Associations between plasma levels of vitamins and cataract in the Italian-American Clinical Trial of Nutritional Supplements and Age-Related Cataract (CTNS): CTNS Report #2.

PURPOSE: To investigate the association at baseline between plasma levels of selected vitamins and the presence and type of cataract in the participants in The Italian-American Trial of Nutritional Supplements and Age-related Cataract. METHODS: At baseline, the participants (1020, 710 with "early cataract" and 310 with "no cataract," 55-75 years of age) received an ocular examination, photographic lens grading, and measurement of plasma levels of vitamins A, C, E, beta-carotene, and of red blood cell glutathione reductase activity. RESULTS: In multiple logistic models adjusted for potential confounders, high vitamin C levels were associated with a protective effect on nuclear (N) [OR: 0.54; 95% CI: 0.30, 0.97] and posterior subcapsular (PSC) cataract (OR: 0.37; 95% CI: 0.15, 0.93). High vitamin E levels were associated with increased prevalence of cortical cataract (C) (OR: 1.99; 95% CI: 1.02-3.90), PSC (OR: 3.27; 95% CI: 1.34, 7.96) and of any cataract (OR: 1.86; 95% CI: 1.08, 3.18). CONCLUSIONS: In agreement with some earlier studies, we found higher plasma levels of vitamin C to be associated with reduced prevalence of N and PSC cataracts. The finding of an increased prevalence of some types of cataract with higher levels of vitamin E was unexpected, has not been previously reported, and could be due to unadjusted confounding.

Transcriptional up-regulation of the protooncogenes c-fos and c-jun following vitreous removal and short-term in vitro culture of bovine lenses.

Chemical (mainly oxidative) and mechanical (anterior capsule injury) stresses have been reported to up-regulate the expression of the protooncogenes c-fos and c-jun in the lens. Another potentially stressful, yet largely unexplored condition, inherent to all experiments requiring the in vitro culturing of isolated lenses, is vitreous removal. Based on the results of an extensive RNA gel blot analysis conducted on epithelial/capsule preparations isolated from calf lenses dissected and cultured under different conditions, we show, here, that lens isolation and short-term culture (1-2.5 hr, without any significant GSH depletion) result in a strong and time-dependent up-regulation of the c-jun and c-fos mRNAs. This response, which relies on transcriptional protooncogene activation and is more intense for c-fos than for c-jun, is in part prevented by the preservation of the lens-vitreous contact, but not by the culture of vitreous-stripped lenses on a vitreous bed. Supplementation of the culture medium with the antioxidant N -acetyl-cysteine slightly reduced the c-jun, but not the c-fos response. Protooncogene up-regulation thus appears to be mainly determined by the disruption of critical lens-vitreous interactions. Since this response takes place in the epithelial cells, these data also point to the existence of a communication mechanism whereby a posteriorly applied mechanical stress is transmitted to, and perceived by, the anterior lens surface.

Retroillumination versus reflected-light images in the photographic assessment of posterior capsule opacification.

PURPOSE: To investigate the relative merit of retroillumination and of reflected light slit-lamp-derived photographs in the assessment of the opacification of the posterior lens capsule. METHODS: Retroillumination and slit-lamp-derived reflected-light photographs were taken on 23 consecutive eyes with posterior capsule opacification (PCO) in uncomplicated pseudophakia. Subjective grading was performed on both types of photographs to evaluate the extent and density of posterior capsular opacification. Best-corrected visual acuity (BCVA) before and after YAG laser capsulotomy was used to assess the impact of capsular opacification on visual function. RESULTS: After capsulotomy all patients attained a BCVA > or = 46 letters (> or =20/32) with a mean increase of 25 letters, indicating that PCO was the cause of visual impairment in these patients. The relative capacity of retroillumination and of reflected-light photographs to adequately capture the extent and the severity of posterior capsule opacification varied considerably. Reflected-light images, in addition to frequently producing higher severity scores for the opacity than retroillumination photographs, in 4 of 23 eyes (17.4%) proved to be the only technique able to document the presence of PCO. CONCLUSIONS: Our results indicate that, with respect to retroillumination images, reflected-light photography has an increased ability to adequately capture the presence and the severity of PCO and that the use of only retroillumination images may lead to its underestimation. This may be relevant to clinical studies aiming to evaluate incidence and progression of this condition.

The ageing lens.

