Dystrophin gene deletions in South Indian Duchenne muscular dystrophy patients.

66 unrelated patients from Southern India with Duchenne Muscular Dystrophy (DMD) were studied for intragenic deletion in 18 exons and Pm region of the DMD gene using multiplex PCR. Of these 41 (62.1%) showed intragenic deletions. 78% of the deletions were located at the distal hotspot region (44-55 exons) and 22% of the deletions were located at the proximal region (exon 2-19). Exon 50 is most frequently deleted. Deletions in isolated cases were significantly more compared to familial cases. The lower incidence reported from South India compared to North India, is suggestive of variations in the Southern and Northern population.

Translocation Down syndrome.

Cytogenetic investigations carried out on 1021 cases of Down syndrome revealed translocation in 46 cases. The most frequent was of t(14;21) and t(21;21) types. Most of the translocation DS cases (n = 31) were born to younger mothers (< 25 years), when compared to pure trisomy 21 DS cases. Parental karyotypes, family history and parental ages has helped us greatly in offering genetic counseling, prenatal diagnosis and estimating the risk for the next conception.

Translocation Down syndrome.

Cytogenetic investigations carried on 1021 cases of Down syndrome revealed translocation in 46 cases. The most frequent was of t(14;21) and t(21;21) types. Most of the translocation DS cases (n = 31) were born to younger mother's (< 25 years), when compared to pure trisomy 21 DS cases. Parental karyotypes, family history and parental ages has helped us greatly in offering genetic counseling, prenatal diagnosis and estimating the risk for the next conception.

Evaluation of immunoperoxidase staining technique in the diagnosis of Acanthamoeba keratitis.

PURPOSE: We describe a simple procedure of Immunoperoxidase (IP) technique, using indigenously raised antibody, to screen corneal scrapings for Acanthamoeba cysts and trophozoites. This study sought to determine the utility of this test in the diagnosis of Acanthamoeba keratitis. METHODS: A high titre polyclonal antibody against a local clinical isolate (axenic) of Acanthamoeba species (trophozoite lysate antigen) was raised in rabbits and used for standardization of IP technique for corneal scrapings. Twenty two smears of corneal scrapings, collected from patients showing Acanthamoeba cysts in corneal scrapings stained with calcofluorwhite (pool-1) and patients showing no cysts in similar scrapings (pool-2), were coded and stained by IP technique by a masked technician. All 22 patients had also been tested for bacteria, fungus, and Acanthamoeba in their corneal scrapings by smears and cultures. IP stained smears were examined for organisms including cysts and trophozoites of Acanthamoeba and background staining by two observers masked to the results of other smears and cultures. The validity of the IP test in detection of Acanthamoeba cysts and trophozoites was measured by sensitivity, specificity, positive predictive value and negative predictive value in comparison (McNemar test for paired comparison) with calcofluor white staining and culture. RESULTS: Based on the readings of observer 1 and compared to calcofluor white staining, the IP test had a sensitivity of 100%, a specificity of 94%, positive predictive value of 80% and negative predictive value of 100%. When compared to culture, the values were 83%, 100%, 100% and 94% respectively. Trophozoites missed in calcofluor white stained smears, were detected in 2 out of 6 cases of culture-positive Acanthamoeba keratitis. The Kappa coefficient of interobserver agreement was determined as fair (30.4%). CONCLUSION: The immunoperoxidase technique is a simple and useful test in the diagnosis of Acanthamoeba keratitis. This can supplement the culture results.

Cytogenetic studies of 1001 Down syndrome cases from Andhra Pradesh, India.

Cytogenetic data obtained from investigating 1001 patients of Down syndrome (DS) and their parents over a period of 20 years (January 1979-January 1999) are presented. The frequency of pure trisomy, mosaicism and translocation was 87.92, 7.69 and 4.39 per cent respectively. The origin of the extra chromosome 21 due to meiotic non-disjunction was 79.24 per cent maternal and 20.76 per cent paternal. A high frequency of acrocentric chromosome associations was also observed in mothers of children of Down syndrome, this might have predisposed to an enhanced risk for non-disjunction. Birth order of DS showed a higher number of first and second borns. Reproductive performances of the parents indicated a high rate of abortions, compared to controls. Cytogenetic investigations carried out over these years greatly helped in the management of these children and for counseling the affected families.

