence of this knowledge on attitude and practice. RESULTS: Of 1938 individuals, 966 (49.9%) had knowledge of DM and 718 (37.1%) had knowledge of DR. Knowledge about DM was more in women (OR=1.93; 95% CI: 1.55-2.39), in subjects who followed the Christian faith (OR=1.48; 95% CI: 1.07-2.04) and in those who belonged to the upper socioeconomic strata (OR=2.60; 95% CI: 1.84-3.67). The knowledge of DR was significantly higher among subjects who spoke the Malayalam language (OR=3.80; 95% CI: 2.03-7.13), who followed the Christian faith (OR=1.73; 95% CI: 1.27-2.35), and in those who belonged to the upper socioeconomic strata (OR=1.85; 95% CI: 1.32-2.58). Compared with those who had no knowledge of DR (n = 1220), significant percentages of individuals with knowledge (n = 718) had the right attitude - to go for regular eye examinations - (65.9% vs 93.3%) (p<0.0001) ). Regarding practice patterns, only 36.5% of individuals with knowledge about DR believed that if they controlled their blood sugar, they could avoid a visit to an ophthalmologist, compared with 55.5% with no knowledge (p<0.0001). CONCLUSION: The results suggest that we need to propagate aggressive and comprehensive awareness models to educate rural populations on DM and DR.
Abstract: PURPOSE: The study was undertaken to evaluate the prevalence of retinitis pigmentosa (RP) in rural and urban South India. METHODS: Seven thousand seven hundred and seventy four subjects aged 40 years or more from rural and urban Tamil Nadu, underwent comprehensive ophthalmic examination out of 9576 enumerated (81.17%). After a thorough ophthalmic examination, fundus photographs were taken for documentation. Statistical analysis was done using SPSS for Windows (ver 14). RESULTS: 7461 (95.9%) subjects had fundus details seen in both eyes. Thirteen subjects (0.17%; 4 males, 9 females) were diagnosed as retinitis pigmentosa. Retinitis pigmentosa in the urban population was seen in approximately 1 in 930 persons, while 1 in 372 of rural subjects had the disorder. This figure is greater than other reports from the western populations and that of the conservative estimate of 1 in 4000. The age and gender adjusted prevalence rate of retinitis pigmentosa to national census 2001 was 0.155%. Eight subjects (61.53%) had visual acuity less than 3/60. CONCLUSION: Prevalence of RP in South India appears to be alarmingly higher in comparison to those seen in other parts of the world.
Abstract: Autosomal recessive congenital hereditary endothelial dystrophy (CHED2) is a severe and rare corneal disorder that presents at birth or shortly thereafter, characterized by corneal opacification and nystagmus. Recently the gene for CHED2 was identified and seven different mutations in the SLC4A11 gene were reported. Here, we report seven novel mutations and two previously identified mutations in families from India and the United Kingdom with recessive CHED. The novel changes include two nonsense (p.Trp240X; p.Gln800X) three missense (p.Glu143Lys; p.Cys386Arg; p.Arg755Trp) and two splice site mutations (c.2240+1G>A; c.2437-1G>A). Interestingly, the c.2398C>T (p.Gln800X) and c.2437-1G>A identified in two affected siblings represent the first compound heterozygous mutations in the SLC4A11 gene.
Abstract: The genetic background of congenital glaucoma in a consanguineous south Indian family was examined by homozygosity analyses. Significant evidence for the homozygosity of alleles was detected for markers D2S177 and D2S1346 that are tightly linked to the CYP1B1 gene, and further involvement of this gene was confirmed by the co-segregation of a novel truncating mutation (Q110X) in exon 2 with the disease in all affected members. Newborn genetic screening and carrier identification were also performed in the family. The role of consanguinity and the risk of autosomal recessive disease were discussed and genetic counseling was given.
Abstract: A 27-bp variable number tandem repeat (VNTR) in intron 4 of endothelial nitric oxide synthase (eNOS) gene has been associated with the risk for developing diabetic retinopathy (DR) in various ethnic populations. Hundred and eighty seven patients with retinopathy (cases; DR+) and 188 patients without retinopathy (controls: DR-) from southern India who had type 2 diabetes mellitus (T2DM) for more than 10 years, were included in the study. We could neither find significant allelic association with clinical severity of DR nor with macular edema. Our results suggest lack of association of intron 4 VNTR of eNOS gene with DR in southern India.
Abstract: OBJECTIVES: This study was conducted with two objectives. The first was to estimate the frequency of loss of heterozygosity (LOH) of the RB1 gene as a mechanism in disease causation in tumors of patients from India. The second objective was to employ RB1 molecular deletion and microsatellite-based linkage analysis as laboratory tools, while counseling families with a history of retinoblastoma (RB). METHODS: DNA was extracted from peripheral blood and tumors of 54 RB patients and their relatives. Eight fluorescent microsatellite markers, both intragenic and flanking the RB1 gene, were used. After PCR amplification, samples were run on an ABI PRISM 310 genetic analyzer for LOH, deletion detection, and haplotype generation. RESULTS: LOH was found in conjunction with tumor formation in 72.9% of RB patients (39/54 patients; p=0.001; 95% CI 0.6028, 0.8417); however, we could not associate various other clinical parameters of RB patients with the presence or absence of RB1 LOH. Seven germline deletions (13% of RB patients) were identified, and the maternal allele was more frequently lost (p=0.01). A disease co-segregating haplotype was detected in two hereditary autosomal dominant cases. CONCLUSION: LOH of the RB1 gene could play an important role in tumor formation. Large deletions involving RB1 were observed, and a disease co-segregating haplotype was used for indirect genetic testing. This is the first report from India where molecular testing has been applied for RB families in conjunction with genetic counseling. In tertiary ophthalmic practice in India, there is an emerging trend towards the application of genetical knowledge in clinical practice.
Abstract: BACKGROUND/AIMS: Growth factors have been implicated in the pathogenesis of diabetic retinopathy (DR). IGF-1 is known to trigger a critical cascade of molecular events that initiate retinal angiogenesis. Increased vitreous IGF-1 levels have been correlated with the severity of ischemia-associated diabetic retinal neovascularization. In the present study, a cytosine-adenine (CA)(n) repeat in the promoter of the IGF-1 gene is studied for association with DR. METHODS: A total of 127 patients with retinopathy (cases: DR+) and 81 patients without retinopathy (controls: DR-) who had type 2 diabetes were recruited for the study. Patients underwent detailed clinical examination and DR was graded based on stereoscopic digital fundus photographs. Frequencies of alleles and genotypes between the two groups were analyzed for significance using relevant statistical tests. (CA)(17) and (CA)(18) repeats were the more frequent alleles. RESULTS: The frequency of the 18-repeat genotype was significantly higher in DR+ patients when compared to DR- patients and found to confer a 2.4 times (95% CI: 1.2-5.0) and 2.8 times (95% CI: 1.1-7.5) higher risk for developing DR and proliferative DR, respectively, when compared to <18-repeat genotypes. CONCLUSIONS: Our study suggests that the 18-repeat genotype is a susceptibility genotype for DR and its clinical severity in a Southern Indian cohort.