The human lens grows by a process of epithelial cell division at its equator and the formation of generations of differentiated fibre cells. Despite the process of continuous remodelling necessary to achieve growth within a closed system, the lens can retain a high level of light transmission throughout the lifetime of the individual, with the ability to form sharp images on the retina. Continuous growth of the lens solves the problem imposed by terminal differentiation within a closed, avascular system, from which cells cannot be shed. The lens fibre tips arch over the equator to meet anteriorly and posteriorly and form branching sutures of increasing complexity. The stages of branching may create the optical zones of discontinuity seen on biomicroscopy. The lens is exposed to the cumulative effects of radiation, oxidation and postranslational modification. These later proteins and other lens molecules in such a way as to impair membrane functions and perturb protein (particularly crystallin) organisation, so that light transmission and image formation may be compromised. Damage is minimised by the presence of powerful scavenger and chaperone molecules. Progressive insolublisation of the crystallins of the lens nucleus in the first five decades of life, and the formation of higher molecular weight aggregates, may account for the decreased deformability of the lens nucleus which characterises presbyopia. Additional factors include: the progressive increase in lens mass with age, changes in the point of insertion of the lens zonules, and a shortening of the radius of curvature of the anterior surface of the lens. Also with age, there is a fall in light transmission by the lens, associated with increased light scatter, increased spectral absorption, particularly at the blue end of the spectrum, and increased lens fluorescence. A major factor responsible for the increased yellowing of the lens is the accumulation of a novel fluorogen, glutathione-3-hydroxy kynurenine glycoside, which makes a major contribution to the increasing fluorescence of the lens nucleus which occurs with age. Since this compound may also cross-link with the lens crystallins, it may contribute to the formation of high-molecular-weight aggregates and the increases in light scattering which occur with age. Focal changes of microscopic size are observed in apparently transparent, aged lenses and may be regarded as precursors of cortical cataract formation.

Oxidative stress and age-related cataract.

The authors review the available evidence supporting the possible role of oxidative stress in cataract formation from an epidemiological and a clinical point of view. They discuss in more detail what is presently known about the molecular mechanisms of response of the mammalian lens to an oxidative insult and report unpublished data on gene modulation upon oxidative stress in a bovine lens model. Main research endeavors that seem to be a most promising source of new insights into the problem of age-related cataract formation are briefly discussed.

Influence of cataract surgery on progression of lens opacities in the fellow eye.

OBJECTIVE: To evaluate whether, in the Italian-American natural history study, cataract surgery in one eye influences the incidence/progression rate of lens opacities in the fellow eye. DESIGN: Follow-up study of age-related cataract. PARTICIPANTS: A total of 1399 participants with age-related cataracts were regularly followed for 5 years and cataract status evaluated by the Lens Opacities Classification System II on slit-lamp and retroillumination lens photographs. A total of 228 participants had cataract surgery in 1 eye during the study period, and 192 had sufficient follow-up visits to be included in the analysis. MAIN OUTCOME MEASURE: The incidence/progression of cataract was defined as two or more consecutive visits with a severity grade greater than the baseline grade. RESULTS: When entered into a Cox model, incidence/progression of specific cataract types was not associated with cataract surgery on the fellow eye and was not significantly different from that observed in participants who had no cataract surgery. CONCLUSIONS: Cataract extraction does not influence incidence/progression rate of specific lens opacities in the fellow eye.

Self-assessment of the quality of vision: association of questionnaire score with objective clinical tests.

PURPOSE: To investigate the association of a quality of life-visual function questionnaire with an objective clinical test of visual function. METHODS: We have developed a questionnaire to assess self-reported visual satisfaction in ophthalmic patients suffering from chronic eye conditions causing visual impairment. The questionnaire was administered to 120 patients suffering from age-related cataract, chronic open angle glaucoma, age-related macular degeneration, branch retinal vein occlusion, and presbyopia or minor refractive defects. All the participants also underwent determination of visual acuity, contrast sensitivity, glare, and visual field. RESULTS: The questionnaire has a good reproducibility, a high internal consistency, and is able to discriminate between the different groups of patients. The total questionnaire score is significantly associated with the results of all visual function tests with the exception of glare. When entered into a multiple linear regression model, near visual acuity and contrast sensitivity are still considerably associated with the total questionnaire score. The psychological attitude of the patient towards his/her health problem is also associated with the total average score. CONCLUSIONS: Overall, the model explains 49% of the variance in the average questionnaire score.

An independent evaluation of the Age-Related Eye Disease Study (AREDS) cataract grading system.