Adaptation of WHOQOL as health-related quality of life instrument to develop a vision-specific instrument.

The WHOQOL instrument was adapted as a health-related QOL instrument for a population-based epidemiologic study of eye diseases in southern India, the Andhra Pradesh Eye Disease Study (APEDS). A follow-up question was added to each item in WHOQOL to determine whether the decrease in QOL was due to any health reasons including eye-related reasons. Modifications in WHOQOL and translation in local language were done through the use of the focus groups including health professionals and people not related to health care. The modified instrument has 28 items across 6 domains of the WHOQOL and was translated into the local language, Telugu, using the pragmatic approach. It takes 10-20 minutes to be administered by a trained interviewer. Reliability was within acceptable range. This health-related QOL instrument is being used in the population-based study APEDS to develop a vision-specific QOL instrument which could potentially be used to assess the impact of visual impairment on QOL across different cultures and for use in evaluating eye-care interventions. This health-related QOL instrument could also be used to develop other disease-specific instruments as it allows assessment of the extent to which various aspects of QOL are affected by a variety of health problems.

Unilateral visual impairment in an urban population in southern India.

This study assessed the prevalence and causes of unilateral visual impairment in the urban population of Hyderabad city as part of the Andhra Pradesh Eye Disease Study. Stratified, random, cluster, systematic sampling was used to select 2,954 subjects from 24 clusters representative of the population of Hyderabad. Eligible subjects underwent detailed eye examination including logMAR visual acuity, refraction, slitlamp biomicroscopy, applanation tonometry, gonioscopy, dilatation, cataract grading, and stereoscopic evaluation of fundus. Automated threshold visual fields and slitlamp and fundus photography were done when indicated by standardised criteria. Unilateral visual impairment was defined as presenting distance visual acuity < 6/18 in the worse eye and > or = 6/12 in the better eye, which was further divided into unilateral blindness (visual acuity < 6/60 in the worse eye) and unilateral moderate visual impairment (visual acuity < 6/18-6/60 in the worse eye). A total of 2,522 subjects (85.4% of eligible), including 1,399 > or = 30 years old, participated in the study. In addition to the 1% blindness and 7.2% moderate visual impairment (based on bilateral visual impairment criteria) reported earlier in this sample, 139 subjects had unilateral visual impairment, an age-gender-adjusted prevalence of 3.8% (95% confidence interval 2.7-4.9%). The major causes of this visual impairment 39.9% were refractive error (42.9%), cataract (14.4%), corneal disease (11.5%), and retinal disease (11.2%). Of this unilateral visual impairment was blindness. The major causes of unilateral blindness were corneal disease (23.2%), cataract (22.5%), retinal disease (18%), and optic atrophy (12.9%). On the other hand, the predominant cause of unilateral moderate visual impairment was refractive error (67%) followed by cataract (9%). Of the total unilateral visual impairment, 34.3% was present in those < 30 years old and 36.2% in those 30-49 years old. Unilateral visual impairment afflicts approximately 1 in 25 persons in this urban population. A large proportion of this unilateral visual impairment is present in younger age groups. The causes of unilateral visual impairment, like those of bilateral visual impairment in this population, are varied, suggesting therefore, that in addition to the current focus of eye care in India predominantly on cataract, other causes of visual impairment need to be addressed as well.

HPLC analysis of closed, open, and reflex eye tear proteins.

Changes in the closed, open and reflex eye tear proteins of normal subjects were compared and analysed. Tear proteins were resolved by high-performance liquid chromatography (HPLC) utilising both gel filtration (P-300 SW) and reverse-phase (C-18) columns and the HPLC fractions were further analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing and non-reducing conditions. The protein composition of the closed-eye tear was significantly different from that of the open and reflex-eye tear. Secretory IgA (sIgA) was the predominant protein in closed eye tears constituting 49% of the total protein compared to 11% in reflex tears, whereas lysozyme was the predominant protein (53%) in reflex tears. Levels of lactoferrin, lipocalin and lysozyme were relatively constant in both open and reflex tears. HPLC profiles of the closed-eye tears, upon continuous stimulation of lacrimal glands indicated that sIgA was significantly reduced whereas lactoferrin, lipocalin, and lysozyme were significantly increased. These results indicate that the tear composition upon waking attains that of the open eye within 4 to 5 minutes, and upon continuous stimulation this reflects the reflex-eye tear composition. It also indicates that mechanisms responsible for changes in concentration of constitutive and regulated tear protein with stimulus can be studied successfully using non-invasive methods to collect human tears.