Abstract: PURPOSE: KIF14 a mitotic kinesin gene plays an important role in cytokinesis. Deregulation of KIF14 may be a pathway of tumor progression and results in decreased patient survival as seen in breast tumors. Recently, KIF14, a possible gene that drives gain of chromosome arm 1q (the most commonly gained chromosomal region in retinoblastoma), has been shown to be a strong oncogene candidate overexpressed by more than two orders of magnitude in retinoblastoma. This study was conducted to quantify the expression of KIF14 in human retinoblastoma tumors and correlate it with disease phenotype. METHODS: KIF14 expression was examined by using real-time RT-PCR in 30 retinoblastoma tumors with age at diagnosis between 3 and 68 months. Two 18-month-old, three adult (55-62 years), and three fetal (one 18 weeks' and another pooled retina of 18 and 20 weeks' gestation) retinas were used as the control. KIF14 expression was normalized to the housekeeping control gene TBP and compared with that in an 18-month-old control retina. The protein expression was confirmed in tumor cells by immunohistochemistry and phenotypic correlation was performed. RESULTS: KIF14 was expressed between 3- and 207-fold greater than 18-month-old retina in 30 retinoblastoma tumors (P < 0.0001). Immunohistochemistry revealed KIF14 localization to both nucleus and cytoplasm of tumor cells. KIF14 mRNA overexpression correlated significantly with older age at diagnosis (P = 0.006). There was no association with differentiation, invasion, or duration of the disease with KIF14 overexpression. CONCLUSIONS: Overexpression of KIF14 was confirmed in primary human retinoblastoma and showed that patients with an older age at diagnosis express significantly higher levels of KIF14.
Abstract: AIM: To present an analysis of a screening model for diabetic retinopathy and compare the results of screening between rural and urban populations. METHODS: Between June 2003 and September 2004, 51 diabetic retinopathy screening camps (rural, 25; urban, 26) were conducted in three southern districts of India. The target population, aged 30 years and above, underwent comprehensive eye evaluation and those with referable diabetic retinopathy (proliferative diabetic retinopathy, severe non-proliferative diabetic retinopathy, severe diabetic macular oedema, or a combination of these) were referred to the base hospital for further treatment. RESULTS: Among 7716 diabetic subjects, the age and sex adjusted prevalence of diabetic retinopathy was 18% in the rural areas and 17% in the urban areas. The prevalence of referable retinopathy was 6.8% in rural areas and 4.6% in urban areas (p<0.001). Around 63% of individuals in rural areas and 75% in urban areas had never previously had their eyes examined for diabetic retinopathy. Multivariate analysis revealed the following risk factors for diabetic retinopathy: age more than 50 years, known diabetes, prolonged duration of diabetes, and eyes with moderate or severe visual impairment (p<0.0001). CONCLUSIONS: The study describes a comprehensive diabetic retinopathy screening model which can identify sight threatening retinopathy and provide necessary treatment for rural and urban populations.
Abstract: BACKGROUND: Extracellular matrix (ECM) accumulation in the trabecular meshwork tissues of glaucoma patients has been demonstrated as one of the factors that contribute to glaucoma pathology. Transforming growth factor-beta (TGFbeta) has its fundamental function in regulating the ECM molecules and has been implicated in glaucoma pathology. In this study, the association of the TGFB1-509C>T single nucleotide polymorphism (SNP) with primary open angle glaucoma (POAG) in patients from India is analyzed. METHODS: One-hundred and six POAG patients and 104 controls were selected after comprehensive ophthalmic examinations. TGFB1 alleles were typed by restriction enzyme digestion with the isoschizomer Eco81I of Bsu36I, whose site is altered by the -509C>T SNP, and statistically analyzed for any significant association. Two clinical variables, vertical cup disc ratio (CDR) and intraocular pressure (IOP), were compared at diagnosis by the Mann-Whitney test for any significant association with the polymorphism. RESULTS: Statistical analysis between the two groups did not suggest any significant difference in the distribution of allele and genotype frequencies. The Mann-Whitney test did not show any significant p value for the clinical parameters IOP (p = 0.29 and 0.59) and CDR (p = 0.26 and 0.17). CONCLUSIONS: The current study shows that the TGFB1-509C>T polymorphism might not be associated with POAG. Analysis of the other polymorphisms in the regulatory region of the TGFB1 gene could give a better understanding of the role of TGFbeta in POAG pathogenesis.
Abstract: AIM: To study the prevalence and causes of blindness in a rural south Indian population. METHODS: 3924/4800 enumerated (81.75%) subjects, aged 40 years or more from rural Tamil Nadu, underwent comprehensive ophthalmic examination-visual acuity, refraction, intraocular pressure, gonioscopy, cataract grading (LOCS II), retinal examination, and SITA Standard where indicated. Blindness was defined using WHO criteria as best corrected visual acuity of less than 3/60 and/or visual field of less than 10 degrees in the better eye. The influence of age, sex, literacy, and occupation was assessed using multiple logistic regression. RESULTS: 753 subjects (19.2%; 321 males, 432 females) presented with a visual acuity of <3/60; 132 subjects (3.36%, 95% CI: 2.80 to 3.93) were diagnosed to be blind. Cataract was responsible in 74.62% of eyes; glaucoma, cystoid macular oedema, optic atrophy, and corneal scars accounted for 3.79% each. Bilateral causes of blindness were cataract (78.63%), glaucoma (4.29%), optic atrophy (3.42%), cystoid macular oedema, and corneal scars (2.56% each). In 19 eyes (7.2%) the blindness was probably related to cataract surgery. Blindness was positively associated with increasing age (p<0.0001). CONCLUSION: 3.36% of the studied rural population was bilaterally blind, with cataract being the single most important cause.
Abstract: PURPOSE: Molecular genetic diagnostics for retinoblastoma are prerequisite for accurate risk prediction and effective management. Developing a retinoblastoma diagnostic model to establish a flow for laboratory tests is thus a necessity for tertiary ophthalmic institutions. An efficient diagnostic model could reduce the overall health care costs, redirect the resources to the high risk group and also avoid unnecessary worry for families. To the best of our knowledge there has hitherto been no comprehensive diagnostic model for retinoblastoma implemented in any institution in India. METHODS AND DISCUSSION: The diagnostic model demonstrates the logical and practical flow of various genetics tests like karyotyping, loss of heterozygosity analysis, molecular deletion, linkage analysis (familial cases), mutation screening of -CGA exons first and then non-CGA exons, methylation screening of RB1 and essential promoter regions screening in a laboratory. Conclusions: The diagnostic model proposed offers acomprehensive methodology to identify the causative two-hits for retinoblastomas that could be used while genetic counseling families. This model is applicable in tertiary hospitals in India and neighboring countries, which have the highest incidence of retinoblastoma and fertility rates in the world. We suggest that this diagnostic model could also be applied with modification for other cancers.
Abstract: BACKGROUND: Genetic analysis has a beneficial impact on retinoblastoma management enabling definite risk assessment. However, information regarding genotype-phenotype correlation in retinoblastoma is limited. AIM: To analyze the retinoblastoma susceptibility gene for mutations in retinoblastoma patients and correlate the genotypes the phenotypes. METHODOLOGY: Eleven retinoblastoma patients, who underwent molecular genetic studies were classified into high, moderate or low disease severity groups based on phenotype. RESULTS: Seven patients had high disease severity and four moderate disease severity. Eleven truncating mutations were detected; six were in the N-terminus region of the retinoblastoma protein and two in the A/B pocket (p=0.03). CONCLUSIONS: No significant association between mutation type and disease severity could be established in the present study. However a positive correlation between location of the mutations in certain domains of the retinoblastoma protein and disease severity was observed. To the best of our knowledge this is the first genotype-phenotype correlation study in retinoblastoma patients from India.