PURPOSE: To assess intra- and interobserver reproducibility of the Age-Related Eye Disease Study (AREDS) system for grading lens opacities and to provide data on its capacity to reliably detect changes in lens status. METHODS: Independent and replicate grading of 40 sets of lens photographs (one slit-lamp and two retroillumination photographs) were performed by three experienced observers. Patients were participants in the Collaborative Italian-American Clinical Trial of Nutritional Supplements which is testing the effect of a mineral-multivitamin supplement on age-related cataract (CTNS). Scatterplots and intraclass correlation were used to assess measurement error. RESULTS: Analysis revealed good intra- and interobserver reproducibility of the system. Greatest intraobserver measurement error showed 100% of pairs within 10% areal difference for cortical cataract, 97.5% within 15% areal difference for posterior subcapsular cataract, and 100% within 1 density unit difference for nuclear opacity. Greatest interobserver measurement error showed 95% of pairs within 10% areal difference for cortical cataract, 97.5% within 15% areal difference for posterior subcapsular cataract, and 97.5% within 1.5 density unit difference for nuclear opacity. CONCLUSIONS: The AREDS lens opacities grading system appears to be sufficiently reliable to detect changes of at least 10% areal involvement for cortical, 15% areal involvement for posterior subcapsular, and 1.0 units for nuclear opacities. It therefore seems sufficiently sensitive to adequately monitor progression of lens opacities in a longitudinal study of patients with early cataract. Its applicability in a population with advanced or complex mixed opacities must await further testing.

Molecular basis of open-angle glaucoma in Italy.

Glaucoma is a group of ocular diseases characterized by an optic neuropathy in which degeneration of retinal ganglion cells leads to a characteristic excavation of the optic nerve head. Primary open-angle glaucoma (POAG) can be subdivided into two groups according to age of onset:- 1. the more common middle- to late-age onset, chronic open-angle glaucoma (COAG) diagnosed after the age of 40 years; 2. the rarer juvenile open-angle glaucoma (JOAG), which is diagnosed between the age of 3 years and early adulthood. Recently, the gene coding for the trabecular meshwork-induced glucocorticoid response protein (TIGR), located in chromosome 1 (1q23-25), was found mutated in patients affected by POAG. In this work we describe the clinical and molecular genetic features of several Italian families affected by autosomal dominant POAG, collected in various regions of Italy.

Vitreous fluorophotometry and changes in blood-retinal barrier permeability induced by bendazac lysine.

Vitreous fluorophotometry was used to investigate the effect of Bendazac lysine on the blood retinal barrier in 12 insulin-dependent diabetics with mild background retinopathy. The study was a randomized, double blind, cross-over trial, drug versus Placebo. Each treatment period was of 4 months. The vitreous penetration coefficient was reduced by 21% (95% c.i. 12, 30; p = 0.001) by treatment with respect to Placebo.

Location and severity of cortical opacities in different regions of the lens in age-related cataract.

PURPOSE: To examine the distribution of cortical opacities across the lens in the Italian-American Natural History Study of Age-Related Cataract and to study the association between an index of sunlight exposure and the location of cortical cataract within the lens. METHODS: Lens photographs of one eye of 731 persons with cortical opacities (503 with pure and 228 with mixed types of opacity) were included in the analysis. A radial grid superimposed on the photographs was used to assess presence, location, and severity of wedge-shaped cortical opacities. RESULTS: Both the prevalence and the extent of cortical opacities were highest in the inferior-nasal quadrant and lowest in the superior-nasal quadrant of the lens. In polychotomous logistic regression, persons with the greatest excess areal involvement in the inferior half of the lens were more likely to have high exposure to sunlight, as measured by a sunlight index, than persons with excess involvement in the superior half of the lens (odds ratio, 1.73; 95% confidence interval 1.03, 2.93). Excess areal involvement of the inferior lens also was associated with the pure type of cortical cataract and with the total extent of the opacity. CONCLUSIONS: Age-related cortical opacities occur more frequently inferiorly than superiorly and, to a lesser extent, nasally than temporally. Possibly higher exposure of these lens segments to sunlight may explain this preferential location of cortical opacities.

Glutathione S-transferase M1 genotype and age-related cataracts. Lack of association in an Italian population.

