Protective association of A-T-T haplotype of DMT1 gene against risk of human age-related nuclear cataract.

BACKGROUND: Age-related cataract (ARC) is profoundly associated with oxidative stress. Iron plays a pivotal role in generating oxidative stress and promoting deleterious irreversible damage to the macromolecules. Divalent metal transporter 1 (DMT1) mediates the uptake of iron into the cell. Aberrant transcript expression of DMT1 gene in lenses of human ARC was reported. The present investigated the genetic association between DMT1 gene polymorphisms and risk of ARC. METHODS: DNA from peripheral blood of ARC subjects (n = 764) and age-matched controls (n = 794) was isolated. Genotyping of three single-nucleotide polymorphisms (SNPs) - rs224589 (C/A), rs1048230 (T/C), and rs2285230 (T/C) - of DMT1 gene was performed by polymerase chain reaction (PCR)-restriction fragment length polymorphism technique. Level of DMT1 transcript expression was determined by quantitative real-time PCR analysis using RNA from lens epithelial and fiber cells. RESULTS: Nuclear cataract showed a higher frequency of CC genotypes (OR = 1.40; 95%CI = 1.01-1.95; p = 0.04) of SNP rs224589 and a significantly lower frequency of A-T-T haplotype (OR = 0.63; 95%CI = 0.42-0.92; p = 0.02) than that of controls. The A-T-T haplotype demonstrated a dominant protective effect against disease risk when compared to the more common haplotype (C-T-T) (p = 0.01). The haplotype pairs C-T-T/C-T-T and A-C-C/A-C-C showed higher level of transcript expression of DMT1 than C-T-T/A-T-T haplotype pair (p < 0.05). Further, a novel genetic variation (c.1328A>G; p.N443S) in exon 3 of DMT1 gene was observed in a subject with nuclear cataract. CONCLUSIONS: The results highlighted a protective association of A-T-T haplotype against the risk of ARC.

Genetic investigation of ocular developmental genes in 52 patients with anophthalmia/microphthalmia.

BACKGROUND: Mutation in eye developmental genes has been reported to cause anophthalmia and microphthalmia. However, in India, especially in the Western Indian population, such reports are scarce. Hence, the present study aims to investigate mutations in 15 ocular developmental genes in patients with anophthalmia and microphthalmia in the western region of India. MATERIALS AND METHODS: Genomic DNA was isolated from the blood of 52 individuals affected with microphthalmia and anophthalmia, and 50 healthy normal controls. Polymerase chain reaction (PCR) was carried out for 15 genes including BMP4, CRYBA4, FOXE3, GDF6, GJA3, GJA8, MITF, OTX2, PAX6, PITX3, RAX, SIX3, SIX6, SOX2, and VSX2 using gene-specific primers spanning the exon-intron boundaries and part of a promoter region. The amplified PCR products were purified and then subjected to Sanger's bi-directional sequencing. Nucleotide variations were examined using a basic local alignment search tool (BLAST). RESULTS: Bi-directional sequencing identified 8 novel and 14 known variations. Out of this, the variations GJA3-c.92T>A; p.Ile31Asn, SOX2-c.542C>A; p.Pro181Gln and SOX2-c.541_542delinsGA; p.Pro181Glu were found to be deleterious by in silico analysis. The GJA3-p.Ile31Asn mutation was identified in a patient with bilateral microphthalmia, microcornea, and membranous cataract. The SOX2-p.Pro181Gln and SOX2-p.Pro181Glu mutations were identified in patients with isolated bilateral microphthalmia and microphthalmia with microcornea, respectively. A novel nondeleterious missense variation was identified in the GJA8 gene in a patient with anophthalmia. CONCLUSION: These results support the crucial role of GJA3 and SOX2 in eye development and indicate a detailed functional study to understand the molecular mechanisms underlying the disease pathology.

Association of FOXE3-p.Ala170Ala and PITX3-p.Ile95Ile Polymorphisms with Congenital Cataract and Microphthalmia.

PURPOSE: To investigate the association of FOXE3-p.Ala170Ala (rs34082359) and PITX3-p.Ile95Ile (rs2281983) polymorphisms with congenital cataract and microphthalmia in a western Indian population. METHODS: FOXE3-p.Ala170Ala (c.510C>T) and PITX3-p.Ile95Ile (c.285C>T) polymorphisms were genotyped in 561 subjects consisting of 242 cases with congenital cataract, 52 with microphthalmia, and 267 controls using polymerase chain reaction-restriction fragment length polymorphism. Approximately 10% of samples were randomly sequenced for each single nucleotide polymorphism to confirm the genotypes. The prediction of mRNA secondary structure for polymorphism FOXE3-p.Ala170Ala and PITX3-p.Ile95Ile was performed. RESULTS: A significantly high frequency of T allele and a borderline significance in the frequency of TT genotype of FOXE3-p.Ala170Ala was observed in microphthalmia cases, as compared to controls [T allele: OR: [CI] = 1.8 [1.15-2.72], P = 0.0115; TT: OR [CI] = 2.9 [1.14-7.16], P = 0.0291). The frequency of CC genotype was significantly low in microphthalmia cases when compared to controls (CC: OR [CI] = 0.5 [0.24-0.86, P = 0.0150). There was no significant difference in the allele and genotype frequencies of PITX3-p.Ile95Ile between cases and controls. A slight free energy change was observed in the secondary structure of mRNA between the FOXE3-p.Ala170Ala C-allele (-917.60 kcal/mol) and T-allele (-916.80 kcal/mol) and between PITX3-p.Ile95Ile C-allele (-659.80 kcal/mol) and T-allele (-658.40 kcal/mol). CONCLUSION: The present findings indicate that FOXE3-p.Ala170Ala 'T' allele and 'TT' genotype could be predisposing factors for microphthalmia while 'CC' genotype might play a protective role against it. A reduction in the free energy change associated with FOXE3-p.Ala170Ala 'T' allele could further contribute towards disease risk.