Abstract: OBJECTIVE: To estimate the prevalence of primary angle-closure glaucoma, primary angle closure (PAC), and primary angle-closure suspect (PACS) and its associated risk factors in a rural population in southern India. METHODS: Three thousand and nine hundred thirty-four (81.95%) of 4800 enumerated subjects aged 40 years or older underwent a complete ophthalmic examination, including compression gonioscopy. Glaucoma was diagnosed using International Society of Geographical and Epidemiological Ophthalmology classification. RESULTS: Data were analyzed for 3924 subjects (81.75%). Primary angle-closure glaucoma was diagnosed in 34 subjects (0.87%; 95% confidence interval [CI], 0.58 to 1.16) (27 women, 7 men). The mean intraocular pressure was 20.71 +/- 9.24 mm Hg. One subject (2.94%) was blind. Twenty-eight subjects (0.71%; 95% CI, 0.45 to 0.98) were diagnosed to have PAC (21 women, 7 men). Eleven subjects (39.3%) had an intraocular pressure greater than 21 mm Hg, 13 subjects (46.43%) had peripheral anterior synechiae, and 4 subjects (14.29%) had both. Two hundred forty-six subjects (6.27%; 95% CI, 5.51 to 7.03) had PACS (168 women, 78 men). Primary angle closure and primary angle-closure glaucoma were more common in women (age-adjusted odds ratio, 3.02; 95% CI, 1.66 to 5.51) with an increasing prevalence with age. Increasing intraocular pressure was associated with the disease (odds ratio, 1.14; 95% CI, 1.09 to 1.19). There was no association with hypertension and hyperopia. Axial length and anterior chamber depth were longer in the normal group than in the 3 groups with angle closure (P<.05). Women had shorter axial lengths than men (P<.001) in the angle closure groups. CONCLUSIONS: The overall prevalence of primary angle closures (PAC and primary angle-closure glaucoma) in a rural population of southern India was 1.58%. There was a female preponderance, and the disease tends to be asymptomatic.
Abstract: INTRODUCTION: The aim of this study was to compare the occurrence of diabetic retinopathy in targeted screening diabetic patients (Group I) with newly diagnosed diabetic patients in general practice (Group II). MATERIALS AND METHODS: This was an observational cross-sectional study. Data were obtained from 25,313 subjects who participated in the diabetic screening camps, and 128 newly diagnosed diabetes who presented to the diabetic retinopathy screening camps in general practice in rural and urban south India. The study variables were collected from all patients who underwent eye examination from the target screening detected diabetics [(n = 173) Group I] and those newly diagnosed in general practice [(n = 128) Group II]. The variations in prevalence of diabetic retinopathy and sight-threatening diabetic retinopathy in Group I and Group II and the factors affecting it were identified. RESULTS: The occurrence of diabetic retinopathy was 6.35% (95% CI, 2.5-9.5) in Group I and 11.71% (95% CI, 5.6-16.4) in Group II. No significant difference was observed on occurrence of diabetic retinopathy, including sightthreatening retinopathy, in rural versus urban population and in Group I versus Group II. Patients diagnosed in general practice (Group II) with systolic blood pressure (BP) >140 were more likely to have retinopathy (P = 0.02). CONCLUSIONS: Diabetic retinopathy including sightthreatening complications was found at the time of diagnosis of diabetes in the targeted screening group as well as in newly diagnosed diabetics in the general practice group.
Abstract: PURPOSE: Optineurin gene (OPTN) mutations are reported in primary open angle glaucoma patients (POAG) from different populations. The coding and noncoding regions of OPTN were screened for mutations in 100 Indian high tension glaucoma patients (HTG). The frequency of the OPTN M98K mutation in an additional 120 patients (70 HTG and 50 normal tension glaucoma [NTG]) was analyzed by restriction enzyme digestion. METHODS: The HTG patients (about 40 years of age) were characterized by open angles on gonioscopy, with raised intraocular pressure (IOP) more than 21 mmHg (<21 mmHg on office diurnal phasing for NTG), and typical glaucomatous disc changes with corresponding visual field defects in the absence of any secondary cause. One hundred HTG patients and controls were screened for OPTN mutations by direct sequencing using an ABI prism 310/3100 Avant genetic analyzer. The M98K status was analyzed by restriction enzyme digestion with StuI. A genotype/phenotype correlation was also attempted for OPTN sequence alterations with clinical parameters such as age at diagnosis, intraocular pressure, cup:disc ratio, etc. The putative change in the transcription factor binding site for the IVS7 +24G>A polymorphism was attempted with AliBaba software (version 2.1). RESULTS: Six sequence alterations were observed in the 100 POAG patients by direct sequencing. The M98K substitution was observed in a total of 10 patients (7/170 HTG and 3/50 NTG) contributing to 4.1% in HTG and 6% in the NTG group and not in the controls. The IVS7+24G>A nucleotide change showed a significant difference in the HTG group (7/100) when compared to the control group (0/100) and found to be associated with increased IOP at diagnosis (p=0.03). The IVS7+24G>A polymorphism resulted in the creation of binding sites for transcription factors NF-1 and CPE that were not present in the wild type. CONCLUSIONS: The current study suggests a possible role of SNPs rather than mutations in OPTN in POAG pathology in the Indian population.
Abstract: PURPOSE: A four-generation family containing eight affected males who inherited X-linked developmental lens opacity and microcornea was studied. Some members in the family had mild to moderate nonocular clinical features suggestive of Nance-Horan syndrome. The purpose of the study was to map genetically the gene in the large 57-live-member Asian-Indian pedigree. METHODS: PCR-based genotyping was performed on the X-chromosome, by using fluorescent microsatellite markers (10-cM intervals). Parametric linkage analysis was performed by using two disease models, assuming either recessive or dominant X-linked transmission by the MLINK/ILINK and FASTLINK (version 4.1P) programs (http:www.hgmp.mrc.ac.uk/; provided in the public domain by the Human Genome Mapping Project Resources Centre, Cambridge, UK). The NHS gene at the linked region was screened for mutation. RESULTS: By fine mapping, the disease gene was localized to Xp22.13. Multipoint analysis placed the peak LOD of 4.46 at DSX987. The NHS gene mapped to this region. Mutational screening in all the affected males and carrier females (heterozygous form) revealed a truncating mutation 115C-->T in exon 1, resulting in conversion of glutamine to stop codon (Q39X), but was not observed in unaffected individuals and control subjects. conclusions. A family with X-linked Nance-Horan syndrome had severe ocular, but mild to moderate nonocular, features. The clinical phenotype of the truncating mutation (Q39X) in the NHS gene suggests allelic heterogeneity at the NHS locus or the presence of modifier genes. X-linked families with cataract should be carefully examined for both ocular and nonocular features, to exclude Nance-Horan syndrome. RT-PCR analysis did not suggest nonsense-mediated mRNA decay as the possible mechanism for clinical heterogeneity.