PURPOSE: To investigate possible associations between the gene number and allelic forms of glutathione S-transferase M1 (GSTM1) and the occurrence of nucleic and cortical age-related cataracts. METHODS: Patients with cortical cataract, nuclear cataract, mixed and cortical cataract, and no cataract were sytematically selected from subjects evaluated in the Italian-American Study of the Natural History of Age-Related Cataract. The patients were typed for the A, B, and null alleles of GSTM1 using a variation of the amplification refractory mutation system. RESULTS: Forty-nine percent of patients (50/102) with cortical cataracts, 45% (13/29) with nuclear cataracts, 51% (36/71) with mixed nuclear and cortical cataracts, and 50% of controls (49/98) were homozygous for the null GSTM1 allele. Twenty-five percent of patients (26/102) with cortical cataracts, 24% (7/29) with nuclear cataracts, 31% with mixed nuclear and cortical cataracts, and 27% of controls (26/98) displayed only the A allele for GSTM1. Twenty-four percent of patients (24/102) with cortical cataract, 24% (7/29) with nuclear cataracts, 14% (10/71) with mixed nuclear and cortical cataract, and 18% of controls showed only the B allele for GSTM1. Two percent of patients (2/102) with cortical cataracts, 7% (2/29) with nuclear cataracts, 4% (3/71) with mixed nuclear and cortical cataracts, and 5% of controls (5/98) showed both A and B alleles for GSTM1. CONCLUSIONS: No associations between the GSTM1 alleles, including the null allele, and cataracts were detected in this study.

Distribution of cataract types in the Italian-American case-control study and at surgery in the Parma area.

PURPOSE: To compare the distribution of cataract types at surgery with the distribution detected in an ophthalmology clinic-based case-control study in the same geographic area (Parma, Italy). METHODS: The distribution of cataract type assessed according to the Lens Opacities Classification System I in 284 consecutive patients 45 years of age or older, who were admitted for cataract surgery to the Institute of Ophthalmology in Parma during 1994, was compared with the distribution assessed in 1008 participants in the Italian-American case-control study of age-related cataract in the Parma metropolitan area from 1987 to 1989. RESULTS: Analysis of cataract distribution indicates in the surgical group, compared with the case-control population, a significant increase of nuclear (N) and posterior subcapsular (PSC) opacities (any), a reduction of pure forms of cortical and N cataracts, and a marked increase of mixed types of opacities with a simultaneous N and PSC component. CONCLUSION: Although cortical opacities are probably the most prevalent type of age-related lens change in the general population of Parma metropolitan area, the type of cataract most frequently responsible for the decision of patients to undergo cataract surgery is a mixed type of opacity with an N-PSC component.

A dose-response effect between a sunlight index and age-related cataracts. Italian-American Cataract Study Group.

To explore the existence of a dose-response relationship between sunlight exposure and risk of age-related cataracts, we analyzed data collected from 1008 patients with cataracts and 469 control subjects enrolled in the Italian-American Case-Control Study of Age-Related Cataracts. Fourteen variables related to sunlight exposure history were included in the questionnaire administered to the study participants. A sunlight index was constructed and its relationship to the presence of cataracts was modeled by logistic regression. After adjustments for potential confounding variables and for age and sex, a significant dose-response effect (P = 0.01) was detected between the sunlight exposure index and the presence of pure cortical cataracts. With the exception of corticonuclear cataracts, all the other mixed types of opacity also showed a dose-response association with the sunlight index. These data support the hypothesis that sunlight exposure is a risk factor in the development of cortical cataracts, and demonstrate the existence of a dose-response relationship in this association.

Influence of type and severity of pure forms of age-related cataract on visual acuity and contrast sensitivity. Italian American Cataract Study Group.

PURPOSE: To compare the relationship between logMAR visual acuity (VA) and cataract severity and between contrast sensitivity (CS) and cataract severity in pure types of age-related lens opacities. METHODS: Analysis included patients followed in the ongoing Italian-American Study of the Natural History of Age-Related Cataract. Lens opacities were classified and graded according to the Lens Opacities Classification System II (LOCS II). Visual acuity was measured with the Early Treatment Diabetic Retinopathy Study Chart. Contrast sensitivity was measured with the Pelli-Robson chart. RESULTS: Data from 1,076 eyes were used for the analysis (366 clear lenses; 550, 124, and 36 eyes with cortical, nuclear and posterior subcapsular cataract, respectively). In age-adjusted analyses, increasing severity of all three cataract types was associated with progressively higher logMAR VA, which translates into poorer acuity, and lower CS scores. For both VA and CS, the effect of increasing severity was greatest for nuclear and least for cortical opacities. After adjusting for age and VA, CS scores were no longer associated with cataract type and severity, with the exception of advanced cortical opacities. CONCLUSIONS: Increased cataract severity, as determined by LOCS II grading, is strongly associated with both VA and CS scores. Contrast sensitivity scores obtained from testing at low spatial frequency do not seem to offer additional information over standard VA testing in early cortical and posterior subcapsular opacities nor in nuclear cataracts.

Lens sodium channels are inactivated by anti-MIP26 antibodies.