Developing a model to reduce blindness in India: The International Centre for Advancement of Rural Eye Care.

With the continuing high magnitude of blindness in India, fresh approaches are needed to effectively deal with this burden on society. The International Centre for Advancement of Rural Eye Care (ICARE) has been established at the L.V. Prasad Eye Institute in Hyderabad to develop such an approach. This paper describes how ICARE functions to meet its objective. The three major functions of ICARE are design and implementation of rural eye-care centres, human resource development for eye care, and community eye-health planning. ICARE works with existing eye-care centres, as well as those being planned, in underserved areas of India and other parts of the developing world. The approach being developed by ICARE, along with its partners, to reduce blindness is that of comprehensive eye care with due emphasis on preventive, curative and rehabilitative aspects. This approach involves the community in which blindness is sought to be reduced by understanding how the people perceive eye health and the barriers to eye care, thereby enabling development of strategies to prevent blindness. Emphasis is placed on providing good-quality eye care with attention to reasonable infrastructure and equipment, developing a resource of adequately trained eye-care professionals of all cadres, developing a professional environment satisfactory for patients as well as eye-care providers, and the concept of good management and financial self-sustainability. Community-based rehabilitation of those with incurable blindness is also part of this approach. ICARE plans to work intensively with its partners and develop these concepts further, thereby effectively bringing into practice the concept of comprehensive eye care for the community in underserved parts of India, and later in other parts of the developing world. In addition, ICARE is involved in assessing the current situation regarding the various aspects of blindness through well-designed epidemiologic studies, and projecting the eye-care needs for the future with the help of reliable information. With balanced attention to infrastructure, manpower, financial self-sustenance, and future planning, ICARE intends to develop a practical model to effectively reduce blindness in India on a long-term basis.

Causes of corneal graft failure in India.

The success of corneal grafting in visual rehabilitation of the corneal blind in India depends on survival of the grafts. Understanding the causes of graft failure may help reduce the risk of failure. We studied these causes in a series of 638 graft failures at our institution. Multivariate logistic regression analysis was used to evaluate the association of particular causes of graft failure with indications for grafting, socioeconomic status, age, sex, host corneal vascularization, donor corneal quality, and experience of surgeon. The major causes of graft failure were allograft rejection (29.2%), increased intraocular pressure (16.9%), infection excluding endophthalmitis (15.4%), and surface problems (12.7%). The odds of infection causing graft failure were significantly higher in patients of lower socioeconomic status (odds ratio 2.45, 95% CI 1.45-4.15). Surface problems as a cause of graft failure was significantly associated with grafts done for corneal scarring or for regrafts (odds ratio 3.36, 95% CI 1.80-6.30). Increased intraocular pressure as a cause of graft failure had significant association with grafts done for aphakic or pseudophakic bullous keratopathy, congenital conditions or glaucoma, or regrafts (odds ratio 2.19, 95% CI 1.25-3.84). Corneal dystrophy was the indication for grafting in 12 of the 13 cases of graft failure due to recurrence of host disease. Surface problems, increased intraocular pressure, and infection are modifiable risk factors that are more likely to cause graft failure in certain categories of patients in India. Knowledge about these associations can be helpful in looking for and aggressively treating these modifiable risk factors in the at-risk categories of corneal graft patients. This can possibly reduce the chance of graft failure.

Economic burden of blindness in India.

Economic analysis is one way to determine the allocation of scarce resources for health-care programs. The initial step in this process is to estimate in economic terms the burden of diseases and the benefit from interventions for prevention and treatment of these diseases. In this paper, the direct and indirect economic loss due to blindness in India is calculated on the basis of certain assumptions. The cost of treating cataract blindness in India is estimated at current prices. The economic burden of blindness in India for the year 1997 based on our assumptions is Rs. 159 billion (US$ 4.4 billion), and the cumulative loss over lifetime of the blind is Rs. 2,787 billion (US$ 77.4 billion). Childhood blindness accounts for 28.7% of this lifetime loss. The cost of treating all cases of cataract blindness in India is Rs. 5.3 billion (US$ 0.15 billion). Similar estimates for causes of blindness other than cataract have to be made in order to develop a comprehensive approach to deal with blindness in India.