Abstract: PURPOSE: To study whether the difference in the demographic characteristics of participants and non participants could result in biased prevalence estimates and associations. AIM: To compare the non-participant and participant characteristics, and to ascertain if non-response bias is present in the rural population of the Chennai Glaucoma Study (CGS). METHODS: Rural participants and non-participants were compared with regard to socio-demographic variables (age, gender, religion, mother tongue, literacy and employment). RESULTS: 4800 subjects aged 40 years or over were enumerated, 82% (3934: 45% male and 55% female) responded. Gender did not influence participation (adjusted OR-1.11, CI: .91-1.36). Subjects in the 70-79 year age group were more likely to respond (OR-1.76; CI-1.31-2.38). Hindus had a higher participation rate than Christians or Muslims (adjusted OR-2.63, CI: 1.80-3.84). The other predictors of participation were illiteracy (adjusted OR-1.44, CI: 1.22-1.70), unemployment (OR-1.28, CI: 1.04-1.58), place of residence (main villages) (OR-6.66, 95% CI: 4.6-9.64). CONCLUSION: Based on our study findings, it does not seem likely that participation bias will affect the study results.
Abstract: PURPOSE: Karyotype analysis in hereditary retinoblastoma is considered to be of marginal value in risk prediction due to uncertainties in the assessment of 13q14 deletions. However, it is a low cost genetic test for retinoblastoma in developing countries. In the present study, the results of karyotype analysis were refined by a statistical method to overcome limitations. METHODS: Karyotype analysis was performed by trypsin-Giemsa banding and naked eye karyotyping for 33 bilateral, 25 unilateral and one regressed retinoblastoma patients. The percentage of metaphases with 13q14 deletions in each case was plotted on a scatter diagram. Normalization of the data was achieved by log transformation and the results were statistically analyzed by one-sample 't' test using SPSS version 9.0. RESULTS: Seven samples had 13q14 deletion percentages above the cutoff value. One-sample 't' test showed significance (p< 0.001). By this method, two unilateral and five bilateral patients had 13q14 deletions, constituting 11.8 % of cases. CONCLUSION: For accuracy, statistical analysis should be considered as an adjunct in karyotyping.
Abstract: PURPOSE: To determine the prevalence of primary open-angle glaucoma (POAG) and the associated risk factors in a rural population in southern India. METHODS: Subjects aged 40 years or more (n = 3934) underwent a complete ophthalmic examination. Glaucoma was diagnosed according to the International Society of Geographical and Epidemiologic Ophthalmology classification. RESULTS: Complete data were available for 3924 subjects (response rate, 81.75%). In eyes with normal suprathreshold visual fields, the mean intraocular pressure was 14.29 +/- 3.32 mm Hg (97.5th and 99.5th percentiles, 21 and 25 mm Hg, respectively). The mean vertical cup-to-disc ratio was 0.39 +/- 0.17 (97.5th and 99.5th percentiles, 0.7 and 0.8, respectively). Sixty-four subjects had definite POAG (1.62%, 9.5% CI 1.42-1.82); 30 were men and 34 were women. Subjects with POAG (59.85 +/- 10.43 years) were older (P < 0.001) than the study population (53.78 +/- 10.71 years). In only one (1.5%) person was POAG diagnosed before the study. Two (3.12%) subjects were blind due to POAG; 21 (32.81%) subjects had a presenting IOP >21 mm Hg, and 43 (67.19%) had an IOP <21 mm Hg. The mean central corneal thickness in subjects with POAG (502.82 +/- 35.29 microm) was not different from that of the normal study population (505.93 +/- 31.11 microm). No association was found with diabetes mellitus, systemic hypertension, gender, and myopia. Increasing IOP (per mm Hg) was associated with the disease (OR 1.12; 95% CI, 1.08-1.16). The odds for POAG increased with advancing age after adjustment for gender. CONCLUSIONS: The prevalence of POAG in this population was 1.62%. The prevalence increased with age, and 98.5% were not aware of the disease.
Abstract: PURPOSE: Congenital microcoria is a rare autosomal dominant developmental disorder of the iris associated with myopia and juvenile open angle glaucoma. Linkage to the chromosomal locus 13q31-q32 has previously been reported in a large French family. In the current study, a three generation Asian Indian family with 15 congenital microcoria (pupils with a diameter <2 mm) affected members was studied for linkage to candidate microsatellite markers at the 13q31-q32 locus. METHODS: Twenty-four members of the family were clinically examined and genomic DNA was extracted. Microsatellite markers at 13q31-q32 were PCR amplified and run on an ABI Prism 310 genetic analyzer and genotyped with the GeneScan analysis. Two point and multipoint linkage analyses were performed using the MLINK and SUPERLINK programs. RESULTS: Peak two point LOD scores of 3.5, 4.7, and 5.3 were found co-incident with consecutive markers D13S154, DCT, and D13S1280. Multipoint analysis revealed a 4 cM region encompassing D13S1300 to D13S1280 where the LOD remains just over 6.0 Thus we confirm localization of the congenital microcoria locus to chromosomal locus 13q31-q32. In addition, eight individuals who had both microcoria and glaucoma were screened for glaucoma genes: myocilin (MYOC), optineurin (OPTN) and CYP1B1. Using direct sequencing a point mutation (144 G>A) resulting in a Q48H substitution in exon 1 of the MYOC gene was observed in five of the eight glaucoma patients, but not in unaffected family members and 100 unrelated controls. CONCLUSIONS: We have confirmed the localization of the congenital microcoria locus (MCOR) to 13q31-q32 in a large Asian Indian family and conclude that current information suggests this is a single locus disorder and genetically homogeneous. When combined with the initial linkage paper our haplotype and linkage data map the MCOR locus to a 6-7 cM region between D13S265 and D13S1280. The DCT locus, a member of the tyrosinase family involved in pigmentation, maps within this region. Data presented here supports the hypothesis that congenital microcoria is a potential risk factor for glaucoma, although this observation is complicated by the partial segregation of MYOC Q48H (1q24.3-q25.2), a mutation known to be associated with glaucoma in India. Fine mapping and candidate gene analysis continues with the hope that characterizing the micocoria gene will lead to a better understanding of microcoria and glaucoma causation. The relationship between microcoria, glaucoma, and the MYOC Q48H mutation in this family is discussed.