Sodium-specific channels can be functionally identified in phosphatidylcholine liposomes incorporating detergent-solubilized membrane proteins from pig lens epithelium and outer cortex. The transport of sodium is saturable, specific and protease-sensitive. MIP26 was identified in the solubilized membrane fraction and in the liposomes by means of Western blot analysis. Pre-treatment of liposomes with anti-MIP26 antiserum abolished the transport of sodium. These data indicate that MIP26 is associated to a sodium selective transport activity.

Enzyme activities in human lens epithelium of age-related cataract.

PURPOSE: To investigate associations between enzyme activity of glutathione reductase (GR) with and without added flavin adenine dinucleotide (FAD), glutathione peroxidase (GPX), and 6-phosphogluconic dehydrogenase (6PGDH) in the lens epithelium collected at surgery, and some nutritional and biochemical variables determined in the same individuals during the Italian-American Case-Control Study of age-related cataract. METHODS: One hundred eighty-three epithelium capsule samples were collected from 174 patients undergoing surgery. Data on enzyme activity were obtained from 52 samples for 6-PGDH and from 53 samples for GR and for GPX. The Lens Opacity Classification System II was used to classify and grade cataracts. RESULTS: No correlation was found between enzyme activity in lens epithelium and the same enzymatic activity in erythrocytes (with the exception of a negative correlation between lens and erythrocyte 6PGDH activity), or the type and severity of cataract. No correlation was found between lens GPX activity and plasma selenium and between lens GR activation coefficient (GRAC) and riboflavin intake. Lens GR with added FAD and lens GRAC were significantly correlated to plasma vitamin E level. Lens GRAC was positively correlated to a nutritional vitamin index. CONCLUSIONS: Present data stress the difficulty in verifying the assumption that biochemical indices collected on plasma and on erythrocyte actually reflect the status of these factors in the lens itself.

Sugar alcohols in the lens epithelium of age-related cataract.

We have measured the levels of glucose and sugar alcohols in the lens epithelium of 24 non-diabetic and 21 diabetic patients undergoing extracapsular cataract extraction for age-related cataract. Lens status was assessed preoperatively according to the Lens Opacities Classification System II. In comparison to non-diabetics, the lens epithelia of diabetic patients have increased levels of glucose and sorbitol, and lower content of myo-inositol. Both myo-inositol decrease and sorbitol accumulation are strictly related to the fasting blood sugar level. No correlation or trend was detected in diabetic subjects between myo-inositol or sorbitol level and cataract type. In non-diabetic patients nuclear opacification was associated with a significant increase of myo-inositol level in lens epithelium.

The effect of cataract severity and morphology on the reliability of the Lens Opacities Classification System II (LOCS II).

Data collected from 3646 eyes in the Italian-American Natural History Study of Age-Related Cataract were used to investigate whether the reliability of the Lens Opacities Classification System II (LOCS II) by the severity of the opacity that is being graded or is influenced by the presence and severity of coexisting opacities. Reliability was assessed by comparing the slit-lamp gradings of two clinical examiners (346 eyes) and the gradings performed at the slit lamp with gradings of photographs (3646 eyes). The severity of cortical and nuclear opacities did not affect the reproducibility of slit-lamp gradings, but clinical grading of posterior subcapsular opacities became more reliable as the severity of the posterior subcapsular opacities increased. More advanced coexisting opacities decreased the agreement in the slit-lamp diagnosis of nuclear, but not cortical or posterior subcapsular, opacities. Comparisons of clinical and photographic gradings showed very good to excellent agreement for nuclear and cortical opacities, regardless of the severity of the specific opacity or the severity of the coexisting opacities. Agreement in diagnosing posterior subcapsular opacities was decreased in eyes with milder posterior subcapsular opacities and in eyes with more severe coexisting nuclear and/or cortical opacities. The effect of the severity of the opacity being graded and the severity of coexisting opacities on the reliability of the LOCS II must be considered in studies that use the system to classify and grade cataracts.

Increased ion traffic through non-specific cation pathways in the ageing human lens. Evidence from radiotracer fluxes studies.

86Rb efflux studies were carried out on normal human lenses in vitro. The data confirmed previous studies showing that 86Rb efflux increases with age. Removal of Ca2+ from the lens perifusate increased 86Rb efflux at all ages. The fractional increase above baseline was highest in the younger lenses, while the net increase of the 86Rb efflux induced by a Ca(2+)-free medium increased with age. This study supports the idea that Ca(2+)-sensitive, non-specific cation channels are present in the human lens and that their contribution to membrane permeability increases as the lens ages.