CDC Kerala--The Untold Story.

This article is our life time experience in conceptualizing and systematically developing Child Development Centre (CDC) Kerala in the last 25 years, from a research project to a national training centre in child and adolescent development and premarital counseling. CDC Kerala's major contribution was in creating a 'conceptual framework' of a valid link between childhood disability, low birth weight, adolescent girls' nutrition and fetal onset adult lifestyle diseases. It all started with a randomized controlled trial (RCT) proving beyond doubt that early stimulation is effective in improving the neurodevelopmental status of high risk babies at one and two years and the same cohort was followed-up in detail at 5, 13, 16, 19 and 24 completed years. The process of establishing CDC Kerala is being presented under (i) clinical child development, (ii) adolescent care counseling, (iii) young adults and premarital counseling and (iv) institution building.

Scintigraphic scoring system for grading severity of gastro-esophageal reflux on 99mTc sulfur colloid gastro-esophageal reflux scintigraphy: A prospective study of 39 cases with pre and post treatment assessment.

AIM: The study aimed at developing a scoring system for scintigraphic grading of gastro-esophageal reflux (GER), on gastro-esophageal reflux scintigraphy (GERS) and comparison of clinical and scintigraphic scores, pre- and post-treatment. MATERIALS AND METHODS: A total of 39 cases with clinically symptomatic GER underwent 99mTc sulfur colloid GERS; scores were assigned based on the clinical and scintigraphic parameters. Post domperidone GERS was performed after completion of treatment. Follow up GERS was performed and clinical and scintigraphic parameters were compared with baseline parameters. RESULTS: Paired t-test on pre and post domperidone treatment clinical scores showed that the decline in post-treatment scores was highly significant, with P value < 0.001. The scintigraphic scoring system had a sensitivity of 93.9% in assessing treatment response to domperidone, specificity of 83.3% i.e., 83.3% of children with no decline in scintigraphic scores show no clinical response to Domperidone. The scintigraphic scoring system had a positive predictive value of 96.9% and a negative predictive value of 71.4%. CONCLUSION: GERS with its quantitative parameters is a good investigation for assessing the severity of reflux and also for following children post-treatment.

A Rare Scenario of Stenosed Type IV Dual LAD with Normal Myocardial Perfusion Scan.

Dual left anterior descending artery (LAD) is a rare variation in the coronary artery anatomy having 4 different subtypes. We report a rare scenario of type IV dual LAD, with short LAD critical stenosis, compensated by a long LAD perfusing the same territory, which was missed on initial angiographic evaluation. Myocardial perfusion scan (MPS) of this patient showed preserved perfusion. A review of angiogram revealed this anomaly, indicating that a meticulous correlation of anatomic and functional modalities is necessary as it could change further management of patient.

Fourier phase analysis on equilibrium gated radionuclide ventriculography: Range of phase spread and cut-off limits in normal individuals.

AIM: To define the range of phase spread on equilibrium gated radionuclide ventriculography (ERNV) in normal individuals and derive the cut-off limit for the parameters to detect cardiac dyssynchrony. MATERIALS AND METHODS: ERNV was carried out in 30 individuals (age 53±23 years, 25 males and 5 females) who had no history of cardiovascular disease. They all had normal left ventricular ejection fraction (LVEF 55-70%) as determined by echocardiography, were in sinus rhythm, with normal QRS duration (≤120 msec) and normal coronary angiography. First harmonic phase analysis was performed on scintigraphic data acquired in best septal view. Left and right ventricular standard deviation (LVSD and RVSD, respectively) and interventricular mechanical delay (IVMD), the absolute difference of mean phase angles of right and left ventricle, were computed and expressed in milliseconds. Mean + 3 standard deviation (SD) was used to derive the cut-off limits. RESULTS: Average LVEF and duration of cardiac cycle in the study group were 62.5%±5.44% and 868.9±114.5 msec, respectively. The observations of LVSD, RVSD and right and left ventricular mean phase angles were shown to be normally distributed by Shapiro-Wilk test. Cut-off limits for LVSD, RVSD and IVMD were calculated to be 80 msec, 85 msec and 75 msec, respectively. CONCLUSION: Fourier phase analysis on ERNV is an effective tool for the evaluation of synchronicity of cardiac contraction. The cut-off limits of parameters of dyssynchrony can be used to separate heart failure patients with cardiac dyssynchrony from those without. ERNV can be used to select patients for cardiac resynchronization therapy.