Abstract: PURPOSE: To describe the methodology of the Sankara Nethralaya-Diabetic Retinopathy Epidemiology and Molecular Genetic Study (SN-DREAMS 1), an ongoing population-based study to estimate the prevalence of diabetes and diabetic retinopathy in urban Chennai, Tamil Nadu, South India, and also to elucidate the clinical, anthropometric, biochemical and genetic risk factors associated with diabetic retinopathy. METHODS: In this ongoing study, we anticipate recruiting a total of 5830 participants. Eligible patients, over the age of 40 years, are enumerated using the multistage random sampling method. Demographic data, socioeconomic status, physical activity, risk of sleep apnea, dietary habits, and anthropometric measurements are collected. A detailed medical and ocular history and a comprehensive eye examination, including stereo fundus photographs, are taken at the base hospital. Biochemical investigations (total serum cholesterol, high-density lipoproteins, serum triglycerides, hemoglobin, glycosylated hemoglobin HbA1c) and genetic studies of eligible subjects are conducted. A computerized database is created for the records. CONCLUSION: The study is expected to result in an estimate of the prevalence of diabetes and diabetic retinopathy and a better understanding of biochemical and genetic risk factors associated with diabetic retinopathy in an urban South Indian population. Worldwide, the prevalence of diabetes mellitus, in particular type II diabetes, is rising at an alarming rate. The World Health Organization (WHO) and International Diabetes Federation (IDF) have predicted that the number of cases of adult-onset diabetes would more than double by 2030 from the present level of 171 million to 366 million-an increase of 214%.1 In developed countries, this increase in diabetic population would be around 42% and in developing countries, particularly in India, it is even higher; i.e. 150%.1 In India, the prevalence of diabetes mellitus in the urban population is around 12.1%, as reported by the national urban diabetes study2 conducted in six major cities. Studies have shown the prevalence of diabetes to be higher among the high-income groups (25.5%) as compared to low-income groups (12.6%).3,4,5 The assessment of socioeconomic status was based on income,6, 7 education,2, 7 occupation2 or caste6-which are not representative of the actual socioeconomic status. In the present study, however, the sample was stratified on socioeconomic scoring. This scoring was calculated on the basis of several parameters such as the residence being rented or owned, the number of rooms in the house, the highest educational status, the highest salary, the highest occupation, material possessions (cycle, TV, audio, car, etc.) and house/land value. To the best of our knowledge, this kind of comprehensive socioeconomic scoring has not been done before for prevalence studies on diabetic retinopathy in the general population.
Abstract: AIM: To elucidate the rate of non-response among diabetics (recently diagnosed in rural diabetic screening camps) who were referred for eye examination to detect diabetic retinopathy. METHODS: At diabetic retinopathy screening camps, all patients underwent dilated fundus examination using binocular indirect ophthalmoscopy. RESULTS: Of the 4,111 known diabetics, only 2231(55%) patients attended the diabetic retinopathy screening camps. Likewise, of the 1076 newly detected diabetics, only 125 (11.6%) attended the diabetic retinopathy screening camps. CONCLUSIONS: Non-response of such a magnitude calls for creating greater awareness among masses on diabetes and its microvascular complications. Second, conducting simultaneous diabetes screening and diabetic retinopathy screening camps could minimize the dropout rate.
Abstract: CONTEXT: Worldwide, the prevalence of diabetic retinopathy is increasing at an alarming rate. WHO has predicted that in India the number of adults with diabetes will be the highest in the world: from 19 million in 1995 to 80 million in 2030. Although originally thought to be a disease of an urban population, the prevalence of diabetes mellitus is increasing in rural areas as well. The socioeconomic burden resulting from visual impairment or blindness caused by diabetic retinopathy, particularly in the working age group, is a serious concern. ISSUE: In order to combat diabetic retinopathy related blindness, Sankara Nethralaya, the premier eye institute of India, in collaboration with the Lions Clubs International Foundation (LCIF) and the RD Tata Trust, Mumbai, India launched a major diabetic retinopathy screening program in the rural community of South India. The objectives were to create awareness among the rural population of diabetic retinopathy with emphasis on early detection, to conduct diabetes and diabetic retinopathy screening camps, and to bring to the base hospital patients who have sight-threatening diabetic retinopathy, for ancillary investigations such as fluorescein angiography, ultrasound and to perform laser photocoagulation or vitreous surgery, or both. Other objectives included training general ophthalmologists and general physicians in order to develop an integrated diabetic retinopathy model. To address the question as to why certain individuals run the risk of developing sight threatening diabetic retinopathy, biochemical and genetic factors were also studied. The program was launched in June 2003 and 3 rural districts have been screened. To the time of writing, 128 screening camps had been organized, 103 awareness meetings conducted, 23 ophthalmologists trained and 43 general physicians attended the continuing medical education program on diabetic retinopathy. LESSONS: The key elements in the successful implementation of this program have been a team approach, involvement of community leaders and voluntary organizations, and support of the district and state administrators.
Abstract: INTRODUCTION: Genetic testing is increasingly being used to evaluate susceptibility to hereditary diseases because it is a cost effective screening method. Predictive testing for retinoblastoma can help to save the vision and avoid unnecessary (and invasive) eye examinations for probands and their close relatives. This study was undertaken to evaluate the cost effectiveness of the retinoblastoma genetic screening strategy established in our hospital. STUDY DESIGN: Cytogenetic study of peripheral blood, mutational, and methylation analyses from the tumor DNA of 25 patients with retinoblastoma was undertaken. The cost for retinoblastoma (RB1) gene screening was calculated based on the cost of the chemicals and consumables used and the clinical examination charges at our hospital. A comparison was made between the cost of genetic screening and clinical testing for retinoblastoma. Retinoblastoma patients underwent clinical management and genetic testing at Sankara Nethralaya, Chennai, India. RESULTS: By adopting a genetic screening strategy, a 3.5-fold cost saving was seen for a proband while a 6-fold saving was seen for a family with two sibs compared to the cost of clinical examination. The clinical examination fee and cost of genetic screening for a proband was dollarUS536 and dollarUS152, respectively, while for a nuclear family with two sibs the costs were dollarUS1071 and dollarUS175, respectively. DISCUSSION: Savings for a family will be higher if indirect costs, such as savings in travel times to and from the hospital and labor savings, were taken into account. Cost will be a major factor in determining the implementation of genetic screening for RB1 gene in the clinical practice. CONCLUSION: In our study in India, genetic screening for retinoblastoma was cheaper than conventional screening and was useful in the genetic counseling of the families.
Abstract: PURPOSE: To report the prevalence of refractive errors in a rural south Indian population. METHODS: Four thousand eight hundred subjects (age, >39 years) from rural south India were enumerated for a population-based study. All participants underwent complete ophthalmic evaluation. Subjects who were phakic in the right eye with best corrected visual acuity of 20/40 or better were included for analysis. Association of refractive errors with age, sex, cataract, and diabetes mellitus were analyzed. RESULTS: Of the 3924 responders, 2508 were eligible. The unadjusted prevalence of emmetropia (spherical equivalent [SE], -0.50 to +0.50 diopter sphere [DS]), myopia (SE < -0.50 DS), high myopia (SE < -5.00 DS), and hyperopia (SE > 0.50 DS) were 50.60%, 26.99%, 3.71%, and 18.70% and age and gender adjusted for the rural Tamil Nadu population were 46.77%, 30.97%, 4.32%, and 17.94%, respectively. The prevalence of emmetropia decreased significantly with age (P < 0.0001), and the prevalence of myopia and high myopia increased significantly with age (P < 0.001) and were significantly associated with nuclear sclerosis (P < 0.001). The prevalence of hyperopia increased until 60 years of age and then decreased. Hyperopia was more common among women than men (P < 0.001) and was negatively associated with nuclear sclerosis (P < 0.001) and positively with diabetes mellitus (P = 0.008). Of the participants with astigmatism (cylindrical error greater than 0.50 DC), 9.80% had with-the-rule (WTR) and 77.44% against-the-rule (ATR) astigmatism. The prevalence of WTR and ATR astigmatism significantly decreased (P < 0.001) and increased (P = 0.006) with age, respectively. CONCLUSIONS: The pattern of refractive errors in this rural south Indian population is similar to those reported in other tropical regions of the world.