Planning to reduce childhood blindness in India.

Reduction of blindness in children assumes particular significance since a blind child suffers from more blind-years than a blind adult. Estimates of the number of children blind in India and the causes of blindness are relatively crude as there are little reliable epidemiologic data. As a result of this, there is no organised approach to the control of childhood blindness in India. In order to address this issue, a workshop on childhood blindness was held at the L.V. Prasad Eye Institute, Hyderabad in November 1996. The aim of this workshop was to review available data, consider possible strategies, and make recommendations concerning the control of childhood blindness in India. These recommendations along with background information about childhood blindness are presented in this paper.

Ophthalmic antiviral chemotherapy: an overview.

Antiviral drug development has been slow due to many factors. One such factor is the difficulty to block the viral replication in the cell without adversely affecting the host cell metabolic activity. Most of the antiviral compounds are analogs of purines and pyramidines. Currently available antiviral drugs mainly inhibit viral nucleic acid synthesis, hence act only on actively replicating viruses. This article presents an overview of some of the commonly used antiviral agents in clinical ophthalmology.

Indications for penetrating keratoplasty in India.

Indications for penetrating keratoplasty (PK) in the developing world from a large series are not well documented. This study was done to evaluate the indications for PK in a major eye care institution in India. The records for a consecutive series of 1,964 PKs were analysed and multiple logistic regression was used to study the effect of age, socioeconomic status and sex on the indications for PK. The indications for PK were corneal scarring in 551 (28.1%) including adherent leukoma in 147 (7.5%), regrafts in 336 (17.1%), active infectious keratitis in 239 (12.2%), aphakic bullous keratopathy in 231 (11.8%), pseudophakic bullous keratopathy in 209 (10.6%), corneal dystrophies in 165 (8.4%) including Fuchs' dystrophy in 23 (1.2%), keratoconus in 118 (6%), and miscellaneous in 115 (5.9%). The odds that the patient belonged to lower socioeconomic status were significantly higher if the PK was done for active infectious keratitis (odds ratio 2.73, p < 0.0001), corneal scarring (odds ratio 1.72, p = 0.0009) or regraft (odds ratio 1.44, p = 0.047). Corneal scarring, including adherent leukoma, and active infectious keratitis are relatively more common indications whereas keratoconus, pseudophakic bullous keratopathy and Fuchs' dystrophy are less common indications for PK in India than reported from the developed world. Indications for PK which carry a poorer prognosis for graft survival are relatively more common in India than in the developed world.

Gram negative bacteria and contact lens induced acute red eye.

Two patients using hydrogel contact lenses on a daily wear schedule slept overnight with the lenses and woke up with a Contact Lens Induced Acute Red Eye (CLARE). The contact lenses recovered aseptically at the time of the event grew significant colonies of Pseudomonas aeruginosa and Aeromonas hydrophila in patient A and Pseudomonas aeruginosa and Serratia liquefaciens from patient B. Similar organisms from the contact lenses were recovered from the lens case and lens care solutions of patient B. In both the patients the condition resolved on discontinuation of lens wear. Patient compliance as a requirement for successful contact lens wear is highlighted with the illustration of these cases.

Cyanoacrylate adhesive with conjunctival resection and superficial keratectomy in Mooren's ulcer.

Seventeen eyes of thirteen patients with Mooren's ulcer were treated with a combination therapy of local and systemic steroids, conjunctival resection, superficial keratectomy and application of cyanoacrylate tissue adhesive. The pathology was classified as acute, subacute and chronic. Ulcers were graded based on the extent of corneal thinning, degree and extent of ulceration, and amount of inflammation. Fourteen eyes (82.4%) healed completely with formation of a vascularised scars, while three eyes (17.6%) failed to respond to treatment and either went into phthisis bulbi or healed with gross tissue distortion. Our study suggests an early intervention of this therapy with cyanoacrylate tissue adhesive application for effective control of Mooren's ulceration.