Abstract: Retinoblastoma can arise due to mutational inactivation or methylation of RB1 gene promoter. A 600-bp CpG island consisting of the essential promoter is present at the 5' end of RB1 gene. Hypermethylation of the CpG island within the RB1 promoter region has been described in unilateral retinoblastoma. In vitro and in vivo studies have suggested that methylation of the RB1 promoter dramatically reduces gene activity. In the present study methylation status of the CpG island within the promoter region of RB1 gene has been evaluated by methylation specific polymerase chain reaction to define the molecular mechanism responsible for retinoblastoma in Indian patients. One unilateral and two bilateral nonhereditary patients had methylation of the RB1 promoter region in which 6.6% of our patients had complete methylation of the RB1 promoter region. This study shows methylation of RB1 promoter is not a major mechanism for retinoblastoma patients in India. Methylation analysis is used in genetic counseling of the family.
Abstract: Glaucoma is one of the major causes of blindness in the Indian population. Mutations in the myocilin (MYOC) gene have been reported in different populations. However, reports on MYOC mutations in Indian primary open-angle glaucoma (POAG) patients and juvenile open-angle glaucoma (JOAG) patients are sparse. We therefore screened 100 unrelated POAG/JOAG patients for MYOC mutations. Patients with POAG/JOAG were clinically diagnosed. Genomic DNA from such patients was collected and studied for MYOC mutations by direct sequencing. Nucleotide variations were compared with unrelated healthy controls by restriction enzyme digestion. Secondary structure prediction for the sequence variants was performed by Chou-Fasman method. A novel mutation in exon 1 (144 G-->Alpha) resulting in Gln48His substitution was observed in 2% of the patients. Four other polymorphisms were also observed. The novel mutation was seen in four other affected family members of a JOAG patient. The novel mutation was found to alter the secondary structure in the glycosaminoglycan initiation site of the protein. MYOC mutations were found in 2% of the population studied. MYOC gene may not be playing a significant role in causing POAG in the Indian population.
Abstract: Effective counselling and management of retinoblastoma families using genetic information is presently practised in many parts of the world. We studied histopathological, chromosomal and molecular-genetic data of two retinoblastoma patients from India. The two patients, one with bilateral and the other with unilateral retinoblastoma, underwent complete ophthalmic examination, cytogenetic study, retinoblastoma gene (RB1) mutational analysis and RB1 promoter region methylation screening. In the bilateral retinoblastoma patient deletion of chromosome region 13q14 in peripheral blood lymphocytes and a hemizygous novel 8-bp deletion in exon 4 of RB1 in tumour sample were observed. In the unilaterally affected patient CGA to TGA transition protein truncation mutations were observed in exons 8 and 14 of RB1.
Abstract: PURPOSE: To describe the methodology of a population-based study to estimate the prevalence of glaucoma in a rural and urban South Indian population and to study the genetics of glaucoma in this population. METHODS: A sample size of 4758 each for rural and urban populations in the Indian state of Tamil Nadu was calculated. Eligible subjects aged 40 years and above from the rural study area covering 32 contiguous villages and the urban area comprising five random clusters in Chennai city are enumerated. Demographic data are collected in the field. A detailed clinical examination, including glaucoma diagnostic procedures, is conducted at the examination centre. Pedigree ascertainment and genetic studies are performed for subjects with occludable angles or glaucoma. Data are recorded in a computerised database. CONCLUSIONS: This study is expected to result in an estimation of the prevalence and a better understanding of the genetics of glaucoma in this region.
Abstract: Genetic factors have been identified that regulate the severity and the rapidity of onset of retinopathy in diabetic patients. Polymorphisms in (CA)( n) present upstream of the promoter of the aldose reductase (ALR2 ) gene have been shown to be associated with retinopathy in different ethnic populations. We aimed to study the association between the (CA)( n) polymorphism and type 2 diabetic patients with and without retinopathy in the Asian Indian population. We screened 105 diabetic patients with retinopathy (DR) and 109 diabetic patients without retinopathy (DNR) for the (CA)( n) polymorphism and compared the results with those of an unrelated healthy control group (CT). We identified 13 alleles in our diabetic population. The Z-2 allele (136 bp) showed an association with the DR group (13.81%) with a significant p value (p = 0.029) when compared with the DNR group (7.34%). The Z-2 allele also showed a significant association with those DR patients who had proliferative retinopathy (PDR) and maculopathy (MAC) (p = 0.004). The Z-2 allele is, therefore, a high-risk allele for diabetic retinopathy in the Asian Indian patients.
Abstract: Nitric oxide, a signal transduction molecule, when modulated causes various diseases including diabetic retinopathy. In diabetes, allelic polymorphism of the inducible nitric oxide synthase (iNOS) gene is associated with retinopathy in the Northern Irish population. In the present study we investigated the Asian Indian population. One hundred and ninety-nine unrelated Asian Indian patients with 15 or more years of type 2 diabetes were divided into two groups: (a) diabetic retinopathy (DR) and (b) diabetic nonretinopathy (DNR) subjects. In these groups the pentanucleotide microsatellite repeat located 2.5 kb upstream of the transcription start site of the iNOS gene was amplified by polymerase chain reaction and analyzed. Eleven alleles, 175-225 bp, were identified. Allele 210 bp was significantly associated with retinopathy (p = 0.044). Individuals carrying this allele had twice the risk of developing retinopathy compared with those who did not carry this allele [odds ratio (OR) - 2.03; 95% CI 0.96-4.35]. Alleles 200 and 220 bp were also significantly associated with no retinopathy and no serious retinopathy complications, respectively. In the Asian Indian population, allele 210 bp of the iNOS gene is a high-risk allele for developing retinopathy and alleles 200 and 220 bp protect an individual from developing retinopathy or its complications.
Abstract: AIM/HYPOTHESIS: The binding of advanced glycation end products (AGE) to the receptor induces cellular oxidative stress and vascular dysfunction and this is implicated in the pathogenesis of diabetic retinopathy (DR). This study aims to investigate the frequency of Gly82Ser polymorphism in exon 3 of the receptor for AGE (RAGE) gene and its association with DR in Asian Indian patients who have type II diabetes. METHODS: 200 Asian Indian patients with at least 15-year duration of type II diabetes were identified. This group included (1) 100 patients with retinopathy (DR) and (2) 100 patients without retinopathy (DNR). Fifty unrelated healthy controls (CT) were also included in the study. Genotype frequencies of Gly82Ser polymorphism were studied by polymerase chain reaction (PCR) amplification and restriction fragment length polymorphism analysis using AluI enzyme. Later, the nucleotide change was confirmed by DNA sequencing. RESULTS: The frequency of the Ser82 allele was significantly higher, 18% in the DNR group compared to 7% in the DR group (P=.03). The same genotype was 2% in the CT group. CONCLUSION/INTERPRETATION: Our result suggests that Ser82 allele in the receptor for AGE gene is a low-risk allele for developing DR in Asian Indian patients who have type II diabetes.
Abstract: OBJECTIVE: Consanguineous marriage is a widely practised social custom in Asia and northern Africa. In south India, Dravidian Hindus have contracted consanguineous marriages for over 2,000 years. In the present study, the influence of consanguinity on the prevalence of visual disorders was examined in patients attending a specialist genetic eye clinic. SUBJECTS AND METHODS: A total of 2,335 patients attending Sankara Nethralaya, Chennai, India, were screened for genetic eye disorders over a five-year period. The patients were drawn from all parts of India and from neighbouring countries in south Asia. RESULTS AND DISCUSSION: Six hundred and seventy-three (28.8%) of the patients tested for ophthalmic genetic disorders reported a family history of consanguinity. The majority (n = 574) of these families were from south India. In the patient group as a whole, the most common form of consanguineous union was between first cousins (n = 367), followed by uncle/niece marriage (n = 177), equivalent to a mean coefficient of inbreeding alpha = 0.0202. Among the consanguineous families, 430 of 673 (63.9%) had retinitis pigmentosa, 167 of these cases were autosomal recessive and 199 were isolated cases. The public in regions such as south India should be made aware of the merits and demerits of consanguineous marriages.
Abstract: We used multiplex PCR follwed by sequencing to screen for mutations in the 14 exons of the RPE65 gene in early-childhood-onset autosomal recessive retinitis pigmentosa (arRP) and Leber's congenital amaurosis (LCA) patients. The RPE65 protein is believed to play an important role in the metabolism of vitamin A in the visual cycle and mutations identified in the gene could have implications for vitamin A-based therapeutic intervention. We were able to identify a homozygous mutation (AAT --> AAG) in exon 9 in an arRP patient and a heterozygous missense transversion (AAT --> AAG) also in exon 9 of an LCA patient. We also identified a polymorphism in exon 10 (GAG --> GAA) in an arRP as well as an LCA patient. Mutation screening would be greatly facilitated by multiplex PCR which could cut down costs, labour and time involved. The nucleotide changes observed in this study could be de novo. Though a larger study has been undertaken, from the preliminary results it appears that in India the RPE65 gene seems to be less involved in causation of LCA.
Abstract: Saethre-Chotzen syndrome is an autosomal dominant disease characterized by craniosynostosis, ptosis, and limb and external ear abnormalities. Variable expressivity is a well-known phenomenon in this disorder. A large Indian family has been recently identified as carrying a nonsense TWIST mutation (Q28 X) in 17 members, of whom 16 were examined in detail. Only 4 (25%) of the patients showed patent craniostenosis, namely, oxycephaly. The penetrance of craniosynostosis in this family is lower than previously reported in the literature. Fifteen patients (93%) had moderate to severe ptosis. Minor limb and external ear abnormalities were present in most patients. Eyelid features were the hallmark of the disease for 12 members of the family, suggesting that mutations in TWIST may lead to a phenotype with mainly palpebral features and no craniostenosis. The clinical analysis of this large family clearly illustrates the significant variable expressivity, probably related to haploinsufficiency because of the TWIST mutation. This phenotypic variability remains unclear but could be the result of modifier genes and/or genetic background effect, as noticed previously in the transgenic twist-null heterozygous mice.
Abstract: PURPOSE: To determine chromosomal abnormalities and inheritance pattern in patients with retinoblastoma from a referral hospital in southern India. MATERIALS AND METHODS: Eighty-one retinoblastoma patients from 78 families were included in this study. Peripheral venous blood was taken for chromosomal analysis and pedigree was ascertained for segregation analysis. RESULTS: Male to female ratio was 1.7:1, 55.56% were bilateral retinoblastoma, the mean age of onset was 12.37 months in bilateral and 33.07 months in unilateral cases (p=0.048). Majority (90.12%) had sporadic inheritance and 6.17% had autosomal dominant inheritance. In chromosomal abnormalities, 8.33% had 13q14 deletion, three cases had de novo balanced translocations. CONCLUSION: The age of onset of the disease was much earlier in the bilateral cases compared to unilateral cases. Sporadic inheritance was predominant while only a small percentage of patients had autosomal dominant inheritance. The percentage of patients with 13q14 deletion was higher than reported in the literature and three novel chromosomal translocations were observed. This is one of the largest series of cases reported from India.
Abstract: Two cases of retinitis pigmentosa (RP) with associated sickle cell disease in one patient, and situs inversus totalis in the other are reported. To our best knowledge, these associations have never been reported in RP.
Abstract: The association of tumor necrosis factor (TNF) with diabetic retinopathy (DR) has been described previously. A total of 207 Asian Indian patients of 15-year duration of type 2 diabetes were identified. This group included (i) 100 patients with DR and (ii) 107 patients without retinopathy (DNR). In this study, we correlated the length of the (GT)n microsatellite di-nucleotide repeat upstream to the promoter region of TNF gene with susceptibility for the development of retinopathy. The microsatellite was polymerase chain reaction amplified and electrophoresed on polyacrylamide gel and silver stained. In our study population, there were 18 alleles ranging from 97 to 131 base pairs (bp). Allele 4 (103 bp) had a higher prevalence (9.81%) in the DNR group compared to that (2.5%) in the DR group (P=0.002). Patients with retinopathy and allele 8 (111 bp) had a tendency to develop proliferative diabetic retinopathy (PDR). In this study of Indian subjects, it is suggested that allele 4 is a low risk allele for developing retinopathy and allele 8 (111 bp) shows an association with PDR.
Abstract: The disks of vertebrate photoreceptors are produced by outgrowths of the plasma membrane. Hence genes that encode retinal proteins targeted to plasma membrane protrusions represent candidates for inherited retinal degenerations. One such candidate is the gene encoding human prominin (mouse)-like 1 (PROML1, previously known as AC133 antigen) which belongs to the prominin family of 5-transmembrane domain proteins. Murine prominin (prom) shows a strong preference for plasma membrane protrusions in a variety of epithelial cells whereas PROML1 is expressed in retinoblastoma cell lines and adult retina. In the present study, molecular genetic analyses of a pedigree segregating for autosomal recessive retinal degeneration indicated that the affected individuals were homozygous for a nucleotide 1878 deletion in PROML1. This alteration is predicted to result in a frameshift at codon 614 with premature termination of translation. Expression of a similar prom deletion mutant in CHO cells indicated that the truncated protein does not reach the cell surface. Immunocytochemistry revealed that prom is concentrated in the plasma membrane evaginations at the base of the outer segments of rod photoreceptors. These findings suggest that loss of prominin causes retinal degeneration, possibly because of impaired generation of the evaginations and/or impaired conversion of the evaginations to disks.
Abstract: Retinitis pigmentosa (RP) is a group of clinically and genetically heterogeneous disorders characterised by night blindness, constriction of visual field, and dystrophic changes of the retina. Previous genetic studies have shown extensive allelic and non-allelic genetic heterogeneity of RP. Here we describe an Indian family with multiple consanguineous marriages and a total of four patients with autosomal recessive (AR) RP. The homozygosity mapping strategy was successfully used and indicated close linkage between the disease locus and D2S380, D2S441, D2S291, and D2S1394 with maximum lod scores between 1.51-3.07 at theta=0.00. The analysis of multiply informative meioses maps the locus (RP28) for ARRP in this family between D1S1337 and D2S286 on 2p11-p15. The involvement of visinin (VSNL1), a promising candidate gene assigned to chromosome 2p by previous studies, has been excluded by the absence of linkage.
Abstract: Autosomal recessive retinitis pigmentosa (arRP) is a genetically and clinically heterogeneous and progressive degenerative disorder of the retina, leading usually to severe visual handicap in adulthood. To date, disease loci/genes have been mapped/identified only in a minority of cases. DNA samples were collected from 20 large consanguineous Indian families, in which arRP segregated and that were suitable for homozygosity mapping of the disease locus. After excluding linkage to all known arRP loci, a genome-wide scan was initiated. In two families, homozygosity mapping, haplotype analysis, and linkage data mapped the disease locus (RP22) in an approximately 16-cM region between D16S287 and D16S420 on the proximal short arm of chromosome 16. No mutation has been found by direct sequencing in the gene (CRYM) encoding micron crystallin, which maps in the critical region.
Abstract: PURPOSE: Because corneal tissue with familial subepithelial corneal amyloidosis (FSCA; gelatinous drop-like dystrophy of the cornea) contains lactoferrin the possibility that the FSCA gene was the human lactoferrin (hLF) gene was investigated. Due to contradictory published information we also mapped the hLF gene. METHODS: We mapped the hLF gene using a genomic clone of the entire hLF gene as a probe by fluorescence in situ hybridization (FISH). Utilizing PCR primers that are specific to the hLF gene, we also mapped the hLF via radiation somatic cell hybrid analysis. Linkage of the FSCA gene to the hLF gene was evaluated by genetic linkage analysis using polymorphic markers within and in the vicinity of the hLF gene. RESULTS: The hLF gene mapped to the short arm of chromosome 3 at 3p21. Linkage analysis using polymorphic markers for hLF and haplotype analysis of the 3p21 loci indicates that the FSCA gene is not linked to the 3p21 locus. CONCLUSIONS: The gene for FSCA is not the hLF gene in these families.
Abstract: Adult-onset cataract (AOC) is a major ocular health problem and is the number one cause of blindness in the world. It is interesting to note that if the development of cataract is delayed by 10 years, the number of cataract surgeries needed would decrease by 45%. To prevent or delay cataract, the molecular pathological mechanisms underlying the lens change have to be understood, and this requires that the genes involved in such mechanisms should be identified. Hence, in this study we aim to identify AOC families which show a clear mendelian inheritance pattern, as only these families would be ideal for mapping the genes responsible. Over a period of 8 months, from September 1995-April 1996, 17 families with two or more affected members were identified. Segregation analysis showed autosomal dominant inheritance in multiple affected families. We propose to map the genes responsible for cataract in these families by linkage analysis and mutational screening of candidate genes.
Abstract: Autosomal recessive childhood-onset severe retinal dystrophy (arCSRD) designates a heterogeneous group of disorders affecting rod and cone photoreceptors simultaneously. The most severe cases are termed Leber congenital amaurosis (LCA), while the less aggressive forms are usually considered juvenile retinitis pigmentosa. Recently, mutations in the retinal-specific guanylate cyclase gene were found in patients with LCA. Disease genes implicated in other forms of arCSRD are expected to encode proteins present in the neuroretina or in the retinal pigment epithelium (RPE). The RPE, a monolayer of cells separating the vascular-rich choroid and the neuroretina, is in intimate contact with the outer segments of rods and cones via the microvilli surrounding the photoreceptors. The RPE expresses a tissue-specific and evolutionarily highly conserved 61 kD protein (RPE65) present at high levels in vivo. Although the function of RPE65 is not yet known, an important role in the RPE/photoreceptor vitamin-A cycle is suggested by the fact that RPE65 associates both with serum retinol-binding protein and with the RPE-specific 11-cis retinol dehydrogenase, an enzyme active in the synthesis of the visual pigment chromophore 11-cis retinal. Here we report that the analysis of RPE65 in a collection of about 100 unselected retinal-dystrophy patients of different ethnic origin revealed five that are likely to be pathogenic mutations, including a missense mutation (Pro363Thr), two point mutations affecting splicing (912 + 1G-->T and 65 + 5G-->A) and two small re-arrangements (ins144T and 831del8) on a total of nine alleles of five patients with arCSRD. In contrast to other genes whose defects have been implicated in degenerative retinopathies, RPE65 is the first disease gene in this group of inherited disorders that is expressed exclusively in the RPE, and may play a role in vitamin-A metabolism of the retina.
Abstract: Blepharophimosis syndrome (BPES) is an autosomal dominant disorder involving abnormal eyelid development. Cytogenetic and linkage analyses have previously implicated the chromosome 3q23 region in multiple cases of this syndrome. However, in a few cases cytogenetic analyses have implicated other chromosomal regions in this condition. Here we report linkage of BPES in a large Indian pedigree to chromosome 7p13-p21; affected only two-point and multipoint analyses using D7S488, D7S2551 and D7S2562 both showed peak lod scores of 3.61 coincident with D7S2562. Recombinations in affected individuals placed the critical region between D7S488 and D7S629. When both affected and unaffected individuals were considered, a maximum two-point lod score of 3.38 at theta = 0.08 was obtained with D7S2551 while a peak multipoint lod score of 3.64 was obtained between D7S488 and D7S2551. Segregation analysis revealed two unaffected individuals carrying the affected haplotype accounted for the difference in peak, relative to the affected only analysis. The chromosome 7p candidate genes inhibin beta A and epidermal growth factor receptor map outside this region whereas the HOX1 gene cluster may map inside this region. Although BPES is sometimes associated with female infertility due to premature ovarian failure, in the current family affected females were fertile. The current finding together with the previous evidence implicating chromosome 3q2 provides strong evidence that BPES involves locus heterogeneity; this point should be considered when counselling affected families.
Abstract: Seventy-eight families with retinitis pigmentosa, presenting at the genetic clinic of Sankara Nethralaya, Madras, over a period of 6 months (from April to September 1993), were assessed to determine the different genetic types: 9% were autosomal dominant; 36%, autosomal recessive; 3%, X-linked recessive; 44%, isolated cases and 8%, undetermined genetic type. A high incidence of consanguinity was observed in autosomal recessive (57%) and isolated cases (37%). Segregation analysis showed good agreement in autosomal dominant (chi 2 = 0.864) and recessive families (p = 0.222). The high proportion of autosomal recessive and isolated cases in this study, when compared with other similar studies, is due to the high incidence of consanguineous marriages in the Indian subcontinent.
Abstract: Oguchi disease is a rare autosomal recessive form of congenital stationary night blindness. The condition is associated with fundus discolouration and abnormally slow dark adaptation. Earlier studies suggested that the 48 kD protein S antigen may be involved in the recovery phase of light transduction. Previous cytogenetic and linkage studies have localised the S antigen gene (SAG) to chromosome 2q37.1. In the present study markers which map to distal chromosome 2q were typed in an inbred Oguchi pedigree. The segregation data obtained suggested that the affected subjects are homozygous by descent for a region between D2S172 and D2S345. An intragenic SAG polymorphism was homozygous in all affected people and a recombination event suggested that SAG maps proximal to D2S345. Collectively, these findings support the hypothesis that a defect in S antigen may be responsible for Oguchi disease.
